AU5671799A - Heteroaryl amidines, methylamidines and guanidines as protease inhibitors - Google Patents

Heteroaryl amidines, methylamidines and guanidines as protease inhibitors Download PDF

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AU5671799A
AU5671799A AU56717/99A AU5671799A AU5671799A AU 5671799 A AU5671799 A AU 5671799A AU 56717/99 A AU56717/99 A AU 56717/99A AU 5671799 A AU5671799 A AU 5671799A AU 5671799 A AU5671799 A AU 5671799A
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carboxamidine
methylthiothiophene
amino
thiazol
methylthio
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James B. Hoffman
Carl R. Illig
Juan Jose Marugan
M. Jonathan Rudolph
Nalin L. Subasinghe
Kenneth J. Wilson
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3 Dimensional Pharmaceuticals Inc
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3 Dimensional Pharmaceuticals Inc
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    • C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
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    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
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    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

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Description

PCT/US99/180 6 5 WO 00/47578 5 HETEROARYL AMIDINES, METHYLAMIDINES AND GUANIDINES AS PROTEASE INHIBITORS Background of the Invention 10 Field of the Invention The present invention relates to novel heteroaryl compounds that function as enzyme inhibitors, and particularly to a new class of non-peptidic inhibitors of proteolytic enzymes such as urokinase (uPa). Related Art 15 Proteases are enzymes that cleave proteins at single, specific peptide bonds. Proteases can be classified into four generic classes: serine, thiol or cysteinyl, acid or aspartyl, and metalloproteases (Cuypers et al., J. Biol. Chem. 257:7086 (1982)). Proteases are essential to a variety of biological activities, such as digestion, formation and dissolution of blood clots, reproduction and the 20 immune reaction to foreign cells and organisms. Aberrant proteolysis is associated with a number of disease states in man and other mammals. The human neutrophil proteases, elastase and cathepsin G, have been implicated as contributing to disease states marked by tissue destruction. These disease states include emphysema, rheumatoid arthritis, corneal ulcers and glomerular nephritis. 25 (Barret, in Enzyme Inhibitors as Drugs, Sandler, ed., University Park Press, Baltimore, (1980)). Additional proteases such as plasmin, C- 1 esterase, C-3 convertase, urokinase and tissue-type plasminogen activators, acrosin, and kallikreins play key roles in normal biological functions of mammals. In many WO 00/47578 PCT/US99/1 8 0 6 5 -2 5 instances, it is beneficial to disrupt the function of one or more proteolytic enzymes in the course of therapeutically treating a mammal. Serine proteases include such enzymes as elastase (human leukocyte), cathepsin G, plasmin, C-1 esterase, C-3 convertase, urokinase and tissue-type plasminogen activators, acrosin, chymotrypsin, trypsin, thrombin, factor Xa and 10 kallikreins. Human leukocyte elastase is released by polymorphonuclear leukocytes at sites of inflammation and thus is a contributing cause for a number of disease states. Cathepsin G is another human neutrophil serine protease. Compounds with the ability to inhibit the activity of these enzymes are expected to have an 15 anti-inflammatory effect useful in the treatment of gout, rheumatoid arthritis and other inflammatory diseases, and in the treatment of emphysema. Chymotrypsin and trypsin are digestive enzymes. Inhibitors of these enzymes are useful in treating pancreatitis. Inhibitors of urokinase plasminogen activator are useful in treating excessive cell growth disease states, such as benign prostatic hypertrophy, 20 prostatic carcinoma and psoriasis. Urokinase (urinary-type plasminogen activator or uPA; International Union of Biochemistry Classification Number: EC3.4.21.31) is a proteolytic enzyme which is highly specific for a single peptide bond in plasminogen. It is a multidomain serine protease, having a catalytic "B" chain (amino acids (aa) 25 144-411), and an amino-terminal fragment ("ATF", aa 1-143) consisting of a growth factor-like domain (4-43) and a Kringle domain (aa 47-135). The uPA Kringle domain appears to bind heparin, but not fibrin, lysine, or aminohexanoic acid. The growth factor-like domain bears some similarity to the structure of epidermal growth factor (EGF) and is thus also referred to as "EGF-like" domain. 30 The single chain pro-uPA is activated by plasmin, cleaving the chain into a two chain active form that is stabilized by a disulfide bond. Cleavage of the peptide bond in plasminogen by urokinase ("plasminogen activation") results in the formation of a potent general protease, plasmin. Many cell types use urokinase as a key initiator of plasmin-mediated proteolytic 35 degradation or modification of extracellular support structures (e.g., the WO 00/47578 PCT/US99/180 6 5 -3 5 extracellular matrix (ECM) and the basement membrane (BM)). Cells exist, move, and interact with each other in tissues and organs within the physical framework provided by the ECM and BM. Movement of cells within the ECM or across the BM requires local proteolytic degradation or modification of these structures, allowing cells to "invade" into adjacent areas that were previously 10 unavailable. Central to the ability of urokinase to mediate cellular migration and invasiveness is the existence of specific high affinity urokinase receptors (uPARs) which concentrate urokinase on the cell surface, leading to the generation of locally high plasmin concentrations between cells and ECM or BM (Blasi, F., et 15 al., Cell Biol. 104:801-804 (1987); Roldan, A.L., et al., EMBO J. 9:467-74 (1990)). The binding interaction is apparently mediated by the EGF-like domain (Rabbani, S.A., et al., J. Biol. Chem. 267:14151-56 (1992)). Cleavage of pro-uPA into active uPA is accelerated when pro-uPA and plasminogen are receptor-bound. Thus, plasmin activates pro-uPA, which in turn activates more 20 plasmin by cleaving plasminogen. This positive feedback cycle is apparently limited to the receptor-based proteolysis on the cell surface, since a large excess of protease inhibitors is found in plasma, including
U
2 antiplasmin, PAl-1 and PAI-2. High plasmin concentrations between invasive cells and ECM or BM are necessary in order to overcome inhibitory effect of these ubiquitous plasmin 25 inhibitors. Thus, it is cell surface receptor-bound urokinase, and not simply free urokinase secreted by cells, which plays the predominant role in initiating cellular invasiveness. Plasmin can activate or degrade extracellular proteins such as fibrinogen, fibronectin, and zymogens, including matrix metalloproteinases. Plasminogen 30 activators thus can regulate extracellular proteolysis, fibrin clot lysis, tissue remodeling, developmental cell and smooth muscle cell migration, inflammation, and metastasis. Cellular invasiveness initiated by urokinase is central to a wide variety of normal and disease-state physiological processes (reviewed in Blasi, F., etal.,J. Cell Biol. 104:801-804 (1987); Dano, K., etal.,Adv. Cancer Res. 44:139 35 266 (1985); Littlefield, B.A., Ann. N. Y. Acad. Sci. 622:167-175 (1991); Saksela, WO 00/47578 PCT/US99/180 65 -4 5 0., Biochim. Biophys. Acta 823:35-65 (1985); Testa, J.E., and Quigley, J.P., Cancer Metast. Rev. 9:353-367 (1990)). Such processes include, but are not limited to, angiogenesis (neovascularization), bone restructuring, embryo implantation in the uterus, infiltration of immune cells into inflammatory sites, ovulation, spermatogenesis, tissue remodeling during wound repair, restenosis 10 and organ differentiation, fibrosis, local invasion of tumors into adjacent areas, metastatic spread of tumor cells from primary to secondary sites, and tissue destruction in arthritis. Inhibitors of urokinase therefore have mechanism-based anti-angiogenic, anti-arthritic, anti-inflammatory, anti-restenotic, anti-invasive, anti-metastatic, anti-osteoporotic, anti-retinopathic (for angiogenesis-dependent 15 retinopathies), contraceptive, and tumoristatic activities. Inhibitors of urokinase are useful agents in the treatment of a variety of disease states, including but not limited to, benign prostatic hypertrophy, prostatic carcinoma and psoriasis. Beneficial effects of urokinase inhibitors have been reported using anti urokinase monoclonal antibodies and certain other known urokinase inhibitors. 20 For instance, anti-urokinase monoclonal antibodies have been reported to block tumor cell invasiveness in vitro (Hollas, W., et al., Cancer Res. 51:3690-3695, (1991); Meissauer, A., et al., Exp. Cell Res. 192:453-459 (1991)), tumor metastasis and invasion in vivo (Ossowski, L., J. Cell Biol. 107:2437-2445 (1988); Ossowski, L., et al., J. Cancer Res. 51:274-81 (1991)), and angiogenesis 25 in vivo (Jerdan, J. A., et al., J. Cell Biol. 115[3 Pt 2]:402a (1991)). In addition, amiloride, a known urokinase inhibitor of only moderate potency, has been reported to inhibit tumor metastasis in vivo (Kellen, J.A., et al., Anticancer Res. 8:1373-1376 (1988)) and angiogenesis/capillary network information in vitro (Alliegro, M.A., et al., J. Cell Biol. 115[3 Pt 2]:402a (1991)). 30 Urokinase plays a significant role in vascular wound healing and arterial neointima formation after injury, most likely affecting cellular migration. Urokinase mediates plasmin proteolysis, which in turn promotes vascular wound healing and associated neointima formation (Carmeliet et al., Circ. Res. 81:829 839 (Nov. 1997), Lupu et al., Fibrinolysis 10 Supp 2:33-35 (1996)). A viral 35 serine proteinase inhibitor, SERP-1, has been employed to reduce plaque WO 00/47578 PCT/US99/180 6 5 -5 5 formation after primary balloon angioplasty in rabbits. This activity has been attributed to the inhibition by SERP-1 of cellular proteinases, such as plasmin or urokinase (Lucas et al., Circulation 94:2890-2900 (1996)). A need continues for non-peptidic compounds that are potent and selective urokinase inhibitors, and which possess greater bioavailability and fewer 10 side-effects than currently available urokinase inhibitors. Accordingly, new classes of potent urokinase inhibitors, characterized by potent inhibitory capacity and low toxicity, are potentially valuable therapeutic agents for a variety of conditions. Summary of the Invention 15 The present invention is broadly directed to the use of heteroaryl amidines, methylamidines and guanidines having Formula I (below) as protease inhibitors, preferably as urokinase inhibitors. Compounds of the present invention exhibit anti-urokinase activity via direct, selective inhibition of urokinase, or are intermediates useful for forming 20 compounds having such activity. Compounds of the present invention inhibit urokinase and are, therefore, useful anti-angiogenic, anti-arthritic, anti inflammatory, anti-restenotic, anti-invasive, anti-metastatic, anti-osteoporotic, anti-retinopathic (for angiogenesis-dependent retinopathies), contraceptive, and tumoristatic treatment agents. For example, such treatment agents are useful in 25 the treatment of a variety of disease states, including but not limited to, benign prostatic hypertrophy, prostatic carcinoma, tumor metastasis and psoriasis. Also provided are methods to inhibit extracellular proteolysis, methods to treat benign prostatic hypertrophy, prostatic carcinoma, tumor metastasis, psoriasis, and other conditions by administering the compound of Formula I. 30 A number of the heteroaryl compounds described herein are novel compounds. Therefore, the present invention is also directed to novel compounds of Formula I.
WO 00/47578 PCT/US99/180 6 5 -6 5 Further provided are pharmaceutical compositions comprising a compound of Formula I and one or more pharmaceutically acceptable carriers or diluents and said pharmaceutical compositions further comprising athrombolytic agent such as tissue plasminogen activator and streptokinase. Further provided are methods of synthesizing compounds of Formula I. t0 Detailed Description of the Preferred Embodiments The present invention is broadly directed to a method of inhibiting proteases, particularly serine proteases, by contacting a serine protease with a compound of the general Formula I: 2 RI NR4 15 or a solvate, hydrate or pharmaceutically acceptable salt thereof; wherein: X is O, S orNR 7 , where R 7 is hydrogen, alkyl, aralkyl, hydroxy(C 2
-
4 )alkyl, or alkoxy(C 2
-
4 )alkyl; Y is a direct covalent bond, CH 2 or NH; Z is NR'R 6 , hydrogen or alkyl, provided that Y is NH whenever Z is 20 hydrogen or alkyl;
R
1 is hydrogen, amino, hydroxy, halogen, cyano, Cl- 4 alkyl or -CH 2 R, where R is hydroxy, amino or C,_ 3 alkoxy;
R
2 and R 3 are independently: i. hydrogen; 25 ii. halogen; iii. hydroxy; iv. nitro; v. cyano; WO 00/47578 PCT/US99/18065 -7 5 vi. amino, monoalkylamino, dialkylamino, monoarylamino, diarylamino, monoalkylmonoarylamino, monoaralkylamino, diaralkylamino, monoalkylmonoaralkylamino, monoheterocycleamino, diheterocycleamino, monoalkylmonoheterocycleamino, alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, alkylsulfonylamino, 0 aralkylsulfonylamino, aralkenylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, di(aralkylsulfonyl)amino, di(aralkenylsulfonyl)amino, di(arylsulfonyl)amino, or di-(heteroarylsulfonyl)amino, formylamino, alkanoylamino, alkenoylamino, alkynoylamino, aroylamino, aralkanoylamino, aralkenoylamino, heteroaroylamino, heteroaralkanoylamino, H(S)CNH-, or 5 thioacylamino, wherein any of the aryl or heteroaryl containing groups can be optionally substituted on the aromatic ring and wherein any of the heterocycle containing groups can be optionally ring substituted; vii. aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl, acyl, aminoacyl, monoarylaminocarbonyl, diarylaminocarbonyl or .0 monoalkylmonoarylaminocarbonyl; viii. aminothiocarbonyl, monoalkylaminothiocarbonyl, dialkylaminothiocarbonyl, thioacyl or aminothioacyl; ix. aminocarbonylamino, mono- and dialkylaminocarbonylamino, mono- and diarylaminocarbonylamino, or mono- and :5 diaralkylaminocarbonylamino; x. aminocarbonyloxy, mono- and dialkylaminocarbonyloxy, mono and diarylaminocarbonyloxy, mono- and diaralkylaminocarbonyloxy; xi. aminosulfonyl, mono- and dialkylaminosulfonyl, mono- and diarylaminosulfonyl, or mono- and diaralkylaminosulfonyl; 30 xii. alkoxy, or alkylthio, wherein the alkyl portion of each group may be optionally substituted, xiii. aralkoxy, aryloxy, heteroaryloxy, aralkylthio, arylthio, or heteroarylthio, wherein the aryl portion of each group can be optionally substituted; WO 00/47578 PCT/US99/18065 -8 5 xiv. alkylsulfonyl, wherein the alkyl portion can be optionally substituted; xv. aralkylsulfonyl, aralkenylsulfonyl, arylsulfonyl or heteroarylsulfonyl, wherein the aryl portion of each group can be optionally substituted; 10 xvi. alkenyl, or alkynyl; xvii. optionally substituted aryl; xviii. optionally substituted alkyl; xix. optionally substituted aralkyl; xx. optionally substituted heterocycle; or 15 xxi. optionally substituted cycloalkyl; and
R
4 , R 5 and R 6 are independently hydrogen, C- 4 alkyl, aryl, hydroxyalkyl, aminoalkyl, monoalkylamino(C 2
-
1 0 )alkyl, dialkylamino(C 2
-
1 0 )alkyl, carboxyalkyl, cyano, amino, alkoxy, or hydroxy, or -CO 2 Rw, where Rw is alkyl, cycloalkyl, phenyl, benzyl, 20 Rf 0 JR Rh Sor R O 0 Rd
R
e where R d and R e are independently hydrogen, C- 6 alkyl,
C
2
-
6 alkenyl or phenyl, R' is hydrogen, C 1
-
6 alkyl, C 2 -6 alkenyl or phenyl, R; is hydrogen, C 1
_
6 alkyl, C 2
-
6 alkenyl or phenyl, and Rh is aralkyl or C,- 6 alkyl. 25 The present invention is also directed to novel compounds of Formula I, where X, Y and R 1
-R
6 are as defined above; provided that at least one of R 2 or R 3 is selected from the group consisting of: (a) an optionally substituted alkyl group, preferably Cl-C 6 alkyl, more preferably Ci-C 3 ; 30 (b) alkoxy, aryloxy, alkylthio or arylthio, any of which is optionally substituted; WO 00/47578 PCT/US99/18065 -9 5 (c) optionally substituted C 6
-C
1 4 aryl, or optionally substituted aralkyl, except that R 3 is not nitrophenyl or aminophenyl, when R' and R 2 are both hydrogen or methyl; (d) optionally substituted heterocycle; and (e) optionally substituted cycloalkyl. 10 When an alkyl-containing group, heterocyclic-containing group or aryl containing group of R 2 or R 3 is optionally substituted, the optional substituents can be 1 to 4 non-hydrogen substituents, provided that the resulting compound is stable. Values of optional substituents on alkyl groups are halogen, hydroxy, thiol, amino, monoalkylamino, dialkylamino, formylamino, aminoiminomethyl, 15 acylamino, aminoacyl, mono- or di- alkylaminocarbonyl, thiocarbonylamino, thioacylamino, aminothiocarbonyl, alkoxy, aryloxy, aminocarbonyloxy, mono or di-alkylaminocarbonyloxy, mono- or diarylaminocarbonyloxy, mono- or diaralkylaminocarbonyloxy, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, alkylsulfonylamino, arylsulforiylamino, aralkylsulfonylamino, 20 alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, mono- or di- alkylaminothiocarbonyl, aralkoxy, carboxy, carboxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, nitro, cyano, trifluoromethyl, alkylthio and arylthio. Preferred values of optional substituents on an alkyl group are chloro, hydroxy, amino, mono(Cl.
4 )alkylamino, di(C 1
-
4 )alkylamino, formylamino, C 2
.
6 25 acylamino, aminocarbonyl, C 2
-
8 aminoacyl, C 1
-
6 alkoxy, C 6
.
1 4 aryloxy, carboxy, carboxy(Cl- 6 )alkyl, C 2
.
8 alkoxycarbonyl, nitro, cyano, trifluoromethyl, C- 6 alkylthio, C 6
-
1 4 arylthio, C 1
-
6 alkylsulfonylamino, C 7
.
15 aralkylsulfonylamino, C 6
-
1 0 arylsulfonylamino, mono- or di(Cl.
6 )alkylaminocarbonyloxy, mono- or di
(C
6 .I)arylaminocarbonyloxy, mono- or di(C 7
.
15 )aralkylcarbonyloxy, CI.-6 30 alkoxycarbonylamino, C 7
-C
5 aralkoxycarbonylamino, and C 6 -Co 1 0 aryloxycarbonylamino. Preferred values of optional substituents on aryl-containing and heterocyclic-containing groups include chloro, hydroxy, amino, mono(Cl.
4 ) alkylamino, di(Cl-4)alkylamino, formylamino, C 2
-
6 acylamino, aminocarbonyl, C 2 -8 35 aminoacyl, C 3
.
7 cycloalkyl, Cl.
6 alkyl, Cl.
6 alkoxy, C 6
.-
14 aryloxy, carboxy, WO 00/47578 PCT/US99/18065 -10 5 carboxy(Ci- 6 )alkyl, C 2
-
8 alkoxycarbonyl, nitro, cyano, trifluoromethyl, C-6 alkylthio, C 6
-
1 4 arylthio, C 6
-
4 aryl, substituted phenyl, tetrazolyl, thienyl (further optionally substituted by one, two or three of chloro, hydroxy, C,.
4 alkyl, Cl-4 alkoxy, amino or carboxy), 3,4-methylenedioxy, 3,4-ethylenedioxy, 3,4-propylenedioxy, C, 6 alkylsulfonylamino, C 7
-
5 aralkylsulfonylamino, C1-6 10 arylsulfonylamino, C- 6 alkyl/sulfonyl, C 6 -1 0 arylsulfonyl, mono- or di(C 1
.
6 )alkylaminocarbonyloxy, mono- or di- C 6 o 10 arylaminocarbonyloxy, mono or di-(C 71 5 )aralkylcarbonyloxy, C 1
-
6 alkoxycarbonylamino, C 7
-C
1 5 aralkoxycarbonylamino, C 6
-C
10 aryloxycarbonylamino, C 2
-
6 thioacylamino, aminothiocarbonyl, and C 2
-
8 aminothioacyl. 15 Preferred values of R' include hydrogen, amino, hydroxy and fluoro. A preferred value of R 2 is Formula II: Ar , RH .R9 where Ar is phenyl, thiazolyl, thiazolinyl, oxazolyl, isothiazolyl, isoxazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, thienyl (thiophenyl), pyrrolyl, 20 oxazolinyl and benzothienyl. Preferred values ofR 3 include Cl-4 alkyl (optionally substituted), halogen, amino, acylamino, C, 6 _ alkylthio (such as methylthio or ethylthio), Cl- 6 alkoxy (such as methoxy and ethoxy), trifluoromethyl, methylsulfonyl, and benzylthio. A preferred value of X is divalent sulfur (S). Z5 Preferred values of Y are a covalent bond or -NH-, most preferably a covalent bond. Preferred values of R 4 , R 5 and R 6 in Formula I are hydrogen, hydroxy, cyano, C 1
.-
6 alkyl, or C 1
_
6 alkoxy. Suitable values of R 4 , R 5 and R 6 include hydrogen, methyl, ethyl, propyl, n-butyl, hydroxy, methoxy, and ethoxy. In the 0 most preferred embodiments, R 4 , R 5 and R 6 are each hydrogen. Preferred values of R 4 , R 5 and R 6 in Formula I also include prodrugs such as -CO 2 Rw, where Rw, in each instance, is preferably one of C 14 alkyl,
C
4
.
7 cycloalkyl or benzyloxycarbonyl. Suitable values of R 4 , R' and R 6 include WO 00/47578 PCT/US99/18065 -11 5 hydrogen, methyl, ethyl, propyl, n-butyl, hydroxy, methoxy, ethoxy, cyano,
-CO
2
CH
3 , -CO 2
CH
2
CH
3 and -CO 2
CH,CH
2
CH
3 . In the most preferred embodiments, R 4 , R 5 and R 6 are each hydrogen. Also preferred at R 4 , R 5 and R 6 is the group -CO 2 Rw, where Rw is one of Rf0 _' Rh 0 or 0 Rg 0 Rd Re 0 where Rd-Rh are defined as above. When R 4 , R 5 and R 6 are -CO 2 Rw, where Rw is one of one of these moieties, the resulting compounds are prodrugs that possess desirable formulation and bioavailability characteristics. A preferred value for each of Rd, R e and R9 is hydrogen, R f is methyl, and preferred values for Rh include benzyl and tert-butyl. 5 Preferred values of R 7 include hydrogen, C- 6 alkyl, C 6
-
10 ar(Cl.
4 )alkyl, and
C
2
-
6 hydroxyalkyl. Suitable values are hydrogen, methyl, ethyl, and benzyl. The term "alkyl" as employed herein by itself or as part of another group refers to both straight and branched chain radicals of up to 12 carbons, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, 0 heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl. The term "alkenyl" is used herein to mean a straight or branched chain radical of 2-20 carbon atoms, unless the chain length is limited thereto, including, but not limited to, ethenyl, 1-propenyl, 2-propenyl, 2-methyl-l-propenyl, 5 1-butenyl, 2-butenyl, and the like. Preferably, the alkenyl chain is 2 to 10 carbon atoms in length, more preferably, 2 to 8 carbon atoms in length most 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 chain length is limited thereto, wherein 0 there is at least one triple bond between two of the carbon atoms in the chain, WO 00/47578 PCT/US99/18065 -12 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, most preferably from 2 to 4 carbon atoms in length. In all instances herein where there is an alkenyl or alkynyl moiety as a substituent group, the unsaturated linkage, i.e., the vinylene or acetylene linkage ) is preferably not directly attached to a nitrogen, oxygen or sulfur moiety. The term "alkylthio" as employed herein by itself or as part of another group refers to a straight or branched chain radical of 1 to 20 carbon atoms, unless the chain length is limited thereto, bonded to a sulfur atom, including, but not limited to, methylthio, ethylthio, n-propylthio, isopropylthio, and the like. 5 Preferably the alkylthio chain is 1 to 10 carbon atoms in length, more preferably 1 to 8 carbon atoms in length. The term "alkoxy" as employed herein by itself or as part of another group refers to a straight or branched chain radical of 1 to 20 carbon atoms, unless the chain length is limited thereto, bonded to an oxygen atom, including, but not ) limited to, methoxy, ethoxy, n-propoxy, isopropoxy, and the like. Preferably the alkoxy chain is 1 to 10 carbon atoms in length, more preferably 1 to 8 carbon atoms in length. The term "cycloalkyl" as employed herein by itself or as part of another group refers to cycloalkyl groups containing 3 to 9 carbon atoms. Typical i examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and cyclononyl. The term "halogen" or "halo"as employed herein by itself or as part of another group refers to chlorine, bromine, fluorine or iodine with chlorine being preferred. ) The term "acyl" as employed herein by itself or as part of another group refers to the group -C(O)R g where R9 is alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, heteroaryl, heteroarylalkyl or heteroarylalkenyl. Preferred acyl groups are alkanoyl, aralkanoyl and aroyl groups (-C(O)R g where R9 is C, 8 alkyl, C 6
-
1 0 aryl(C,4)alkyl or C 6 10 aryl).
WO 00/47578 PCT/US99/18065 -13 5 The term "thioacyl" as employed herein by itself or as part of another group refers to the group -C(S)R g where R9 is alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, heteroaryl, heteroarylalkyl or heteroarylalkenyl, preferably C1., alkyl. The term "thiocarbonyl" as employed herein by itself or as part of another group refers to the group -C(S)-. D The term "monoalkylamine" as employed herein by itself or as part of another group refers to an amino group which is substituted with one alkyl group having from 1 to 6 carbon atoms. The term "dialkylamine" as employed herein by itself or as part of another group refers to an amino group which is substituted with two alkyl groups, each 5 having from 1 to 6 carbon atoms The term "aryl" as employed herein by itself or as part of another group refers to monocyclic or bicyclic aromatic groups containing from 6 to 14 carbons in the ring portion, preferably 6-10 carbons in the ring portion, such as phenyl, naphthyl or tetrahydronaphthyl. 3 The term "aralkyl" or "arylalkyl" as employed herein by itself or as part of another group refers to Ci- 6 alkyl groups as discussed above having an aryl substituent, such as benzyl, phenylethyl or 2-naphthylmethyl. The terms "heterocyclic," "heterocyclo" or "heterocycle" as employed herein by themselves or as part of larger groups refers to a saturated or wholly or 5 partially unsaturated 3-7 membered monocyclic, or 7-10 membered bicyclic ring system, which consists of carbon atoms and from one to four heteroatoms independently selected from the group consisting of O, N, and S, wherein the nitrogen and sulfur heteroatoms can be optionally oxidized, the nitrogen can be optionally quaternized, and including any bicyclic group in which any of the ) above-defined heterocyclic rings is fused to a benzene ring, and wherein the heterocyclic ring can be substituted on carbon or on a nitrogen atom if the resulting compound is stable. Especially useful are rings containing one oxygen or sulfur, one to three nitrogen atoms, or one oxygen or sulfur combined with one or two nitrogen atoms. Examples of such heterocyclic groups include piperidinyl, 5 piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2- WO 00/47578 PCT/US99/18065 -14 5 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, indanyl, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazoyl, 0 benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, and oxadiazolyl. Morpholino is the same as morpholinyl. The term "heteroatom" is used herein to mean an oxygen atom ("O"), a 5 sulfur atom ("S") or a nitrogen atom ("N"). It will be recognized that when the heteroatom is nitrogen, it may form an NRYRz moiety, wherein R Y and Rz are, independently from one another, hydrogen or C, to C 8 alkyl, or together with the nitrogen to which they are bound, form a saturated or unsaturated 5-, 6-, or 7 membered ring. 0 The term "heteroaryl" as employed herein refers to groups having 5 to 14 ring atoms; 6, 10 or 14 x electrons shared in a cyclic array; and containing carbon atoms and 1, 2 or 3 oxygen, nitrogen or sulfur heteroatoms (where examples of heteroaryl groups are: thienyl, benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, chromenyl, xanthenyl, 5 phenoxathiinyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinazolinyl, cinnolinyl, pteridinyl, 4cH-carbazolyl, carbazolyl, P-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, ) phenothiazinyl, isoxazolyl, furazanyl and phenoxazinyl groups). The expression "prodrug" denotes a derivative of a known direct acting drug, which derivative has enhanced delivery characteristics and therapeutic value as compared to the drug, and is transformed into the active drug by an enzymatic or chemical process. Useful prodrugs are those where R 4 , R 5 and/or R 6 are WO 00/47578 PCT/US99/18065 -15 5 -CO 2 Rw, where Rw is defined above. See, U.S. PatentNo. 5,466,811 and Saulnier et al., Bioorg. Med Chem. Lett. 4:1985-1990 (1994). The term "substituted", as used herein, means that one or more hydrogens of the designated moiety are replaced with a selection from the indicated group, provided that no atom's normal valency is exceeded, and that the substitution 0 results in a stable compound. When a substituent is keto (i.e., =0), then 2 hydrogens attached to an atom of the moiety are replaced. By "stable compound" or "stable formula" is meant herein a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture and formulation into an efficacious therapeutic agent. 5 A first preferred group of compounds falling within the scope of the present invention include compounds of Formulalwherein X is sulfur or oxygen; Y is a covalent bond or -NH-; R' is hydrogen, amino, hydroxy or halogen; R 4 , R 5 and R 6 are independently hydrogen, C-.
4 alkyl, amino, cyano, C-4 alkoxy or hydroxy, and are preferably all hydrogen; one of R 2 or R 3 is hydrogen, C- 6 alkyl 0 (optionally substituted with hydroxy, amino, carboxy or aminocarbonyl), C,-6 alkylthio or C,- 6 alkoxy; and the other of R 2 or R' is aminoacyl, acylamino, aminosulfonyl, sulfonylamino, aminocarbonylamino, alkoxycarbonylamino, optionally substituted oxazolyl, optionally substituted isoxazolyl, optionally substituted benzothienyl, optionally substituted furanyl, optionally substituted 5 pyrazolyl or optionally substituted pyridyl. Specific compounds within the scope of the invention include the compounds described in the Examples, such as the following: 4-[4-(4-chlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-phenyl-5-methylthiothiophene-2-carboxamidine; 0 4-[4-(2,4-dichlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-(4-methylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; methyl 4-[4-(4-phenylphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxylate; 4-[4-(3-methoxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine, 4 -[4-(3-hydroxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine, 5 4-(4-phenylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine, WO 00/47578 PCT/US99/18065 -16 5 4
-[
4 -(4-nitrophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine, 4- [4-(3,4-ethylenedioxyphenyl)thiazol-2-yl]-5 -methylthiothiophene-2 carboxamidine, 4- [4-(3,4-propylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine, 1o 4-[4-(4-(naphth-2-yl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine, 4-isopropylsulfonyl-5-methylthiothiophene-2-carboxamidine; 4-phenyl-5-methylthiothiophene-2-carboxamidine; 4
-[
4 -(4-chlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[ 4
-(
4 -phenylphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; .5 4
-[
4
-(
4 -methoxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-(2-naphthylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-[4-(4-chloro-3-methylphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-(5-methyl-4-phenylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; :0 4
-[
4
-(
4 -chloro-3-nitrophenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4 -(5-phenyloxazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4- [4-(3 -fluoro-5-trifluoromethylphenyl)-5-methylthiazol-2-yl] -5 methylthiothiophene-2-carboxamidine; .5 4-[4-(3,5-bis(trifluoromethyl)phenyl)-5-methyl-thiazol-2-yl]-5 methylthiothiophene-2-carboxamidine; 4-[4-(3-fluoro-5-trifluoromethylphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-[4-(3-bromophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 0 4
-[
4
-(
3
,
4 -methylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4
-[
4
-(
4 -methylphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(3,5-bis(trifluoromethyl)phenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 5 4
-[
4
-(
2 -methoxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; WO 00/47578 PCT/US99/18065 -17 5 4-(4-phenylimidazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-[4-(2,4-dimethoxyphenyl)thiazol-2-yl-5 -methylthiothiophene-2 carboxamidine; 4-(4-benzylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-[4-(3,4-dichlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 0 4-[4-(3-methylphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4- [4-(3, 5 -dimethoxyphenyl)thiazol-2-yl] -5 -methylthiothiophene-2 carboxamidine; 4-[4-(2-methylphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4- [4-(2,5 -dimethoxyphenyl)thiazol-2-yl] -5 -methylthiothiophene-2 5 carboxamidine; 4-(4,5-diphenylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-(2-phenyl)thiazol-4-yl-5-methylthiothiophene-2-carboxamidine; 4-[4-(2-chloro-3-pyridyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(phenoxymethyl)thiazol-2-yl]-5-niethylthiothiophene-2-carboxamidine; 0 4-(4-cyclohexylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-[4-(4-chlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(2-hydroxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(3 -trifluoromethoxyphenyl)thiazol-2-yl] -5 -methylthiothiophene-2 carboxamidine; 5 4-[4-(2-chloro-4-pyridyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-(5-phenyl-2-pyridyl)-5-methylthiothiophene-2-carboxamidine; 4- [2-(2-chlorophenylamino)thiazol-4-yl] -5 -methylthiothiophene-2 carboxamidine; 4- [2-(3 -methoxyphenylamino)thiazol-4-yl] -5 -methylthiothiophene-2 0 carboxamidine; 4-[2-(phenylamino)thiazol-4-yl]-5-methylthiothiophene-2-carboxamidine; 4- [2-(2,5-dimethoxyphenylamino)thiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; 4-(2-aminothiazol-4-yl)-5-methylthiothiophene-2-carboxamidine; WO 00/47578 PCT/US99/1 8065 -18 5 4- [ 2
-(
4 -chloro-2-methylphenylamino)thiazol4yl] -5-methylthiothiophene-2 carboxamidine; 4- [ 2
-(
4 -dimethylaminophenylamino)thiazol4yl] -5 -methylthiothiophene-2 carboxamidine; 4- [ 2
-(
4 -methoxyphenylamino)thiazol-4.yl] -5 -methylthiothiophene-2 10 carboxamidine; 4- [4-(4-hydroxy-3 -methoxyphenyl)thiazol-2-yI] -5 -methylthiothiophene-2 carboxamidine; 4- [4-(3 -hydroxy-4-methoxyphenyl)thiazol.2 .yl] -5 -methylIthiothiophene-2 carboxamidine; 5 4- [ 2 -(2-fluorophenylamino)thiazo 4-yl] - -methylthiothiophene-2 carboxamidine; 4- [2-(2,4,5 -trimethylphenyl)aminothiazol-4-y1]-5 -methylthiothiophene-2 carboxamidine; 4- [2-(3 -chloro-2-methylphenyl)amindthiazol4yl] -5 -methylthiothiophene-2 0 carboxamidine; 4- [ 2
-(
2 -isopropylphenyl)aminothiazol4-y I] -5 -methylthi othiophene-2 carboxamidine; 4- [ 2 -(4-benzyloxyphenyl)aminothiazoI -4yl] -5-methylthiothiophene-2 carboxamidine; 5 4
-[
2
-(
2 -bromophenyl)aminothiazo I 4y1> 5 methylthiothiophene-2 carboxamidine; 4- [ 2
-(
2 ,5 -dichlorophenyl)aminothiazo I-4-y1] -5 -methylthiothiophene-2 carboxamidine; 4- [ 2
-(
2 -bromo-4-methylphenyl)aminothiazol-4yl] -5 -methylthiothiophene-2 0 carboxamidine; 4- [2-(2,3 -dichlorophenyl)aminothiazol4-y1] -5 -methylthiothiophene-2 carboxamidine; 4- [2-(3 , 4 ,5 -trimethoxyphenyl)aminothiazol.4-y1] -5 -methylthiothiophene-2 carboxamidine; WO 00/47578 PCT/US99/18065 -19 5 4
-[
2
-(
2 -piperidinylethyl)aminothiazol-4yl]-5-methylthiothiophene-2 carboxamidine; 4- [2-(4-methylphenyl)aminothiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; 4-(4-phenyloxazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 0 4-[2-(diphenylmethyl)aminothiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; and 4-[2-(3-phenylpropyl)aminothiazol-4-yl]-5-methylthiothiophene-2 carboxamidine, as well as pharmaceutically acceptable salts thereof, for example the 5 hydrochloride, hydrobromide and acetate salts thereof, or a prodrug thereof. A second preferred group of compounds falling within the scope of the present invention include compounds ofFormula I wherein X is sulfur or oxygen; Y is a covalent bond or -NH-; Z is NRR 6 ; R' is hydrogen, amino, hydroxy or halogen; R 4 , R 5 and R 6 are independently hydrogen, C,- 4 alkyl, amino, C 1 4 alkoxy 0 or hydroxy, and are preferably all hydrogen; one of R 2 or R 3 is hydrogen, C, 6 alkylthio, C 6 alkyl optionally substituted with OH, NH 2 , COOH or aminocarbonyl, or C,- 6 alkoxy; and the other of R 2 or R 3 is: I Ar l RH ,I where: 5 Ar is a group selected from the group consisting of phenyl, thiazolyl, thiazolinyl, oxazolyl, isothiazolyl, isoxazolyl, furanyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, thienyl (thiophenyl), tetrazolyl, pyrrolyl, pyrazolyl, oxadiazolyl, oxazolinyl, isoxazolinyl, imidazolinyl, triazolyl, pyrrolinyl, benzothiazolyl, benzothienyl, benzimidazolyl, 1,3-oxazolidin-2-onyl, imidazolin 0 2-onyl (preferably phenyl, thiazolyl, thiazolinyl, oxazolinyl, isothiazolyl, isoxazolyl, imidazolyl, pyridyl, pyrimidinyl, thienyl, pyrrolyl, oxazolinyl and benzothienyl), any of which can optionally include an exocyclic = O (keto) or = WO 00/47578 PCT/US99/18065 -20 5 NR' (imino) group, where R' is alkyl, aryl, aralkyl, alkylamino, arylimino or aralkylimino; and
R
8 and R 9 are independently selected from the group consisting of hydrogen, halogen, amino, mono(C 1
.-
4 )alkylamino, di(C .- 4 )alkylamino, arylamino, mono- and di- (C 6
-
1 4 )arylamino, mono- and di-(C 6
-
1 4 )ar(C..- 6 )alkylamino, 0 formylamino, C 2
-
6 acylamino, aminocarbonyl, C 2 -8 aminoacyl, C 2
.-
6 thioacylamino, aminothiocarbonyl, C 2
.-
8 aminothioacyl, C 1
-
6 alkyl, C 3
.
8 cycloalkyl, C,.
6 alkoxy, carboxy, carboxy(C.- 6 )alkyl, C 2
.-
8 alkoxycarbonyl, nitro, cyano, trifluoromethyl, thiazolyl, thiazolinyl, oxazolyl, isothiazolyl, isoxazolyl, furanyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, thienyl (thiophenyl), tetrazolyl, pyrrolyl, 5 pyrazolyl, oxadiazolyl, oxazolinyl, isoxazolinyl, imidazolinyl, triazolyl, pyrrolinyl, benzothiazolyl, benzothienyl, benzimidazolyl, 1,3-oxazolidin-2-onyl, imidazolin-2-onyl, C 6
-
14 aryloxy, C.- 6 alkylthio, C 6
-
1 4 arylthio, C 6
-
14 aryl, or C 6
.-
1 4 ar(C,-_ 6 )alkyl, wherein the aforementioned heteroaryl groups and the aryl portions of C 6
.
1 4 aryloxy, mono- and di C6.
14 aryl amino, mono- and di- C 6
.
14 0 ar(Cl- 6 )alkylamino, C 6
-
1 4 arylthio, C 6
.-
14 ar(Cl.
6 )alkyl, and C 6
_
4 aryl can be further optionally substituted, preferably by one, two or three of halogen, hydroxy, amino, mono(C 1 l.
4 )alkylamino, di(C- 1
.
4 )alkylamino, formylamino, C 1
.
4 acylamino,
C,-
4 aminoacyl, mono- or di-(C,.- 4 )alkylaminocarbonyl, thiocarbonylamino,
C
1
.-
4 thioacylamino, aminothiocarbonyl, C-.
4 alkoxy, C 6
.-
1 0 aryloxy, 5 aminocarbonyloxy, mono- or di(C.- 4 )alkylaminocarbonyloxy, mono- or di(C 6 -io)arylaminocarbonyloxy, mono- or di(C.
1 5 )aralkylaminocarbonyloxy,
C
1 _ 4 alkylsulfonyl, C 6
.
1 0 arylsulfonyl, (C 7
-
1 is)aralkylsulfonyl, Cl- 4 alkylsulfonylamino,
C
6
-,
10 arylsulfonylamino, (C 7 .- )aralkylsulfonylamino, aminosulfonyl, mono- and di-alkylaminosulfonyl, mono- and di-arylaminosulfonyl, mono- and di 0 aralkylaminosulfonyl, C- 4 alkoxycarbonylamino, C 7
-
1 5 aralkoxycarbonylamino,
C
6 _,aryloxycarbonylamino, mono- or di-(C 1
-.
4 )alkylaminothiocarbonyl, C 7 ,saralkoxy, carboxy, carboxy(C.- 4 )alkyl, Cl- 4 alkoxycarbonyl, C 1 , 4 alkoxycarbonylalkyl, carboxy(C,.
4 )alkoxy, alkoxycarbonylalkoxy, nitro, cyano, trifluoromethyl, C 1
.-
4 alkylthio and C 6
.-
1 0 arylthio, or by 3,4-methylenedioxy, 3,4 5 ethylenedioxy, and 3,4-propylenedioxy.
WO 00/47578 PCT/US99/18065 -21 5 Preferred values of R' and R 9 are halogen, C 1 6 alkyl, Cl- 6 alkoxy, hydroxy, nitro, trifluoromethyl, C 6 -1 0 aryl (further optionally substituted by one or two of chloro, halogen, C- 6 alkyl, C 1
.
6 alkoxy, hydroxy, nitro, trifluoromethyl, carboxy, 3,4-methylenedioxy, 3,4-ethylenedioxy, 3,4-propylenedioxy, or amino), 4-phenylphenyl (biphenyl), C,.
6 aminoalkyl, carboxy, C, 6 _ alkyl, 0 3,4-methylenedioxy, 3,4-ethylenedioxy, 3,4-propylenedioxy, amino, C_-6 alkanoylamino, C 6
-
14 aroylamino, C 1
-
6 hydroxyalkyl, thienyl (further optionally substituted by one or two of chloro, amino, methyl, methoxy, or hydroxy) and tetrazolyl. More preferably, R 2 is thienyl, oxazolyl, or thiazolyl, optionally substituted by any of the aforementioned groups. 5 Examples of preferred R' and R 9 groups include 4-chlorophenyl, 2,4-dichlorophenyl, methyl, 4-nitrophenyl, 3-nitrophenyl, 4-methoxyphenyl, 3-methoxyphenyl, 2-methoxyphenyl, 3-(2,4-dimethylthien-5-yl)phenyl, 3-hydroxyphenyl, 5-(carboxymethyl)thien-2-yl, phenyl, 3,4-ethylenedioxyphenyl, 3,4-propylenedioxyphenyl, naphth-'-yl, 3-phenyl-4-(tetrazol-5-yl)phenyl, 0 2,4-dichlorophenyl), 4-phenylphenyl, 3-methoxyphenyl, 3-hydroxyphenyl, 3-phenylphenyl, phenylthiomethyl, 2-chloro-4,5-dimethoxyphenyl, 4-chloro-3 methylphenyl, 5-methyl-4-phenyl, 4-chloro-3-nitrophenyl, 3-fluoro-5-trifluoromethylphenyl, 3,5-bis(trifluoromethyl), 3-fluoro-5 trifluoromethylphenyl, 3-bromophenol, 3,4-methylenedioxyphenyl, 5 4-methylphenyl, 3-methylphenyl, 3,5-bis(trifluoromethyl)phenyl, 2-methoxyphenyl, 6-phenyl-2-pyridyl, 2,4-dimethoxyphenyl, 3,4 dimethoxyphenyl, benzyl, 3,4-dichlorophenyl, 3-methylphenyl, 3,5 dimethoxyphenyl, 2-methylphenyl, 2,5-dimethoxyphenyl, 2-chloro-3-pyridyl, phenoxymethyl, cyclohexyl, 2-hydroxyphenyl, 3-trifluoromethoxyphenyl, 2 0 chloro-4-pyridyl, 3-chloro-4-pyridyl, 2-chlorophenylamino, 3 methoxyphenylamino, phenylamino, 2,5-dimethoxyphenylamino, amino, 4 chloro-2-methylphenylamino, 4-dimethylaminophenylamino, 4-methoxyphenylamino, 4-hydroxy-3-methoxyphenyl, 3-hydroxy-4 methoxyphenyl, 2-fluorophenylamino, 2,4,5-trimethylphenylamino, 3-chloro-2 5 methylphenylamino, 2-isopropylphenylamino, 4-benzyloxyphenylamino, 2- WO 00/47578 PCT/US99/18065 -22 5 bromophenylamino, 2 ,5-dichlorophenylamino, 2-bromo-4-methylphenylamino, 2,3-dichlorophenylamino, 3,4,5-trimethoxyphenylamino, 2-piperidinylethylamino, 4-methylphenylamino, 2-thienyl, 2-5,6,7,8 tetrahydronaphthyl, 3-(2-phenoxyacetic acid)phenyl, 2-(2-phenoxyacetic acid)phenyl, diphenylmethylamino, 3-phenylpropylamino, 3-phenylphenyl, 0 phenylthiomethyl, 2-chloro-4,5-dimethoxyphenyl, and isopropyl. A third preferred group of compounds are those of Formula I wherein: X is sulfur; Y is a covalent bond; Z is NRR 6 ; 5 R' is hydrogen;
R
3 is methylthio or methyl;
R
4 , R 5 and R 6 are all hydrogen; and
R
2 is Formula II, where Ar is phenyl, thiazolyl, oxazolyl, benzothienyl, pyridyl, or imidazolyl; aiid R 8 and R 9 are independently hydrogen, 0 or C 6
.
1 0 aryl or heterocycle, optionally substituted by one, two or three of chloro, hydroxy, Cl- 4 alkyl, C 3
-
6 cycloalkyl, C1-4 alkoxy, amino, carboxy, phenyl, naphthyl, biphenyl, hydroxyphenyl, methoxyphenyl, dimethoxyphenyl, carboxyalkoxyphenyl, alkoxycarbonylalkoxy, carboxyethoxy, alkylsulfonylaminophenyl, arylsulfonylaminophenyl, acylsulfonylaminophenyl, 5 aralkylsulfonylaminophenyl, heteroarylsulfonylaminophenyl where the heteroaryl portion is optionally halo or C, 6 alkyl substituted, chlorophenyl, dichlorophenyl, aminophenyl, carboxyphenyl, nitrophenyl, or by 3,4-methylenedioxy, 3,4 ethylenedioxy, and 3,4-propylenedioxy. A fourth preferred group of compounds are those of Formula I wherein: 0 X is sulfur; Y is a direct covalent bond; Z is NR'R 6 ; R' is hydrogen;
R
2 is alkyl, ar(alkyl), alkylsulfonyl, -SO 2 -alkyl, amido, amidino, 5 or WO 00/47578 PCT/US99/18065 -23 Ar 5 R 9 where Ar is an aromatic or heteroaromatic group selected from the group consisting of phenyl, thiazolyl, oxazolyl, imidazolyl and pyridyl; R' and R 9 are independently selected from the group consisting of 0 hydrogen, carboxy, phenyl, naphthyl, alkyl, pyridyl, oxazolyl, furanyl, cycloalkyl and amino, any of which may be optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, alkyl, haloalkyl, alkaryl, heteroaryl, phenyl, naphthyl, alkoxy, aryloxy, hydroxy, amino nitro, thiophenyl, benzothiophenyl, fluorenyl, 3,4-ethylenedioxy, 3,4-methylenedioxy, 5 3,4-propylenedioxy, arylsulfonamido, alkylsulfonamido and aryloxy, each of said 1 to 3 substituents may be further optionally substituted with one or more groups selected from alkoxy, haloalkyl, halogen, alkyl, amino, acetyl, hydroxy, dialkylamino, dialkylamino acyl, monoalkylaminoacyl, -SO 2 -heteroaryl,
-SO
2 -aryl, or aryl; 0 R' is -SO 2 -alkyl, trifluoromethyl, S(O)-alkyl, hydrogen, alkoxy, alkylthio, alkyl, aralkylthio; and
R
4 , R 5 , R 6 are hydrogen. Preferred compounds of this embodiment are those where Ar is a thiazolyl, preferably thiazol-2-yl or thiazol-4-yl, and at least one of R' and R 9 is 5 substituted phenyl, most preferably on the 4-position of the thiazol-2-yl group. Also preferred are compounds where R 2 is a 4-phenylthiazol-2-yl group wherein said phenyl is further optionally substituted. and R 3 is methylthio. A fifth preferred group of compounds are those of Formula III: WO 00/47578 PCT/US99/18065 -24 0 O R 7 N 5 /III NH A S
NH
2 or a pharmaceutically acceptable salt or prodrug thereof, where A is methylthio or methyl; G' is -0-, -S-, -NH-, or a covalent bond; n is an integer from 1-10, preferably from 1-6; 0 m is an integer from 0-1; and R' and R" are independently selected from hydrogen, alkyl, aryl or aralkyl, or R' and R" are taken together with the N atom to which they are attached form a 3-8 membered heterocyclic ring, optionally containing an additional O, N, or S atom, and when said 3-8 membered heterocyclic ring 5 contains an additional N atom, said additional N atom is optionally substituted by hydrogen, Cl 14 alkyl, C 6
-
1 0 aryl, C 6 -o 10 ar(Cl_ 4 )alkyl, acyl, alkoxycarbonyl or benzyloxycarbonyl. Most preferred compounds of Formula III are those for which R' and R", taken together with the N atom to which they are attached, form a ring selected 10 from piperazinyl, pyrrolidinyl, piperidinyl or morpholinyl, which are further optionally substituted with 1 to 4 non-hydrogen substituents selected from halogen, hydroxy, amino, monoalkylamino, dialkylamino, formylamino, acylamino, aminoacyl, mono- or di- alkylaminocarbonyl, thiocarbonylamino, thioacylamino, aminothiocarbonyl, alkoxy, aryloxy, aminocarbonyloxy, mono .5 or di-alkylaminocarbonyloxy, mono- or diarylaminocarbonyloxy, mono- or diarakylaminocarbonyloxy, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, WO 00/47578 PCT/US99/18065 -25 5 alkylsulfonylamino, arylsulfonylamino, arakylsulfonylamino, alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, mono- or di- alkylaminothiocarbonyl, aralkoxy, carboxy, carboxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, nitro, cyano, trifluoromethyl, alkylthio and arylthio, where each of these substituents has the preferred values set forth for Formulae I and II 10 above. Examples of preferred compounds of Formula III include: 5-methylthio-4-[4-(3- { [N-(2-morpholin-4 ylethyl)carbamoyl]methoxy} phenyl)( 1,3 -thiazol-2-yl)]thiophene-2 carboxamidine, 15 5-methylthio-4- {4-[3 -(2-morpholin-4-yl-2-oxoethoxy)phenyl](1,3-thiazol-2 yl) } thiophene-2-carboxamidine, 5-methylthio-4- {4-[3-(2-oxo-2-piperazinylethoxy)phenyl](1,3-thiazol-2 yl) }thiophene-2-carboxamidine, 4-[4-(3- { [N-(2-aminoethyl)carbamoyl]methoxy } phenyl)(1,3-thiazol-2-yl)]-5 20 methylthiothiophene-2-carboxamidine, 4-(4- { 3- [2-(4-acetylpiperazinyl)-2-oxoethoxy]phenyl} (1,3 -thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine, 4-(4- { 3- [2-(4-methylpiperazinyl)-2-oxoethoxy]phenyl } (1,3 -thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine, 25 the compound described in Example 151, 5-methylthio-4-[4-(3- {2-oxo-2-[4-benzylpiperazinyl]ethoxy}phenyl)(1,3-thiazol 2-yl)]thiophene-2-carboxamidine, (D,L)-4-(4- { 3- [2-(3-aminopyrrolidinyl)-2-oxoethoxy]phenyl } (1,3-thiazol-2-yl)) 5-methylthiothiophene-2-carboxamidine, 30 5-methylthio-4- {4-[3-(2-oxo-2-piperidylethoxy)phenyl](1,3-thiazol-2 yl) } thiophene-2-carboxamidine, (D,L)-ethyl 1-(2- { 3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy } acetyl)piperidine-2-carboxylate, 5-methylthio-4- {4-[3-(2-oxo-2-pyrrolidinylethoxy)phenyl](1,3-thiazol-2 35 yl)}thiophene-2-carboxamidine, WO 00/47578 PCT/US99/18065 -26 5 5-methylthio-4-[4-(3-{2-oxo-2-[4-benzylpiperidyl]ethoxy}phenyl)(1,3-thiazol-2 yl)]thiophene-2-carboxamidine, (D,L)-4-(4-{3-[2-(3-methylpiperidyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine, 4-(4-{3-[2-(4-methylpiperidyl)-2-oxoethoxy]phenyl } (1,3-thiazol-2-yl))-5 0 methylthiothiophene-2-carboxamidine, 4-(4-{3-[2-(2-azabicyclo[4.4.0]dec-2-yl)-2-oxoethoxy]phenyl }(1,3-thiazol-2-yl)) 5-methylthiothiophene-2-carboxamidine, (D,L)-ethyl 1-(2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetyl)piperidine-3-carboxylate, 5 5-methylthio-4- { 4-[3-(2-oxo-2-(1,2,3,4-tetrahydroquinolyl)ethoxy)phenyl](1,3 thiazol-2-yl) } thiophene-2-carboxamidine, ethyl 1-(2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy } acetyl)piperidine-4-carboxylate, 4-(4- {3-[2-((3R)-3-hydroxypiperidyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 0 methylthiothiophene-2-carboxamidine, D,L-4-(4- {3-[2-(2-ethylpiperidyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine, 4-(4- { 3-[2-((3 S)-3-hydroxypyrrolidinyl)-2-oxoethoxy]phenyl } (1,3-thiazol-2-yl)) 5-methylthiothiophene-2-carboxamidine, 5 D,L-4-[4-(3- {2-[3-(hydroxymethyl)piperidyl]-2-oxoethoxy }phenyl)(1,3-thiazol-2 yl)]-5-methylthiothiophene-2-carboxamidine, 4- {4-[3 -(2- {(2R)-2-[(phenylamino)methyl]pyrrolidinyl}-2 oxoethoxy)phenyl](1,3-thiazol-2-yl)}-5-methylthiothiophene-2-carboxamidine, 4-[4-(3- {2-[(3R)-3-(methoxymethyl)pyrrolidinyl]-2-oxoethoxy }phenyl)(1,3 0 thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine, 1-(2- { 3- [2-(5-amidino-2-methylthio-3-thienyl)- 1,3-thiazol-4 yl]phenoxy } acetyl)piperidine-3-carboxamide, and 2- {3-[2-(5- { [(tert-butoxy)carbonylamino]iminomethyl }-2-methyl-3-thienyl)-l1,3 thiazol-4-yl]phenoxy } acetic acid; 5 or pharmaceutically acceptable salts or prodrugs thereof.
WO 00/47578 PCT/US99/18065 -27 5 A sixth preferred group of compounds are those of Formula IV: R"' HN IV NH A S
NH
2 or a pharmaceutically acceptable salt or prodrug thereof, where A is methylthio or methyl; and R" is hydrogen, C 6
-
14 aryl, C 1
-
6 alkyl, Cl- 6 alkoxy (C 6
-
14 )aryl, amino(C 6
-
1 4 )aryl, 0 monoalkylamino(C 6
-
1 4 )aryl, dialkylamino(C 6
-
1 4 )aryl, C 6 -oar(C,- 6 )alkyl, heterocycle(C 2
-
6 )alkyl such as morphdlinoalkyl, piperazinylalkyl and the like,
C,
6 alk(C 6
-
14 )aryl, amino(C- 6 )alkyl, mono(C 1
.
6 )alkylamino(Cl.
6 )alkyl, di(C - 6 )alkylamino(C _ 6 )alkyl, hydroxy(C 6 -1 4 )aryl, or hydroxy(C - 6 )alkyl, where the aryl and heterocyclic rings can be further optionally substituted by 1-4 non 5 hydrogen substituents selected from halogen, hydroxy, amino, mono(C- 6 )alkylamino, di(Cl- 6 )alkylamino, formylamino, (C,.
6 )acylamino, amino(C 1
_
6 )acyl, mono- or di-(C,- 6 )alkylaminocarbonyl, thiocarbonylamino, (C - 6 )thioacylamino, aminothiocarbonyl, (CI-)alkoxy, (C 6 o- 10 )aryloxy, aminocarbonyloxy, mono- or di-(C- 6 )alkylaminocarbonyloxy, mono- or di 0 (C 6 -o)arylaminocarbonyloxy, mono- or di(C 6
-
1 0 )ar(C 1
-
6 )alkylaminocarbonyloxy,
(C
1
_
6 )alkylsulfonyl, (C 6 .o 1 )arylsulfonyl, (C 6
-
10 )ar(C 1
-
6 )alkylsulfonyl, (C 1 6 )alkylsulfonylamino, C 6
.,
0 arylsulfonylamino,
(C
6 . .o)ar(Ci .
6 )alkylsulfonyl amino,(C 6 )alkoxycarbonylamino,
(C
6 o-io)ar(C 1
-
6 )alkoxycarbonylamino, C 6
-
1 0 aryloxycarbonylamino, mono- or di- (C 1 . 5 6 )alkylaminothiocarbonyl, (C 6
-
1 0 )ar(Cl- 6 )alkoxy, carboxy, (Cl- 6 )carboxyalkyl, C - 6 alkoxycarbonyl, (C - 6 )alkoxycarbonyl(C 1 6 )alkyl, nitro, cyano, trifluoromethyl,
(C
1
-
6 )alkylthio and C 6
.
1 0 arylthio. Examples of preferred compounds of Formula IV include: WO 00/47578 PCT/US99/18065 -28 5 4- {2-[(3-methoxyphenyl)amino](1,3-thiazol-4-yl) }-5-methylthiothiophene-2 carboxamidine, 4- { 2-[(4-methoxyphenyl)amino](1,3-thiazol-4-yl) } -5-methylthiothiophene-2 carboxamidine, 4-(2- { [4-(dimethylamino)phenyl]amino}(1,3-thiazol-4-yl))-5 10 methylthiothiophene-2-carboxamidine, 4- {2-[(4-chloro-2-methylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine, 4- {2-[(diphenylmethyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2 carboxamidine, 15 5-methylthio-4- {2-[(3-phenylpropyl)amino](1,3-thiazol-4-yl) } thiophene-2 carboxamidine, 5-methylthio-4- {2-[(2,4,5-trimethylphenyl)amino](1,3-thiazol-4-yl)}thiophene-2 carboxamidine, 4- { 2-[(2-fluorophenyl)amino](1,3-tlilazol-4-yl) } -5-methylthiothiophene-2 20 carboxamidine, 4- {2-[(3-chloro-2-methylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine, 4-(2- { [2-(methylethyl)phenyl]amino } (1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxamidine, 25 5-methylthio-4-(2- {[4-(phenylmethoxy)phenyl]amino} (1,3-thiazol-4 yl))thiophene-2-carboxamidine, 4-{2-[(2-bromophenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2 carboxamidine, 4- {2-[(2,6-dichlorophenyl)amino](1,3-thiazol-4-yl) }-5-methylthiothiophene-2 30 carboxamidine, 4- {2-[(2-bromo-4-methylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine, 5-methylthio-4- {2-[(2-morpholin-4-ylethyl)amino](1,3-thiazol-4-yl)}thiophene-2 carboxamidine, WO 00/47578 PCT/US99/18065 -29 5 4- { 2- [(2,3-dichlorophenyl)amino](1,3-thiazol-4-yl) }-5-methylthiothiophene-2 carboxamidine, 5-methylthio-4- {2-[(3,4,5-trimethoxyphenyl)amino](1,3-thiazol-4-yl)}thiophene 2-carboxamidine, 5-methylthio-4- {2-[(2-piperidylethyl)amino](1,3-thiazol-4-yl)}thiophene-2 10 carboxamidine, 4-(2- { [(4-methylphenyl)methyl]amino}(1,3-thiazol-4-yl))-5-methylthiothiophene 2-carboxamidine, 4-(2- { [4-(4-chlorophenoxy)phenyl]amino } (1,3 -thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine, 15 4-(2- { [4-phenoxyphenyl]amino } (1,3 -thiazol-4-yl))-5 -methylthiothiophene-2 carboxamidine, 5-methylthio-4-(2-{ [4-(phenylamino)phenyl]amino } (1,3-thiazol-4-yl))thiophene 2-carboxamidine, 5-methylthio-4-(2-{ [4-benzylphenylfamino } (1,3-thiazol-4-yl))thiophene-2 20 carboxamidine, 5-methylthio-4-(2-{ [4-(piperidylsulfonyl)phenyl]amino } (1,3-thiazol-4 yl))thiophene-2-carboxamidine, 5-methylthio-4-[2-(3-quinolylamino)( 1,3-thiazol-4-yl)]thiophene-2 carboxamidine, 25 5-methylthio-4-[2-(2-naphthylamino)(1,3 -thiazol-4-yl)]thiophene-2 carboxamidine, 4-[2-(2H-benzo[3,4-d] 1,3-dioxolan-5-ylamino)(1,3-thiazol-4-yl)]-5 methylthiothiophene-2-carboxamidine, 4- {2-[(7-bromofluoren-2-yl)amino](1,3-thiazol-4-yl) } -5-methylthiothiophene-2 30 carboxamidine, 4-{ 2-[(4-cyclohexylphenyl)amino](1,3-thiazol-4-yl) }-5-methylthiothiophene-2 carboxamidine, 5-methylthio-4-(2- { [4-(phenyldiazenyl)phenyl]amino } (1,3-thiazol-4 yl))thiophene-2-carboxamidine, WO 00/47578 PCT/US99/18065 -30 5 5-methylthio 4-(2- { [3-(hydroxymethyl)phenyl]amino } (1,3-thiazol-4-yl)) thiophene-2-carboxamidine, 4-[2-({3-[(3-methylpiperidyl)methyl]phenyl } amino)(1,3-thiazol-4-yl)]-5 methylthiothiophene-2-carboxamidine, 4- { 2-[(3-hydroxyphenyl)amino](1,3-thiazol-4-yl) } -5-methylthiothiophene-2 0 carboxamidine, 4-(2-{ [4-(carbamoylmethoxy)phenyl] amino } (1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine, 5-methyl-4- {2-[(3,4,5-trimethoxyphenyl)amino](1,3-thiazol-4-yl) } thiophene-2 carboxamidine, 5 5-methyl-4- {2-[(4-phenoxyphenyl)amino]( 1,3-thiazol-4-yl)}thiophene-2 carboxamidine, 5-methyl-4-[2-(phenylamino)(1,3-thiazol-4-yl)]thiophene-2-carboxamidine, and 4-(4-isoxazol-5-yl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine; as well as pharmaceutically acceptable" salts and prodrugs thereof. 0 A seventh preferred group of compounds are compounds of Formula I, or a pharmaceutically acceptable salt or prodrug thereof, wherein: X is sulfur or oxygen, preferably sulfur; Y is a covalent bond or -NH-, preferably a covalent bond; Z is NR'R 6 ; 5 R' is hydrogen, amino, hydroxy or halogen, preferably hydrogen;
R
4 , R 5 and R' are independently hydrogen, C 1
-
4 alkyl, amino, C 1
-
4 alkoxy or hydroxy, and are preferably all hydrogen; R' is hydrogen, C,- 6 alkylthio, Cl- 6 alkyl optionally substituted with OH,
NH
2 , COOH or aminocarbonyl, or C, 6 alkoxy, preferably methylthio or methyl; 0 and
R
2 is alkylsulfonylamino, aralkylsulfonylamino, aralkenylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, di(aralkylsulfonyl)amino, 5 di(aralkenylsulfonyl)amino, di(arylsulfonyl)amino, or di- WO 00/47578 PCT/US99/18065 -31 (heteroarylsulfonyl)amino, wherein any of the aryl or heteroaryl containing groups can be optionally substituted on the aromatic ring; or amino, monoalkylamino, dialkylamino, monoarylamino, diarylamino, monoalkylmonoarylamino, Smonoaralkylamino, diaralkylamino, monoalkylmonoaralkylamino, monoheterocycleamino, diheterocycleamino, monoalkylmonoheterocycleamino, wherein any of the aryl or heteroaryl containing groups can be optionally substituted on the aromatic ring and 5 wherein any of the heterocycle containing groups can be optionally ring substituted; or alkanoylamino, alkenoylamino, alkynoylamino, aroylamino, aralkanoylamino, aralkenoylamino, heteroaroylamino, heteroarylalkandylamino, any of which is optionally 0 substituted on the aromatic ring; or alkoxy and alkylthio, either of which is optionally substituted, or aryloxy, aralkoxy, arylthio, aralkylthio, arylsulfonyl, aralkylsulfonyl, aralkenylsulfonyl, any of which is optionally substituted on the aromatic ring; or 5 alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, wherein any of the aryl containing groups can be optionally substituted on the aromatic ring; or formylamino, H(S)CNH-, or thioacylamino. 0 Preferred optional substituents are halogen, C- 6 alkyl, CI- 6 alkoxy, hydroxy, nitro, trifluoromethyl, C 6
-
1 0 aryl, C 6
-
1 0 aryloxy, C 6
.
10 arylmethoxy (wherein the aryl groups on these aryl-containing substituents are further optionally substituted by one or two of chloro, halogen, C- 6 alkyl, C, 6 _ alkoxy, phenyl, hydroxy, nitro, trifluoromethyl, carboxy, 3,4-methylenedioxy, 5 3,4-ethylenedioxy, 3,4-propylenedioxy, or amino), C 1
_
6 aminoalkyl, carboxy, WO 00/47578 PCT/US99/18065 -32 5 alkyl, 3,4-methylenedioxy, 3,4-ethylenedioxy, 3,4-propylenedioxy, amino, mono ordi- (CI- 6 )alkylamino, mono- or di- C 6 -1 0 arylamino, C 1
.
6 alkylsulfonylamino, C 6 . 1 0 arylsulfonylamino, C 1
.
8 acylamino, C, 1
.
8 alkoxycarbonyl, Cl- 6 alkanoylamino, C 6 1 4 aroylamino, C1.6 hydroxyalkyl, methylsulfonyl, phenylsulfonyl, thienyl (further optionally substituted by one or two of chloro, amino, methyl, methoxy, or 0 hydroxy) and tetrazolyl. In one aspect of this embodiment, R 2 is preferably C-.
6 alkylsulfonylamino, C 6
.
10 ar(C 1
-
6 )alkylsulfonylamino, C 6
.
1 0 o ar(C2- 6 )alkenylsulfonylamino, C 6
.
10 arylsulfonylamino, heteroarylsulfonylamino, di(C 6 1 0 ar(Cl- 6 )alkylsulfonyl)amino, di(C 6 o- 1 0 ar(C 2
-
6 )alkenylsulfonyl)amino, di(C 6 . 5 1 0 arylsulfonyl)amino, or di-(heteroarylsulfonyl)amino, wherein any of the aryl or heteroaryl containing groups can be optionally substituted on the aromatic ring. Especially preferred R2 groups in this embodiment of the invention include C 6
.
10 arylsulfonylamino, di-(C 6 o- 10 arylsulfonyl)amino, C 6
-
10 ar(C _ 3 )alkylsulfonylamino, di-(C 6
.
10 ar(C 1
.
3 )alkylsulfonyl)amino, 0 thienylsulfonylamino, any of which is optionally substituted on the aromatic ring. Useful values of R2, when R 2 is a substituted sulfonylamino group include biphenylsulfonylamino, bis(biphenylsulfonyl)amino, naphth-2-ylsulfonylamino, di(naphth-2-ylsulfonyl)amino, 6-bromonaphth-2-ylsulfonylamino, di(6 bromonaphth-2-ylsulfonyl)amino, naphth-1-ylsulfonylamino, di(naphth- 1 .5 ylsulfonyl)amino, 2-methylphenylsulfonylamino, di-(2 methylphenylsulfonyl)amino, 3-methylphenylsulfonylamino, di-(3 methylphenylsulfonyl)amino, 4-methylphenylsulfonylamino, di-(4 methylphenylsulfonyl)amino, benzylsulfonylamino, 4 methoxyphenylsulfonylamino, di-(4-methoxyphenylsulfonyl)amino, 4 30 iodophenylsulfonylamino, di-(4-iodophenylsulfonyl)amino, 3,4 dimethoxyphenylsulfonylamino, bis-(3,4-dimethoxyphenylsulfonyl)amino, 2-chlorophenylsulfonylamino, di-(2-chlorophenylsulfonyl)amino, 3-chlorophenylsulfonylamino, di-(3-chlorophenylsulfonyl)amino, 4-chlorophenylsulfonylamino, di-(4-chlorophenylsulfonyl)amino, 35 phenylsulfonylamino, di-(phenylsulfonyl)amino, 4-tert- WO 00/47578 PCT/US99/18065 -33 butylphenylsulfonylamino, di-(4-tert-butylphenylsulfonyl)amino, 2 phenylethenylsulfonylamino, and 4-(phenylsulfonyl)thien-2-ylsulfonylamino. In another aspect of this embodiment, R 2 is preferably amino, mono(Cl- 6 )alkylamino, di(C- 6 )alkylamino, mono(C 6
-
10 )arylamino, di(C 6 -o 1 0 )arylamino, mono(C .
6 )alkylmono(C 6
-
1 0 o)arylamino, Imonoar(CI- 6 )alkylamino, di(C 6
-
1 0 )ar(C1- 6 )alkylamino, mono(Cl- 6 )alkylmono(C 6 . io)ar(Cl- 6 )alkylamino, monoheteroarylamino, diheteroarylamino, mono(Cl- 6 )alkylmonoheteroarylamino, wherein any of the aryl or heteroaryl containing groups can be optionally substituted on the aromatic ring. Especially preferred R2 groups in this embodiment of the invention 5 include mono(C 6
-
1 0 )arylamino, mono(C .
6 )alkylmono(C6- 10 )arylamino, mono(C-I 0 )ar(C.
3 )alkylamino, mono(C 6 _)alkylmono(C6- 1
I
0 )ar(C .- 3 )alkylamino, monoheteroarylamino, and mono(Cl- 6 )alkylmonoheteroarylamino. Examples of suitable heteroarylamino groups include 1,3-thiazol-2-ylamino, imidazol-4 ylamino, quinolin-2-ylamino and quin6lin-6-ylamino. ) Useful values of R 2 , when R 2 is a substituted amino group include anilino, naphth-2-ylamino, naphth-1-ylamino, 4-(biphenyl)thiazol-2-ylamino, 4-(phenyl)thiazol-2-ylamino, 4-phenyl-5-methylthiazol-2-ylamino, 4-hydroxy-4 trifluoromethylthiazol-2-ylamino, 3-phenylphenylamino, pyrimidin-2-ylamino, 4-isopropylphenylamino, 3-isopropylphenylamino, 4-phenylphenylamino, 3 5 fluoro-4-phenylphenylamino, 3,4-methylenedioxyphenylamino, n butylphenylamino, N-methyl-N-(2-methylphenyl)amino, 3-nitrophenylamino, 4 methoxyphenylamino, 3-methoxyphenylamino, 2-methoxyphenylamino, 2 methylphenylamino, 3-methylphenylamino, 3,4-dimethylphenylamino, 3 chlorophenylamino, 4-chlorophenylamino, 4-(3-fluoro-4-methylphenyl)amino, 0 4-(indan-5-yl)amino, benzylamino, indanylmethylamino, 2,3 dihydrobenzofuranylmethyl, 2-phenylimidazol-5-yl, 3-hydroxybenzyl, 3 phenoxyphenylamino, 4-phenoxyphenylamino, 3-benzyloxyphenylamino, 4 benzyloxyphenylamino, quinolin-6-ylamino, quinolin-3-ylamino, 4 (phenylamino)phenylamino, 4-(4-ethylphenyl)phenylamino, 4- WO 00/47578 PCT/US99/18065 -34 5 (dimethylamino)phenylamino, 4-cyclohexylphenylamino, 4-(9-ethylcarbazol-3 yl)amino, 4-(t-butyl)phenylamino, and 4-methylthiophenyl amino. In another aspect of this embodiment, R 2 is preferably an acylamino group, such as alkanoylamino, alkenoylamino, aroylamino, aralkanoylamino, aralkenoylamino, heteroaroylamino, heteroarylalkanoylamino, any of which is 0 optionally substituted on the aromatic ring. Especially preferred R 2 groups in this embodiment of the invention include (C 6
.
1 0 )arylcarbonylamino, C 6
-
0 ar(Cl- 3 )alkylcarbonylamino, C 6
-
1 0 ar(C 2 -3)alkenylcarbonylamino, C 6
-
1 0 aryloxy(Cl- 3 )alkylcarbonylamino, C 3
.
8 cycloalkylcarbonylamino, C 1.6 alkylcarbonylamino, and heteroarylcarbonylamino, .5 such as furanylcarbonylamino, and quinolinylcarbonylamino. Useful values of R 2 , when R 2 is an acylamino group include 3-hydroxyphenylcarbonylamino, 2-phenylethenylcarbonylamino, phenylcarbonylamino, cyclohexylcarbonylamino, 4-methyl-3 nitrophenylcarbonylamino, furan-2-yldarbonylamino, tert-butylcarbonylamino, O 5-(3,5-dichlorophenoxy)furan-2-ylcarbonylamino, naphth- 1 -ylcarbonylamino, quinolin-2-ylcarbonylamino, 4-ethoxyphenylcarbonylamino, phenoxymethylcarbonylamino, and 3-methylphenylcarbonylamino. In another aspect of this embodiment, R 2 is preferably C 6
-
1 0 aryloxy, C 6
-,
10 ar(C,.
6 )alkoxy, C 6
-
0 arylsulfonyl, C 6 10 ar(Cl- 6 )alkylsulfonyl, or C6-10 Z5 ar(C 2
.
6 )alkenylsulfonyl, any of which is optionally substituted on the aromatic ring. Especially preferred R 2 groups in this embodiment of the invention include C 6 -1 0 aryloxy, and C 6
-
10 arylsulfonyl. Useful values of R 2 , when R 2 is an aryloxy or arylsulfonyl group include phenoxy, naphthyloxy, phenylsulfonyl, and naphthylsulfonyl. 30 Representative compounds within the scope of this seventh embodiment of the invention include: 5-methylthio-4-(6-quinolylamino)thiophene-2-carboxamidine 5-methylthio-4-[(3-phenylphenyl)amino]thiophene-2-carboxamidine 5-methylthio-4-(3-quinolylamino)thiophene-2-carboxamidine 35 5-methylthio-4-(pyrimidin-2-ylamino)thiophene-2-carboxamidine WO 00/47578 PCT/US99/18065 -35 5 4-[(4-cyclohexylphenyl)amino]-5 -methylthiothiophene-2-carboxamidine methyl 4-amino-5-methylthiothiophene-2-carboxylate methyl 4- [(aminothioxomethyl)amino]-5 -methylthiothiophene-2-carboxylate 5-methylthio-4- [(4-phenyl( 1,3 -thiazol-2-yl))amino]thiophene-2-carboxamidine 5 -methylthio-4- { [4- (4-pheny lphenyl)( 1,3 -thiazol-2-yl)] amino)} thiophene-2 0 carboxamidine 4- [(5 -methyl-4-phenyl( 1,3 -thiazol-2-yl))amino] -5-methylthiothiophene-2 carboxamidine 4-f{ [4-hydroxy-4-(trifluoromethyl)(1I,3 -thiazolin-2-yl)] amino)} -5 methylthiothiophene-2-carboxamidine 5 5-methylthio-4-(2-naphthylamino)thiophene-2-carboxamidine 4- [(4-chlorophenyl)amino] -5 -methylthiothiophene-2-carboxamidine 4- [(3 -methylphenyl)amino]j-5-methylthiothiophene-2-carboxamidine 4- [(3 -methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4- { [3 -(methylethyl)phenyl] amino I -5-niiethylthiothiophene-2-carboxamidine 0 5-methylthio-4- [(3-nitrophenyl)amino]thiophene-2-carboxamidine 4- { [4-(methylethyl)phenyl] amino I -5-methylthiothiophene-2-carboxamidine 4- [(3 ,4-dimethylphenyl)amino] -5-methylthiothiophene-2-carboxamidine 5-methylthio-4-[(4-phenylphenyl)amino]thiophene-2-earboxamidine 4- [(3 -fluoro-4-phenylphenyl)amino]-5-methylthiothiophene-2-caboxamidine 5 4-(2H-benzo [d] 1 ,3-dioxolen-5-ylamino)-5-methylthliothiophene-2-carboxamidine 4-[(4-butylphenyl)amino]-5 -methylthiothiophene-2-carboxamidine 5-methylthio-4- [benzylamino]thiophene-2-carboxamidine 4-(indan-5-ylamino)-5-methylthiothiophene-2-carboxamidine 4-(2,3-dihydrobenzo[b]furan-5 -ylamino)-5-methylthiothiophene-2-carboxamidine 0 5-methylthio-4-[(2-phenylimidazol-4-yl)amino]thiophene-2-carboxamidine 5-methylthio-4- [(2-quinolylmethyl)aminojthiophene-2-carboxamidine 4-1{[(3 -hydroxyphenyl)methyl] amino } -5 -methylthiothiophene-2-carboxamidine 5-methylthio-4-(phenylcarbonylamino)thiophene-2-carboxamidine 4-((2E)-3 -phenylprop-2-enoylamino)-5-methylthiothiophene-2-carboxamidine 5 4-[(4-chlorophenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine WO 00/47578 PCT/US99/18065 -36 5 4 -(cyclohexylcarbonylamino)-5-methylthiothiophene-2-carboxamidine methyl 4-[(4-methyl-3-nitrophenyl)carbonylamino]-5-methylthiothiophene-2 carboxylate 4-(2-furylcarbonylamino)-5-methylthiothiophene-2-carboxamidine 4-(2,2-dimethylpropanoylamino)-5-methylthiothiophene-2-carboxamidine 10 4- { [5-(3,5-dichlorophenoxy)(2-furyl)]carbonylamino}-5-methylthiothiophene-2 carboxamidine 5-methylthio-4-(naphthylcarbonylamino)-thiophene-2-carboxamidine 5-methylthio-4-(2-quinolylcarbonyl-amino)thiophene-2-carboxamidine 4-[(3-methoxyphenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine 5 4-[2-(2-hydroxy-5-methoxyphenyl)acetylamino]-5-methylthiothiophene-2 carboxamidine 4-[(4-ethoxyphenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine 5-methylthio-4-(2-phenoxyacetylamino)-thiophene-2-carboxamidine 4-[( 3 -methylphenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine z0 5-methylthio-4- { [3-(phenylmethoxy)phenyl]amino }thiophene-2-carboxamidine 5-methylthio-4-[(3-phenoxyphenyl)amino]thiophene-2-carboxamidine 5-methylthio-4-[(4-phenoxyphenyl)amino]thiophene-2-carboxamidine 4-[(2-methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4-[(2-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 15 4-[(3-chlorophenyl)amino]-5-methylthiothiophene-2-carboxamidine 4-(methylphenylamino)-5-methylthiothiophene-2-carboxamidine 5-methyl-4-(phenylamino)thiophene-2-carboxamidine 4- { [4-(dimethylamino)phenyl]amino } -5-methylthiothiophene-2-carboxamidine 4 -[(4-ethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 0 5-methylthio-4- { [4-(phenylmethoxy)phenyl]amino}thiophene-2-carboxamidine 5-methylthio-4- { [4-(phenylamino)phenyl]amino } thiophene-2-carboxamidine 4
-[(
4 -methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4-[(3-fluoro-4-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4 -(indan-5-ylamino)-5-methylthiothiophene-2-carboxamidine 5 4 -[(9-ethylcarbazol-3-yl)amino]-5-methylthiothiophene-2-carboxamidine WO 00/47578 PCT/US99/18065 -37 5 5-methylthio-4- { [( 4 -phenylphenyl)sulfonyl]amino } thiophene-2-carboxamidine 4- {bis[(4-phenylphenyl)sulfonyl]amino } -5-methylthiothiophene-2-carboxamidine 5-methylthio- 4 -[(2-naphthylsulfonyl)-amino]thiophene-2-carboxamidine 4-[bis( 2 -naphthylsulfonyl)amino]-5-methylthiothiophene-2-carboxamidine 4- { [(6-bromo(2-naphthyl))sulfonyl]amino } -5-methylthiothiophene-2 0 carboxamidine 4- { bis[(6-bromo(2-naphthyl))sulfonyl]amino } -5-methylthiothiophene-2 carboxamidine 5-methylthio- 4 -[(naphthylsulfonyl)-amino]thiophene-2-carboxamidine 4-[bis(naphthylsulfonyl)amino]-5-methylthiothiophene-2-carboxamidine 5 4- { [(2-methylphenyl)sulfonyl]amino } -5-methylthiothiophene-2-carboxamidine 4- {bis[(2-methylphenyl)sulfonyl]amino}-5-methylthiothiophene-2-carboxamidine 4- { [( 3 -methylphenyl)sulfonyl]amino } -5-methylthiothiophene-2-carboxamidine 4- {bis[(3-methylphenyl)sulfonyl]amino} -5-methylthiothiophene-2-carboxamidine 4- { [(4-methylphenyl)sulfonyl]amino } -3-methylthiothiophene-2-carboxamidine 0 4- {bis[(4-methylphenyl)sulfonyl]amino}-5-methylthiothiophene-2-carboxamidine 5-methylthio-4- { [benzylsulfonyl]amino } -thiophene-2-carboxamidine 5-methylthio-4-phenoxythiophene-2-carboxamidine 5-methylthio-4-(phenylsulfonyl)thiophene-2-carboxamidine as well as salts thereof, such as hydrochloride or trifluoracetate salts and prodrugs 5 thereof. Methods of Use and Pharmaceutical Compositions For medicinal use, the pharmaceutically acceptable acid addition salts, those salts in which the anion does not contribute significantly to toxicity or pharmacological activity of the organic cation, are preferred. The acid 0 addition salts are obtained either by reaction of an organic base of Formula I with an organic or inorganic acid, preferably by contact in solution, or by any of the standard methods detailed in the literature available to any practitioner skilled in the art. Examples of useful organic acids are carboxylic acids such WO 00/47578 PCT/US99/18065 -38 5 as maleic acid, acetic acid, tartaric acid, propionic acid, fumaric acid, isethionic acid, succinic acid, cyclamic acid, pivalic acid and the like; useful inorganic acids are hydrohalide acids such as HCI, HBr, HI; sulfuric acid; phosphoric acid and the like. Preferred acids for forming acid addition salts include HCI and acetic acid. 0 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 compounds within the scope of the invention include leukocyte neutrophil elastase, a proteolytic enzyme implicated in the pathogenesis of emphysema; chymotrypsin and trypsin, digestive enzymes; 5 pancreatic elastase, and cathepsin G, a chymotrypsin-like protease also associated with leukocytes; thrombin and factor Xa, proteolytic enzymes in the blood coagulation pathway. Inhibition of thermolysin, a metalloprotease, and pepsin, an acid protease, are also contemplated uses of compounds of the present invention. The compounds of the present invention are preferably 0 employed to inhibit trypsin-like proteases. Compounds of the present invention that inhibit urokinase plasminogen activator are potentially useful in treating excessive cell growth disease state. Compounds of the present that inhibit urokinase are, therefore, useful as anti-angiogenic, anti-arthritic, anti-inflammatory, anti-invasive, anti 5 metastatic, anti-restenotic, anti-osteoporotic, anti-retinopathic (for angiogenesis-dependent retinopathies), contraceptive, and tumoristatic treatment agents. For example, such treatment agents are useful in the treatment of a variety of disease states, including but not limited to, benign prostatic hypertrophy, prostatic carcinoma, tumor metastasis, restenosis and 0 psoriasis. Also provided are methods to inhibit extracellular proteolysis, methods to treat benign prostatic hypertrophy, prostatic carcinoma, tumor metastasis, restenosis and psoriasis by administering the compound of Formula I. For their end-use application, the potency and other biochemical parameters of the enzyme inhibiting characteristics of compounds of the present invention 5 are readily ascertained by standard biochemical techniques well known in the WO 00/47578 PCT/US99/18065 -39 5 art. Actual dose ranges for this application will depend upon the nature and severity of the disease state of the patient or animal to be treated as determined by the attending diagnostician. It is to be expected that a general dose range will be about 0.01 to 50 mg, preferably 0.1 to about 20 mg per kg per day for an effective therapeutic effect. 10 An end use application of the compounds that inhibit chymotrypsin and trypsin is in the treatment of pancreatitis. For their end-use application, the potency and other biochemical parameters of the enzyme-inhibiting characteristics of the compounds of the present invention is readily ascertained by standard biochemical techniques well known in the art. Actual dose ranges 15 for their specific end-use application will, of course, depend upon the nature and severity of the disease state of the patient or animal to be treated, as determined by the attending diagnostician. It is expected that a useful dose range will be about 0.01 to about 50 mg, preferably about 0.1 to about 20 mg per kg per day for an effective therapeutic effect. 20 Compounds of the present invention that are distinguished by their ability to inhibit either factor Xa or thrombin may be employed for a number of therapeutic purposes. As factor Xa or thrombin inhibitors, compounds of the present invention inhibit thrombin production. Therefore, these compounds are useful for the treatment or prophylaxis of states characterized 25 by abnormal venous or arterial thrombosis involving either thrombin production or action. These states include, but are not limited to, deep vein thrombosis; disseminated intravascular coagulopathy which occurs during septic shock, viral infections and cancer; myocardial infarction; stroke; coronary artery bypass; fibrin formation in the eye; hip replacement; and 30 thrombus formation resulting from either thrombolytic therapy or percutaneous transluminal coronary angioplasty (PCTA). By virtue of the effects of both factor Xa and thrombin on a host of cell types, such as smooth muscle cells, endothelial cells and neutrophils, the compounds of the present invention find additional use in the treatment or 35 prophylaxis of adult respiratory distress syndrome; inflammatory responses; WO 00/47578 PCT/US99/18065 -40 5 wound healing; reperfusion damage; atherosclerosis; and restenosis following an injury such as balloon angioplasty, atherectomy, and arterial stent placement. The compounds of the present invention may be useful in treating neoplasia and metastasis as well as neurodegenerative diseases, such as Alzheimer's disease and Parkinson's disease. 10 When employed as thrombin or factor Xa inhibitors, the compounds of the present invention may be administered in an effective amount within the dosage range of about 0.1 to about 500 mg/kg, preferably between 0.1 to 30 mg/kg body weight, on a regimen in single or 2-4 divided daily doses. Human leucocyte elastase is released by polymorphonuclear leukocytes 15 at sites of inflammation and thus is a contributing cause for a number of disease states. 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 leucocyte elastase inhibitory properties of compounds of the present invention 20 are determined by the method described below. Cathepsin G has also been implicated in the disease states of arthritis, gout and emphysema, and in addition, glomerulonephritis and lung infestations caused by infections in the lung. In their end-use application the enzyme inhibitory properties of the compounds of Formula I is readily ascertained by standard biochemical 25 techniques that are well-known in the art. The Cathepsin G inhibitory properties of compounds within the scope of the present invention are determined by the following method. A preparation of partially purified human Cathepsin G is obtained by the procedure of Baugh et al., Biochemistry 15: 836 (1979). Leukocyte granules 30 are a major source for the preparation of leukocyte elastase and cathepsin G (chymotrypsin-like activity). Leukocytes are lysed and granules are isolated. The leukocyte granules are extracted with 0.20 M sodium acetate, pH 4.0, and extracts are dialyzed against 0.05 M Tris buffer, pH 8.0 containing 0.05 M NaCl overnight at 4oC. A protein fraction precipitates during dialysis and is 15 isolated by centrifugation. This fraction contains most of the chymotrypsin- WO 00/47578 PCT/US99/18065 -41 5 like activity of leukocyte granules. Specific substrates are prepared for each enzyme, namely N-Suc-Ala-Ala-Pro-Val-p-nitroanilide and Suc-Ala-Ala-Pro Phe-p-nitroanilide. The latter is not hydrolyzed by leukocyte elastase. Enzyme preparations are assayed in 2.00 mL of 0.10 M Hepes buffer, pH 7.5, containing 0.50 M NaCl, 10% dimethylsulfoxide and 0.0020 M Suc-Ala-Ala 10 Pro-Phe-p-nitroanilide as a substrate. Hydrolysis of the p-nitroanilide substrate is monitored at 405 nm and at 25oC. Useful dose range for the application of compounds of the present invention as neutrophil elastase inhibitors and as Cathepsin G inhibitors depend upon the nature and severity of the disease state, as determined by the 5 attending diagnostician, with a range of 0.01 to 10 mg/kg body weight, per day, being useful for the aforementioned disease states. Additional uses for compounds of the present invention include analysis of commercial reagent enzymes for active site concentration. For example, chymotrypsin is supplied as a standard reagent for use in clinical 0 quantitation of chymotrypsin activity in pancreatic juices and feces. Such assays are diagnostic for gastrointestinal and pancreatic disorders. Pancreatic elastase is also supplied commercially as a reagent for quantitation of a l antitrypsin in plasma. Plasma a,-antitrypsin increases in concentration during the course of several inflammatory diseases, and a,-antitrypsin deficiencies are 15 associated with increased incidence of lung disease. Compounds of the present invention can be used to enhance the accuracy and reproducibility of these assays by titrametric standardization of the commercial elastase supplied as a reagent. See, U.S. Patent No. 4,499,082. Protease activity in certain protein extracts during purification of 0 particular proteins is a recurring problem which can complicate and compromise the results of protein isolation procedures. Certain proteases present in such extracts can be inhibited during purification steps by compounds of the present invention, which bind tightly to various proteolytic enzymes.
WO 00/47578 PCT/US99/18065 -42 5 The pharmaceutical compositions of the invention can be administered to any animal that can experience the beneficial effects of the compounds of the invention. Foremost among such animals are humans, although the invention is not intended to be so limited. The pharmaceutical compositions of the present invention can be 0 administered by any means that achieve their intended purpose. For example, administration can be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, or ocular routes. Alternatively, or concurrently, administration can be by the oral route. The dosage administered will be dependent upon the age, health, and weight of the recipient, kind of 5 concurrent treatment, if any, frequency of treatment, and the nature of the effect desired. In addition to the pharmacologically active compounds, the new pharmaceutical preparations can contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the 0 active compounds into preparations that can be used pharmaceutically. The pharmaceutical preparations of the present invention are manufactured in a manner that is, itself, known, for example, by means of conventional mixing, granulating, dragee-making, dissolving, or lyophilizing processes. Thus, pharmaceutical preparations for oral use can be obtained by 5 combining the active compounds with solid excipients, optionally grinding the resulting mixture and processing the mixture of granules, after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as saccharides, for example, lactose or sucrose, mannitol or sorbitol, cellulose preparations and/or 0 calcium phosphates, for example, tricalcium phosphate or calcium hydrogen phosphate, as well as binders, such as, starch paste, using, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone. If desired, disintegrating agents can be added, 5 such as, the above-mentioned starches and also carboxymethyl-starch, cross- WO 00/47578 PCT/US99/18065 -43 5 linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as, sodium alginate. Auxiliaries are, above all, 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. Dragee cores are provided with suitable coatings that, if desired, are resistant to gastric 10 juices. For this purpose, concentrated saccharide solutions can be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol, and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. In order to produce coatings resistant to gastric juices, solutions of suitable cellulose preparations, such as, acetylcellulose phthalate 15 or hydroxypropylmethyl-cellulose phthalate, are used. Dye stuffs or pigments can be added to the tablets or dragee coatings, for example, for identification or in order to characterize combinations of active compound doses. Other pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of 20 gelatin and a plasticizer, such as, glycerol or sorbitol. The push-fit capsules can contain the active compounds in the form of granules that may 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, 25 such as, fatty oils or liquid paraffin. In addition, stabilizers may be added. Suitable formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form, for example, water soluble salts, alkaline solutions and cyclodextrin inclusion complexes. Especially preferred salts are hydrochloride and acetate salts. One or more 30 modified or unmodified cyclodextrins can be employed to stabilize and increase the water solubility of compounds of the present invention. Useful cyclodextrins for this purpose are disclosed in U.S. Patent Nos. 4,727,064, 4,764,604, and 5,024,998. In addition, suspensions of the active compounds as appropriate oily 35 injection suspensions can be administered. Suitable lipophilic solvents or WO 00/47578 PCT/US99/18065 -44 5 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 can contain substances that increase the viscosity of the suspension, for example, sodium carboxymethyl cellulose, sorbitol, and/or dextran. Optionally, the 0 suspension may also contain stabilizers. Methods of Making Many synthetic methods used to form compounds of the present invention generally involve the formation of an amidine from a carboxylic acid derivative, such as an ester or a nitrile. In the process a Lewis acid, such as 5 trimethylaluminum, is added to a source of ammonia, such as ammonium chloride in an aprotic solvent, such as a toluene, under an inert atmosphere (e.g., under an atmosphere of nitrogen or argon gas) at a temperature between -15oC and 5oC, preferably at 0 0 C. An appropriate carboxylic acid derivative is added to the mixture and the mixture is heated at reflux for a predetermined 0 period of time, preferably between 1 hr. and 24 hrs., and most preferably between 1 hr. and 4 hrs. The resulting solution is allowed to cool to room temperature and the amidine product isolated by known methods.
WO 00/47578 PCT/US99/18065 -45 5 Description of Syntheses The chemical schemes appear after the description of the schemes. Scheme la Scheme la illustrates a general approach to compounds of Formula I 10 where X = O or S, R 3 = alkylthio, aralkylthio, arylthio, alkyloxy, aralkyloxy or aryloxy, Y = bond and Z = NR'R 6 . When R 22 and R 21 of compounds 2 and 3 are retained in the final product, they correspond to R 2 and R 3 of Formula I, respectively. Otherwise R 22 and R 21 represent groups which, after further transformations, will become R 2 and R 3 of Formula I. 15 Starting with the heterocycle where X = O or S appropriately substituted by two leaving groups, the leaving groups can be sequentially displaced by appropriate nucleophiles (preferably the anion of the group R 21 or
R
22 to be substituted) to produce the mono- or disubstituted heterocycles. Examples of leaving groups include halogens (chlorine, bromine or iodine), 20 sulfonates (methanesulfonate, toluenesulfonate or trifluoromethanesulfonate) or sulfones (methylsulfonyl). Preferable nucleophiles include anions of thiols or alcohols having as the counterion an alkali or alkali earth metal such as sodium, lithium, potassium, magnesium or cesium, or in some cases, a transition group metal such as zinc, copper or nickel. In certain cases where 25 the nucleophile used contains an anion on carbon, catalysis of the displacement may be useful for this transformation. Examples of catalysts would include compounds containing palladium, silver or Ni salts. Scheme lb Scheme lb illustrates approaches to providing the functionality of 30 Y(CNR 4 )Z in compounds of Formula I where X = N, O or S, R 22 and R 21 are defined as in Scheme la. Depending on the nature of the group W in 3, several methods may be employed in the transformation of W to Y(CNR 4
)Z.
WO 00/47578 PCT/US99/18065 -46 5 When W in 3 is a cyano group (CN), primary amide (CONH 2 ) or ester
(COR
23 ), direct conversion to an unsubstituted amidine 5 (i.e. Formula I where Y = bond, Z = NR 5
R
6 and R 4 , R 5 , R 6 = H) can be effected by treatment with a reagent consisting of a Lewis acid complexed to ammonia. This complex is produced by treatment of ammonia or an ammonium salt, 10 preferably an ammonium halide and most preferably ammonium chloride or bromide, with an appropriate Lewis acid, preferably a trialkylaluminum and most preferably trimethyl- or triethylaluminum in a solvent inert to the Lewis acid employed. For example, when a trialkylaluminum Lewis acid is employed with an ammonium halide, reaction occurs with loss of one 15 equivalent of alkane to produce the dialkylhaloaluminum complex of ammonia (see for example Sidler, D.R., et al, J. Org. Chem., 59:1231 (1994)). Examples of suitable solvents include unsaturated hydrocarbons such as benzene, toluene, xylenes, or mesitylene, preferably toluene, or halogenated hydrocarbons such as dichloroethane, chlorobenzene or dichlorobenzene. The 20 amidination reaction is generally carried out at elevated temperatures, preferably 40-200 oC, more preferably 80-140 oC, and most preferably at the reflux temperature of a solvent in the range of 80-120 oC. When W is a cyano group (CN), direct conversion to a mono- or disubstituted amidine 5 (R 4 , R 5 , R 6 = H) is also possible by treatment with a 25 reagent consisting of a Lewis acid, preferably a trialkylaluminum, complexed to a mono- or disubstituted amine H 2
NR
5 or HNR'R 6 (Garigipati, R., Tetrahedron Lett. 31: 1969 (1990)). Alternatively the same addition of a mono- or disubstituted amine may catalyzed by a copper salt such as Cu(I) chloride (Rousselet, G., et al, Tetrahedron Lett. 34: 6395 (1993)). 30 When W in 3 is a carboxyl group (CO 2 H), indirect conversion to an unsubstituted amidine 5 can be carried out by initial esterification to 4 by any of a number of well-known dehydrating agents (for example, dicyclohexylcarbodiimide) with an alcohol (R 23 0H). More preferably 4 can be made by initial formation of an acid chloride by treatment of 3 with any of a 35 number of anhydrides of HCI and another acid, such as thionyl chloride, WO 00/47578 PCT/US99/18065 -47 5 POC1 3 , PC1 3 , PC1 5 , or more preferably oxalyl chloride, with or without an added catalyst such as N,N-dimethylformamide (DMF), followed by the alcohol R 2 3 0H. Conversion to the unsubstituted amidine 5 (R 4 , R 5 , R 6 = H) can be carried out by treatment with a Lewis acid complexed to ammonia. Amidines 5 also can be produced indirectly by conversion of 3 (W = 10 CN) to iminoethers 6 by exposure to a strong acid such as a hydrogen halide,
HBF
4 or other non-nucleophilic acid, preferably gaseous HCI in the presence of an alcohol R 23 0H (R 23 = alkyl, branched alkyl or cycloalkyl, preferably Me or Et) and most preferably with the alcohol as solvent. Alternatively when W = CONH 2 , conversion to an iminoether can be carried out by treatment with a 15 trialkyloxonium salt (Meerwein's salts). In either case, treatment of the iminoether 6 with ammonia (R', R 6 = H) or a mono- or disubstituted amine
(HNR'R
6 ) provides the corresponding unsubstituted or substituted amidines 5 (i.e. via classical Pinner synthesis: Pinner, A., Die Iminoaether und ihre Derivate, Verlag R. Oppenheim, Berlin (1892)). 0 When W = NH 2 in 3, treatment with a reagent Z(CNR 4 )L where Z = alkyl and L is a leaving group such as O-alkyl and preferably OMe, provides the subclass of amidines 135 (Z = alkyl ) which are isomeric to 5 (Formula I, where Y = NH, Z = H or alkyl). Examples of reagents for this reaction include methyl or ethyl acetimidate hydrochloride. Alternatively treatment of 3 (W = 5 NH 2 ) with a trialkyl orthoformate ester, preferably trimethyl- or triethyl orthoformate, followed by an amine R 4
NH
2 affords the corresponding formidines 135 (Z = H) (Formula I, where Y = NH, Z = H). Also, when W = NH 2 , 3 can be treated with a reagent Z(CNR 4 )L where
R
4 = H and Z = NR'R 6 and L is a leaving group such as pyrazole, 0 methylpyrazole, S03H, S-alkyl, S-aryl, trifluoromethanesulfonate (OTf) or trifluoromethanesulfonamide (NHTf), preferably pyrazole, SO 3 H or trifluoromethanesulfonamide (NHTf). Examples of these reagents include aminoiminosulfonic acid (Miller, A.E. and Bischoff, J.J., Synthesis, 777 (1986) and 1H-pyrazole-l1-carboxamidine hydrochloride (Bernatowicz, M.S., 5 et al., J. Org. Chem. 57:2497 (1992)). Such treatment provides guanidines WO 00/47578 PCT/US99/18065 -48 5 136 directly (Formula I where Y = NH, Z = NR 5
R
6 ). Alternatively a reagent
Z(CNP
1 )L may be also used where Z = NHP 2 and L again a leaving group such as pyrazole, methylpyrazole, SO3H, S-alkyl, S-aryl, trifluoromethanesulfonate (OTf) or trifluoromethanesulfonamide (NHTf), to provide protected guanidines (P', p2 = alkoxylcarbonyl, aralkoxycarbonyl or polymer-bound 0 alkoxylcarbonyl similar to those described below in Scheme 4a) where the protecting groups P' and P 2 can then be removed to give unsubstituted 136 (R 4 ,
R
5 and R 6 = H). Protected guanidines are advantageous when further transformations are required after introduction of the guanidine functionality where an unprotected guanidine would not be stable. Examples of these 5 protected reagents include reagents such as N,N'-bis(tert-butoxycarbonyl)-S methylthiourea (Bergeron, R.J. and McManis, J.S, J. Org. Chem. 52:1700 (1987)), N,N'-bis(benzyloxycarbonyl)- 1 H-pyrazole- 1 -carboxamidine or N,N' bis(tert-butoxycarbonyl)- 1 H-pyrazole- 1-carboxamidine (Bernatowicz, M.S., et al., Tetrahedron Letters, 34: 3389 (1993)), N,N'-bis(benzyloxycarbonyl) 0 N"-trifluoromethanesulfonylguanidine, and N,N'-bis(bis(tert-butoxycarbonyl) N"-trifluoromethanesulfonylguanidine (Feichtinger, K., et al, J. Org. Chem. 63:3804 (1998)). Detailed descriptions and examples of these protecting groups and their use as protection for amidines are further outlined in Schemes 4a, 4b and 5. 5 When W in 3 is an ester (CO 2
R
23 ) or carboxyl group (CO 2 H), indirect conversion to an N-substituted or unsubstituted methylamidine (Formula I where Y = CH 2 , Z= NR 5
R
6 ) can be carried out by initial reduction of the ester or carboxyl by any of a number of well-known reducing agents. When W in 3 is an ester (CO 2
R
23 ), examples of reducing agents include reducing agents such 0 lithium aluminum hydride (LAH) and lithium borohydride. When W in 3 is a carboxyl group (CO 2 H), examples of reducing agents include LAH and borane complexed to THF, dimethyl sulfide, dimethylamine or pyridine. The resulting hydroxymethyl derivative (W = CH 2 OH) is converted to a cyanomethyl derivative (W = CH 2 CN) by initial formation of a leaving group 5 (W = CH 2 L) where the leaving group L is a halogen (chlorine, bromine or WO 00/47578 PCT/US99/18065 -49 5 iodine) or sulfonate ester (for example methanesulfonate, toluenesulfonate or trifluoromethanesulfonate). Displacement of L by cyanide can then be performed by treatment with a metal cyanide such as LiCN, NaCN, KCN or CuCN in a polar solvent such as DMF and with or without a catalyst such as a crown ether, to afford the cyanomethyl derivative (see for example Mizuno, 10 Y., et al, Synthesis, 1008 (1980)). More preferably, the conversion of W =
CH
2 OH to W = CH 2 CN may be effected by a Mitsunobu reaction (Mitsunobu, 0., Synthesis, 1 (1981)) using an azodicarboxylate ester such as diethyl azodicarboxylate or diisopropyl azodicarboxylate, Ph 3 P and a source of cyanide such as HCN or more preferably acetone cyanohydrin (Wilk, B. .5 Synthetic Commun. 23:2481 (1993)). Treatment of the resulting cyanomethyl intermediate (W = CH 2 CN) under the conditions described for the conversion of 3 (W = CN) to 5 (either directly or indirectly via 6) provides the corresponding amidinomethyl products. Scheme Ic 0 When not commercially available, alkylthiothiophenes (3, X = S, R' = OH or NH 2 , R21 = SR 54 , W = CN, CO 2
R
2 3 , CONH 2 ) can be synthesized by the methods illustrated in Scheme 1c. Condensation of carbon disulfide and a malonic acid derivative (R 5 2
CH
2
R
2 2 ) in the presence of two alkylating agents
R
54 L and WCH 2 L and a base in a suitable medium provide 3 (Dolman, H., 5 European Patent Application No. 0 234 622 Al (1987)). When R 22 = R 52 = CN, the resulting R 1 will be NH 2 ; when R 2 2 = R 5 2 = CO 2
R
23 , the resulting R' will be OH; and when R 22 and R 52 = CN, CO 2
R
23 , the resulting R' can be selected to be OH or NH 2 (and R 2 2 = CN or CO 2
R
23 ) depending on the reaction conditions and order of reagent addition. Examples of malonic acid 0 derivatives suitable for this transformation include but are not limited to malonate diesters such as dimethyl malonate or diethyl malonate (R 5 2 , R 22
CO
2
R
2 3 , R 2 3 = Me or Et), malononitrile (R 52 , R 2 2 = CN), or methyl or ethyl cyanoacetate (R 52 = CO 2
R
23 , R 22 = CN, R 2 3 = Me or Et). Leaving groups L include halides such as chloride, bromide or iodide, preferably bromide or WO 00/47578 PCT/US99/18065 -50 5 iodide, or sulfonates such as toluenesulfonate, benzenesulfonate, methanesulfonate or trifluoromethanesulfonate. Examples of alkylating agent
R
5 4 L include primary or secondary alkyl, allyl or aralkyl halides or sulfonates, such as methyl iodide, isopropyl bromide, allyl bromide, benzyl chloride or methyl trifluoromethanesulfonate, or a 2-haloacetate ester such as tert-butyl 2 10 bromoacetate. Examples of alkylating agents WCH 2 L include 2 chloroacetonitrile, methyl 2-bromoacetate or 2-bromoacetamide. Suitable media are generally polar aprotic solvents, for example, N,N dimethylformamide (DMF), N,N-dimethylacetamide (DMA), N methylpyrrolidinone (NMP) or dimethylsulfoxide (DMSO), preferably DMF. 5 Alternatively compounds 3 (R 22 = CN) can be synthesized from precursors 138 (derived from malononitrile, R 54 L and carbon disulfide), a thioglycolate WCHSH and a base in a suitable polar solvent, preferably methanol (Tominaga, Y., et al, J. Heterocyclic Chem. 31:771 (1994)). When 3 contains an amino group at R', it can be diazotized with .0 subsequent loss of nitrogen to give 3, R' = H by treatment with a nitrosating agent in suitable solvent. Nitrosating agents include nitrosonium tetrafluoroborate, nitrous acid or, more preferably and alkyl nitrite ester such as tert-butyl nitrite. Suitable solvents are those which are stable to the nitrosating agents, preferably DMF, benzene or toluene. 5 Scheme Id When not commercially available, heterocyclic precursors 1 or 2 (X = O, S; W = CO 2
R
23 , COOH; L = halogen) used in Scheme la can be synthesized by the methods illustrated in Scheme ld. Depending on the conditions used, treatment of compounds such as 139 with elemental halogen 0 (Cl 2 , Br 2 or 12, preferably Br 2 ) or an N-halosuccinimide reagent, preferably N bromosuccinimide (NBS), affords either 1 or 2 directly. Description of suitable solvents and conditions to selectively produce 1 or 2 are found in Karminski-Zamola, G. et al, Heterocycles 38:759 (1994); Divald, S., et al, J. Org. Chem. 41:2835 (1976); and Bury, P., et al, Tetrahedron 50:8793 (1994).
WO 00/47578 PCT/US99/18065 -51 5 Scheme 2a Scheme 2a illustrates the synthesis of compounds 12 representing the subclass of compounds for which R 2 is Formula II, where Ar = 2-thiazolyl, Y = bond and Z = NRR 6 . Starting with compound 1 (L = Br) and using the sequential displacement methodology discussed for Scheme la, R 21 can be 10 first introduced to give 7. This is followed by a second displacement with a metal cyanide such as copper (I) cyanide, sodium cyanide or lithium cyanide and most preferably copper (I) cyanide at a temperature of 80-200 oC and preferably at 100-140 oC, in a polar aprotic solvent, preferably DMF or DMSO, to give 8. After esterification by any of the means described for the 15 conversion of 3 to 4, conversion to the thioamide is carried out by treatment of the nitrile with any of the methods well known in the art (see for example Ren, W., et al., J. Heterocyclic Chem. 23:1757 (1986) and Paventi, M. and Edward, J.T., Can. J. Chem. 65:282 (1987)). A preferable method is treatment of the nitrile with hydrogen sulfide in the presence of a base such as a trialkyl or 20 heterocyclic amine, preferably triethylamine or pyridine, in a polar solvent such as acetone, methanol or DMF and preferably methanol. Conversion to the thiazole can be executed by classical Hantzsch thiazole synthesis followed by amidine formation as discussed in Scheme lb. Scheme 2b 25 Scheme 2b illustrates the synthesis of compounds representing the subclass of compounds for which R 2 is Formula II where, in addition to being an alternate route to Ar = 2-thiazolyl (20) (see 12, Scheme 2a) also provide compounds of Formula II where Ar = 2-oxazolyl (16) or 2-imidazolyl (18) (Y = bond and Z = NR'R 6 ). Starting with compound 9, a selective hydrolysis of 30 the nitrile with a tetrahalophthalic acid, preferably tetrafluoro- or tetrachlorophthalic acid, can be used to give 7 according to the method of Gribble, G.W. et al., Tetrahedron Lett. 29: 6557 (1988). Conversion to the acid chloride can be accomplished using the procedures discussed for conversion of 3 to 4, preferably with oxalyl chloride in dichloromethane in the WO 00/47578 PCT/US99/18065 -52 5 presence of a catalytic amount of DMF. Coupling of the acid chloride to an aminoketone (R 2 6
COCH(R
2 7
)NH
2 ) can be performed in the presence of an acid scavenger, preferably N,N-diisopropylethylamine (DIEA) or pyridine in a suitable solvent such as DMF, dichloromethane or tetrahydrofuran (THF) to afford the common intermediate 14. Alternatively coupling of the acid 10 chloride to a less-substituted aminoketone (R 26
COCH
2
NH
2 ) can be used followed by optional alkylation with alkylating agent R 27 L in the presence of a base, preferably NaH or t-BuOK. Transformation of 14 to the corresponding 2-oxazolyl (15), 2-imidazolyl (17) or 2-thiazolyl (19) esters can carried out by the methodology of Suzuki, M., et al., Chem. Pharm. Bull. 34:3111 (1986) 15 followed by amidination according to Scheme lb. In addition, direct conversion of ketoamide 14 to imidazolyl derivative 18 is possible under the same conditions for conversion of 17 to 18 when conducted for extended periods, preferably greater than 2 h. Scheme 2c 20 Scheme 2c describes a general route to the synthesis of oxazoles, imidazoles and thiazoles of structure 27, 29 and 31 respectively. Acid 2 (see Scheme la) is converted to the ester by methods that are well known in the art (Theodora W. Greene and Peter G. M. Wuts, John Wiley and Sons, Inc. 1991). For example methyl ester 21 is formed by treating the acid in an appropriate 25 solvent such as methanol with trimethylsilyldiazomethane. Alternatively the acid is treated with oxalyl chloride and catalytic amounts of dimethylformamide (DMF) in an appropriate solvent such as dichloromethane to form the acid chloride, which is then treated with methanol to give the methyl ester. Ester 21 is treated with a palladium (0) catalyst such as palladium 30 tetrakistriphenylphosphine, and an alkylstannane such as hexa-n butyldistannane or tri-n-butyltin chloride in an appropriate solvent such as DMF at elevated temperatures (50 oC - 120 oC) to give the arylstannane of general structure 22 (Stille, J.K., Angew. Chem. Int. Ed. Engl. 25:508-524 (1986)). The stannane 22 is then treated with acid chlorides in the presence of WO 00/47578 PCT/US99/18065 -53 5 a palladium(0) catalyst to give ketone 23. The ketone is treated with ammonia/ammonium chloride to give amine 24. Alternatively the ketone is reacted with an azide such as sodium azide in a suitable solvent such as DMF, and the resulting azidoketone is reduced to amine 23 with a suitable reducing agent such as catalytic hydrogenation in the presence of palladium on carbon 0 and an acid such as HC1 (Chem. Pharm. Bull. 33:509-514 (1985)). Ketoamides 25 are formed by coupling the ketoamine 24 with a variety of suitably functionalized acid chlorides. Alternatively amide coupling may be performed using any of a number of peptide coupling reagents such as 1,3 dicyclohexylcarbodiimide (Sheehan, J. C. et al., J. Am. Chem. Soc., 77:1067 5 (1955)) or Castro's reagent (BOP, Castro, B., et al., Synthesis 413 (1976)). In another approach, amides 25 are formed directly from ketones 23 by reacting with various amide salts in an appropriate solvent such as DMF. The amide salts are generated by treating the amides with a suitable base such as sodium hydride (NaH). For example acetamide is treated with NaH in DMF at 0 oC to 0 give sodium acetamide. Keto amide 25 is cyclized to the oxazole 26, imidazole 28 and thiazole 30 using procedures similar to that shown in scheme 2b. Oxazole 26, imidazole 28 and thiazole 30 are treated with trimethylaluminum and ammonium chloride in refluxing toluene to give the amidines 27, 29 and 31 respectively. 5 Scheme 2d Scheme 2d illustrates to the preparation of compounds of Examples 42-43, where R 21 and R 43 correspond in Formula Ito groups R 3 and R 2 , respectively. The acids 2 can be converted to the stannane by treatment with base, such as n-butyl lithium or sec-butyl lithium, followed by trimethyltin D chloride. The resulting acid can be then converted to the ester 22 by methods that are well known in the art (Theodora W. Greene and Peter G. M. Wuts, Protective Groups in Organic Chemistry, John Wiley and Sons, Inc. 1991). For example the methyl ester can be made by treating the acid 2 in a suitable solvent such as methanol with trimethylsilyldiazomethane. The stannane 22 WO 00/47578 PCT/US99/18065 -54 5 can be reacted with suitable halides in the presence of catalytic amounts of a palladium catalyst, such as palladium tetrakistriphenylphosphine, to give the esters 32 (Stille, J.K., Angew. Chem. Int. Ed. Engl. 25:508-524 (1986)). These esters are then treated with trimethylaluminum and ammonium chloride in refluxing toluene to give the amidines 33. In the case where R 4 3 Ln (n = 2), this 10 can be cross-coupled to an aryl, heteroaryl or vinyl boronic acid or ester to give compounds 34 (Miyaura, N. and Suzuki, A., Chem. Rev. 95:2457-2483 (1995)). This can usually be done in the presence of catalytic amounts of a palladium (0) catalyst such as tetrakistriphenylphosphine palladium and a base such as potassium carbonate in DMF at 90 0 C. Similar cross-coupling reactions 15 can also be achieved by using aryl, heteroaryl and vinyl stannanes instead of boronic acids or esters. These esters are converted to the amidines 35 in the manner previously described. Scheme 2e Scheme 2e represents a modification to the methodology outlined in 20 Scheme 2b which allows synthesis of compounds of Formula II where Ar = 2 thiazolyl, 2-oxazolyl or 2-imidazolyl (Y = bond and Z = NR'R 6 ) but which are regioisomeric to 16, 18 or 20 in the relative positions of substituents R 26 and
R
27 . This is illustrated in Scheme 2b by the synthesis of 2-oxazolyl derivative 39. Thus, acid 13 can be coupled to an hydroxy-containing amine 25 R 27
CH(NH
2
)CH(R
26 )OH to give amide 36 by any of a number of amide coupling reagents well known in the art (see Bodanszky, M. and Bodanszky, A., The Practice ofPeptide Synthesis, Springer-Verlag, New York (1984)). More preferably 13 can be converted to the corresponding acid chloride using any of the procedures mentioned for conversion of 3 to 4 followed by 30 treatment with the R 27
CH(NH
2
)CH(R
26 )OH in the presence of an acid scavenger, preferably N,N-diisopropylethylamine (DIEA) or pyridine in a suitable solvent such as DMF, dichloromethane or tetrahydrofuran (THF) to give 36. Oxidation of the alcohol 36 to the aldehyde 37 (R 2 6 = H) or ketone 37
(R
26 = alkyl, aryl, aralkyl, heterocycle) can be effected by any of a number of WO 00/47578 PCT/US99/18065 -55 5 common methods known in the art (see for example F. Carey, F.A., Sundberg, R.J. Advanced Organic Chemistry, Part B: Reactions and Synthesis, 3rd Edition, Plenum Press, New York (1990)), preferably by a mild Moffatt-type oxidation such as a Swern oxidation (Mancuso, A.J., Huang, S.L. and Swern, D., J. Org. Chem. 3329 (1976)) or more preferably using Dess-Martin reagent 10 (Dess, D.B. and Martin, J.C., J. Org. Chem. 48:4155 (1983)). Conversion to the heterocycle (in this case the oxazole) is effected with any of a number of reagents including phosphorus oxychloride, P 2 0 5 or thionyl chloride (see Moriya, T., et al., J Med Chem. 31:1197 (1988) and references therein). Alternatively closure of 37 with either Burgess reagent or under Mitsunobu 15 conditions affords the corresponding oxazolinyl derivatives (Wipf, P.and Miller, C.P., Tetrahedron Lett. 3: 907 (1992)). Final amidination to 39 as in Scheme lb completes the synthesis. Scheme 2f Scheme 2f illustrates a general approach to the synthesis of thiazoles of 20 structure 43 (Formula II, X = S, Ar = thiazolyl). Nitriles of structure 40 can be treated with hydrogen sulfide (H 2 S) in a suitable solvent such as methanol, or pyridine in the presence of a base such as triethylamine to give thioamides 41 (Ren,W. et al., J Heterocyclic Chem. 23:1757-1763 (1986)). Thioamides 41 can be then treated with various haloketones 42 preferably bromoketones 25 under suitable reaction conditions such as refluxing acetone or DMF heated to 500 C - 800 C to form the thiazoles 43 (Hantzsch, A. R. et al., Ber. 20:3118 (1887)). Scheme 2g Scheme 2g illustrates one synthetic route to 2-haloketones of structure 30 42 which are employed in the synthesis of thiazolyl derivatives as in Schemes 2a and 2f. 2-Bromoketones 42 (L = Br) are prepared by treating the ketone 44 with a suitable brominating agent such as Br 2 or N-bromosuccinimide in a suitable solvent such as chloroform or acetic acid (EP 0393936 Al).
WO 00/47578 PCT/US99/180 65 -56 5 Alternatively, the ketone 44 is treated with a polymer-supported brominating agent such as poly(4-vinyl)pyridinium bromide resin (Sket, B., et al., Synthetic Communications 19:2481-2487 (1989)) to give bromoketones 42. In a similar fashion 2-chloroketones are obtained by treating 44 with copper (II) chloride in a suitable solvent such as chloroform (Kosower, E. M., et al., J. Org. Chem. 10 28:630 (1963)).
WO 00/47578 PCT/US99/1 8 0 6 5 -57 5 Scheme 2h Scheme 2h illustrates another synthetic route to 2-haloketones of structure 42 which is particularly useful in that it employs acids 45 or activated carbonyl compounds such as 46 as precursors which are more readily available than the ketones 44. The acid 45 is converted to the acid halide 46 (L = Cl, Br 10 or OCOR 3 9 ) by treating with a suitable halogenating reagent. For example, an acid chloride is formed by treating 45 with oxalyl chloride and catalytic amounts of DMF in dichloromethane. The acid chloride is converted to a diazoketone by treatment with trimethysilyldiazomethane (Aoyama, T. et al., Tetrahedron Lett. 21:4461-4462 (1980)). The resulting diazoketone is 15 converted to a 2-haloketone of structure 42 by treatment with a suitable mineral acid. For example a bromoketone is formed by treating the diazoketone in a suitable solvent such as acetonitrile
(CH
3 CN) with a solution of 30% hydrogen bromide (HBr) in acetic acid (Organic Synthesis Collective Vol III, 119, John Wiley and Sons, New York, Ed. Horning E. C.). In an 20 alternative approach the acid 45 is converted to the mixed-anhydride 46 by treatment with a suitable chloroformate such as isobutyl chloroformate or tert butyl chloroformate in a suitable solvent, such as tetrahydrofuran or dichloromethane, in the presence of a base such as N-methylmorpholine. The mixed anhydride 46 is converted to a diazoketone by treatment with 25 trimethylsilyldiazomethane and the resulting diazoketone is converted to a haloketone in the manner described above. Scheme 2i When amide coupling as described in Scheme 2e is followed directly by amidination, compounds of Formula I where R 2 or R 3 is aminoacyl or 30 aminoiminomethyl can be derived. Thus, coupling of acid 13 (or the corresponding acid chloride as previously described) with an amine R"R 5 2 NH can afford 130 which can be carried on to the amidine 131. Upon either longer or more vigorous additional treatment (for example, higher temperatures) with WO 00/47578 PCT/US99/18065 -58 5 a Lewis acid-ammonia reagent as described in Scheme lb, the amide group can be converted to an aminoiminomethyl group to give a bisamidine compound 132. Scheme 3a Acid 13 can also be converted to an amine 47 from which 10 sulfonamides, ureas and urethanes can be formed (Formula I where R 2 or R =
NR
3 2
SO
2
R
1 , NHCONRIR 2 or NHCOR', respectively). Scheme 3a illustrates this methodology for introduction of these three groups at R 2 of Formula I. Conversion of the acid 13 to an intermediate acyl azide can be followed by heating of such azide in the presence of an alcohol under Curtius 15 rearrangement conditions to form the carbamate ester of the alcohol. Subsequent carbamate ester hydrolysis yields amine 47. The intermediate acyl azide may be synthesized by coupling the acid 13 to hydrazine through the acid chloride or by any of the amide coupling procedures discussed for Scheme 2e followed by nitrosation of the resulting hydrazide by any of the 20 nitrosating agents discussed for conversion of 3 (R' = NH 2 ) to 3 (R' = H) in Scheme 1c. More preferably conversion of 13 to 47 is carried out through treatment of acid 13 with diphenylphosphoryl azide in the presence of an alcohol, preferably tert-butanol, and a base, preferably triethylamine or DIEA, as shown in Scheme 3a, to give a tert-butylcarbamate that is readily 25 decomposed to the salt of amine 47 on exposure to an acid, preferably HCI or trifluoroacetic acid in a suitable solvent such as CH 2 C1 2 . Further treatment with a base such as NaOH or preferably K 2 C0 3 or NaHCO 3 provides the free base 47. Treatment of amine 47 with a sulfonyl chloride R 31S O 2 C1 in the presence of an acid scavenger, such as pyridine or DIEA, followed by optional 30 alkylation on nitrogen with an alkylating agent R 32 L in the presence of a base such as K 2 C0 3 , DIEA or more preferably sodium hydride, in a solvent such as THF, MeCN or CH 2 C1 2 affords the sulfonylamine functionality at R 2 (48). When necessary, this transformation can be catalyzed by the presence of 4 dimethylaminopyridine for less reactive sulfonyl chlorides. Similar treatment WO 00/47578 PCT/US99/1 8 0 6 5 -59 5 of amine 47 with an isocyanate
R
51 NCO or carbamyl chloride
R
51
R
5 2 COC1 affords the aminocarbonylamine functionality at R 2 (50). Similar treatment of amine 47 with an acid chloride
R
31 COCI affords the carbonylamine functionality at R 2 (52). Conversion of the esters in 48, 50 and 52 to amidines as previously mentioned gives the products 49, 51 and 53. Further conversion 10 of the acylamino group of 53 as discussed for synthesis of 132 also provides access to the iminomethylamino group at R 2 (54). Scheme 3b Introduction of an aminosulfonyl group (including monoalkylaminosulfonyl and dialkylaminosulfonyl groups) for R 2 of Formula 15 I can be carried out starting from amine such as 47 as well. Conversion to a sulfonyl chloride by the method of Gengnagel, et al. (U.S. Patent No. 3,947,512 (1976)) and treatment with an amine R 34
NH
2 followed by optional alkylation on nitrogen with R 35 L (under the sulfonylation and alkylation conditions described in Scheme 3a) provides 56 which is further converted to 20 amidines 57 as previously described. Scheme 3c In addition to the synthesis outlined in Scheme 3a, amine 47 may also be produced as illustrated in Scheme 3c. A nitrothienyl ester 122 (Dell'Erba, C. and Spinelli, D., Tetrahedron, 21: 1061 (1965), Dell'Erba, C. et al., J. 25 Chem. Soc, Perkin Trans 2, 1779 (1989)) with a suitable leaving group L may be substituted with an anion of R 21 to give intermediate 123. Amine 47 is then derived from reduction of the nitro group. Appropriate reagents to effect reduction of the nitro functionality include hydrogen gas in the presence of a catalyst such as palladium or platinum metal deposited on carbon or barium 30 sulfate in any of a number of solvents such as methanol, ethanol, ethyl acetate, DMF or THF. More preferably, tin (II) chloride may be employed as a reductant in solvents such as DMF or THF, or in the presence of HCI in a solvent such as methanol or ethanol. Alternatively, metals such as zinc or iron WO 00/47578 PCT/US99/18065 -60 5 (Stanetty, P. and Kremslehner, M., Heterocycles 48: 259 (1998)) may also be used. Scheme 4a Scheme 4a illustrates the preparation of the compounds of Formula III and Examples 48-59 and 61-77. The amidine moiety of compounds of 0 structure 60 can be protected with a protecting group P' that can be readily removed from 62 and 64 using methods known to those skilled in the art (Theodora W. Greene and Peter G. M. Wuts, John Wiley and Sons, Inc. 1991). For example, a tert-butoxycarbonyl (BOC) protecting group can be removed by exposure to strongly acidic medium such as hydrogen chloride in a suitable 15 solvent such as dioxane, or by trifluoroacetic acid in a suitable solvent such as methylene chloride. Benzyloxycarbonyl (Cbz) protecting groups can be removed by catalytic hydrogenation using palladium on carbon as a catalyst in solvents such as ethanol or tetrahydrofuran. In some cases, P' can be a solid support such as polystyrene or 20 polyethyleneglycol-grafted polystyrene which can be attached to the amidine moiety via a cleavable linker such as 4-(benzyloxy)benzyloxy-carbonyl (using carbonate Wang resin). Attaching an amidine to a solid support can be achieved by treating a solid support having a linker containing an appropriately activated functional group with the amidine under suitable conditions. For 25 example, an amidine can be attached to Wang resin by treating para nitrophenylcarbonate Wang resin with the amidine and a suitable base such as DBU in a suitable solvent such as DMF. When D is OH or SH the protected amidines 61 can be alkylated with carboxy-protected (protecting group is R 36 ) haloaliphatic acids, such as bromoacetic acid or bromopropionic acid in the 30 presence of a suitable base such as cesium carbonate or DIEA, in a suitable solvent such as DMF with heating when necessary to give compounds of structure 62. When D is NO 2 , the nitro group can be reduced prior to alkylation using an appropriate reducing agent, such as tin (II) chloride, in a suitable solvent such as DMF, or by catalytic hydrogenation using palladium on carbon WO 00/47578 PCT/US99/1806 5 -61 5 as a catalyst in solvents such as ethanol or tetrahydrofuran. Other useful carboxy protecting groups are well known in the art (Theodora W. Greene and Peter G. M. Wuts, Protective Groups in Organic Chemistry, John Wiley and Sons, Inc. 1991). For example, tert-butyl ester can be removed by exposure to strongly acidic medium such as hydrogen chloride in a suitable solvent such as 0 dioxane, or such as trifluoroacetic acid in a suitable solvent such as methylene chloride. Benzyl ester can be removed by catalytic hydrogenation using palladium on carbon as a catalyst in solvents such as ethanol or tetrahydrofuran or by base hydrolysis. When protecting groups P' and R 3 6 in compounds 62 are orthogonal (as 15 defined by the ability to remove one protecting group preferentially in the presence of the other), R 3 6 can be preferentially removed to give acids 63. For example when P' is BOC and R 3 6 is OMe, the methyl ester can be removed by treating with a base such as sodium hydroxide in a suitable solvent such as aqueous tetrahydrofuran leaving the BOC group intact. When protecting 20 groups P' and R 3 6 in compounds 62 are not orthogonal, both protecting groups are removed, and the amidine can be protected with a suitable protecting group such as BOC or a suitably functionalized resin. The protected amidine 63 can be treated with various amines under suitable amide coupling conditions, such as in the presence 1-hydroxy-7-azabenzotriazole (HOAt), O-(7 25 azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU) and DIEA to form amides of structure 64. The amidine protecting group can be then removed, for example by treating with an acid, such as trifluoroacetic acid in a suitable solvent such as methylene chloride, when a BOC protecting group is employed, to give amidines 65. 30 Scheme 4b Scheme 4b illustrates a specific example which utilizes the method described in Scheme 4a. The amidine moiety of 66 can be monoprotected with a tert-butyloxycarbonyl group. The monoprotected phenoxyamidine 67 can be alkylated on the phenolic hydroxy group with an ester of 2-bromoacetic WO 00/47578 PCT/US99/18065 -62 5 acid to give 68. In the case where the ester can be removed by base, it can be hydrolyzed with aqueous base, such as NaOH, to give the acid 69. This acid can be treated with various amines in the presence of 1-hydroxy-7 azabenzotriazole (HOAt), O-(7-azabenzotriazol- 1 -yl)-1,1,3,3 tetramethyluronium hexafluorophosphate (HATU) and DIEA to form amides 10 of structure 70. The amines are unsubstituted, di- or mono-substituted aliphatic or aromatic amines. In some cases the amines are cyclic-amines such as piperazine and piperidine. The amides 70 are then treated with trifluoroacetic acid to give the amidines 71. In the case where the ester 68 is acid-labile, it can be treated with trifluoroacetic acid to give the amidino-acid 15 72. This amidine can be loaded on to an insoluble support, such as polystyrene or polyethyleneglycol-grafted polystyrene via a cleavable linker, such as Wang, which is functionalized as an activated carbonate such as p nitrophenylcarbonate or succinimidyl carbonate. Generally this can be done by treating the activated carbonate resin with the amidine and a suitable base such 20 as DBU in a suitable solvent such as DMF. The support-bound acid 73 can be treated with various amines in the presence of 1-hydroxy-7-azabenzotriazole (HOAt), O-(7-azabenzotriazol- 1 -yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU) and DIEA to form amides. These amides are then cleaved from the solid support by treating with trifluoroacetic acid to give 25 compounds of structure 71. Scheme 5 Scheme 5 illustrates a synthetic route to amidines containing di substituted thiazoles represented by compounds for which R 2 is Formula II and both R' and R 9 are non-hydrogen substituents . The ketoamide 74 can be 30 converted to the mono-bromoketoamide by treating with bromine in acetic acid. Thiazoles 76 are formed by reacting the bromoketoamide with 10 under suitable conditions, preferably by heating the mixture in DMF or acetone. Amidines 77 are formed by heating 76 in toluene with trimethylaluminum and ammonium chloride. The amidines 77 are treated with strong acid such as HCl WO 00/47578 PCT/US99/18065 -63 5 to give the acids 78. The amidines 78 are in one route protected with a suitable protecting group such as BOC to give 79. The protected amidines 79 are treated with various amines under suitable coupling conditions, such as in the presence of HOAt, HATU, and DIEA to form various amides. The amidine protecting group can be then removed, for example by treating with 0 trifluoroacetic acid in a suitable solvent such as methylene chloride, when a BOC protecting group is employed to give amidines 80. In a second route, the amidines 78 can be loaded onto an insoluble support, such as polystyrene or polyethyleneglycol-grafted polystyrene via a cleaveable linker, such as Wang resin, which is functionalized as an activated carbonate ester, such as p 5 nitrophenylcarbonate or succinimidyl carbonate, to give a resin-bound scaffold 81. The resin-bound acid 81 can be treated with various amines under suitable coupling conditions such as in the presence of HOAT, HATU and DIEA to form amides. These amides are then cleaved from the solid support by treating with trifluoroacetic acid to give amidines 80. O Scheme 6a Scheme 6a illustrates the preparation of compounds of Examples 144, 145, 146, 147, 148, 149, 150 and 151. Compounds of this invention correspond to those of Formula I where R 2 is Formula II and where Ar is thiazole and R 37 and R" 8
(R
8 and R 9 of Formula II) are phenyl, which can be 5 additionally substituted. Starting from 2,5-dibromothiophene 90, treatment with lithium diisopropylamide followed by R 21 L, where L is a leaving group, preferably a halogen, mesylate, tosylate, or methyl sulfate, and more preferably iodomethane or methyl sulfate, according to the procedure of Kano, et al., Heterocycles 20(10):2035 (1983), gives 91. Compound 91 can be treated with 0 an appropriate base, preferably a lithium alkyl like n-butyllithium, sec butyllithium, or t-butyllithium, and more preferably n-butyllithium, followed by carbon dioxide gas and conversion of the resulting carboxylate salt to the free acid with a mineral acid, preferably hydrochloric acid. Conversion to ester 21 can be carried out by preparation of the acid chloride using oxalyl WO 00/47578 PCT/US99/18065 -64 5 chloride and treatment of this intermediate acid chloride with an alcohol R 23 in an appropriate solvent, preferably dichloromethane, with an appropriate base, preferably pyridine. Compound 21 can be treated with copper (I) cyanide in refluxing dimethylformamide to give compound 9. Compound 9 can be treated with hydrogen sulfide gas in an appropriate solvent, preferably 0 methanol, containing an appropriate base, preferably triethylamine to give compound 10. Compound 10 can be treated with an appropriate ketone where L is a leaving group, preferably halogen, mesyl, or tosyl, and most preferably bromo, refluxing in a suitable solvent, preferably, acetone, dimethylformamide, dimethyl acetamide, methyl ethyl ketone, or other polar 5 aprotic solvents, and most preferably acetone to give compound 92. Compound 92 is treated with an appropriate reagent, preferably the aluminum amide reagent to give amidine 93. Scheme 6b Scheme 6b illustrates the preparation of the compound of Example 0 144, which corresponds to a compound for which R 2 is Formula II, and where Ar is thiazole and R 8 and R 9
(R
37 and R 38 in Scheme 6b) are phenyl, which can be optionally substituted. Starting from 2,5-dibromothiophene 90, treatment with n-butyllithium produces an anion which undergoes a rearrangement (Kano, S., et al, Heterocycles 20:2035 (1983)). Quenching with carbon 5 dioxide gas and conversion of the resulting carboxylate salt to the free acid with a mineral acid, preferably hydrochloric acid, gives acid 94. Conversion to ester 95 can be carried out by preparation of the acid chloride using oxalyl chloride and treatment of this intermediate acid chloride with an alcohol R 2 3 OH in an appropriate solvent, preferably dichloromethane, with an appropriate 0 base, preferably pyridine. Compound 95 can be treated with copper (I) cyanide in refluxing dimethylformamide to give compound 96. Compound 96 can be treated with hydrogen sulfide gas in an appropriate solvent, preferably methanol, containing an appropriate base, preferably triethylamine to give compound 97. Compound 97 can be treated with an appropriate ketone where WO 00/47578 PCT/US99/18065 -65 5 L is a leaving group, preferably halogen, mesyl, or tosyl, and most preferably bromo, refluxing in a suitable solvent, preferably, acetone, dimethylformamide, dimethyl acetamide, methyl ethyl ketone, or other polar aprotic solvents, and most preferably acetone to give compound 98. Compound 98 is treated with an appropriate reagent, preferably the aluminum 0 amide reagent (Al(CH 3
)
3
/NH
4 C1) to give amidine 99. Scheme 7a Scheme 7a illustrates the preparation of compounds for which R 2 is Formula H and Ar is thiazol-4-yl. As illustrated, the acids 13 can be converted to their acid chlorides by treatment with oxalyl chloride with 5 dimethylformamide catalysis in methylene chloride, or by using thionyl chloride, either neat or in an organic solvent, at ambient or elevated temperature. Compounds are then homologated to the desired a-haloketones 100 by sequential treatment with trimethylsilyldiazomethane and hydrogen bromide. An alternative would be to substitute diazomethane (generated from :0 Diazald®, Aldrich Chemical Co., Milwaukee, WI) for the trimethylsilyldiazomethane. Also, the conversion of 13 to 100 can be effected using the procedure derived for the synthesis of compound 42 from compound 46. The alpha-haloketones 100 are then allowed to react with the 5 appropriate thiourea (Scheme 7b) or thioamide derivative in an organic solvent, preferably acetone or dimethylformamide at 70 'C to give 2 aminothiazoles or thiazoles 101. The thiazoles 101 can be treated with the aluminum amine reagent (Al(CH 3
)
3
/NH
4 CI) formed at ambient temperature by the reaction of 0 trimethylaluminum with ammonium chloride in an organic solvent, preferably toluene. The ester can then be converted to the amidines 102 at elevated temperatures, preferably higher than 80 oC.
WO 00/47578 PCT/US99/18065 -66 5 Scheme 7b As shown in Scheme 7b, amines 110 (or their hydrochloride salts) can be converted to their respective mono-substituted thioureas (methan-1 thiones) 112 by treatment with thiophosgene to form the intermediate isothiocyanates 111. Preferred conditions include treating the amine with 10 thiophosgene in a biphasic solvent system composed of a halogenated solvent such as chloroform and an aqueous phase of saturated sodium bicarbonate. Alternatively, the reaction may be effected by treatment of 110 with a hindered amine and thiophosgene such as triethylamine or diisopropylethylamine in an organic solvent such as tetrahydrofuran or methylene chloride. Another 15 alternative to forming isothiocyanates 111 is the direct treatment of primary amines and carbon disulfide in pyridine with dicyclohexylcarbodiimide (Jochims, Chem. Ber. 101:1746 (1968)). Isothiocyanates 111 can be converted to thioureas 112 by treatment with an ammonia-alcohol solution, preferably a 2M ammonia in methanol or 20 ethanol solution, at room temperature or elevated temperatures (>70 0 C). Alternatively, the thioureas 112 can be prepared directly form the appropriate urea (or thioamide from the appropriate amide when R 8 =alkyl or aryl)) by treatment with Lawesson's reagent (Lawesson, S.-O., et. al. Bull. Soc. Chim. Belg. 87:223, 293 (1978)). 25 Scheme 8 Scheme 8 illustrates the preparation of compounds of this invention where R 2 is Formula II and Ar is thiazole and R 37 and R 3 8 are phenyl which is further substituted by a sulfonylamino or carbonylamino group. Starting from thioamide 10, treatment with a nitro substituted 2-halo-acetophenone, where 30 the halogen is chloro, bromo, or iodo, preferably bromo, refluxing in a suitable solvent, preferably acetone, dimethylformamide, dimethyl acetamide, methyl ethyl ketone, or other polar aprotic solvents, and most preferably acetone. The reduction of nitroaryl compound 113 can be carried out with a suitable reducing agent, preferably tin (II) chloride, titanium (II) chloride, iron (III) WO 00/47578 PCT/US99/18065 -67 5 chloride, lithium metal, sodium metal, catalytic hydrogenation over platinum or palladium catalyst, and most preferably 20% aqueous solution of titanium (III) chloride. The acylation of aniline 114 can be carried out with an appropriate acyl compound R 42 L where L is a halogen, preferably chloro, in an appropriate solvent, preferably dichloromethane, containing a base, preferably 0 pyridine, N-methylmorpholine, or diisopropylethylamine. Alternatively, the acylation of aniline 114 is carried out with an activated carboxylic acid compound R 42 COL where L is hydroxy activated with dicyclohexylcarbodiimide, ethyl-3-(diethylamino)propylcarbodiimide (EDAC), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium 5 hexafluorophosphate (HATU), orpentafluorophenyl. The sulfonylation of aniline 114 can be carried out with and appropriate sulfonyl chloride compound R 41
SO
2 L in an appropriate solvent, preferably dichloromethane, containing a base, preferably N-methyl morpholine, diisopropylethylamine, or pyridine, most preferably N-methyl morpholine, with or without a !0 condensation catalyst, preferable dimethylaminopyridine (DMAP). The amidinylation of compounds 115 and 117 can be carried out with an appropriate reagent, preferably the aluminum amide reagent (Al(CH 3
)
3
/NH
4 C). Scheme 9 5 Scheme 9 illustrates the preparation of compounds of Formula I, for which one of R 5 and R 6 is a non-hydrogen substituent. The amidines 5 are converted to the amidoximes 119 by heating with hydroxylamine in a suitable solvent such as ethanol. The cyanoamidines 120 are prepared by heating the amidines 5 with cyanamide in a suitable solvent such as ethanol. (Huffman, 0 K.R. and Schaeffer, F., J. Amer. Chem. Soc. 28:1812 (1963). Alternatively 5 can be heated with an amine such as methylamine to give the N-alkylated amidines 121.
WO 00/47578 PCT/US99/18065 -68 5 Scheme 10 Scheme 10 illustrates an approach to compounds of Formula I where X = S or O, R 2 = arylamino, R 3 = alkylthio, aralkylthio, arylthio, alkyloxy, aralkyloxy, aryloxy, alkylamino, dialkylamino, aralkylamino, diaralkylamino, arylamino or diarylamino, Y = bond and Z = NR'R 6 0 Aminothiophenes 47 (Formula I where X = S, or O; R 2
=NH
2 ) can be reacted with an arylboronic acid (R 5 6
B(OR
8
)
2 , R 58 = H) or arylboronic ester
(R
5 6
B(OR
58
)
2 , R 5 8 = alkyl) in the presence of a copper catalyst, preferably copper (II) acetate, and an amine base such as triethylamine or pyridine (Chan, D. M. T. et al., Tetrahedron Lett 39: 2933 (1998)) to give a thienylarylamine 5 124. Conversion of the ester to amidine 125 is carried out in the manner previously described in Scheme lb. Scheme 11 Another route to compounds of this invention, where R2 = arylamino or alkylarylamino and alkylamino and where R', Y and Z are as described in 0 Scheme 10, is shown in Scheme 11 where intermediate 2 (R 21 = R 3 , L = leaving group) is aminated using conditions well known in the art. (See for example: Ahman, J. and Buchwald, S. L., Tetrahedron Lett. 38: 6363 (1997) and Wolfe, J. P. and Buchwald, S. L., Tetrahedron Lett. 38: 6359 (1997). For reviews see: Frost, C. G and Mendonca, P., J. Chem. Soc, Perkin Trans 1: 5 2615 (1998) and Wolfe, J. P. etal.,Acc. Chem. Res. 31: 805 (1998).) Thus, 2 may be treated with an aniline R 56
R
57 NH (R 5 6 = aryl, R 57 =H or alkyl) in the presence of a palladium catalyst, a suitable palladium ligand and a base to give 127. Suitable catalysts include any of a number of Pd(0) or Pd(II) salts, such as tetrakis(triphenylphosphino) palladium (0), 0 dichlorobis(acetonitrile)palladium (II) or preferably palladium (II) acetate or tris(dibenzylideneacetone)dipalladium. The most appropriate ligands for any given reaction are often compound-dependent and are discussed in detail in the aforementioned references but may include 1,1 ' bis(diphenylphosphino)ferrocene (DPPF), 1-[2- WO 00/47578 PCT/US99/18065 -69 5 (diphenylphosphino)ferrocenyl]ethyl methyl ether (PPF-OMe), or preferably 2,2'-bis(diphenylphosphino)- 1,1'-binaphthyl (BINAP). Appropriate bases include sodium t-butoxide or preferably cesium carbonate or potassium phoshate. Useful solvents include DMF, dioxane, dimethoxyethane or preferably toluene. Conversion of the ester to amidine 128 is carried out in the 10 manner previously described in Scheme lb. Scheme 12 The corresponding compounds of Formula I where R 2 = alkylamino and R 3 , Y and Z are as described in Scheme 10 are produced as shown in Scheme 12 by initial reductive alkylation of amine 47 with an aldehyde 5 R 59 CHO or ketone R 59
COR
6 " in the presence of any of a number of suitable reducing agents including sodium borohydride, sodium cyanoborohydride or, more preferably, sodium or tetraalkylammonium salts of triacetoxyborohydride to give 129. Depending on the reducing agent employed, suitable solvents may include an alcohol, such as methanol, ethanol or isopropanol, or solvents 10 such as THF or dichloromethane. Conversion of the ester to the amidine 130 is again carried out in the manner previously described in Scheme lb. Scheme 13 Scheme 13 illustrates an approach to compounds of Formula I where
R
2 is a 2-thiazolylamino and R 3 , Y and Z are as described in Scheme 10. In this 5 method, amine 47 can be first converted to a thiourea 131 using the various procedures outlined in Scheme 7b. Further reaction of the thiourea with a leaving group-substituted ketone R 37
COCH(L)R
8 , preferably a 2-haloketone as described in Schemes 2f, 2g or 2h, can provide thiazolylaminothiophenes 132 which are then converted to the corresponding amidines 133 by the 0 previously described methodology of Scheme lb.
WO 00/47578 PCT/US99/18065 -70 Scheme ]a L R 21 R2 L Br R22
R
1 ~RI 2 R' x =0, S W =CN, COOH, CONH 2 , C0 2 R 23 L =Br Schzemeilb
R
4 XV = R~ NH 2
W=NH
2
R
4 N X 22R Z/1 135 R 1 3 z ' N Z H R 1 136 Z = H, alkyl Z =NR 6
R
6 Z = NR 5
R
6 WCO W= CN, CONH, or W=CN (COd) 2 Ithen3 =NCl 20
R
23 OH C 2
R
23 W= HCI I R 2 H (or CH 2
N
2
R
5 = R6= H R 6 = alkyl, aryl, aralky R3=Me)/ A4(CH 3
)
3 / NH4CI AI(CH 3
)
3 I/ R 5
R
6 NH Toluene /IA Toluene I A R 21 ~R 21 R2
AI(CH
3
)
3 R3X NH 4 CI R x 23 8 N R X Toluene A R6*-HCI 0 HId 1R R'N N 0 R4R5=RHNH 5 NH 6 WO 00/47578 PCT/US99/18065 -71 Scheme Jc R21 SR-4
CS
2 + R1 2
CH
2
R
22 + R4L + WCH 2 L base X R2- base R 54 SS + R2 w- WH R22 R2= CN, C0 2
R
23 , CONH 2 Rl3R 22 = CN 138 R2= CN, C0 2
R
23 W = CN, C0 2
R
23 , CONH 2 X =S L = Cl, Br, 1, OTs, OMs, OTf R 2 1 = SR64 RI OH, NH 2 W =CN, CONH 2 , C0 2
R
23 diazotization X R22 R = NH 2 w* S Scheme Id L L W* w w*
R
1 2 139 R R 1=HX0,S L=Br L= Br W=CN, COOH, CON H 2 , C0 2 R 23 WO 00/47578 PCT/US99/18065 -72 Scheme 2a L
R
21 21 X L R 2 1 M X Br MCN X CN M = Li, Na, K 0O
H.
O
1 W 1 X= O,S O 7 8 O W= COOH L = Br
(COCI)
2 then R 23 0H or CH 2
N
2
(R
23 = Me)
R
21 R21 R21 SXRM S X R 2 4
COCH(R
25 )L X H 2 S Ra X C A I CN R O4 R - 0 NH 2 0 O 11 O 10 O
AI(CH
3
)
3 L = CI, Br, I, OMs, OTs AI(CH3)3
NH
4 Cl Toluene IA R21 S R2
H
2 N N R24 NH 12 WO 00/47578 PCT/US99/18065 -73 Scheme 2b F F COOH R21 R21 1) (COCI) 2 X F COOH X 2) R 26
COCH(R
27
)NH
2 CN COOH o OCN F Oor R - R23 1) (COCI) 2 O 9 O 13 2) R 26
COCH
2
NH
2 3) Optional alkylation X =O,S R 21 with base R 27 L X R27 L = CI, Br, I, OMs, OTs O 14 R26 0 POCla NH 4 0Ac P 2 s 5 HOAc or Lawesson's reagent
RR
21
R
21 X RX H R R R-'Rz7 Rn' - N RZ7 R3" 2 O 15 O 17 O 19
AI(CH
3
)
3 / NH 4 CI
AI(CH
3
)
3 / NH 4 CI
AI(CH
3
)
3 / NH 4 CI Toluene / A Toluene / Toluene I A
H
2 X N RH 2 X8 H 2 N NH NH NH NH16 18 20 WO 00/47578 PCT/US99/18065 -74 Scheme 2c
CH
2
N
2
R
21 R21
(R
23 = Me) x Pd(0) catalyst X , SnR 2 - - Br a SnR 2 or (COCI) 2 R23- (SnR283) 2 R23O then R 23 0H O 21 O 22 0 21 0 2 X = O, S R 2 8-nBu W = COOH L = CI, Br, I, OTf R21 Pd(0) catalyst x O
NH
3 / NH 4 CI
R
12 CH(L)COCI 23 0 L or R 1) MN 3 Cl, Br, , OMs, OTs 0 R 2) H 2 / Pd (cat) LCI, Br, I, OMs, OTs 23 HCI R21
R
21 RMCONHM X 0 R3COCI X 0 R2
NH
2 R23O NH 0 R 0 R3 O R HCI O 0 24 25
POC
3
NH
4 OAc I \ PS HOAc or Lawesson's reagent
R
2 1 R29 R 21 R29 R 21 R29 X N X\ X N R231 O R0 R2- O O R3 0 02 R0 R 0H SR3 R33 o 26 O 28 O 30
AI(CH
3
)
3 I NH 4 CI AI(CH 3
)
3 I NH 4 CI AI(CH 3
)
3 / NH 4 CI Toluene A I Toluene I A Toluene / A
R
21 R29 R 21 R29 R 21 R29 XN N NX N
H
2 N O R H 2 N R H2S / N 0 H NH N H 27 NH 29 N H 31 WO 00/47578 PCT/US 99/180 65 -75 Scheme 2d R21 Br 1. base R1 S(2)
(CH
3
)
3 SnCI R 43 1-n (n = 1-2) X 2. (CH) 3 SiCHN 2 S ~'Pd(PPh 3
)
4 R3or DMF, 120 0 C W R 2 L/base OR1 23 L = Br, 1, OTf W ,OO 22 43 8 Me aryl, heteroaryl, allyl, vinyl 2
R
21
R
43 L-n-i When n = 1 S NH- 4 01
AI(CH
3
)
3 32 TolueneA ,OMe When n =2 R44B(OH) 2
R
21
R
43 R44~ ~ ~ ~DF arl 0 0 Coayl P(Ph) aryl, ~ DMF h9e0-ry P( Ch
K
2 C0 3 S R 21
R
43
R
21
R
43 HN NH 2 R- R44 NH- 4 CI - R R4"N 3 S 7AI(CH 3
)
3 S7 TolueneIA H NH- 2 OMe 35 34 WO 00/47578 PCT/US99/1 8065 -76 Scheme 2e 1) (COdI) 2 0 X \ COOH 2) R 27
CH(NH
2 )CH(R 26 )OH X R 27 0 03 R 21 H 0 oxidation X R2 POCd 3 0 37 0 R 27
AI(CH
3
)
3
INH
4 CI R 27 X N.. TolueneA X
H
2 N R 21-r : 3 (R 2639 R2 0 38NH Scheme 2f 0 42R8 CN NHs 21 -2IM O 21 31_____ _ IN N 1 w S R- triethylamine S Rw s 40 41 43 WO 00/47578 PCT/US99/18065 -77 Scheme 2g 0 0 R37 R38 halogenation
R
37 '
R
38 L 44 42 L = halogen Scheme 2h 0 1)TMSCHN 2
R
37
R
37 COOH R 3 7 COL 2)HL L 2) H L 38 42 45 46 L = CI, Br
R
38 = H L = CI, Br, OCOR 39 L = CI, Br
(R
39 = iBu, tBu) 5 Scheme 2i R21
R
2 1 0 AI(CH 3
)
3
/NH
4 CI X 1) (COCI)2 X Toluene/A COOH >
R
2 3 O 2) R 5 s 1
RS
2 NH R2 3 0 N-R 13 2 0 0 130 X =N, 0, S
R
21 R21 O NH X AI(CH 3
)
3
/NH
4 CI NH SToluene/A XH2N
H
2 N 51 N - 51
R
52 R52 NH 131 NH 132 WO 00/47578 PCT/US99/1 8065 -78 Scheme 3a1 1) (PhO) 2
P(O)N
3 X X\ COOH ET 3 N/t-BuOH 0 NH 2 R302)
CF
3
COOHICH
2
CI
2
R
23 R 23 13 3) aq NaHCO 3 4 00 1) R 31 S0 2 C1 2) R 32 Lbase Rs 1 NCO R 31 COCI (optional) or R 51 R1 2 NCOCI R1R 21
R
2 1 R2-0 R3SO 2 R" R 2 3-
HON
2 R5 R 23 05 FCR3 48 050 00
AI(CH
3
)
2 INHPC AI(CH,) 3
INH
4 CI AI(CH 3
)
3
/NH
4 CI Toluene/A Toluene/A TolueneIA
HNNR
22 S0 2 R 31 H2 \ NHCONR" 1
R
52 XY 53 NHCOR 31 49 y 51 5 NH NH NH
R
2 1
AI(CH,),/NH
4 CI Toluene/A
H
2 Hx
NHH
WO 00/47578 PCT/US99/18065 -79 Scheme 3b R21 HCI/NaNO 3 / X 1) R4NH 2 47 CuSO4/NaHSO 3 23 - 2) R Lbase (optional) 55 O R21
R
21 X AI(CH 3
)
3
/NH
4 CI X
SO
2 NR34 3 5 Toluene/A -
SO
2 N R34R 3 5 R23 01
H
2 56 57 O NH Scheme 3c L R21 R 2 1 X R 2 1 X NO 2 Reduction X NH
NO
2 NH 2 R 23/O" ") R23/ \ 2/ \ 0 2 R 23/ 23 O 122 o 123 0 47 X =0, S L = CI, Br, I, OTs, OMs WO 00/47578 PCT/US99/18065 -80 Scheme 4a 21 21 Ar & D Ar 1. optional reduction X 2. Br(CH 2 )nCOR 36 SR protecting (P 1 ) group X R Cs,CO. introduction DMF, 700C p i n = 1-4 H NH 2 H H H D = OH, SH, NO 2 X = S,O 61 60 When P1 and R 36 21 are ortho ona 21 Ar LE--(Cn R 36 removal Ar --E-(C X R R X R OH When
P
1 and
R
36 Hi 62 are not orthogonal H 63 Hh N E = O,S, NH HP 63 H 1. P 1 and R 36 removal H 2. protecting (P 1 ) group introduction HOAT HOAT * HATU HN DMF R" 21 21 Ar LE_.(C2)n ArR,-R-2C ArA X R - R P 1 removal X R 9 H N 2 65 H P i 64 H SUBSTITUTE SHEET (RULE 26) WO 00/47578 PCT/US99/18065 -81 Scheme 4b / - / N :TO N y~O BrCHCOR 36 DIEA 0 CS 2
CO
3
H
2 DMF lo DMF, 70 0 C 2H R 36 08u~, OMe 66 H 67 / / ~N \/When R 3 6 OMe ~ N '/ S 0 6 NaOH, THF S 0 OH H H-1 69 H H 68 IWhen R 36 =OBul HOAT /R' 50% TFAIDCM HATU IHN 2% H,0 DMF /S\ S\ N N S 0 1 OH A'0 1 0 70 0 "R H 2 TFA salt H H 7 72
NO
2 j~ ~ ~ ~ ~ ~ ~~~~0 TFA.LJeIJIslbl upr ~, ~gDCM DBU, DMF /S 1. /lHOA T /' / S\ / N DMF "R N S0 O 2.50% TFAIDCM S A'1 O OH *2% H,0 \R0 0 0 H H 2 71 ~ ~e 7Z Isolule ~TFA salt 73 WO 00/47578 PCT/US99/18065 -82 Scheme 5 0 0Br 2 /AcOH Acetone reflux H H R 21 -0 Br __ NI-I 2 OR 23 10 (R 23 =Me) NH NHA S AIMe3/NH 4 CI S \HCIIH 2 0 S\ R 21 ~N / toluene, reflux -0-R 21 ~N R / 21 ~N ' S ~S OMe HAN NH- 2 H NH 2 76777 H R 0_____ 0 / 2 1. HOAT ,R' NIE HATU HAN\2 DIEA DMF R' N DFS 2. 50% TFAIDCMN 2% H 2 0 H
S
7 H H9 H NH- 2 80 1. 50% TFAIDCM 2% H 2 01.HA 2. NO 2 1.HAT HN DMF R 2. 50% TFAIDCM Linke Insoluble SUpport H 2% H 2 0 DBU, DMF S R 21 N /
S"
H 81 WO 00/47578 PCT/US99/18065 -83 Scheme 6a Br 1) LDA, -780C. B S 2) R 21 -L r rL = CI, Br, I, OMs, OTs, OSO 2 OMe Br S R 21 90 91 1) base, -780C 2) CO 2 (g) 3) 6N HCI (aq) L 1) base,-78 0 C S L 2) R 21 -L S R21 L = CI, Br, I, OMs, OTs, OSO 2 OMe 1 HO 7 W= COOH W= COOH L=Br L = Br 1) (COCI) 2 , CH 2
CI
2 , DMF 2) R 23 -OH, Pyridine CN o /\ ____ _/ \ S R 21 CuCN, DMF, reflux R 21 R2WL- R 2 -O 21
R
23 = Me, i-Pr R 23 = Me, i-Pr L= Br
H
2 S, TEA, MeOH
R
37 ' L= Br 38
NH
2 L = CI, Br, I, OTs, OMs 'o /\ O / \ Acetone, reflux R 21 S
R
2 1 R231R2. 92
R
2 - 10 92
R
23 = Me, i-Pr AI(CH 3
)
3 , NH 4 CI toluene, reflux 3 37 N HN H S R21 H2 93 WO 00/47578 PCT/US99/18065 -84 Scheme 6b 1) Butyllithium, -78 0 C. 2) C0 2 (g) r 3) 6N HCI (aq) , O Br S r B S 90 94 OH 1) (COCI) 2 , CH 2
CI
2 , DMF 2) R 2 3 -OH, Pyridine NC S O CuCN, DMF, reflux BrO Br s 96 O-R 23 95 O-R23
R
2 3 = Me, i-Pr
H
2 S, TEA, MeOH
SR
3 8 23 L =ACletlr , r eiOTsOMs R 3 R S Acetone, reflux I / S
H
2 N O-R 2 3 R O-R 23 97 98
AI(CH
3
)
3 , NH 4 CI toluene, reflux
R
3 S NH I, s
R
37 :N
NH
2 99 SUBSTITUTE SHEET (RULE 26) WO 00/47578 PCT/US99/18065 -85 Scheme 7a
R
23
R
23 0 1. oxalyl chloride O X / O 2. TMSCHN 2 , then X OH 30% HBr-acetic acidBr R21
R
21
R
9 13 100 R 9 = H 13 100
R
2 3 = CH 3 S
H
2 N R8 acetone or DMF 70 0 C
NH
2
R
9
OR
23
R
9 R SR AIMe 3 , NH 4 CI HN
-
S toluene, 118 0 C O S
R
2 1 R21 102 101 Scheme 7b s S thiophosgene S
H
2 N R 40 CHCl 3 , sat'd. NaHCO 3 C NH 3 -solvent H2NR 1 N
H
2
NHR
40
R
4 0 110 111 112 WO 00/47578 PCT/US99/18065 -86 Scheme 8 Br N NO,
NH
2 0 2 N S R21 S R21 Acetone, reflux
R
23 -O 20% Ti 113
R
23 -O 10 113
R
23 = CH 3 C3(aq) 20% TiCI 3 (aq) THF, rt N
NH
2 1141 S 21
R
23 -0 114
CH
2
CI
2
CH
2
CI
2 N-methyl morpholine (1.5 eq) Pyridine DMAP (0.1 eq) (1.0 eq)R4-CI (1.0 eq) R4t'kL 1 00 N N-C- R 4 N N-S0 2 - R 41 7o R21 5\ / 21 SS R2 R23-O 115 R 23 -0 117
AI(CH
3
)
3 , NH 4 CI AI(CH 3
)
3 , NH 4 CI toluene, reflux toluene, reflux 0
H
1 H N N- C- R42 N N- S0 2 - R 4 1 HN, / S HN / \ S 21 116 s R2 118 H2N H118
H
2 N H 2
N
WO 00/47578 PCT/US99/18065 -87 Scheme 9 R21 R22 x Rso HN H 5 (when R 50 = H)
NH
2 OH NH 2 CN MeNH 2 EtOH, A EtOH, A MeOH, A
R
21 R22 R 21 R22 R 2 1 22 x OH CN Me HHN HN H HN H H 119 120 121 Scheme 10
R
21 R21 R 21 Ai(CH 3
)
3 / X R 56 B(ORs 8 ) X 6 AI(CR NH 4 X
N-
2 NH4CI X" Rs 0 Cu(II) catalyst N 57 Toluene/A NR 57 R23/ H 0 47 124 NH 125 X = O, S
R
57 = H
R
5 7 = H WO 00/47578 PCT/US99/18065 -88 Scheme 11
R
21 R21 AI(CH3)z R21 RS6RS7H X' 'AI(CH 3
)
3 /I X R 56
R
57 NH X NR NH 4 CI xR Pd catalyst Toluene / A N O ligand O R7 H R 7 R23/ base R23/ H 2 0 2 127 NH 128 X =O,S L = CI, Br, I, OSO 2
CF
3 Scheme 12
R
21
R
21
R
59
R
60 R1R9 6 R R21 59 R AI(CH 3
)
3 / R 21
R
59 R6 X RS 9
COR
6 0 Xy NHI4CI X /
NH
2 reducing Toluene / A N agent O H H2 H R23/ R / 2 o 47 O 129 NH 130 X = O, S 5Scheme 13
R
37 1)CSCI 2
R
2 1 S
R
21 N -R R 38
CH
2
CI
2 X -NH 2 S 47 aq. NaHCO 3 NH R 37 COCH(L)Rj NH 2)NH 3 -MeOH 0 R 0 R23/ 23 0 131 R o 132 X = O, S
R
37
R
21 N
R
38 Al(CH 3
)
3 S
NH
4 CI X N Toluene / A
H
2 N NH 133 WO 00/47578 PCT/US99/18065 -89 Example 1 4-J4-(4-chlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine Trimethylaluminum (2.0 M in toluene, 2 mL) was added dropwise over 10 min to a suspension of ammonium chloride (216 mg) in toluene (2 mL), stirred under 5 N 2 at 0 oC. When gas evolution moderated, the mixture was stirred at 25 0 C for 30 min, when most of the solid had dissolved, methyl 4-[4-(4-chlorophenyl)(1,3-thiazol 2-yl)]-5-methylthiothiophene-2-carboxylate (100 mg, Maybridge Chemical Co., Cornwall, U.K.) was added in one portion. This solution was heated to reflux in stages over 1 h. After 2.5 h of reflux, the reaction mixture was allowed to cool to 25 0 C, and 10 was poured on to a vigorously stirred slurry of silica gel (2 g) in CHC13 (20 mL). After 20 min the solids were collected by suction filtration, and washed with MeOH (3x1 0 mL). The combined filtrates were evaporated to dryness, and the residual yellow solid was subjected to preparative thin-layer chromatography to obtain 77 mg of 4 [(4-chlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine as a yellow 15 solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 2.80 (s, 3H), 7.55-7.59 (m, 1H), 8.04-8.13 (m, 1H), 8.31 (s, 1H), 8.69 (s, 1H),), 9.2 (broad s, 4H). Mass spectrum (MALDI TOF, m/z): Calcd. for C 15
H
12 C1N 3
S
3 , 365.9 (M+H), found 366.9. Example 2 20 5-Methylthiothiophene-2-carboxamidine 5-(Methylthio)thiophene-2-carbonitrile (100 mg, Maybridge Chemical Company, Cornwall, UK) was taken in a dry 2 dram vial. To this a solution of saturated HCI in anhydrous MeOH (4 mL) was added. The vial was tightly capped and the mixture was stirred for 24 h. The vial was cooled in an ice bath, uncapped 25 and N 2 was bubbled through the solution to remove dissolved HC1. The solvent was removed under reduced pressure and the resulting residue was dried under high vacuum for 24 h. A solution of methanolic ammonia (2M NH 3 in MeOH) was added to the vial, and the mixture was stirred for 3 days. Methanol was removed under vacuum and the resulting residue was subjected to preparative thin-layer 30 chromatography to obtain 5-(methylthio)thiophene-2-carboxamidine as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 5 2.64 (s, 3H), 7.22 (d, J=3.8 Hz, 1H), 7.95 (broad d, WO 00/47578 PCT/US99/18065 -90 J=3.33 Hz, 1H), 9.4 (broad s, 4H). Mass spectrum (MALDI-TOF, m/z): Calcd. for
C
6 HsN2S 2 , 172.3 (M+H), found 173.0. Example 3 5 5-Methylthio-4-phenylthiophene-2-carboxamidine Methyl 5-methylthio-4-phenylthiophene-2-carboxylate (100 mg, Maybridge Chemical Company, Cornwall, UK) was treated in a manner similar to that for Example 1, to give 50 mg of 4-phenyl-5-methylthiothiophene-2-carboxamidine as an off-white solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 5 2.65 (s, 3H), 7.39-7.60 (m, 5H), 10 8.27 (s, 1H), 9.2 (broad s, 4H). Mass spectrum (MALDI-TOF, m/z): Calcd. for
C
12
H
1 2
N
2
S
2 , 248.4 (M+H), found 249.0. Example 4 4-[4-(2,4-Dichlorophenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 15 carboxamidine Methyl 4-[4-(2,4-dichlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate (100 mg, Maybridge Chemical Company, Cornwall, UK) was treated in a manner similar to that for Example 1, to give 60 mg of 4-[4-(2,4 dichlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine as a yellow 20 solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 2.77 (s, 3H), 7.6 (dd, J=2.2 and 8.5 Hz, 1H), 7.79 (d, J=2.2 Hz, 1H), 8.09 (d, J=8.5 Hz, 1H), 8.3 (s, 1H), 8.6 (s, 1H). Mass spectrum (MALDI-TOF, m/z): Calcd. for C IHiN 3
S
3 C1 2 , 400.0 (M+H), found 400.1. Example 5 25 4
-(
4 -Methyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine Methyl 4-(4-methyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxylate (100 mg, Maybridge Chemical Company, Cornwall, UK) was treated in a manner similar to that for Example 1, to give 40 mg of 4-(4-methylthiazol-2-yl)-5 methylthiothiophene-2-carboxamidine as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 30 MHz) 8 2.43 (s, 3H), 2.7 (s, 3H), 7.38 (s, 1H), 8.28 (s, 1H). Mass spectrum (MALDI TOF, m/z): Calcd. for CIoHIIN 3
S
3 , 270.0 (M+H), found 270.1.
WO 00/47578 -91- PCT/US99/18065 Example 6 a) Methyl 5-methylthio-4-(4-(2-naphthyl)(1,3-thiazol-2-yl))thiophene-2 carboxylate: Methyl 4-(aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (40 mg, Maybridge Chemical Company, Cornwall, UK) was reacted with 2-bromo-2' 5 acetonaphthone (1.1 eq) in a manner similar to Example 13 step (a) to give 40 mg of methyl 5-methylthio-4-(4-(2-naphthyl)(1,3-thiazol-2-yl))thiophene-2-carboxylate.
'H
NMR (CDC1 3
/CD
3 OD; 300 MHz) 8 3.71 (s, 3H), 3.94 (s, 3H), 7.47-7.55 (mn, 2H), 7.67 (s, 1H), 7.84-7.99 (m, 3H), 8.08 (dd, J=1.75 Hz and 8.6 Hz, 1H), 8.3 (s, 1H), 8.5 (s, 1H). 10 b) 5-Methylthio-4-(4-(2-naphthyl)(1,3-thiazol-2-yl))thiophene-2 carboxamidine: Methyl 5-methylthio-4-(4-(2-naphthyl)(1,3-thiazol-2-yl))thiophene 2-carboxylate, (40 mg) as prepared in the previous step was treated in a manner similar to that for Example 1, to give 30 mg of 4-[4-(naphth-2-yl)thiazol-2-yl]-5 methylthiothiophene-2-carboxamidine. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 2.83 (s, 15 3H), 7.52-7.69 (min, 2H), 7.95-8.01 (min, 2H), 8.05 (d, J=8.6 Hz, 1H), 8.24 (dd, J=1.69 Hz and 8.6 Hz, 1H), 8.4 (s, 1H), 8.65 (s, 1H), 8.74 (s, 1H). Mass spectrum (MALDI TOF, CHCA matrix, m/z): Calcd. for C 19
H,
5
N
3
S
3 , 382.1 (M+H), found 382.0. Example 7 20 5-Methylthio-4-J4-(4-phenylphenyl)(1,3-thiazol-2-yl)]thliophene-2-carboxamidine hydrochloride a) Methyl 5-methylthio-4-[4-(4-phenylphenyl) (1,3-thiazol-2-yl)]thiophene-2 carboxylate: 27 mg (0.109 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene 25 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 2 mL of reagent grade acetone. 4'-Phenyl-2-bromoacetophenone (33 mg; 0.120 mmol; Aldrich Chemical Co., Milwaukee, WI) was added and the solution was allowed to reflux for 2.5 h. The solution was allowed to cool and solid was filtered and washed with methanol and dried in vacuo to afford 30 mg (65% yield) of methyl 5-methylthio-4-[4-(4-phenylphenyl)(1,3 30 thiazol-2-yl)]thiophene-2-carboxylate. 'H-NMR (DMSO-d 6 , 300 MHz) 6 8.28 (s, 1H), 8.24 (s, 1H), 8.17 (d, J=8.5 Hz, 2H), 7.8 (d, J=8.5Hz, 2H), 7.74-7.77 (mn, 2H), 7.48-7.53 (mn, 2H), 7.40 (min, 1H), 2.78 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for
C
22
H,
6
NO
2
S
3 :423.0 (M+H), found 424.4.
WO 00/47578 PCT/US99/18065 -92 b) 5-Methylthio-4-[4-(4-phenylphenyl) (1,3-thiazol-2-yl)]thiophene-2 carboxamidine hydrochloride: To a stirred suspension of 0.473 mmol ( 25 mg) of ammonium chloride (Fisher Scientific Pittsburgh, PA) in 2 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 237 ptL (0.473 mmol) of 2M 5 trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir at 0 oC for 30 min after which 20 mg (0.0473 mmol) of methyl 5-methylthio-4 [4-(4-phenylphenyl)(1,3-thiazol-2-yl)]thiophene-2-carboxylate was added to solution and allowed to reflux for 2.5 h. The reaction mixture was quenched by pouring over a slurry of 500 mg of silica in 10 mL of chloroform. The silica was poured onto a sintered glass funnel 10 and washed with a 10% methanol/CH 2 C1 2 solution and concentrated. The crude product was purified on a 1 mm silica prep plate eluting with 10% methanol/CH 2 C1 2 to afford 10 mg (53% yield) of 5-methylthio-4-[4-(4-phenylphenyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine hydrochloride. Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for C 21
H,
7
N
3
S
3 : 408.1(M+H), found 408.0. 15 Examples 8 & 9 4-[4-(3-Methoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride and 4-J4-(3-hydroxyphenyl) (1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxamidine hydrochloride 20 a) Methyl 4-[4-(3-methoxyphenyl) (1,3-thiazol-2-yl)]-5-mnethylthiothiophene-2 carboxylate: 32 mg (0.133 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 2 mL of reagent grade acetone. 3'-Methoxy-2-bromo acetophenone (0.155 mmol; 36 mg; Aldrich 25 Chemical Co.) was added and the solution was allowed to reflux for 2.5 h The solution was allowed to cool and a solid was filtered and washed with methanol and dried in vacuo. The solid was purified on 1 mm silica plate eluting with 25% ethyl acetate/hexane to afford 31 mg (63% yield) of methyl 4-[4-(3-methoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate. 30 b) 4-[4-(3-Methoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride and 4-[4-(3-hydroxyphenyl) (1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxamidine hydrochloride: To a stirred suspension of 0.821 mmol (44 mg) of ammonium chloride (Fisher Scientific) in 2 mL of anhydrous toluene WO 00/47578 -93- PCT/US99/18065 (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 'C, was added 411 pL (0.821 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) via syringe over 10 min and then let stir at 0 oC for 30 min after which 31 mg (0.0821 mmol) of methyl 4-[4-(3 methoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was added to 5 solution and allowed to reflux for 2.5 h. The reaction mixture was quenched by pouring over a slurry of 500 mg of silica in 10 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2
C
2 solution and concentrated. The crude product was purified on a 1 mm silica prep plate eluting with 10% methanol/CH 2
C
2 to afford 4.4 mg (15% yield) of 4-[4-(3-methoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 10 carboxamidine hydrochloride and 4.2 mg (15% yield) of 4 -[4-(3-hydroxyphenyl)(1,3-thiazol 2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 4-[4-(3-methoxyphenyl)(1,3 thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride: 1 H-NMR (CD 3 OD; 300 MHz) 6 8.5 (s, 1H), 7.9 (s, 1H), 7. 59-7.65 (m, 2H), 7.33-7.38 (m, 1H), 6.91-6.95 (m, 1H), 3.87 (s, 1H), 2.8 (s, 3H) Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for 15 C 1
,
6 HisN 3 0S3: 361.5(M+H), found 362.2. 4-[4-(3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxamidine hydrochloride: 'H-NMR (CD 3 OD; 300 MHz) 6 8.50 (s, 1H), 7.81 (s, 1H), 7.26-7.51 (m, 2H), 7.24 (m, 1H), 6.79 (m, 1H), 2.8 (s, 3H) Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for C 15
HI
3
N
3 0S 3 : 347.5(M+H), found 348.0. 20 Example 10 5-Methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine hydrochloride 25 a) Methyl 5-methylthio- 4 -(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate: 33 mg (0.133 mmol) methyl 4-(aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 2 mL of reagent grade acetone. 2-Bromoacetophenone (0.133 mmol; 27 mg; Aldrich Chemical Co.) was added and the solution was allowed to reflux for 2.5 h. The solution was allowed to cool and the solid 30 was filtered and washed with methanol and dried in vacuo. The solid was purified on 1 mm silica plate eluting with 25% ethyl acetate/hexane mixture to afford 46 mg (90% yield) of methyl 5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate.
WO 00/47578 PCT/US99/18065 -94 b) 5-Methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine hydrochloride: To a stirred suspension of 1.32 mmol ( 71 mg) of ammonium chloride (Fisher Scientific) in 2 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 662 pL (1.32 mmol) of 2M trimethylaluminum in toluene 5 (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir at 0 oC for 30 min after which 46 mg (0.133 mmol) of methyl 5-methylthio-4-(4-phenyl(1,3-thiazol-2 yl))thiophene-2-carboxylate was added to solution and allowed to reflux for 2.5 h. The reaction mixture was quenched by pouring over a slurry of 500 mg of silica in 10 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% 10 methanol/CH 2 C1 2 solution and concentrated. The crude product was purified on a 2 g silica silica SPE column eluting with 10% methanol/CH 2
C
2 to afford 32.5 mg (75% yield) of 5 methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine hydrochloride. 1
H
NMR (DMSO-d 6 ; 300 MHz) 8 8.70 (s, 1H), 8.25 (s, 1H), 8.07-8.11 (min, 2H), 7.37-7.53 (inm, 3H), 2.8 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for C 15
H
13 3
S
3 : 15 331.5(M+H), found 332.1. Example 11 5-Methylthio-4-J4-(4-nitrophenyl) (1,3-thiazol-2-yl)]thiophene-2-carboxamidine hydrochloride 20 a) Methyl 5-methylthio-4-[4-(4-nitrophenyl)(1,3-thiazol-2-yl)]thiophene-2 carboxylate: 38 mg (0.141 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 2 mL of reagent grade acetone. 2-Bromo-4'-nitroacetophenone (0.155 mmol; 38 mg; Aldrich 25 Chemical Co.) was added and the solution was allowed to reflux for 2.5 h. The solution was allowed to cool and a solid was filtered and washed with methanol and dried in vacuo. The crude product was dissolved in CH 2 C1 2 and 0.141 mmol of N-(2-mercapto)aminoethyl polystyrene resin (Calbiochem, San Diego, CA; 1.28mmol/g; 110 mg) was added and allowed to stir overnight. The solution was filtered, concentrated and dried to afford 60 mg 30 (90% yield) of crude methyl 5-methylthio-4-[4-(4-nitrophenyl)(1,3-thiazol-2-yl)]thiophene-2 carboxylate. b) 5-Methylthio-4-[4-(4-nitrophenyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine hydrochloride: To a stirred suspension of 1.66 mmol (90 mg) of ammonium chloride (Fisher WO 00/47578 PCT/US99/18065 -95 Scientific) in 2 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 830 tL (1.66 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir at 0 oC for 30 min after which 60 mg (0.166 mmol) of 5-methylthio-4-[4-(4-nitrophenyl)(1,3-thiazol-2-yl)]thiophene 5 2-carboxylate was added to solution and allowed to reflux for 2.5 h. The reaction mixture was quenched by pouring over a slurry of 500 mg of silica in 10 mL of chloroform. The silica was poured onto a sintered-glass funnel and washed with a 10% methanol/CH 2 Cl 2 solution and concentrated. The crude product was purified on a 1 mm silica prep plate eluting with 10% methanol/CH 2 C1 2 to afford 12 mg (19% yield) of 5-methylthio-4-[4-(4 10 nitrophenyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine hydrochloride. 1H-NMR (CD 3 OD, 300 MHz) 8 8.58 (s, 1H), 8.32-8.33 (mn, 4H), 8.24 (s, 1H), 2.83 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for C, 5
H
2
N
4 0 2
S
3 : 376.5(M+H), found 377.3. Example 12 15 4-[4-(3,4-Ethylenedioxyphenyl)thiazol-2-yl-5-methylthiothiophene-2 carboxamidine hydrochloride a) Methyl 4-(4-(2H,3H-benzof3,4-e]l,4-dioxin-6-yl) (1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate: 40 mg (0.162 mmol) of methyl 4-(aminothioxomethyl) 20 5-methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 2 mL of reagent grade acetone. 1-(2H,3H-benzo[e] 1,4-dioxin-6-yl)-2 bromoethan-1-one (0.162 mmol; 42 mg; Maybridge Chemical Co. LTD., Cornwall, U.K.) was added and the solution was allowed to reflux for 3 h. The solution was allowed to cool and allowed to stir for 2 days after which the reaction solution was concentrated in vacuo. 25 The crude product was dissolved in 50 mL of CH 2
C
2 and partitioned between 50 mL of 1 N NaOH (aq.). The organic layer was obtained and dried over sodium sulfate and concentrated to afford 60 mg (90% yield) of methyl 4-[4-(3,4-ethylenedioxyphenyl)thiazol-2-yl]-5 methylthiothiophene-2-carboxylate. b) 4
-[
4 -(3,4-Ethylenedioxyphenyl)thiazol-2-ylJ-5-methylthiothiophene-2 30 carboxamidine hydrochloride: To a stirred suspension of 1.62 mmol ( 86 mg) of ammonium chloride (Fisher Scientific) in 2 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 810 gtL (1.62 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir at 0 oC for 30 min WO 00/47578 PCT/US99/18065 -96 after which 60 mg (0.162 mmol) of methyl 4-[4-(3,4-ethylenedioxyphenyl)thiazol-2-yl]-5 methylthiothiophene-2-carboxylate was added to solution and allowed to reflux for 2.5 h. The reaction mixture was quenched by pouring over a slurry of 500 mg of silica in 10 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% 5 methanol/CH 2
C
2 solution and concentrated. The crude product was purified on a 1 mm silica prep plate eluting with 10% methanol/CH 2 C1 2 to afford 47 mg (75% yield) of 4-[4-(3,4 ethylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H-NMR (CD 3 OD; 300 MHz) 6 8.53 (s, 1H), 7.73 (s, 1H), 7.56 (d, J= 2Hz, 1H), 7.50 (dd, J=2.1 Hz and 8.4 Hz, 1H), 6.89 (d, J=8.4 Hz, 1H), 4.28 (s, 4H), 2.8 (s, 3H). Mass Spectrum 10 (MALDI-TOF, CHCA matrix, m/z) Calcd. for C 17
H
15
N
3 0 2
S
3 : 389.5(M+H), found 390.1. Example 13 4-[4-(4-Methoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride 15 a) Methyl 4-J4-(4-methoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 30 mg (0.122 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 1.2 mL of reagent grade acetone. 2-bromo-4'-methoxy acetophenone (0.146 mmol; 28 mg; Aldrich 20 Chemical Co.) was added and the solution was allowed to reflux for 3 h. The solution was allowed to cool and a solid was filtered and washed with methanol and dried in vacuo to afford 46 mg (90% yield) of methyl 4-[4-(4-methoxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate. b) 4-[4-(4-Methoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 25 carboxamidine hydrochloride: To a stirred suspension of 1.22 mmol (66 mg) of ammonium chloride (Fisher Scientific) in 2 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 612 pL (1.22 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir at 0 oC for 30 min after which 46 mg (0.122 mmol) of 4-[4-(4-methoxyphenyl)(1,3-thiazol-2-yl)]-5 30 methylthiothiophene-2-carboxylate was added to solution and allowed to reflux for 2.5 h. The reaction mixture was quenched by pouring over a slurry of 500 mg of silica in 10 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2 C1 2 solution and concentrated. The crude product was purified on a 1 mm silica WO 00/47578 PCT/US99/18065 -97 prep plate eluting with 10% methanol/CH 2
C
2 to afford 32 mg (73% yield) of 4-[4-(4 methoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 1 H-NMR (CD 3 OD; 300 MHz) 8 8.53 (s, 1H), 7.98 (d, J=7 Hz, 2H), 7.75 (s, 1H), 7.01 (d, J=5 Hz, 2H), 3.9 (s, 3H), 2.8 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. 5 for Ci 6
H
1 5
N
3 0S 3 : 362.0 (M+H), found 362.2. Example 14 4
-[
4
-(
3
,
4 -Propylenedioxyphenyl)thiazol-2-ylf-5-methylthiothiophene-2-carbioxamidine hydrochloride 10 a) Methyl 4
-[
4 -(3,4-propylenedioxyphenyl)thiazol-2-ylf-5-methylthiothiophene-2 carboxylate: 42 mg (0.170 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 5 mL of reagent grade acetone. 3',4'-Propylenedioxy-2-bromoacetophenone (0.170 mmol; 28 mg; 15 Maybridge Chemical Co. LTD., Cornwall, U.K.) was added and the solution was allowed to reflux for 3 h. The solution was allowed to cool and a solid was filtered and purified on a 1 mm silica prep plate eluting with 20% ethyl acetate/hexane and dried in vacuo to afford 42 mg (59% yield) of methyl 4-[4-(3,4-propylenedioxyphenyl)thiazol-2-yl]-5 methylthiothiophene-2-carboxylate. 20 b) 4-[4-(3,4-Propylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine hydrochloride: To a stirred suspension of 1.01 mmol (54 mg) of ammonium chloride (Fisher Scientific) in 2 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 510 pL (1.01 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir at 0 oC for 30 min 25 after which 42 mg (0.101 mmol) of methyl 4-[4-(3,4-propylenedioxyphenyl)thiazol-2-yl]-5 methylthiothiophene-2-carboxylate was added to solution and allowed to reflux for 3 h. The reaction mixture was quenched by pouring over a slurry of 500 mg of silica in 20 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2
C
2 solution and concentrated to afford 20 mg (50% yield) of 4-[4-(3,4 30 propylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H-NMR (CD 3 OD; 300 MHz) 8 8.53 (s, 1H), 7.78 (s, 1H), 7.68 (d, J=2.2 Hz, 1H), 7.60 (dd, J=2.2 Hz and 8.4 Hz, 1H), 7.00 (d, J=8.3 Hz; 1H), 4.19-4.28 (min, 4H), 2.77 (s, 3H), 2.18-2.23 WO 00/47578 PCT/US99/18065 -98 (min, 2H). Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for C 1 8
H
17
N
3 0 2
S
3 : 404.1 (M+H), found 404.1. Example 15 5 5-Methylthio-4-(4-(2-thienyl) (1,3-thiazol-2-yl))thiophene-2-carboxamidine acetate a) 2-Bromo-1-(2-thienyl)ethan-1-one: To a solution of 500 mg (3.96 mmol) of 2 acetyl thiophene (Aldrich Chemical Co.) dissolved in 20 mL of CHCl 3 , was added 1 drop of 30% HBr/CH 3 COOH (Aldrich Chemical Co.) followed by 3.96 mmol (633 mg; 204 pL) of 10 bromine (Aldrich Chemical Co.) added dropwise over 30 min. The reaction was allowed to stir for 1 h. The solution was concentrated to an oil and dried in vacuo. The crude product was purified on 1 mm silica prep plates eluting with neat CH 2
C
2 to obtain 300 mg (37% yield) of 2-bromo-1-(2-thienyl)ethan-l-one. 1 H-NMR (CDCl 3 ; 300 MHz) 8 7.80 (mn, 2H), 7.18 (min, 1H), 4.37 (s, 2H). 15 b) Methyl 5-methylthio-4-(4-(2-thienyl)(1,3-thiazol-2-yl))thiophene-2-carboxylate: 44 mg( 0.176 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene-2 carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 3 mL of reagent grade acetone. 2-Bromo-1-(2-thienyl)ethan-1-one (0.176 mmol; 36 mg) was added and the solution was allowed to reflux for 3 h. The solution was allowed to cool and was 20 concentrated. The crude product was dissolved in 20 mL of CH 2 C1 2 and washed with 20 mL of lN HCI (aq.). The organic layer was obtained and dried over sodium sulfate to afford 115 mg (80% yield) of crude methyl 5-methylthio-4-(4-(2-thienyl)(1,3-thiazol-2-yl))thiophene-2 carboxylate. c) 5-Methylthlio-4-(4-(2-thienyl) (1,3-thiazol-2-yl)) thiophene-2-carboxamidine 25 acetate: To a stirred suspension of 2.80 mmol (150 mg) of ammonium chloride (Fisher Scientific) in 5 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 1.5 mL (2.8 mmol) was added 2M trimethylaluminum in toluene (Aldrich Chemical Co.) via syringe over 15 min and then let stir at 0 oC for 25 min. after which 115 mg (0.280 mmol) of methyl 5-methylthio-4-(4-(2-thienyl)(1,3-thiazol-2 30 yl))thiophene-2-carboxylate in 5 mL of anhydrous toluene was added to solution and allowed to reflux for 1.5 h. The reaction mixture was quenched by pouring over a slurry of silica in
CH
2
C
2 . The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2
C
2 solution and concentrated. The crude product was purified on a 1 mm silica WO 00/47578 PCT/US99/18065 -99 prep plate eluting with 10% methanol/CH 2 Cl 2 with 1% CH 3 COOH to afford 40 mg (43% yield) of 5-methylthio-4-(4-(2-thienyl)(1,3-thiazol-2-yl))thiophene-2-carboxamidine acetate. 1 H-NMR (CD 3 OD; 300 MHz) 5 8.52 (s, 1H), 7.74 (s, 1H), 7.59 (dd, J=2 Hz and 5 Hz, 1H), 7.42 (dd, J=2 Hz and 5 Hz, 1H), 7.11 (min, 1H), 2.79 (s, 3H). Mass Spectrum (MALDI-TOF, 5 CHCA matrix, m/z) Calcd. for C 3 HiiN 3
S
4 : 338.0 (M+H), found 337.9. Example 16 4
-[
4
-(
3 -Bromophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride 10 a) Methyl 4-[4-(3-bromophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 99 mg (0.400 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 25 mL of reagent grade acetone. 2-Bromo-3'-bromo acetophenone (0.4 mmol; 111 mg) was added and 15 the solution was allowed to reflux for 3 h. The solution was allowed to cool and a solid was filtered and dissolved in 5 mL of hot tetrahydrofuran (THF), (Aldrich Chemical Co.) and purified on a 1 mm silica prep plate eluting with 20% ethyl acetate/hexane and dried in vacuo to afford 66 mg (40% yield) of methyl 4-[4-(3-bromophenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate. 20 b) 4-J4-(3-Bromophenyl)(1,3-thiazol-2-y)]-5-methlylthiothiophene-2 carboxamidine hydrochloride: To a stirred suspension of 1.55 mmol ( 83 mg) of ammonium chloride (Fisher Scientific) in 10 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 774 pL (1.55 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir at 25 0 C for 20 25 min after which 66 mg (0.155 mmol) of 4-[4-(3-bromophenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate was added to solution and allowed to reflux for 3 h. The reaction mixture was quenched by pouring over a slurry of 5 g of silica in 25 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2 C1 2 solution and concentrated. The crude product was purified on 1 mm silica 30 plates eluting with 10% methanol/CH 2 C1 2 to afford 63 mg (90% yield) of 4-[4-(3 bromophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (CD 3 OD; 300 MHz) 6 8.49 (s, 1H), 8.21 (m, 1H), 7.96 (min, 2H), 7.50 (mn, 1H), 7.5 (inm, WO 00/47578 -100- PCT/US99/18065 1H), 7.34 (min, 1H), 2.8 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for C 15
HI
2 BrN 3
S
3 : 411.9 (M+H), found 411.9. Example 17 5 4-[4-(4-Chloro-3-nitrophenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride a) Methyl 4-[4-(4-chloro-3-nitrophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 50 mg (0.202 mmol ) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene 10 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 10 mL of reagent grade acetone. 2-Bromo-4'-chloro-3'-nitroacetophenone (0.212 mmol; 59 mg) was added and the solution was allowed to reflux for 3 h. The solution was allowed to cool and a solid was filtered and dissolved in hot tetrahydrofuran (THF) and purified on a 1 mm silica prep plate eluting with 20% ethyl acetate/hexane and dried in vacuo to afford 60 mg (70% 15 yield) of methyl 4-[4-(4-chloro-3-nitrophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate. b) 4-[4-(4-Chloro-3-nitrophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride: To a stirred suspension of 1.40 mmol ( 75 mg) of ammonium chloride (Fisher Scientific) in 10 mL of anhydrous toluene (Aldrich Chemical Co.) placed 20 under nitrogen atmosphere at 0 'C, 700 pL (1.40 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir for 20 min after which 60 mg (0.140 mmol) of 4-[4-(4-chloro-3-nitrophenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate was added to solution and allowed to reflux for 3 h. The reaction mixture was quenched by pouring over a slurry of 5 g of silica in 50 mL of 25 chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CHC 2 solution and concentrated. The crude product was purified on 1 mm silica plates eluting with 10% methanol/CH 2 C1 2 to afford 17 mg (32% yield) of 4-[4-(4-chloro-3 nitrophenyl)(1, 3 -thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (CD 3 OD; 300 MHz) 8 8.53-8.58 (mn, 2H), 8.26 (dd, J=2.2 Hz and 8.5 Hz, 1H), 8.16 (s, 30 1H), 7.72 (d, J= 8.5 Hz, 1H), 2.80 (s, 3H).
WO 00/47578 PCT/US99/18065 -101 Example 18 4-[4-(4-Chloro-3-methylphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride 5 a) Methyl 4-[4-(4-chloro-3-methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene 2-carboxylate: 155 mg (0.627 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 10 mL of reagent grade acetone. 2-Bromo-1-(4-chloro-3-methylphenyl)ethan-1 one (0.658 mmol; 163 mg) was added and the solution was allowed to reflux for 3 h. The 10 solution was allowed to cool and the reaction mixture was concentrated and dissolved in 50 mL of CH 2 C1 2 . The organic layer was washed with 50 mL of IN HCI (aq.), dried over sodium sulfate and concentrated. The crude product was purified on a 1 mm silica plate eluting with 20% ethyl acetate/ hexane to afford 168 mg (68% yield) of methyl 4-[4-(4 chloro-3-methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. 15 b) 4-[4-(4-Chloro-3-methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride: To a stirred suspension of 4.24 mmol( 227 mg) of ammonium chloride (Fisher Scientific) in 15 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 2.2 mL (4.24 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and 20 then let stir for 20 min at 25'C after which 168 mg (0.424 mmol) of methyl 4-[4-(4-chloro-3 methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was added to solution and allowed to reflux for 2.5 h. The reaction mixture was quenched by pouring over a slurry of 5g silica in chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2 C1 2 solution and concentrated to afford 117 mg (73% yield) of 4 25 [4-(4-chloro-3-methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H-NMR (CD 3 OD; 300 MHz) 8 8.53 (s, 1H), 8.03 (dd, J= 1.2 Hz and 2.7 Hz, 1H), 7.9 (s, 1H), 7.85 (dd, J= 2 Hz and 8.5 Hz 1H), 7.38 (dd, J= 8.3 Hz and 17.4 Hz, 1H), 2.8 (s, 3H) 2.45 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for
C
16
H,
4 C1N 3
S
3 : 380.0 (M+H), found 380.3. 30 WO 00/47578 -102- PCT/US99/18065 Example 19 4 -(5-Methyl- 4 -phenyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine hydrochloride 5 a) Methyl 4-(5-methyl-4-phenyl(1,3-thiazol-2-y))-5-methylthiothiophene-2 carboxylate: 48 mg (0.194 mmol) of methyl 4 -(aminothioxomethyl)-5-methylthiothiophene 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 5 mL of reagent grade acetone. 2-Bromo-1-phenylpropan-l-one (0.223 mmol; 48 mg) was added and the solution was allowed to reflux for 5 h. The solution was allowed to cool and the reaction 10 mixture was concentrated and dissolved in 50 mL of CH 2 C1 2 . The organic layer was washed with 50 mL of 1N HCI (aq.), dried over sodium sulfate and concentrated. The crude product was purified on a 1 mm silica plate eluting with 20% ethyl acetate/ hexane to afford 53 mg (76% yield) of methyl 4-(5-methyl-4-phenyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate. 15 b) 4-(5-Methyl-4-phenyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine hydrochloride: To a stirred suspension of 1.47 mmol (78 mg) of ammonium chloride (Fisher Scientific) in 5 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 'C, 735gL (1.47 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir for 20 min at 25 0 C then, 20 53 mg (0.147 mmol) of methyl 4-(5-methyl-4-phenyl(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate were added to solution and allowed to reflux for 2.5 h. The reaction mixture was quenched by pouring over a slurry of 5g silica in chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2 C1 2 solution and concentrated to afford 26 mg (51% yield) of 4 -(5-methyl-4-phenyl(1,3-thiazol-2 25 yl))-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H-NMR (CD 3 OD; 300 MHz) 8 8.45 (s, 1H), 7.74-7.77 (m, 2H), 7.44-7.50 (m, 2H), 7.38-7.41 (m, 1H), 2.8 (s, 3H) 2.6 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA matrix, m/z) Calcd. for C, 6
H
15
N
3
S
3 : 346.0 (M+H), found 345.6.
WO 00/47578 -103- PCT/US99/18065 Example 20 4-[4-(4-Methylphenyl)(1,3-thiazol-2-yl)]-5-methlylthiothiophene-2-carboxamidine trifluoroacetate 5 a) Methyl 4-[4-(4-methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 103 mg (0.416 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 5 mL of reagent grade acetone. 2-Bromo-4'-methyl acetophenone (0.416 mmol; 89 mg) was added and the solution was allowed to reflux for 3 h. The solution was allowed 10 to cool and crude product was filtered and washed two times with acetone and purified on a 1 mm silica plate eluting with 20% ethyl acetate/ hexane to afford 104 mg (69% yield) of methyl 4-[4-(4-methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. b) 4-[4-(4-Methylphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine trifluoroacetate: To a stirred suspension of 2.87 mmol ( 154 mg) of 15 ammonium chloride (Fisher Scientific) in 10 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 144tL (2.87 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stirred for 20 min at 25 0 C after which 104 mg (0.287 mmol) of 4-[4-(4 methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was added to solution 20 and allowed to reflux for 3 h. The reaction mixture was quenched by pouring over a slurry of 5 g of silica in 50 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2 C1 2 solution and concentrated. The crude product was then purified on a 1 mm silica prep plate eluting with 10% methanol/CH 2
C
2 with 1% CH 3 COOH. The product was then basified with aq. NaOH and extracted with CHC1 3 and concentrated. 25 TFA was added and the product was crystallized from methanol as 4-[4-(4 methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine trifluoroacetate (20 mg; 30% yield). 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.62 (s, 1H), 8.12 (s, 1H), 7.98 (d, 1H, J= 8.1 Hz) 7.31 (d, 1H, J= 8.1 Hz), 2.8 (s, 3H) 2.5 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for Ci 6
H
1 sN 3
S
3 :346.0 (M+H), found 346.1. 30 WO 00/47578 -104- PCT/US99/18065 Example 21 4-[4-(2-methoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride 5 a) Methyl 4-[4-(2-methoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 105 mg (0.424 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 5 mL of reagent grade acetone. 2-Bromo-2'-methoxy acetophenone (0.467 mmol; 110 mg) was added and the solution was allowed to reflux for 3 h. The solution was 10 allowed to cool and the solution concentrated. The crude product was dissolved in 100 mL of
CH
2 Cl 2 and washed one time with 50 mL of 1N NaOH. The organic layer was obtained, dried over sodium sulfate, concentrated and purified on a 1 mm silica plate eluting with 20% ethyl acetate/ hexane to afford 160 mg (95% yield) of methyl 4-[4-(2-methoxyphenyl)(1,3 thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. 15 b) 4-[4-(2-Methoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride: To a stirred suspension of 4.23 mmol( 227 mg) of ammonium chloride (Fisher Scientific) in 10 mL of anhydrous toluene placed under nitrogen atmosphere at 0 oC, 2.12 mL (4.23 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 10 min and then let stir for 20 min at 25 0 C after 20 which 160 mg (0.287 mmol) of methyl 4-[4-(2-methoxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate in a solution of 5 mL of anhydrous toluene was added to solution and allowed to reflux for 3 h. The reaction mixture was quenched by pouring over a slurry of 5 g of silica in 30 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2 C1 2 solution and concentrated. The crude 25 product was then purified on a 2 mm silica prep plate eluting with 10% methanol/CH 2
C
2 with 1% NH 4 OH. The product was then dissolved in 2 mL of 4N HCl/dioxane and concentrated to afford 45 mg (29% yield) of 4-[4-(2-methoxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxamidine hydrochloride. 1 H-NMR (DMSO-d 6 ; 300 MHz) 8 8.68 (s, 1H), 8.36 (dd, J= 1.6 Hz and 7.74 Hz, 1H), 8.21 (s, 1H), 7.36-7.42 (m, 1H), 7.05-7.22 (m, 30 3 H), 3.97 (s, 3H), 2.8 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C 6
H,
5
N
3
OS
3 : 362.0 (M+H), found 361.7.
PCT/US99/18065 WO 00/47578 -105-CT/US99/8065 Example 22 4-[4-(2,4-Dimethoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene- 2 carboxamidine hydrochloride 5 a) Methyl 4-[4-(2,4-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene- 2 carboxylate: 99 mg (0.424 mmol) of methyl 4 -(aminothioxomethyl)-5-methylthiothiophene 2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was dissolved in 5 mL of reagent grade acetone. 2-Bromo-2',4'-dimethoxyacetophenone (0.440 mmol; 114 mg) was added and the solution was allowed to reflux for 2.5 h. The solution was allowed to cool and 10 the crude product was collected as a solid and washed with methanol and dried yielding 91 mg (56% yield) of methyl 4-[4-(2,4-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate. b) 4-[4-(2,4-Dimethoxyphenyl)
(
1 ,3
-
thiazol-2-yl) ] -5 -methylthiothio p h ene -2 carboxamidine hydrochloride: To a stirred suspension of 2.23 mmol (119 mg) of 15 ammonium chloride (Fisher Scientific) in 10 mL of anhydrous toluene placed under nitrogen atmosphere at 0 oC, 1.1 mL (2.23 mmol) of 2M trimethylaluminum in toluene was added via syringe over 10 min and then let stirred for 20 min at 25oC after which 81 mg (0.223 mmol) of methyl 4-[4-(2,4-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate was added to solution and allowed to reflux for 2.5 h. The reaction mixture was 20 quenched by pouring over a slurry of silica in chloroform. The silica was poured onto a sintered glass funnel and washed with a 10% methanol/CH 2 C1 2 solution and concentrated. The crude product was then purified on a 0.5 mm silica prep plate eluting with 10% methanol/CH 2 C1 2 to afford 32 mg (37% yield) of 4-[4-(2,4-dimethoxyphenyl)(1,3-thiazol-2 yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 1 H-NMR (CD 3 OD; 300 MHz) 8 25 8.49 ( s, 1H), 8.31 (d, J= 8.5 Hz, 1H), 7.93 (s, 1H), 6.64 (min, 2H), 3.97 (s, 3 H), 3.85 (s, 3H), 2.79 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C 1 7
H
17
N
3 0 2
S
3 : 392.1 (M+H), found 392.4.
WO PCT/US99/180 6 5 WO 00/47578 -106 Example 23 4-[4-(3,4-Dichlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride 5 a) Methyl 4-[4-(3,4-dichlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 176 mg (0.712 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was reacted with 2-bromo-3',4'-dichloroacetophenone (0.854 mmol; 330 mg) in a manner similar to Example 22, step (a) to afford 270 mg (91% yield) of methyl 4-[4-(3,4 10 dichlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. b) 4-[4-(3,4-Dichlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride: 270 mg (0.648 mmol) of methyl 4-[4-(3,4 dichlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was treated in a manner similar to Example 22, step (b) to afford 135 mg (52% yield) of 4-[4-(3,4 15 dichlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H-NMR (CD30D; 300 MHz) 8 8.54 (s, 1H), 8.22 (d, J= 2 Hz, 1H), 8.02 (s, 1H), 7.94 (dd, J= 2 Hz and 8.4 Hz, 1H), 7.58 (d, J= 8.5 Hz, 1 H), 2.79 (s, 3H). Mass Spectrum (MALDI TOF, CHCA Matrix, m/z) Calcd. for CisH 11 C1 2
N
3
S
3 : 400.0 (M+H), found 400.6. 20 Example 24 4-[J4-(3-Methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride a) Methyl 4-[4-(3-methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 25 carboxylate: Methyl 4-(aminothioxomethyl)-5-methylthiothiophene-2-carboxylate, 106 mg (0.428 mmol) (Maybridge Chemical Co. LTD., Cornwall, U.K.) was reacted with 2-bromo-3' methylacetophenone (0.428 mmol, 91 mg) in a manner similar to Example 22, step (a) to afford 98 mg (63% yield) of methyl 4-[4-(3-methylphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate. 30 b) 4-[4-(3-Methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride: 4-[4-(3-methylphenyl)( 1.3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate, (98 mg, 0.271 mmol) was treated in a similar manner to Example 22, step (b) to afford 75 mg (80% yield) of 4-[4-(3-methylphenyl)(1,3-thiazol-2 yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H-NMR (CD 3 OD; 300 MHz) 8 PCT/US99/180 6 5 WO 00/47578 -107- PCTUS99/18065 8.56 (s, 1H), 7.88 (s, 1H), 7.86 (d, J= 14 Hz, 2H), 7.33 (m, 1H), 7.19 (m, 1 H), 2.79 (s, 3H), 2.42 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C 1 6
H
1 5
N
3
S
3 : 346.0 (M+H), found 346.7 5 Example 25 5-Methylthio-4-(4-(2-5,6,7,8-tetrahydronaphthyl) (1,3-thiazol-2-yl))thiophene- 2 carboxamidine hydrochloride a) Methyl 5-methylthio-4-(4-( 2 -5,6,7,8-tetrahydronaphthyl)(1,3-thiazol-2 10 yl))thiophene-2-carboxylate: Methyl 4-(aminothioxomethyl)-5-methylthiothiophene- 2 carboxylate, (160 mg, 0.647 mmol) (Maybridge Chemical Co. LTD., Cornwall, U.K.) was reacted with 2-bromo- 1 -(2-5,6,7,8-tetrahydronaphthyl)ethan- 1 -one (0.712 mmol; 180 mg) in a manner similar to Example 22, step (a) to afford 106 mg (41% yield) of methyl 5 methylthio-4-( 4
-(
2 -5,6,7,8-tetrahydronaphthyl)(1,3-thiazol-2-yl))thiophene-2-carboxylate. 15 b) 5-Methylthio-4-(4-(2-5, 6 ,7,8-tetrahydronaphthyl) (1,3-thiazol-2-yl))thiophene- 2 carboxamidine hydrochloride: 106 mg (0.264 mmol) of methyl 5-methylthio-4-( 4
-(
2 5,6,7,8-tetrahydronaphthyl)(1,3-thiazol-2-yl))thiophene-2-carboxylate was treated in a similar manner to Example 22, step (b) to afford 88 mg (80% yield) of 5-methylthio-4-( 4
-(
2 -5, 6
,
7
,
8 tetrahydronaphthyl)(1,3-thiazol-2-yl))thiophene-2-carboxamidine hydrochloride. 'H-NMR 20 (CD 3 OD; 300 MHz) 8 8.49 (s, 1H), 7.78 (s, 1H), 7.73 (m, 2H), 7.11 (m, 1 H), 2.79 (m, 7H), 1.82-1.86 (m, 4H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for
C
19
HI
9
N
3
S
3 : 386.1 (M+H), found 386.2 Example 26 25 4-[4-(3,5-Dimethoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride a) Methyl 4-[4-(3,5-dimethoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 100 mg (0.404 mmol) of methyl 4-(aminothioxomethyl)-5 30 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was reacted with 2-bromo-3',5'-dimethoxy acetophenone (0.444 mmol) in a manner similar to Example 22, step (a) to afford 44 mg (27% yield) of methyl 4-[4-(3,5-dimethoxyphenyl)(1, 3 thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate.
PCT/US99/18065 WO 00/47578 -108-CT/US991 8 6 5 b) 4-[4-(3,5-Dimethoxyphenyl) ( 1,3
-
th ia z o l- 2
-
y l) ] -5m e t hy l th i o t h i o p h e n e - 2 carboxamidine hydrochloride: 44 mg (0.108 mmol) of methyl 4-[4-(3,5 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was treated in a manner similar to Example 22, step (b) to afford 25 mg (60% yield) of 4-[4-(3,5 5 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 1 H-NMR (CD 3 OD; 300 MHz) 8 8.52 (s, 1H), 7.91 (s, 1H), 7,23 (d, J= 2.2 Hz, 1H), 6.50 (t, 1H), 3.85 (s, 6 H), 2.89 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C 1 7
H,
1 7
N
3 0 2 S3: 392.11 (M+H), found 392.4. 10 Example 27 4-[4-(2-Methylphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride a) Methyl 4-[4-(2-methylphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiiophene-2 15 carboxylate: 160 mg (0.647 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was reacted with 2-bromo-2'-methyl acetophenone (0.711 mmol, 152 mg) in a manner similar to Example 22, step (a) to afford 124 mg (53% yield) of methyl 4-[4-(2-methylphenyl)(1, 3 thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. 20 b) 4-[4-(2-Methylphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride: 124 mg (0.343 mmol) of methyl 4-[4-(2-methylphenyl)(1,3 thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was treated in a manner similar to Example 22, step (b) to afford 60 mg (50% yield) of 4-[4-(2-methylphenyl)(1,3-thiazol- 2 yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 1 H-NMR
(CD
3 OD; 300 MHz) 8 25 8.50 (s, 1H), 7.65 (m, 2H), 7.22-7.32 (m, 3H), 2.79 (s, 3H), 2.51 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C, 6 HsN 3
S
3 :346.0 (M+H), found 346.2. Example 28 4-J4-(2,5-Dimethoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 30 carboxamidine hydrochloride a) Methyl 4-J4-(2,5-dimethoxyphenyl)( 1 ,3 -th ia z o l-2 -y l) ]-5 -m e thy lt h i o t h i o p h en e-2 carboxylate: 132 mg (0.534 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was WO 00/47578 -109- PCT/US99/18065 reacted with 2-bromo-2',5'-dimethoxy acetophenone (0.587 mmol; 152 mg) in a manner similar to Example 22, step (a) to afford 97 mg (45% yield) of methyl 4-[4-(2,5 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. b) 4-[4-(2,5-Dimethoxyphenyl)( 1 ,3
-
thiazo l-2 yl) ] - 5methylt hi othiophene -2 5 carboxamidine hydrochloride: 97 mg (0.238 mmol) of methyl 4-[4-(2,5 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was treated in a manner similar to Example 22, step (b) to afford 30 mg (32% yield) of 4-[4-(2,5 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 1 H-NMR (CD 3 OD; 300 MHz) 8 8.46 (s, 1H), 8.10 (s, 1H), 7.99 (d, J= 3.2 Hz, 1H), 7.05 (d, 10 J= 9 Hz, 1H), 6.93 (d, J= 3.2 Hz, 1H), 6.90 (d, J= 3.2 Hz, 1H), 3.94 (s, 3H), 3.83 (s, 3H), 2.51 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C 1 7
H
17
N
3 0 2
S
3 : 392.1 (M+H), found 392.1. Example 29 15 4-[4-(4-Chloro(3-pyridyl)) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride a) Methyl 4-[4-(4-chloro(3-pyridyl)) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 240 mg (0.970 mmol) of methyl 4-(aminothioxomethyl)-5 20 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was reacted with 2-bromo-1-(4-chloro(3-pyridyl))ethan-1-one (1.06 mmol; 250 mg) in a manner similar to Example 22, step (a) to afford 286 mg (77% yield) of methyl 4-[4-(4-chloro(3 pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. b) 4-[4-(4-Chloro(3-pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothIiophene-2 25 carboxamidine hydrochloride: 286 mg (0.747 mmol) of methyl 4-[4-(4-chloro(3 pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was treated in a manner similar to Example 22, step (b) to afford 134 mg (49% yield) of 4-[4-(4-chloro(3 pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C 14 H N 4 ClS 3 : 366.9 (M+H), found 30 366.6 PCT/US99/18065 WO 00/47578 -110- PCT/US99/180 6 5 Example 30 4-(4-(2H-Benzo[d]1,3-dioxolen-5-yl)(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxamidine hydrochloride 5 a) 1-(2H-Benzo[3,4-d]l,3-dioxolen-5-yl) -2 -b r omo eth a n - l o n e: To a solution of 2.5 g (15.23 mmol) of 3,4-methylenedioxy acetophenone in 200 mL of anhydrous methanol was added 61 mmol (20 g) of poly (4-vinylpyridinium tribromide), Aldrich Chemical Co., and allowed to reflux for 2.5 h. The solution was filtered and concentrated. 1-(2H-benzo[3,4 d] 1,3-dioxolen-5-yl)-2-bromoethan-1 -one (1.4 g, 38% yield) was obtained methylene 10 chloride/hexanes as off-white crystals. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.20 (s, 1H), 8.07 (s, 1H), 7.63 (m, 2H), 7.03 (dd, J= 1.2 Hz and 7.1 Hz, 1H), 6.09 (s, 2H), 3.86 (s, 3H), 2.75 (s, 3H). b) Methyl 4-(4-(2H-benzo[d]1,3-dioxolen-5-yl) (1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate: 1.4 g (5.66 mmol) of methyl 4-(aminothioxomethyl)-5 15 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was reacted 1-(2H-benzo[3,4-d]1,3-dioxolen-5-yl)-2-bromoethan-l-one (5.66 mmol, 1.37 g) in a manner similar to Example 22, step (a) to afford 1.55 g (70% yield) of methyl 4-(4-(2H benzo [d]1,3-dioxolen-5-yl)(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxylate. c) 4-(4-(2H-Benzo[d1,3-dioxolen-5-yl)
(
1,3 - th ia z o l-2-y l)) -5- m eth y lt h i o th i o p h en e-2 20 carboxamidine hydrochloride: 1.55 g (3.95 mmol) of methyl 4-(4-(2H-benzo[d]1,3 dioxolen-5-yl)(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxylate was treated in a manner similar to Example 22, step (b) to afford 130 mg (9% yield) of 4-(4-(2H-benzo[d] 1,3 dioxolen-5-yl)(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine hydrochloride.
'H
NMR (CD 3 OD; 300 MHz) 8 8.51 (s, 1H), 7.73 (s, 1H), 7.58 (m, 2H), 6.89 (d, J= 8 Hz, 1H), 25 6.00 (s, 2H), 2.79 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C1 6
H,
3
N
3 0 2
S
3 :376.0 (M+H), found 376.1. Example 31 4-[4-(3,4-Dimethoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 30 carboxamidine hydrochloride a) 1-(3,4-Dimethoxyphenyl)-2-bromoethan-l-one: 2 g of 1-(3,4 dimethoxyphenyl)ethan- 1-one (11.1 mmol) was reacted in a manner similar to Example 15, step (a), to yield 1.2 g (42% yield) of 1-(3,4-dimethoxyphenyl)-2-bromoethan- 1-one.
PCT/US99/18065 WO 00/47578 PCT/US991806 5 -111 b) Methyl 4-J4-(3,4-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate: 105 mg (0.424 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge Chemical Co. LTD., Cornwall, U.K.) was reacted with 1-(3,4-dimethoxyphenyl)-2-bromoethan-1-one (0.467 mmol; 120 mg) in a 5 manner similar to Example 22, step (a) to afford 148 mg (85% yield) of methyl 4-[4-(3,4 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. c) 4-[4-(3,4-Dimethoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride: 148 mg (0.363 mmol) of methyl 4-[4-(3,4 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate was reacted in a 10 manner similar to Example 22, step (b) to afford 70 mg (50% yield) of 4-[4-(3,4 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 1 H-NMR (CD 3 OD; 300 MHz) 8 8.50 (s, 1H), 7.76 (s, 1H), 7.61 (m, 2H), 7.31 (m, 1H), 7.01 (d, J= 8 Hz, 1H), 3.9 (s, 3H) 3.86 (s, 3H), 2.78 (s, 3H). Mass Spectrum (MALDI-TOF, CHCA Matrix, m/z) Calcd. for C, 7
H
17
N
3 0 2
S
3 : 392.1 (M+H), found 392.4. 15 Example 32 4-[4-(2-Chloro(3-pyridyl)) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine 20 a) Methyl 4-J4-(2-chloro(3-pyridyl)) (1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate: 2-Chloropyridine-3-carbonyl chloride (300 mg, 1.7 mmol) was dissolved in anhydrous CH 3 CN (4 mL). While stirring well with a magnetic stirrer, trimethylsilyldiazomethane (4 mL, 2M solution in hexane, 8 mmol) was dripped into the reaction mixture. The resulting yellow solution was stirred for 25 2h at room temperature, at which time the mixture was cooled in an ice bath. To the cold solution, 30% HBr in acetic acid (2 mL) was added dropwise with vigorous evolution of gas. This solution was stirred for lh during which time 2-bromo- 1-(2 chloro(3-pyridyl))ethan-1-one precipitated. This solid was collected by filtration and dried under vacuum. The dry solid (142 mg, 0.6 mmol) was dissolved in acetone (10 30 ml). To this solution 5-(methoxycarbonyl)-2-(methylthio)-thiophene-3 thiocarboxamide (100 mg, 0.4 mmol, Maybridge Chemical Company, Cornwall, UK) was added and heated at reflux for 5 h. At this point the solid that precipitated was filtered off and washed with methanol and dried under vacuum to give 110 mg (71%) WO 00/47578 -12- PCT/US99/18065 of methyl 4-[4-(2-chloro(3-pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate. 1 H-NMR (CDCl 3 ; 300 MHz) 8 2.70 (s, 3H), 3.92 (s, 3H), 7.39 (dd, J=4.7 and 7.7 Hz, 1H), 8.11 (s, 1H), 8.22 (s, 1H), 8.38 (dd, fJ=1.9 and 4.7 Hz, 1H), 8.62 (dd, J=1.9 and 7.7 Hz, 1H). 5 b) 4-[4-(2-Chloro(3-pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine: Methyl 4-[4-(2-chloro(3-pyridyl))(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate (100 mg, 0.26 mmol) as prepared in previous step was treated in a manner similar to that for Example 1, to give 50 mg (52%) of 4-[4-(2 chloro(3-pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine as a 10 solid. 1 H-NMR (DMSO-d 6 ; 300 MHz) 8 2.79 (s, 3H), 7.62 (dd, J=4.9 and 7.4 Hz, 1H), 8.41 (s, 1H), 8.49 (m, 2H), 8.69 (s, 1H), 9.1 (broad s, 2H), 9.4 (broad s, 2H). Mass spectrum (ESI, m/z): Calcd. for Cl 4 H, N 4
S
3 Cl: 367.0 (M+H), found 369.0. Example 33 15 4-(4-Cyclohexyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine a) Methyl 4-(4-cyclohexyl(1,3-thiazol-2-yl))-5-methylthiothliophene- 2 carboxylate: Cyclohexanecarbonyl chloride (300 mg, 2.0 mmol) was treated in a manner similar to that for Example 32 to give 2-bromo-1-cyclohexylethan-1-one. The dry solid (125 mg) was dissolved in acetone (10 ml). To this solution 5 20 (methoxycarbonyl)-2-(methylthio)-thiophene-3-thiocarboxamide (100 mg, 0.4 mmol, Maybridge Chemical Company, Cornwall, UK) was added and heated at reflux for 5 h. At this point the solid that precipitated was filtered off and washed with methanol and dried under vacuum to give 100 mg (70%) of methyl 4-(4-cyclohexyl(1,3-thiazol 2-yl))-5-methylthiothiophene-2-carboxylate which was used without further 25 purification in the following step. b) 4-(4-Cyclohexyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxamidine: Methyl 4-(4-cyclohexyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (100 mg, 0.28 mmol) as prepared in previous step was treated in a manner similar to that for Example 1, to give 60 mg (63%) of 4-(4-cyclohexyl(1,3-thiazol-2 30 yl))-5-methylthiothiophene-2-carboxamidine as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 1.21-1.53 (m, 5H), 1.61-1.78 (m, 3H), 2.05 (min, 2H), 2.7 (s, 3H), 2.74 (inm, WO 00/47578 -113- PCT/US99/18065 -113 1H), 7.33 (s, 1H), 8.32 (s, 1H). Mass spectrum (MALDI-TOF, m/z): Calcd. for
C
15
HI
9
N
3
S
3 , 338.1 (M+H), found 338.1. Example 34 5 4-Phenyl-5-(trifluoromethyl)thiophene-2-carboxamidine Methyl 4-phenyl-5-(trifluoromethyl)thiophene-2-carboxylate (100 mg, 0.37 mmol, Maybridge Chemical Company, Cornwall, UK) was treated in a manner similar to that for Example 1 to give 80 mg (85%) of 4-phenyl-5 (trifluoromethyl)thiophene-2-carboxamidine as a solid. 'H-NMR (DMSO-d 6 ; 300 10 MHz) 8 7.45-7.52 (m, 5H), 7.79 (d, J=1.4 Hz, 1H). Mass spectrum (MALDI-TOF, m/z): Calcd. for C 1 2
H
9
F
3
N
2 S, 271.1 (M+H), found 271.2. Example 35 5-Methylthio-4-(2-phenyl(1,3-thiazol-4-yl))thiophene-2-carboxamidine 15 a) Methyl 4-(2-bromoacetyl)-5-methylthiothiophene-2-carboxylate: 5-(Methoxycarbonyl)-2-methylthiothiophene-3-carboxylic acid (200 mg, 0.86 mmol) as prepared in Example 95 was taken in a round bottomed flask and anhydrous
CH
2 C1 2 (10 mL) was introduced to the flask. This solution was cooled in an ice bath under an argon atmosphere. To this mixture oxalyl chloride (328 mg, 2.6 mmol) was 20 added followed by anhydrous DMF (500 pL). The resulting solution was stirred at 4oC for 30 min and then allowed to warm up to room temperature, while monitoring for the disappearance of the acid by TLC. After 2 h solvents were removed under vacuum and the residual oxalyl chloride was removed azeotropically with toluene. The resulting residue was dried under high-vacuum to give the acid chloride as a gray 25 solid. This solid was dissolved in anhydrous CH 3 CN (8 mL). While stirring well with a magnetic stirrer trimethylsilyldiazomethane (4 mL, 8 mmol, 2M solution in hexane) was dripped into the reaction mixture. The resulting yellow solution was stirred for 2h at room temperature, at which time the mixture was cooled in an ice bath. To the cold solution 30% HBr in acetic acid ( 2 mL) was added dropwise, with 30 vigorous evolution of gas. This solution was stirred for lh, during which methyl 4-(2 bromoacetyl)-5-methylthiothiophene-2-carboxylate precipitates. This solid was WO 00/47578 -114- PCT/US99/18065 collected by filtration and dried under vacuum to give 120 mg (45%). 'H-NMR (CDCl 3 ; 300 MHz) 8 2.64 (s, 3H), 3.91 (s, 3H), 4.27 (s, 2H), 8.10 (s, 1H). b) Methyl 5-methylthio-4-(2-phenyl(1,3-thiazol-4-yl))thiophene-2 carboxylate: 5-(Methoxycarbonyl)-2-(methylthio)-thiophene-3-thiocarboxamide (100 5 mg, 0.4 mmol, Maybridge Chemical Company, Cornwall, UK) was dissolved in acetone (20 ml). To this solution, methyl 4-(2-bromoacetyl)-5-methylthiothiophene 2-carboxylate (112 mg) as prepared in previous step was added and heated at reflux for 3 h. At this point the solid that precipitated was filtered off and washed with acetone and dried under vacuum to give 82 mg (65%) of methyl 5-methylthio-4-(2 10 phenyl(1,3-thiazol-4-yl))thiophene-2-carboxylate. 'H-NMR (CDCl 3 ; 300 MHz) 8 2.67 (s, 3H), 3.91 (s, 3H), 7.44-7.49 (min, 3H), 7.61 (s, 1H), 8.03-8.06 (min, 2H), 8.28 (s, 1H). c) 5-Methylthio-4-(2-phenyl(1,3-thiazol-4-yl))thiophene-2-carboxamidine: Methyl 5-methylthio-4-(2-phenyl(1,3-thiazol-4-yl))thiophene-2-carboxylate (80 mg) as prepared in previous step was treated in a manner similar to that for Example 1, to 15 give 50 mg of 5-methylthio-4-(2-phenyl(1,3-thiazol-4-yl))thiophene-2-carboxamidine as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 2.75 (s, 3H), 7.51-7.60 (min, 3H), 8.02 (s, 1H), 8.06 (min, 2H), 8.70 (s, 1H), 9.06 (broad s, 2H), 9.38 (broad s, 2H). Mass spectrum (MALDI-TOF, m/z): Calcd. for CisH, 3
N
3
S
3 , 332.0 (M+H), found 332.1. 20 Example 36 4-[4-(2-Chloro(4-pyridyl)) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine a) Methyl 4-[4-(2-chloro(4-pyridyl))(1,3-thiazol-2-yl)]-5 25 methylthiothiophene-2-carboxylate: 2-Chloropyridine-4-carbonyl chloride (300 mg, 1.7 mmol) was dissolved in anhydrous CH 3 CN (4 mL). While stirring well with a magnetic stirrer trimethylsilyldiazomethane (4 mL, 8 mmol, 2M solution in hexane) was dripped into the reaction mixture. The resulting yellow solution was stirred for 2 h at room temperature, at which time the mixture was cooled in an ice bath. To the 30 cold solution 30% HBr in acetic acid (2 mL) was added dropwise, with vigorous evolution of gas. This solution was stirred for lh, during which time 2-bromo-1-(2 chloro(4-pyridyl))ethan-1-one precipitates. This solid was collected by filtration and dried under vacuum. The dry solid (142 mg, 0.6 mmol) was dissolved in acetone (10 WO 00/47578 -115- PCT/US99/18065 ml). To this solution 5-(methoxycarbonyl)-2-(methylthio)-thiophene-3 thiocarboxamide (100 mg, 0.4 mmol, Maybridge Chemical Company, Cornwall, UK) was added and heated at reflux for 5 h. At this point the solid that precipitated was filtered off and washed with methanol and dried under vacuum to give 100 mg of 5 methyl 4-[4-(2-chloro(4-pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate. 'H-NMR (CD 3 OD; 300 MHz) 5 2.73 (s, 3H), 3.94 (s, 3H, overlapping H20 peak), 7.92-7.99 (min, 2H), 8.05 (s, 1H), 8.24 (s, 2H), 8.48 (min, 1H). b) 4-[4-(2-Chloro(4-pyridyl)) (1,3-thiazol-2-yl)}-5-methylthiothiophene-2 carboxamidine: Methyl 4-[4-(2-chloro(4-pyridyl))(1,3-thiazol-2-yl)]-5 10 methylthiothiophene-2-carboxylate (100 mg, 0.26 mmol) as prepared in previous step was treated in a manner similar to that for Example 1, to give 50 mg of 4-[4-(2 chloro(4-pyridyl))(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine as a solid. 'H-NMR (CDCl 3
/CD
3 OD; 300 MHz) 8 2.82 (s, 3H), 7.95 (dd, J=1.4 and 5.3 Hz, 1H), 8.08 (d, J=1.0 Hz, 1H), 8.23 (s, 1H), 8.42 (d, J=5.3 Hz, 1H), 8.56 (s, 1H). 15 Mass spectrum (MALDI-TOF, m/z): Calcd. for C 14
HIN
4
S
3 C1, 367.0 (M+H), found 367.1. Example 37 4-[4-(4-Chlorophenyl)(1,3-thiazol-2-yl)]-5-(methlylsulfonyl)thiophene-2 20 carboxamidine 4-[4-(4-Chlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine (35 mg, 0.1 mmol) prepared according to Example 1 was dissolved in a mixture of MeOH and CH 2 C1 2 (1:1, 6 mL). While stirring well, m 25 chloroperoxybenzoic acid (100 mg) was added in portions to this solution over a 3h period. The mixture was stirred for a further 2 h and the solvents were removed under vacuum. The resulting residue was dissolved in MeOH (8 mL). Strong anion exchange resin (AG 1-X8, 5 ml, 1.4 meq/mL) was packed into a disposable chromatography column and washed with H20 (5x5 mL) and MeOH (3x5 mL). The 30 methanolic solution from the reaction was slowly introduced into this column, and the column effluent was collected. The column was washed with MeOH (2x5 mL) and these washings were also collected. The combined effluents were evaporated under vacuum and the residue was subjected to preparative thin layer chromatography (silica WO 00/47578 PCT/US99/18065 -116 gel, 10% MeOH in CH 2
CI
2 with 2% acetic acid). The major band was isolated and suspended in CH 2 Cl 2 and filtered. The filtrate was collected and the residue was washed with 10% MeOH in CH 2 C12, saturated with NH 3 . The washings were combined with the original filtrate and the solvents were removed under vacuum. The 5 resulting solid was dissolved in 10% MeOH in CHCl 3 and filtered through a 0.45 micron filter. The filtrate was collected and evaporated under vacuum to give 20 mg (53%) of an off-white solid. 1 H-NMR (CDCl 3
/CD
3 OD; 300 MHz) 8 3.78 (s, 3H), 7.47 (d, J=8.7 Hz, 2H), 7.96 (d, J=8.7 Hz, 1H), 8.00 (s, 1H), 8.35 (s, 1H). Mass spectrum (MALDI-TOF, m/z): Calcd. for C 15
HI
2 0 2
N
3
S
3 C1, 398.0 (M+H), found 398.0. 10 Example 38 Hydrazino[5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl)) (2-thienyl)]methanimine a) 5-Methylthio-4-(4-phenyl(1,3-thlziazol-2-yl))thiophene-2-carboxamide: 15 Liquid ammonia (5 mL) was condensed into a cold (-78 'C) Teflon-lined steel bomb. Methyl 5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate (0.6 g, 1.7 mmol) as prepared in Example 10 step (a) was introduced in one portion and the bomb was sealed and heated in an oil bath at 80 oC for 48h. The bomb was cooled to -78 oC, opened and the ammonia was allowed to evaporate at room temperature. The 20 residual solid was collected and dried under vacuum to give 0.5 g (88%) of 5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamide. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 2.75 (s, 3H), 7.38 (min, 1H), 7.40-7.51 (m, 2H), 8.04-8.18 (inm, 2H), 8.19 (s, 1H), 8.20 (s, 1H). b) 5-Methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carbonitrile: A 25 slurry of P 2 0 5 (2.7 g, 19 mmol) and hexamethyldisiloxane (6.7 mL) in dichloroethane (13 mL) was heated to 90 oC, while stirring under a N 2 atmosphere. After stirring for 2 h, the resulting clear solution was allowed to cool to 40 'C. 5-methylthio-4-(4 phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamide (0.9 g, 2.7 mmol) as prepared in previous step was added to this solution and the mixture was heated at 75 0 C for 5h. 30 This solution was cooled to room temperature and stirred with aqueous NaCl (6 M, 100 mL) for 10 min. As the aqueous solution is added a yellow solid precipitated. After 10 min this solid was separated by filtration, and dried under vacuum to give WO 00/47578 -117- PCT/US99/18065 (0.5 g, 59%) of 5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carbonitrile as a yellow solid. 'H-NMR(DMSO-d 6 ; 300 MHz) 8 2.76 (s, 3H), 7.38 (min, 1H), 7.48 (m, 2H), 8.07 (min, 2H), 8.22 (s, 1H), 8.51 (s, 1H). c) Hydrazino[5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl)) (2 5 thienyl)]methanimine: 5-Methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2 carbonitrile (100 mg, 0.32 mmol) as prepared in previous step was dissolved in EtOH (10 mL). To this solution hydrazine monohydrate (10 eq) was added and the mixture was heated at reflux for 3h. The EtOH solution was concentrated down to 1 mL and water (2 mL) was added to this solution. This resulted in the formation of a white 10 solid. The solid was collected by filtration washed with a small amount of water and dried under vacuum to give 50 mg (45%) of hydrazino[5-methylthio-4-(4-phenyl(1,3 thiazol-2-yl))(2-thienyl)]methanimine. 'H-NMR (CD 3 OD/CDCl 3 ; 300 MHz) 8 2.69 (s, 3H), 7.39 (min, 1H), 7.47 (min, 2H), 7.52 (s, 1H), 7.98 (min, 2H), 8.10 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C 15
HI
4
N
4
S
3 , 347.04 (M+H), found 347.1. 15 Example 39 {IminoJ[5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl))(2-thienyl)]methyl}methylamine 5-Methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine (20 mg, 0.06 mmol) as prepared in Example 10 step (b) was dissolved in MeOH, and to 20 this solution methylamine (0.6 mL, 2M solution in tetrahydrofuran) was added. This solution was refluxed for 6h, at which time the solvents were removed under vacuum to give a solid. This solid was dissolved in a small amount of MeOH. H20 was added dropwise to the methanolic solution until a precipitate was formed. This solid was isolated, washed with a small amount of water and dried under vacuum to give 15 mg 25 (72%) of { imino[5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl))(2 thienyl)]methyl}methylamine. 1 H-NMR (DMSO-d 6 ; 300 MHz) 5 2.77 (s, 3H), 3.00 (s, 3H), 7.36-7.42 (min, 1H), 7.47-7.52 (min, 2H), 8.07-8.10 (min, 2H), 8.23 (s, 1H), 8.55 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C 16 HsN 3
S
3 , 346.5 (M+H), found 346.2.
WO 00/47578 -118- PCT/US99/18065 Example 40 2-{3-[2-(5-Amidino-2-methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid a) 2-Bromo-1-(3-hydroxyphenyl)ethan-1-one: 2-Bromo-l1-(3 methoxyphenyl)ethan-1-one (2 g, 8.7 mmol) was taken in a round bottomed flask 5 equipped with magnetic stir bar. The flask was put under a N 2 atmosphere and
CH
2 C1 2 was introduced into the flask. The resulting solution was cooled in a dry ice acetone bath and BBr 3 (27 mL, IM in CH 2 C1 2 ) was introduced dropwise. The resulting solution was allowed to warm up to room temperature overnight. The solvents were removed under vacuum and the residue was purified by passing through 10 a short pad of silica gel (50 g) to give 1.3 g (69%) of 2-bromo-1-(3 hydroxyphenyl)ethan-1-one as an oil. 'H-NMR (CDCl 3 ; 300 MHz) 8 4.47 (s, 2H), 6.21 (s, 1H), 7.14 (min, 1H), 7.35 (min, 1H), 7.52-7.82 (mn, 2H). b) Methyl 4-[4-(3-hydroxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene 2-carboxylate: 2-Bromo- 1 -(3-hydroxyphenyl)ethan- 1-one (229 mg, 1.1 mmol) as 15 prepared in previous step was treated in a manner similar to that of Example 13, step (a) to give 225 mg (61%) of methyl 4-[4-(3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 5 2.76 (s, 3H), 3.86 (s, 3H), 6.87 (min, 1H), 7.27 (t, J=7.8 Hz, 1H), 7.49 (mn, 2H), 8.12 (s, 1H), 8.20 (s, 1H). 20 c) (tert-Butoxy)-N-({4-[4-(3-hydroxyphenyl) (1,3-thiazol-2-yl)]-5 methylthio(2-thienyl)}iminomethyl)carboxamide: 4-[4-(3-Hydroxyphenyl)(1,3 thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine (2 g, 5.8 mmol), prepared by treating methyl 4-[4-(3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxylate in a manner similar to that for Example 1, was dissolved in anhydrous 25 DMF (10 mL). To this solution di-tert-butyl dicarbonate (1.38 g, 6.3 mmol) and DIEA (2 mL, 11.5 mmol) was added, and the mixture was stirred at room temperature for 18 h. DMF was removed under vacuum and the residue was purified by silica gel column chromatography to give 1.8 g (70%) of (tert-butoxy)-N-({4-[4-(3-hydroxyphenyl)(1,3 thiazol-2-yl)]-5-methylthio(2-thienyl)}iminomethyl)carboxamide as an oil. 'H-NMR 30 (DMSO-d 6 ; 300 MHz) 8 1.58 (s, 9H), 2.81 (s, 3H), 6.81 (min, 1H), 7.28 (t, J=8.0 Hz, 1H), 7.49-7.52 (m, 2H), 8.09 (s, 1H), 8.71 (s, 1H).
WO 00/47578 PCT/US99/18065 -119 d) tert-Butyl 2-{3-[2-(5-{[(tert-butoxy)carbonylamino]iminomethyl}-2 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetate: (tert-Butoxy)-N-( {4-[4-(3 hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methylthio(2-thienyl) } iminomethyl)carboxamide (23 mg, 0.05 mmol) as prepared in previous step was dissolved in anhydrous DMF (1 5 mL). To this solution tert-butyl 2-bromoacetate (20 mg, 0.1 mmol), Cs 2
CO
3 (33.5 mg, 0.1 mmol) and KI (5 mg) was added and the mixture was heated at 70 oC for 18 h. The solvents were removed under vacuum and the residue was purified by preparative silica gel thin-layer chromatography to give 12 mg (42%) of tert-butyl 2- {3-[2-(5 {[(tert-butoxy)carbonylamino]iminomethyl}-2-methylthio-3-thienyl)- 1,3-thiazol-4 10 yl]phenoxy}acetate which was used in the following step. e) 2-{3-[2-(5-Amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetic acid: tert-Butyl 2- {3-[2-(5- { [(tert butoxy)carbonylamino]iminomethyl } -2-methylthio-3-thienyl)- 1,3-thiazol-4 yl]phenoxy}acetate (12 mg, 0.02 mmol) as prepared in previous step was dissolved in 15 1 ml 50% TFA in CH 2 Cl 2 containing 2% H 2 0 and stirred for 4h. The solvents were removed under vacuum. The residual TFA was removed by azeotroping with toluene to give 8.7 mg (100%) of 2- { 3-[2-(5-amidino-2-methylthio-3-thienyl)- 1,3 -thiazol-4 yl]phenoxy} acetic acid as a buff colored solid. 1 H-NMR (CD 3 OD/CDCl 3 ; 300 MHz) 6 2.77 (s, 3H), 4.74 (S, 2H), 6.93 (min, 1H), 7.35 (t, J=7.9 Hz, 1H), 7.62 (min, 1H), 7.68 20 (M, 1H), 7.84 (s, 1H), 8.46 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for
C
17
H
1 5
N
3 0 3
S
3 , 406.5 (M+H), found 406.3. Example 41 2
-{
2 -[2-(5-Amidino-2-methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid 25 a) tert-Butyl 2-{2-[2-(5-{[(tert-butoxy)carbonylamino]iminomethyl}-2 methlylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetate: 4-[4-(2 Hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine (100 mg, 0.29 mmol) as prepared in Example 196 step (b) was treated in a manner similar to that shown in Example 40 step (c) to give 100 mg (0.22 mmol, 77%) of (tert-butoxy) 30 N-({4-[4-(2-hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methylthio(2 thienyl)}iminomethyl)carboxamide. This compound was treated in a manner similar to that shown in Example 40, step (d) to give 63 mg (50 %) of tert-butyl 2-{2-[2-(5- WO 00/47578 -120- PCT/US99/18065 -120 {[(tert-butoxy)carbonylamino]iminomethyl } -2-methylthio-3-thienyl)- 1,3-thiazol-4 yl]phenoxy}acetate. 1 H-NMR (CDC1 3 ; 300 MHz) 6 1.55 (s, 9H), 1.56 (s, 9H), 2.69 (s, 3H), 4.66 (s, 2H), 6.88 (dd, J=0.8 and 8.3 Hz, 1H), 7.14 (dt, J=1.0 and 7.6 Hz, 1H), 7.30 (m, 1H), 8.08 (s, 1H), 8.48 (dd, J=1.8 and 7.8 Hz, 1H), 8.51 (s, 1H). 5 b) 2-{2-[2-(5-Amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetic acid: tert-Butyl 2-{2-[2-(5-{ [(tert butoxy)carbonylamino]iminomethyl}-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetate (60 mg, 0.12 mmol) as prepared in previous step was treated in a manner similar to that shown in Example 40, step (e) to give 22 mg (50 %) of 2-{2-[2 10 (5-amidino-2-methylthio-3-thienyl)- 1,3-thiazol-4-yl]phenoxy}acetic acid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 2.80 (s, 3H), 4.90 (s, 2H), 7.17 (m, 2H), 7.36 (m, 1H), 8.41 (d, J=6.3 Hz, 1H), 8.60 (s, 1H), 8.62 (s, 1H), 9.00 (broad s, 2H), 9.37 (broad s, 2H). Mass spectrum (ESI, m/z): Calcd. for C, 17 HzN303S3, 406.5 (M+H), Found 406.1. 15 Example 42 5-Methylthio-4-(6-phenyl(2-pyridyl))thiophene-2-carboxamidine a) Methyl 4-(1,1-dimethyl-1-stannaethyl)-5-methylthiothiophene-2 carboxylate: 4-Bromo-5-methylthiothiophene-2-carboxylic acid (EP 0676395 A2) (4.67 g, 18.4 mmol) was dissolved in anhydrous THF (30 mL), taken in a round 20 bottomed flask and cooled to -78 oC under a N 2 atmosphere. To this solution n butyllithium (20.3 mL, 40.6 mmol, 2M in cyclohexane) was introduced in a dropwise manner. The resulting solution was stirred at -78 'C for 45 min and then allowed to warm up to -60 'C. To this solution trimethyltin chloride (40.6 mL, 40.6 mmol, 1M in THF) was added dropwise. This solution was stirred at -60 oC for 30 min and then 25 allowed to warm up to room temperature. The THF was removed under vacuum and the residue was treated with H 2 0 and extracted with hexane. The hexane layer was evaporated and the residue was dissolved in Et 2 0. The Et 2 0 solution was washed with 10% HC1, saturated NaCl and dried over anhydrous MgSO 4 . Et 2 0 was removed under vacuum and the residue was taken in MeOH. The MeOH solution was treated with 30 trimethylsilyldiazomethane (18.5 mL, 2M in hexane) and stirred at room temperature for 1 h. The solvents were removed under vacuum to give 2 g (31%) of methyl 4-(1,1- WO 00/47578 -121- PCT/US99/18065 dimethyl- 1-stannaethyl)-5-methylthiothiophene-2-carboxylate as an oil. 'H-NMR (CDCl 3 ; 300 MHz) 8 0.31 (s, 9H), 2.57 (s, 3H), 3.86 (s, 3H), 6.98 (s, 1H). b) Methyl 4-(6-bromo(2-pyridyl))-5-methylthiothiophene-2-carboxylate: Methyl 4-(1,1-dimethyl- 1-stannaethyl)-5-methylthiothiophene-2-carboxylate (195 mg, 5 0.56 mmol) as prepared in previous step, and 2,6-dibromopyridine (398 mg, 1.7 mmol) were taken in anhydrous DMF (2 mL). To this mixture tetrakistriphenylphosphine-palladium (20 mg) was added and heated at 120 oC for 24 h. DMF was removed under vacuum and the residue was purified by preparative silica gel thin -layer chromatography to give 78 mg (41%) of methyl 4-(6-bromo(2 10 pyridyl))-5-methylthiothiophene-2-carboxylate as a solid. 'H-NMR (CDCl 3 ; 300 MHz) 8 2.60 (s, 3H), 3.78 (s, 3H), 7.19 (s, 1H), 7.47 (dd, J=1.1 and 7.7 Hz, 1H)), 7.58 (t, J=7.7, 1H), 7.65 (dd, J=1.1 and 7.4 Hz, 1H). c) Methyl 5-methylthio-4-(6-phenyl(2-pyridyl))thiophene-2-carboxylate: Methyl 4-(6-bromo(2-pyridyl))-5-methylthiothiophene-2-carboxylate (78 mg, 0.23 15 mmol) as prepared in previous step, phenylboronic acid (33 mg, 0.27 mmol) and tetrakistriphenylphosphine-palladium (10 mg) were taken in DMF (1 mL). To this solution K 2
CO
3 (75 mg, 0.54 mmol) and H20 (0.3 mL) were added and the mixture was stirred and heated at 90 'C for 18h. Solvents were removed under vacuum and the residue was dissolved in EtOAc and extracted with H20, washed with saturated NaCl 20 and dried over anhydrous Na 2
SO
4 . Thin-layer chromatography of the aqueous layer indicated the presence of some hydrolyzed product. Therefore the aqueous layer was separated acidified with 10% HCI and extracted with EtOAc. The EtOAc layer was washed with saturated NaCl and dried over anhydrous Na 2
SO
4 . This second EtOAc fraction was evaporated and the residue was dissolved in MeOH and treated with 25 trimethylsilyldiazomethane (1.2 eq). This methanolic solution and the first EtOAc fraction were combined and evaporated. The residue was subjected to preparative thin-layer chromatography (10% EtOAc in Hexane) to give 40 mg (51%) of methyl 5 methylthio-4-(6-phenyl(2-pyridyl))thiophene-2-carboxylate which was used directly in the next step. 30 d) 5-Methylthio-4-(6-phenyl(2-pyridyl))thiophene-2-carboxamidine: Methyl 5-methylthio-4-(6-phenyl(2-pyridyl))thiophene-2-carboxylate (40 mg, 0.12 mmol) as prepared in previous step was treated in a manner similar to that for Example 1, to WO 00/47578 PCT/US99/18065 -122 give 10 mg of 5-methylthio-4-(6-phenyl(2-pyridyl))thiophene-2-carboxamidine as a solid. 'H-NMR (CD 3 OD; 300 MHz) 8 2.69 (s, 3H), 7.45-7.60 (min, 3H), 7.62 (s, 1H), 7.79 (dd, J=0.9 and 7.8 Hz, 1H), 7.96 (dd, J=0.9 and 8.0 Hz, 1H), 8.03-8.12 (mn, 3H). Mass spectrum (ESI, m/z): Calcd. for C, 1 7
H
15
N
3
S
2 , 326.1 (M+H), found 326.1. 5 Example 43 5-Methylthio-4-(3-phenylphenyl)thiophene-2-carboxamidine a) Methyl 5-methylthio-4-(3-phenylphenyl)thiophene-2-carboxylate: Methyl 4-(1,1-dimethyl- 1 -stannaethyl)-5-methylthiothiophene-2-carboxylate (200 mg, 10 0.57 mmol, as prepared in Example 42, step a) and 1-bromo-3-phenylbenzene (266 mg, 1.14 mmol) were taken in anhydrous D)MF (2 mL). To this mixture tetrakistriphenylphosphine-palladium (20 mg) was added and heated at 120 oC for 24 h. DMF was removed under vacuum and the residue was purified by preparative silica gel thin -layer chromatography to give 39 mg (20 %) methyl 5-methylthio-4-(3 15 phenylphenyl)thiophene-2-carboxylate as a solid. 'H-NMR (CD 3 OD; 300 MHz)8 2.60 (s, 3H), 3.75 (s, 3H), 7.3-7.5 (mn, 6H), 7.60-7.66 (min, 4H). b) 5-Methylthio-4-(3-phenylphenyl)thiophene-2-carboxamidine: Methyl 5 methylthio-4-(3-phenylphenyl)thiophene-2-carboxylate (35 mg, 0.1 mmol) as prepared in previous step was treated in a manner similar to that for Example 1, to 20 give 17 mg of 5-methylthio-4-(3-phenylphenyl)thiophene-2-carboxamidine as a solid. 'H-NMR (CD 3 OD; 300 MHz) 8 2.60 (s, 3H), 7.3-7.6 (mn, 10H). Mass spectrum (ESI, m/z): Calcd. for C, 8
H
6
N
2
S
2 , 325.4 (M+H), found 325.2. Example 44 25 5-Methlylthio-4-[4-(phenylthiomethyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine a) Methyl 5-methylthio-4-J4-(phenylthiomethyl)(1,3-thiazol-2-yl)]thiophene 2-carboxylate: 2-Phenylthioacetyl chloride (1.0 g, 5.4 mmol) was treated in a manner similar to that for Example 32 step (a) to give 2-bromo-1-phenylthiomethylethan-1 one. The dry solid (1.3 g, 5.3 mmol) was dissolved in acetone (25 ml). To this 30 solution 5-(methoxycarbonyl)-2-(methylthio)-thiophene-3-thiocarboxamide (1.32 g, 5.3 mmol, Maybridge Chemical Co.) was added and heated at reflux for 5 h. At this point the solid that precipitated was filtered off and washed with acetone and dried WO 00/47578 -123- PCT/US99/18065 under vacuum to give 1.5 g (71%) of methyl 5-methylthio-4-[4 (phenylthiomethyl)(1,3-thiazol-2-yl)]thiophene-2-carboxylate which was used without further purification in the following step. b) 5-Methylthio-4-[4-(phenylthiomethyl)(1,3-thiazol-2-yl)]thiophene-2 5 carboxamidine: Methyl 5-methylthio-4-[4-(phenylthiomethyl)(1,3-thiazol-2 yl)]thiophene-2-carboxylate (1.5 g, 3.8 mmol) as prepared in previous step was treated in a manner similar to that for Example 1, however the product was purified by crystallizing from methanol to give 0.86 g (60%) 5-methylthio-4-[4 (phenylthiomethyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine as a solid. 'H-NMR 10 (DMSO-d 6 ; 300 MHz) 8 2.72 (s, 3H), 4.38 (s, 2H), 7.18-7.39 (min, 5H), 7.57 (s, 1H), 8.46 (s, 1H). Mass spectrum (MALDI-TOF, m/z): Calcd. for Cl 6 HisN 3
S
4 , 378.0 (M+H), found 378.1. Example 45 15 4-[4-(2-Chloro-4,5-dimethoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine a) Methyl 4-[4-(2-chloro-4,5-dimethoxyphenyl) (1,3-thiazol-2-yl)]-5 methylthiothliophene-2-carboxylate: 2-Chloro-4,5-dimethoxybenzoic acid (0.5 g, 2.3 20 mmol) and PCl 5 (0.54 g, 2.6 mmol) were placed in a round bottomed flask fitted with a reflux condenser. The mixture was heated in an oil bath at 120 oC for 70 min. The mixture was allowed to cool and the formed phosphorus oxychloride was removed under vacuum to give 0.52 g (96%) of 2-chloro-4,5-dimethoxybenzoyl chloride as a solid. 2-Chloro-4,5-dimethoxybenzoyl chloride (0.52 g, 2.2 mmol) was treated in a 25 manner similar to that for Example 32 step (a) to give 2-bromo-l-(2-chloro-4,5 dimethoxyphenyl) ethan-1-one. The dry solid (0.65 g, 2.2 mmol) was dissolved in acetone (25 ml). To this solution 5-(methoxycarbonyl)-2-(methylthio)-thiophene-3 thiocarboxamide (0.55 g, 2.2 mmol) was added and heated at reflux for 5 h. At this point the solid that precipitated was filtered off and washed with acetone and dried 30 under vacuum to give 0.53 g (54%) of methyl 4-[4-(2-chloro-4,5 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 2.73 (s, 3H), 3.83 (s, 3H), 3.84 (s, 3H), 3.85 (s, 3H), 7.13 (s, 1H), 7.69 (s, 1H), 8.13 (s, 1H), 8.17 (s, 1H).
WO 00/47578 -124- PCT/US99/18065 b) 4-[4-(2-Chloro-4,5-dimethoxyphenyl)(1,3-thiazol-2-yl)J-5 methylthiothiophene-2-carboxamidine: Methyl 4-[4-(2-chloro-4,5 dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate (0.53 g, 1.2 mmol) as prepared in previous step was treated in a manner similar to that for 5 Example 1, however the product was purified by crystallizing from methanol to give to give 0.3 g (60%) 4-[4-(2-chloro-4,5-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxamidine as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 2.77 (s, 3H), 3.84 (s, 6H), 7.13 (s, 1H), 7.71 (s, 1H), 8.17 (s, 1H), 8.69 (s, 1H), 9.16 (broad s, 2H), 9.48 (broad s, 2H). Mass spectrum (MALDI-TOF, m/z): Calcd. for 10 C, 7
HI
6
N
3 0 2
S
3 C1, 426.0 (M+H), found 426.6. Example 46 4-[(Methylethyl)sulfonyl]-5-methylthiothiophene-2-carboxamidine Methyl 4-[(methylethyl)sulfonyl]-5-methylthiothiophene-2-carboxylate (100 15 mg, Maybridge Chemical Company, Cornwall, UK) was treated in a manner similar to that for Example 1, to give 50 mg of 4-[(methylethyl)sulfonyl]-5 methylthiothiophene-2-carboxamidine. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 1.21 (d, J=6.77 Hz, 6H), 2.66 (s, 3H), 3.55 (m, 1H), 7.85 (s, 1H). Mass spectrum (MALDI TOF, CHCA matrix, m/z): Calcd. for C 9
H
14
N
2 0 2
S
3 , 279.0 (M+H), found 279.3. 20 Example 47 Methyl 2
-{
3
-[
2 -(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetate trifluoroacetate 25 To a solution of 42 mg (0.094 mmol) of (tert-butoxy)-N-({4-[4-(3 hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methylthio(2-thienyl)}iminomethyl)carboxamide, prepared in a manner similar to Example 40, step (c), in 2 mL of anhydrous NN dimethylformamide (DMF) was added potassium iodide (0.006 mmol, 1 mg, Aldrich Chemical Co.), cesium carbonate (0.187 mmol, 61 mg, Aldrich Chemical Co.), and methyl 30 bromoacetate (0.187 mmol, 18 tL, Aldrich Chemical Co.) and heated to 60 oC overnight. The reaction solution was concentrated and purified on a 1 mm silica prep plate eluting with 3% methanol/CH 2 Cl 2 to afford 11 mg (23% yield) of methyl 2-{3-[2-(5-{ [(tert butoxy)carbonylamino]iminomethyl } -2-methylthio-3-thienyl)-1,3-thiazol-4- WO 00/47578 -125- PCT/US99/18065 yl]phenoxy}acetate which was then subjected to a solution of 50% trifluoracetic acid/CHC1 2 for 1 h then concentrated and triturated with diethyl ether and dried to afford 7 mg (77% yield) of methyl 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetate trifluoroacetate. 1 H-NMR (CD 3 OD; 300 MHz) 8 8.51 (s, 1H), 7.92 (s, 5 1H), 7.66 (m, 2H), 7.34-7.39 (t, 1H), 6.93 (m, 1H), 4.8 (s, 2H) 3.80 (s, 3H), 2.78 (s, 3H). Mass Spectrum (LC-Q ESI, m/z) Calcd. for C 18
H
17
N
3 0 3
S
3 : 419.5 (M+H), found 420.3. Example 48 5-Methylthio-4-J4-(3-{[N-benzylcarbamoyl]methoxy}phenyl)(1,3-thiazol-2-yl)]thiophene-2 10 carboxamidine trifluoroacetate 100 mg (0.197 mmol) of 2-{3-[2-(5-{[(tert butoxy)carbonylamino]iminomethyl}-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy})acetic acid, as prepared in the previous step, were dissolved in 1 mL of 15 anhydrous DMF and PyBOP (0.396 mmol, 206 mg), benzylamine (0.396 mmol, 42 mg), and diisopropylethylamine (0.494 mmol; 86 tL) were added to the solution and stirred for 18 hrs after which the solution was concentrated and purified on a 2 g silica SPE column and deprotected with 50% trifluoroacetic acid/ methylene chloride to afford 60 mg (67% yield) of 5-methylthio-4-[4-(3-{[N 20 benzylcarbamoyl]methoxy} phenyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine trifluoroacetate. 'H-NMR (CDCl 3 /TFA-d; 300 MHz) 8 8.97 (s, 1H), 7.86 (s, 1H), 7.53 (t, 1H), 7.33 (m, 7H), 7.17 (d, 1H), 4.79 (s, 2H) 4.59 (s, 2H), 2.95 (s, 3H). Mass Spectrum (ESI, m/z) Calcd. for C 24
H
22
N
4 0 2
S
3 : 494.6 (M+H), found 495.2. 25 Example 49 4-{4-[3-({N-[(3,4-Dimethoxyphenyl)methyl]carbamoyl}methoxy)phenyl](1,3-thiazol 2-yl)}-5-methylthiothiophene-2-carboxamidine trifluoroacetate Dissolved 100 mg (0.197 mmol) of 2-{3-[2-(5- { [(tert 30 butoxy)carbonylamino]iminomethyl}-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetic acid, prepared in a manner similar to Example 48, step (c), in 1 mL of anhydrous DMF and added PyBOP (0.396 mmol, 206 mg), 3,4-dimethoxybenzylamine (0.396 mmol,66 mg), and diisopropylethylamine (0.494 mmol; 86 IL) and let stir for 18 hrs after which solution was concentrated and purified on a 2 g silica SPE column and WO 00/47578 PCT/US99/18065 -126 deprotected with 50% trifluoroacetic acid/ methylene chloride to afford 45 mg (41% yield) 4 {4-[3-({N-[(3, 4 -dimethoxyphenyl)methyl]carbamoyl}methoxy)phenyl]( 1,3-thiazol-2-yl)}-5 methylthiothiophene-2-carboxamidine trifluoroacetate. 'H-NMR (CDCl 3 /TFA-d; 300 MHz) 8 8.48 (s, 1H), 7.78 (s, 1H), 7.72 (min, 1H), 7.66 (d, 1H), 7.39 (t, 1H), 7.02 (d, 1H) 4.68 (s, 5 2H), 4.43 (s, 2H), 3.75 (s, 3H). 3.56 (s, 3H). 2.78 (s, 3H). Mass Spectrum (LC-Q ESI, m/z) Calcd. for C 2 6
H
26
N
4 0 4
S
3 :554.6 (M+H), found 555.2 Example 50 5-Methlylthio-4-{4-[3-({N-[2-(phenylamino)ethyl]carbamoyl}methoxy)phenyl](1,3 10 thiazol-2-yl)}thiophene-2-carboxamidine trifluoroacetate Dissolved 100 mg (0.197 mmol) of 2- {3-[2-(5- { [(tert butoxy)carbonylamino]iminomethyl } -2-methylthio-3-thienyl)- 1,3-thiazol-4 yl]phenoxy}acetic acid, prepared in a manner similar to Example 48, step (c), in 1 mL of 15 anhydrous DMF and added PyBOP (0.396 mmol, 206 mg), N-phenylethylenediamine (0.396 mmol, 54 mg), and diisopropylethylamine (0.494 mmol; 86 ptL) and let stir for 18 hrs after which solution was concentrated and purified on a 2 g silica SPE column and deprotected with 50% trifluoroacetic acid/ methylene chloride to afford 65 mg (63% yield) 5-methylthio 4- {4-[3-({N-[2-(phenylamino)ethyl]carbamoyl } methoxy)phenyl](1,3-thiazol-2 20 yl)}thiophene-2-carboxamidine trifluoroacetate 'H-NMR (CDCl 3 /TFA-d; 300 MHz) 6 8.50 (s, 1H), 7.82 (s, 1H), 7.77 (s, 1H), 7.66 (d, 1H), 7.39 (t,1H), 7.02 (d, 1H) 4.68 (s, 2H), 4.43 (s, 2H), 3.75 (s, 3H). 3.56 (s, 3H). 2.78 (s, 3H). Mass Spectrum (LC-Q ESI, m/z) Calcd. for
C
25
H
25
N
5 0 2
S
3 : 523.6 (M+H), found 524.1 25 Example 51 5-Methylthio-4-[4-(3-{[N-(2-morpholin-4-ylethyl)carbamoyl]methoxy}phenyl) (1,3 thiazol-2-yl)]thiophene-2-carboxamidine trifluoroacetate 83 mg (0.164 mmol) of 2- {3-[2-(5- { [(tert-butoxy)carbonylamino]iminomethyl}-2 30 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy} acetic acid, prepared in a manner similar to Example 40, step (c), was reacted with 2-morpholin-4-ylethylamine (0.328 mmol, 43 L) in a manner similar to Example 48 to afford 46 mg (54% yield) of 5-methylthio-4-[4-(3-{[N-(2 morpholin-4-ylethyl)carbamoyl]methoxy} phenyl)(1,3-thiazol-2-yl)]thiophene-2 carboxamidine trifluoroacetate. 'H-NMR (DMSO-d 6 , 300 MHz) 8 9.38 (bs, 2H), 9.08 (bs, WO 00/47578 -127- PCT/US99/18065 2H), 8.61 (s, 1H), 8.45 (t, 1H), 8.27 (s, 1H), 7.72 (m, 2H) 7.45 (t, 1H), 7.02 (d, J= 8 Hz, 1H), 4.62 (s, 2H), 3.53-3.64 (m, 5H), 3.24-3.38 (m, 5H), 2.80 (s, 3H), 1.1 (t, 2H). Mass Spectrum (ESI, m/z) Calcd. for C 2 3
H
2 7N 5 0 3
S
3 : 517.6 (M+H), found 518.2. 5 Example 52 5-Methylthio-4-{4-[3-(2-morpholin-4-yl-2-oxoethoxy)phenyl](1,3-thiazol-2-yl)}thiophene 2-carboxamidine trifluoroacetate 73 mg (0.144 mmol) of 2- {3-[2-(5-{[(tert-butoxy)carbonylamino]iminomethyl}-2 10 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid, prepared in a manner similar to Example 48, step (c), was reacted with morpholine (0.288 mmol; 25 ptL) in a manner similar to Example 48 step (b) to afford 50 mg (75% yield) 5-methylthio-4-{4-[3-(2-morpholin-4-yl 2-oxoethoxy)phenyl](1,3-thiazol-2-yl) } thiophene-2-carboxamidine trifluoroacetate. 1 H-NMR (DMSO-d 6 /TFA-d; 300 MHz) 8 9.38 (bs, 1H), 9.08 (bs, 2H), 8.66 (s, 1H), 8.22 (s, 1H), 7.72 15 (m, 2H) 7.42 (t, 1H), 6.98-7.00 (dd, J= 2.3 Hz and 8.2 Hz, 1H), 4.95 (s, 2H), 3.53-3.67 (m, 8H), 2.82 (s, 3H). Mass Spectrum (ESI, m/z) Calcd. for C 21
H
22
N
4 0 3
S
3 :474.6 (M+H), found 475.2. Example 53 20 5-Methlylthliio-4-{4-[3-(2-oxo-2-piperazinylethoxy)phenyl](1,3-thiazol-2 yl)}thiophene-2-carboxamidine trifluoroacetate 100 mg (0.198 mmol) of 2- { 3-[2-(5- { [(tert-butoxy)carbonylamino]iminomethyl }-2 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid, prepared in a manner similar to 25 Example 48, step (c), was reacted with tert-butyl piperazinecarboxylate (0.396 mmol; 74 mg) in a manner similar to Example 48, step (b) to afford 40 mg (43% yield) of 5-methylthio-4 {4-[3-(2-oxo-2-piperazinylethoxy)phenyl](1,3-thiazol-2-yl) }thiophene-2-carboxamidine trifluoroacetate. 'H-NMR (DMSO-d 6 /TFA-d; 300 MHz) 8 8.68 (s, 1H), 8.20(s, 1H), 7.75 (m, 2H) 7.43 (t, 1H), 7.01 (dd, J= 2.3 Hz and 8.1 Hz, 1H), 5.02 (s, 2H), 3.76 (bs, 4H), 3.17-3.26 30 (m, 4H). 2.82 (s, 3H). Mass Spectrum (LC-Q ESI, m/z) Calcd. for C 2 1H 23
N
5 0 2
S
3 : 473.6 (M+H), found 474.2.
WO 00/47578 -128- PCT/US99/18065 Example 54 4-[4-(3-{[N-(2-Aminoethyl)carbamoyl]methoxy}phenyl) (1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxamidine hydrochloride 5 51 mg (0.101 mmol) of 2- { 3-[2-(5- { [(tert-butoxy)carbonylamino]iminomethyl}-2 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid, prepared in a manner similar to Example 48, step (c), was reacted with N-(2-aminoethyl)(tert-butoxy)carboxamide (0.202 mmol; 32 mg) in a manner similar to Example 48, step (b) to afford 80 mg (80% yield) of 4 (4- { 3-[(N- {2- [(tert-butoxy)carbonylamino]ethyl } carbamoyl)methoxy]phenyl } (1,3-thiazol-2 10 yl))-5-methylthiothiophene-2-carboxamidine which was then deprotected with 4N HCI in dioxane to afford 36 mg (68% yield) of 4-[4-(3-{ [N-(2 aminoethyl)carbamoyl]methoxy}phenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride. 1 H-NMR (CD 3 OD; 300 MHz) 8 8.55 (s, 1H), 7.95 (s, 1H), 7.73 (min, 2H) 7.41 (t, 1H), 7.05 (min, 1H), 4.80 (s, 2H), 3.51 (mn, 2H), 3.13-3.31 (min, 2H), 2.83 15 (s, 3H). Mass Spectrum (ESI, m/z) Calcd. for C, 9
H
21
N
5 0 2
S
3 : 447.5 (M+H), found 448.2. Example 55 4-(4-{3-[2-(4-Acetylpiperazinyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate 20 52 mg (0.103 mmol) of 2-{3-[2-(5-{[(tert-butoxy)carbonylamino]iminomethyl}-2 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid, prepared in a manner similar to Example 48, step (c), was reacted with 1-acetyl piperazine (0.154 mmol, 20 mg), 1 -hydroxy 7-azabenzotriazole (HOAt)) (0.154 mmol, 21 mg), O-(7-azabenzotriazol- 1 -yl)-1,1,3,3 25 tetramethyluronium hexafluorophosphate) HATU (0.154 mmol, 58 mg) and diisopropylethylamine (0.258 mmol, 44 jiL) in DMF to afford crude product which was then purified on 1 mm silica prep plates eluting with 3% methanol/methylene chloride to afford 28 mg (53% yield) of N-{[4-(4-{3-[2-(4-acetylpiperazinyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2 yl))-5-methylthio(2-thienyl)]iminomethyl}(tert-butoxy)carboxamide. This was subsequently 30 reacted with a solution oftrifluoroacetic acid: methylene chloride: water (47.5%: 47.5%: 2.5%) for 1 hour, concentrated and purified on a silica SPE column eluting with 15% methanol/methylene chloride to afford 20 mg (80% yield) of 4-(4-{3-[2-(4 acetylpiperazinyl)-2-oxoethoxy]phenyl} (1 ,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxamidine trifluoroacetate. 'H-NMR (CD 3 OD; 300 MHz) 8 8.48 (s, 1H), 7.91 (s, 1H), WO 00/47578 PCT/US99/18065 -129 7.69 (m, 2H) 7.38 (t, 1H), 6.99 (dd, J= 2 Hz and 8.1 Hz, 1H), 4.93 (s, 2H), 3.52-3.67 (m, 8H), 2.78 (s, 3H), 2.12 (s, 3H). Mass Spectrum (ESI, m/z) Calcd. for C 23
H
25
N
5 0 3
S
3 : 515.6 (M+H), found 516.2. 5 Example 56 4-(4-{3-[2-(4-Methylpiperazinyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate 54 mg (0.107 mmol) of 2-{3-[2-(5- { [(tert-butoxy)carbonylamino]iminomethyl}-2 10 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid, prepared in a manner similar to Example 48, step (c), was reacted with N-methyl piperazine (0.128 mmol, 14 gL), 1-hydroxy 7-azabenzotriazole (HOAt) (0.128 mmol, 17 mg), O-(7-azabenzotriazol- 1 -yl)- 1,1,3,3 tetramethyluronium hexafluorophosphate) HATU (0.128 mmol, 49 mg) and diisopropylethylamine (0.268 mmol, 56 pL) in DMF to afford crude product which was then 15 partitioned between methylene chloride and 1N NaOH and washed. The organic layer was obtained and similarly washed with 10 % citric acid and saturated aq. sodium chloride, dried over sodium sulfate and concentrated to a yellow oil. The oil was then purified on 1 mm silica prep plates eluting with 5% methanol/methylene chloride to afford (tert-butoxy)-N { imino [4-(4- { 3- [2-(4-methylpiperazinyl)-2-oxoethoxy]phenyl } (1,3 -thiazol-2-yl))-5 20 methylthio(2-thienyl)]methyl}carboxamide. This was subsequently reacted with a solution of trifluoroacetic acid: methylene chloride: water (47.5%: 47.5%: 2.5%) for 1 hour, concentrated and purified on a silica SPE column eluting with 10-15% methanol/methylene chloride to afford 17 mg (33% yield) of 4-(4- { 3-[2-(4-methylpiperazinyl)-2-oxoethoxy]phenyl } (1,3 thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine trifluoroacetate. 'H-NMR (CD 3 OD; 25 300 MHz) 6 8.52 (s, 1H), 7.91 (s, 1H), 7.69 (m, 2H), 7.38 (t, 1H), 6.98 (dd, J= 2.0 Hz and 8.1 Hz, 1H), 4.90 (s, 2H), 3.66 (t, 4H), 2.78 (s, 3H), 2.49-2.57 (m, 4H), 2.35 (s, 3H). Mass Spectrum (ESI, m/z) Calcd. for C 22
H
25
N
5 0 2
S
3 : 487.6 (M+H), found 488.2 Example 57 30 5-Methylthio-4-[4-(3-{2-oxo-2-[4-benzylpiperazinyl]ethoxy}pheniyl) (1,3-thiiazol-2 yl)]thiophene-2-carboxamidine trifluoroacetate 54 mg (0.107 mmol) of 2-{3-[2-(5- { [(tert-butoxy)carbonylamino]iminomethyl}-2 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid, prepared in a manner similar to WO 00/47578 -130- PCT/US99/18065 Example 48, step (c), was reacted with N-benzylpiperazine (0.128 mmol, 22 pL), 1-hydroxy 7-azabenzotriazole (HOAt) (0.128 mmol, 17 mg), O-(7-azabenzotriazol- 1 -yl)- 1,1,3,3 tetramethyluronium hexafluorophosphate) HATU (0.128 mmol, 48 mg) and diisopropylethylamine (0.267 mmol, 50 pL) in DMF to afford crude product which was then 5 partitioned between methylene chloride and 1N NaOH and washed. The organic layer was obtained and similarly washed with 10% citric acid and saturated aq. sodium chloride, dried over sodium sulfate and concentrated to a yellow oil. The oil was then purified on 1 mm silica prep plates eluting with 5% methanol/methylene chloride to afford (tert-butoxy)-N (imino {5-methylthio-4-[4-(3- {2-oxo-2-[4-benzylpiperazinyl]ethoxy}phenyl)(1,3-thiazol-2 10 yl)](2-thienyl)}methyl)carboxamide. This was subsequently reacted with a solution of trifluoroacetic acid: methylene chloride: water (47.5%: 47.5%: 2.5%) for 1 hour, concentrated and purified on a 5 g silica SPE column eluting with 10-15% methanol/methylene chloride to afford 36 mg (60% yield) of 5-methylthio-4-[4-(3-{2-oxo-2-[4 benzylpiperazinyl]ethoxy }phenyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine 15 trifluoroacetate. 'H-NMR (CD 3 OD; 300 MHz) 5 8.54 (s, 1H), 7.93 (s, 1H), 7.71 (min, 2H), 7.50 (s, 5H) 7.39 (t, 1H), 6.99 (dd, J= 2 Hz and 8.1 Hz, 1H), 4.94 (s, 2H), 4.37 (s, 2H), 3.3 (min, 4H), 2.81 (s, 3H), 2.49-2.57 (min, 4H), 2.35 (s, 3H). Mass (ESI, m/z) Calcd. for
C
28
H
29
N
5 0O 2
S
3 : 563.7 (M+H), found 564.3. 20 Example 58 (D,L)- 4-(4-{3-[2-(3-Aminopyrrolidinyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate 41 mg (0.081 mmol) of 2- { 3-[2-(5-{ [(tert-butoxy)carbonylamino]iminomethyl }-2 25 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid, prepared in a manner similar to Example 48, step (c), was reacted with (D,L) (tert-butoxy)-N-pyrrolidin-3-ylcarboxamide (0.122 mmol, 23 mg), O-(7-azabenzotriazol- 1 -yl)- 1,1,3,3-tetramethyluronium hexafluorophosphate) HATU (0.122 mmol, 46 mg), 1-hydroxy-7-azabenzotriazole (HOAt) (0.122 mmol, 17 mg) and diisopropylethylamine (0.203 mmol, 35 pL) in a manner similar to 30 Example 56 to afford 20 mg (53% yield) of (D,L)- 4-(4-{3-[2-(3-aminopyrrolidinyl)-2 oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine trifluoroacetate. 'H-NMR (CD 3 OD; 300 MHz) 8 8.54 (s, 1H), 7.94 (s, 1H), 7.71 (min, 2H), 7.39 (t, 1H), 6.99 (dd, J= 2.0 Hz and 8.1 Hz, 1H), 4.85 (s, 2H), 4.37 (s, 2H), 3.60-4.01 (inm, WO 00/47578 -131- PCT/US99/18065 5H), 2.81 (s, 3H), 2.15-2.71 (m, 2H). Mass Spectrum (ESI, m/z) Calcd. for C 2 1
H
2 3
N
5 0 2
,S
3 : 473.6 (M+H), found 474.3. Example 59 5 5-Methylthio-4-{4-[3-(2-oxo-2-piperidylethoxy)phenyl](1,3-thiazol-2-yl)}thiophene 2-carboxamidine trifluoroacetate 33 mg (0.065 mmol) of 2-{3-[2-(5-{[(tert-butoxy)carbonylamino]iminomethyl}-2 methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid, prepared in a manner similar to 10 Example 40, step (c), was reacted with piperidine (0.078 mmol, 8 pL), O-(7-azabenzotriazol 1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) HATU (0.078 mmol, 30 mg), 1 hydroxy-7-azabenzotriazole (HOAt) (0.078 mmol, 11 mg) and diisopropylethylamine (0.163 mmol, 56pL) in a manner similar to Example 57 to afford 15 mg (41% yield) of 5 methylthio-4- {4-[3-(2-oxo-2-piperidylethoxy)phenyl](1,3-thiazol-2-yl) } thiophene-2 15 carboxamidine trifluoroacetate. 1 H-NMR (CD 3 OD; 300 MHz) 6 8.54 (s, 1H), 7.92 (s, 1H), 7.69 (m, 2H), 7.35-7.40 (t, 1H), 6.98 (dd, J= 2 Hz and 8.1 Hz, 1H), 4.95 (s, 2H), 3.52-3.60 (m, 4H), 2.80 (s, 3H), 1.57-1.70 (m, 6H). Mass Spectrum (ESI, m/z) Calcd. for
C
22
H
24
N
4 0 2
S
3 :472.6 (M+H), found 473.2. 20 Example 60 2-(3-{2-[5-(Imino{[(4-polystyryloxyphenyl)metlhoxy]carbonylamino}methyl)-2 methylthio-3-thienylJ-1,3-thiazol-4-yl}phenoxy)acetic acid 2 g (1.86 mmol) ofp-nitrophenyl carbonate Wang resin (0.93 mmol/g) (Calbiochem 25 Novabiochem, San Diego, CA) was suspended in 9 mL of a 2:1 mixture of anhydrous DMSO:DMF. 2 g (4.93 mmol) of 2-{3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetic acid was added to suspension followed by the addition of 1 mL of 1,8 diazabicyclo[5.4.0]undec-7-ene, (DBU, 6.69 mmol) and let shake vigorously for 5 days after which resin was washed thoroughly with DMF, MeOH, and diethyl ether and dried in vacuo 30 to afford 2 g of resin-bound 2-{3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy } acetic acid.
WO 00/47578 PCT/US99/18065 -132 Example 61 (D,L)-Ethyl 1-(2-{3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetyl)piperidine-2-carboxylate trifluoroacetate 5 100 mg (0.093 mmol) of resin-bound 2- { 3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was suspended 1 mL of anhydrous DMF. O-(7-azabenzotriazol-1-yl)-1,1,3,3 tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5 M; 68 mg), ethyl piperidine-2-carboxylate (0.5 M; 78 pL) and 10 diisopropylethylamine (0.233 mmol, 40 tL) were added and allowed to shake vigorously for 18 hrs, after which the resin was washed thoroughly with DMF, methanol, methylene chloride, and diethyl ether. After drying, crude product was removed from resin by reaction with a solution of trifluoroacetic acid: methylene chloride: water (47.5%: 47.5%: 2.5%) for 1 hour. The solution was filtered and concentrated to a yellow oil. After purification on a 2 g 15 silica SPE column, eluting with a gradient of 3%-10% MeOH/methylene chloride, 15 mg (30% yield) of (D,L)-ethyl 1-(2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetyl)piperidine-2-carboxylate trifluoroacetate was obtained. Mass Spectrum (ESI, m/z) Calcd. for C 25
H
28
N
4 0 4
S
3 : 544.70 (M+H), found 545.2 20 Example 62 5-Methylthio-4-{4-[3-(2-oxo-2-pyrrolidinylethoxy)phenyl](1,3-thiazol-2 yl)}thiophene-2-carboxamidine trifluoroacetate 100 mg (0.093 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 25 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was suspended 1 mL of anhydrous DMF. O-(7-azabenzotriazol-1-yl)-1,1,3,3 tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5 M; 68 mg), pyrrolidine (0.5 M; 42 pL) and diisopropylethylamine (0.233 mmol, 40 [tL) were added and allowed to shake vigorously for 30 18 hours, after which the resin was washed thoroughly with DMF, methanol, methylene chloride, and diethyl ether. After drying, crude product was removed from resin by reaction with a solution of trifluoroacetic acid: methylene chloride: water (47.5%: 47.5%: 2.5%) for 1 hour. After trituration with diethyl ether and drying, 18 mg (42% yield) of 5-methylthio-4 { 4-[3-(2-oxo-2-pyrrolidinylethoxy)phenyl](1,3-thiazol-2-yl) }thiophene-2-carboxamidine WO 00/47578 PCT/US99/18065 -133 trifluoroacetate was obtained. Mass Spectrum (ESI, m/z) Calcd. for C 21
H
2 2
N
4 0 2
S
3 : 458.6 (M+H), found 459.2 Example 63 5 5-Methylthio-4-[4-(3-{2-oxo-2-[4-benzylpiperidyl]ethoxy}phenyl)(1,3-thiazol-2 yl)]thiophene-2-carboxamidine trifluoroacetate 80 mg (0.074 mmol) of resin-bound 2- { 3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 10 60, was suspended in 1 mL of anhydrous DMF. O-(7-azabenzotriazol-1-yl)-1,1,3,3 tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5 M; 68 mg), 4-benzyl piperidine (0.5 M; 88 gL) and diisopropylethylamine (0.185 mmol, 32 tL) were added and allowed to shake vigorously for 18 hrs, after which the resin was washed thoroughly with DMF, methanol, methylene 15 chloride, and diethyl ether. After drying, crude product was removed from resin by reaction with a solution of trifluoroacetic acid: methylene chloride: water (47.5%: 47.5%: 2.5%) for 1 hour. After trituration with diethyl ether and drying, 17 mg (40% yield) of 5-methylthio-4-[4 (3-{2-oxo-2-[4-benzylpiperidyl]ethoxy}phenyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine trifluoroacetate was obtained. Mass Spectrum (ESI, m/z) Calcd. for C 29
H
30
N
4 0 2
S
3 : 562.7 20 (M+H), found 563.3. Example 64 (D,L)-4-(4-{3-[2-(3-Methylpiperidyl)-2-oxoethoxy]phenyl(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate 25 80 mg (0.074 mmol) of resin-bound 2-{3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with (+/-)-3-methyl piperidine (0.5 M, 59 jtL) and O-(7-azabenzotriazol-1 yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 30 azabenzotriazole (HOAt) (0.5 M; 68 mg) and diisopropylethylamine (0.185 mmol, 32 tL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 10 mg (28% yield) of 4-(4- { 3-[2-(3-methylpiperidyl)-2-oxoethoxy]phenyl } (1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for
C
23
H
26
N
4 0 2
S
3 : 486.6 (M+H), found 487.3.
WO 00/47578 -134- PCT/US99/18065 Example 65 4
-(
4
-{
3
-J
2
-(
4 -Methylpiperidyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate 5 80 mg (0.074 mmol) of resin-bound 2 -{3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with 4-methyl piperidine (0.5 M, 59 pL) and O-(7-azabenzotriazol-1-yl) 1,1,3,3-tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 10 azabenzotriazole (HOAt) (0.5 M; 68 mg) and diisopropylethylamine (0.185 mmol, 32 ptL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 12 mg (33% yield) of 4-(4- { 3
-[
2
-(
4 -methylpiperidyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for
C
23
H
26
N
4 0 2
S
3 :486.6 (M+H), found 487.3. 15 Example 66 4-(4-{3-J2-(2-Azabicyclo[4.4. 0]dec-2-yl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trfluoroacetate 20 80 mg (0.074 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with decahydroquinoline (0.5 M, 75 pL) and O-(7-azabenzotriazol-1-yl) 1,1,3,3-tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5 M; 68 mg) and diisopropylethylamine (0.185 mmol, 32 VtL) in 25 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 16 mg (41% yield) of 4-(4-{ 3-[2-(2-azabicyclo[ 4
.
4 .0]dec-2-yl)-2-oxoethoxy]phenyl} (1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for
C
26
H
3 0
N
4 0 2
S
3 : 526.7 (M+H), found 527.2. 30 Example 67 (D,L)-Ethyl 1-( 2
-{
3
-[
2 -(5-amidino-2-methylthio-3-thlienyl)-1,3-thiazol-4 yl]phenoxy}acetyl)piperidine-3-carboxylate trifluoroacetate 80 mg (0.074 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)- 1,3 35 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example WO 00/47578 -135- PCT/US99/18065 60, was reacted with ethyl nipecotate (0.5 M, 78 pL) and O-(7-azabenzotriazol-1-yl)-1,1,3,3 tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5 M. 68 mg) and diisopropylethylamine (0.185 mmol, 32 tL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 18 mg (45% yield) of 5 ethyl 1-(2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetyl)piperidine-3-carboxylate trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for C 2 5
H
28
N
4 0 4
S
3 :545.7 (M+H), found 545.2. Example 68 10 5-Methliylthio-4-{4-[3-(2-oxo-2-(1,2,3,4-tetrahlydroquinolyl)ethoxy)phenyl](1,3 thiazol-2-yl)}thiophene-2-carboxamidine trifluoroacetate 100 mg (0.093 mmol) of resin-bound 2-{3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 15 60, was reacted with 1,2,3,4-tetrahydroisoquinoline (0.5M) and O-(7-azabenzotriazol-1-yl) 1,1,3,3-tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5 M; 68 mg) and diisopropylethylamine (0.233 mmol, 40 pL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 20 mg (42% yield) of 5-methylthio-4-{4-[3-(2-oxo-2-( 1,2,3,4-tetrahydroquinolyl)ethoxy)phenyl](1,3-thiazol-2 20 yl)}thiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for
C
26
H
24
N
4 0 2
S
3 : 520.7 (M+H), found 521.2. Example 69 Ethyl 1-(2-{3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 25 yl]phenoxy}acetyl)piperidine-4-carboxylate trifluoroacetate 100 mg (0.093 mmol) of resin-bound 2- { 3- [2-(5-amidino-2-methylthio-3-thienyl)- 1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with ethyl isonipecotate (0.5 M, 77 mg) and O-(7-azabenzotriazol-l-yl) 30 1,1,3,3-tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5 M; 68 mg) and diisopropylethylamine (0.233 mmol, 40 tL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 21 mg (42% yield) of ethyl 1-(2- { 3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4- WO 00/47578 -136- PCT/US99/18065 yl]phenoxy}acetyl)piperidine-4-carboxylate trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for C 25
H
28
N
4 0 4
S
3 : 545.7 (M+H), found 545.3. Example 70 5 4-(4-{3-[2-((3R)-3-Hydroxypiperidyl)-2-oxoethoxy]phenyl)(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate 100 mg (0.093 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)- 1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 10 60, was reacted with R-(+)-3-hydroxy piperidine (0.5 M, 69 mg) and O-(7-azabenzotriazol-1 yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5M; 68 mg) and diisopropylethylamine (0.233 mmol, 40 pL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 16 mg (36% yield) of 4-(4- { 3-[2-((3R)-3-hydroxypiperidyl)-2-oxoethoxy]phenyl } (1,3-thiazol-2-yl))-5 15 methylthiothiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for
C
2 2
H
2 3
N
4 0 3
S
3 : 489.7 (M+H), found 489.2. Example 71 D,L-4-(4-{3-[2-(2-Ethylpiperidyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 20 methylthiothiophene-2-carboxamidine trifluoroacetate 100 mg (0.093 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with 2-ethyl piperidine (0.5M) and O-(7-azabenzotriazol-1-yl)-1,1,3,3 25 tetramethyluronium hexafluorophosphate) HATU (0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5M; 68 mg) and diisopropylethylamine (0.233 mmol, 40 pL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 11 mg (23% yield) of D,L-4-(4- {3-[2-(2-ethylpiperidyl)-2-oxoethoxy]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for 30 C 2 4
H
27
N
4 0 2
S
3 : 501.4 (M+H), found 501.4.
WO 00/47578 -137- PCT/US99/18065 Example 72 4-(4-{3-[2-((3S)-3-Hydroxypyrrolidinyl)-2-oxoethoxy]phenyl)(1,3-thiazol-2-yi))-S methylthiothiophene-2-carboxamidine trifluoroacetate 5 100 mg (0.093 mmol) of resin-bound 2-{3-[2-(5-amidino-2-methylthio-3 thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with R-(-)-3-pyrrolidinol (0.5 M, 62 mg) and O (7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (HATU, 0.5 M, 190 mg), 1-hydroxy-7-azabenzotriazole (HOAt) (0.5M; 68 mg) and 10 diisopropylethylamine (0.233 mmol, 40 ptL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 10 mg (23% yield) of 4-(4-{3-[2-((3S)-3 hydroxypyrrolidinyl)-2-oxoethoxy]phenyl } (1,3-thiazol-2-yl))-5-methylthiothiophene 2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for
C
21
H
22
N
4 0 3
S
3 : 475.2 (M+H), found 475.2. 15 Example 73 5-Methylthio-4-(4-{3-J(N-(5,6,7,8 tetrahydronaphthyl)carbamoyl)methoxy]phenyl(1,3-thiazol-2-yl))thiophene-2 carboxamidine trifluoroacetate 20 100 mg (0.093 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with 5,6,7,8-tetrahydro-l-naphthylamine (0.5 M, 73 mg) and 0-(7 azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (HATU, 0.5 M, 190 25 mg), 1-hydroxy-7-azabenzotriazole (HOAt) (0.5 M; 68 mg) and diisopropylethylamine (0.233 mmol, 40 jtL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 15 mg (30% yield) of 5-methylthio-4-(4-{3-[(N-(5,6,7,8 tetrahydronaphthyl)carbamoyl)methoxy]phenyl } (1,3-thiazol-2-yl))thiophene-2 carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for C 27
H
2 6
N
4 0 2
S
3 : 535.2 30 (M+H), found 535.3.
WO 00/47578 -138- PCT/US99/18065 Example 74 D, L- 4
-[
4
-(
3
-{
2
-[
3 -(Hydroxymethyl)piperidyl]-2-oxoethoxy}phenyl)(1,3-thiazol-2-yl)]-S methylthiothiophene-2-carboxamidine trifluoroacetate 5 100 mg (0.093 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3 thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with 3-piperidine methanol (0.5 M, 58 mg) and O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (HATU, 0.5 M, 190 mg), 1-hydroxy-7-azabenzotriazole (HOAt) (0.5M; 68 mg) and 10 diisopropylethylamine (0.233 mmol, 40 pL in 1 mL of anhydrous DMF in a manner similar to Example 40 to afford to 19 mg (40% yield) of D,L-4-[4-(3-{2-[3 (hydroxymethyl)piperidyl]-2-oxoethoxy}phenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for C 23
H
2 5
N
4 0 3
S
3 : 503.2 (M+H), found 503.2. 15 Example 75 4 -{4-[3-(2-{(2R)-2-[(Phenylamino)methyl]pyrrolidinyl}-2-oxoethoxy)phenyl](1,3 thiazol-2-yl)}-5-methylthiothiophene-2-carboxamidine trifluoroacetate 20 100 mg (0.093 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with (S)-(+)-2-anilino methyl pyrrolidine (0.5 M, 88 mg) and O-(7 azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (HATU, 0.5 M, 190 mg), 1-hydroxy-7-azabenzotriazole (HOAt) (0.5M; 68 mg) and diisopropylethylamine (0.233 25 mmol, 40 pL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 13 mg (25% yield) of 4-{4-[3-(2- {(2R)-2-[(phenylamino)methyl]pyrrolidinyl}-2 oxoethoxy)phenyl](1,3-thiazol-2-yl)}-5-methylthiothiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for C 28
H
28
N
5 0 2
S
3 : 563.8 (M+H), found 564.2. 30 WO 00/47578 -1 39- PCT/US99/18065 Example 76 4-J4-(3-{2-[(3R)-3-(Methoxymethyl)pyrrolidinylf-2-oxoethoxy}phenyl)(1,3-thiazol 2-yl)]-5-methylthiothiophene-2-carboxamidine trifluoroacetate 5 100 mg (0.093 mmol) of resin-bound 2-{3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with (S)-(+)-2-methoxymethyl pyrrolidine (0.5 M, 58 mg) and O-(7 azabenzotriazol- 1 -yl)- 1,1,3,3-tetramethyluronium hexafluorophosphate) (HATU, 0.5 M, 190 mg), 1-hydroxy-7-azabenzotriazole (HOAt) (0.5M; 68 mg) and diisopropylethylamine (0.233 10 mmol, 40 tL) in 1 mL of anhydrous DMF in a manner similar to Example 63 to afford 16 mg (35% yield) of 4-[4-(3-{2-[(3R)-3-(methoxymethyl)pyrrolidinyl]-2 oxoethoxy}phenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for C 23
H
2 6
N
4 0 3
S
3 : 503.2 (M+H), found 503.3. 15 Example 77 1-(2-{3-[2-(5-Amidino-2-methylthio-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetyl)piperidine-3-carboxamide trifluoroacetate 20 100 mg (0.093 mmol) of resin-bound 2- {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3 thiazol-4-yl]phenoxy}acetic acid (0.93 mmol/g), as prepared in a manner similar to Example 60, was reacted with nipecotamide (0.5 M, 64 mg) and O-(7-azabenzotriazol-l-yl)-1, 1,3,3 tetramethyluronium hexafluorophosphate) (HATU, 0.5 M, 190 mg), 1-hydroxy-7 azabenzotriazole (HOAt) (0.5M; 68 mg) and diisopropylethylamine (0.233 mmol, 40 ptL) in 1 25 mL of anhydrous DMF in a manner similar to Example 63 to afford 11 mg (23% yield) 1-(2 {3-[2-(5-amidino-2-methylthio-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetyl)piperidine-3 carboxamide trifluoroacetate. Mass Spectrum (ESI, m/z) Calcd. for C 2 3
H
2 5
N
4 0 3
S
3 :516.2 (M+H), found 516.3. 30 Example 78 5-Methlylthio-4-{4-[3-(trifluoromethoxy)phenyl](1,3-thiazol-2-yl)}thliiophene-2 carboxamidine hydrochloride a) Methyl 5-methylthio-4-{4-[3-(trifluoromethoxy)pheniyl](1,3-thiazol-2 35 yl)}thiophene-2-carboxylate: 435 mg (1.76 mmol) of methyl 4-(aminothioxomethyl)-5- WO 00/47578 -140- PCT/US99/18065 methylthiothiophene-2-carboxylate was dissolved in 10 mL of reagent grade acetone. 2 bromo-3'-trifluoromethoxy acetophenone, prepared in a manner similar to Example 95, step (a), (1.76 mmol; 497 mg) was added and the solution was allowed to reflux for 3 h. The solution was allowed to cool and concentrated to an oil which was then dissolved in 150 mL 5 of methylene chloride and washed with 50 mL of 10% HCI (aq.) and 50 mL of 2N NaOH (aq.). The organic layer was obtained and dried over magnesium sulfate and concentrated affording 877 mg (90% yield) of a methyl- 5-methylthio-4-{4-[3 (trifluoromethoxy)phenyl](1,3-thiazol-2-yl)}thiophene-2-carboxylate. b) 5-Methylthiio-4-{4-[3-(trifluoromethoxy)phenyl](1,3-thiazol-2-yl)}thiophene-2 10 carboxamidine hydrochloride: To a stirred suspension of 19.4 mmol( 1.04 g) of ammonium chloride (Fisher Scientific) in 20 mL of anhydrous toluene (Aldrich Chemical Co.) placed under nitrogen atmosphere at 0 oC, 9.7 mL ( 19.4 mmol) of 2M trimethylaluminum in toluene (Aldrich Chemical Co.) was added via syringe over 15 min and then let stir at 0 oC for 30 min after which 837 mg (1.94 mmol) of methyl- 5-methylthio-4-{4-[3 15 (trifluoromethoxy)phenyl](1,3-thiazol-2-yl) } thiophene-2-carboxylate was added to solution and allowed to reflux for 3 h. The reaction mixture was quenched by pouring over a slurry of 10 g of silica in 50 mL of chloroform. The silica was poured onto a sintered glass funnel and washed with ethyl acetate and eluting with a 15% methanol/CH 2 C2, solution and concentrated. The crude product was purified on 1 mm silica prep plates eluting with 15% 20 methanol/CH 2 C1 2 and treated with 4N HCl/dioxane to afford 37 mg (5% yield) of 5 methylthio-4- {4-[3-(trifluoromethoxy)phenyl](1,3-thiazol-2-yl) } thiophene-2-carboxamidine hydrochloride. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 9.43 (bs, 1.9 H), 9.05 (bs, 1.9 H), 8.67 (s, 1H), 8.43 (s, 1H), 8.10 (m, 2H), 7.65 (t, 1H), 7.40 (m, 1H), 2.8 (s, 3H). Mass Spectrum (LCQ-ESI, m/z) Calcd. for C 1 6
HI
2
F
3
N
3
OS
3 : 415.5(M+H), found 416.2. 25 Example 79 S-Methylthio-4-(5-phenyl(1,3-oxazol-2-yl))thiophene-2-carboxamidine hydrochloride 30 a) Methyl 5-methylthio-4-[N-(2-oxo-2-phenylethyl)carbamoyl]thiophene-2 carboxylate: To a stirred suspension of 300 mg (1.29 mmol) of 5-(methoxycarbonyl) 2-methylthiothiophene-3-carboxylic acid (as prepared in Example 95) in 10 mL of anhyd CH 2 C1 2 (under a CaSO 4 drying tube) was added 135 mL (1.55 mmol) of oxalyl WO 00/47578 -141- PCT/US99/18065 chloride followed by 30 mL of anhyd DMF. After stirring for 2 h at room temperature, the mixture was concentrated in vacuo. The resulting yellow solid was dissolved in 10 mL of anhyd CH 2
CI
2 , cooled (0oC) and 266 mg (1.55 mmol) of 2 aminoacetophenone was added. N,N-diisopropylethylamine (DIEA) (756 mL, 4.34 5 mmol) was added dropwise over 3 min and the mixture stirred for 1 h at room temperature. The mixture was concentrated to an oil and partitioned between 125 mL of EtOAc and 80 mL of 1 M HC1. The aqueous layer was extracted with ethyl acetate (2 x 30 mL) and the combined organic phases were washed with 1 M HCI (60 mL), saturated NaHCO 3 (120 mL), and brine (120 mL) and dried over Na 2
SO
4 . After 10 removing the solvent in vacuo, the residue was recrystallized from MeOH to afford the title compound as a cream-colored powder (314 mg, 70%). 'H-NMR (300 MHz, DMSO-d 6 ) 6 8.82 (t, 1H, J=6 Hz), 8.43 (s, 1H), 8.02 (d, 2H, J=7 Hz), 7.69 (t, 1H, J=7 Hz), 7.57 (t, 2H, J=7 Hz), 4.72 (d, 2H, J=6 Hz), 3.84 (s, 3H) and 2.57 (s, 3H). Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) calcd. for 15 Ci 6
H
1 5
NO
4
S
2 : 372.0(M+Na). Found: 372.1. b) Methyl 5-methylthio-4-(S-phenyl(1,3-oxazol-2-yl))thiophene-2 carboxylate: To a cooled (0 oC) solution of 80.1 mg (0.229 mmol) of methyl 5 methylthio-4-[N-(2-oxo-2-phenylethyl)carbamoyl]thiophene-2-carboxylate (as prepared in the previous step) in 2 mL of anhyd DMF was added 26.7 mL (0.286 20 mmol) of phosphorus oxychloride. After stirring for 20 h at room temperature, the mixture was concentrated in vacuo. The resulting yellow solid was recrystallized twice from MeOH to afford the title compound as a beige powder (48.8 mg, 64 %). 1 H-NMR (300 MHz, DMSO-d 6 ) 6 8.26 (s, 1H), 7.88 (s, 1H), 7.86 (d, 2H, J=7 Hz), 7.51 (m, 2H), 7.40 (m, 1H), 3.86 (s, 3H), and 2.79 (s, 3H). Mass spectrum (MALDI 25 TOF, a-cyano-4-hydroxycinnamic acid matrix) calcd. for C 1 6
H,
3 NO3S 2 : 332.0 (M + H). Found: 331.9. c) 5-Methylthio-4-(5-phenyl(1,3-oxazol-2-y))thiophene-2-carboxamidine hydrochloride: Methyl 5-methylthio-4-(5-phenyl(1,3-oxazol-2-yl))thiophene-2 carboxylate (37.0 mg, 0.112 mmol, as prepared in the previous step) was treated 30 according to the procedure in Example 10, step (b) using 59.9 mg (1.12 mmol) of ammonium chloride in 0.50 mL of toluene and 0.560 mL (1.12 mmol) of 2 M trimethylaluminum in toluene. The resulting residue was chromatographed on a 5 g WO 00/47578 -142- PCT/US99/18065 silica SPE column (Waters Sep-Pak) with 10% MeOH-CH 2 C1 2 to elute an impurity followed by 20% MeOH-CH 2 Cl 2 to give 39 mg of a light yellow glass. Crystallization from MeOH-MeCN afforded the title compound as a cream-colored solid (33.4 mg, 85 %). 1 H-NMR (300 MHz, DMSO-d 6 ) 8 9.45 (broad s, 2H), 9.13 5 (broad s, 2H), 8.72 (s, 1H), 7.93 (s, 1H), 7.84 (d, 2H, J=7 Hz), 7.53 (t, 2H, J=7 Hz), 7.42 (t, 1H, J=7 Hz), and 2.80 (s, 3H). Mass spectrum (MALDI-TOF, a-cyano-4 hydroxycinnamic acid matrix) calcd. for C 1 5
H,
3
N
3 0S 2 : 316.1 (M + H). Found: 316.5. 10 Examples 80 and 81 5-Methylthio-4-(4-phenylimidazol-2-yl)thiophene-2-carboxamidine trifluoroacetate and 5-Methylthio-4-[N-(2-oxo-2-phenylethyl)carbamoyl]thiophene-2 carboxamidine trifluoroacetate 15 Methyl 5-methylthio-4-[N-(2-oxo-2-phenylethyl)carbamoyl]thiophene- 2 carboxylate (39.4 mg, 0.100 mmol, as prepared in Example 79, step (a)) was treated according to the procedure in Example 10, step (b) using 64.2 mg (1.20 mmol) of ammonium chloride in 0.2 mL of toluene and 0.600 mL (1.20 mmol) of 2 M trimethylaluminum in toluene. The resulting residue was chromatographed on a 5-g 20 silica SPE column (Waters Sep-Pak) with a gradient of 5-20% MeOH-CH 2 C1 2 to elute an impurity followed by 20% MeOH-CH 2 Cl 2 to give a yellow resin. Crystallization from MeOH-Et 2 0-MeCN afforded 16 mg of a yellow solid consisting of two products by 'H-NMR spectra. A portion of the mixture (11 mg) was purified by reverse-phase HPLC (5m C. column, 4.6 x 100 mm, gradient 5-100% solvent B over 15 min, 25 solvent A = 0.1% TFA/H 2 0, solvent B = 0.1%TFA/MeCN, detection at 215 nm) to afford 6 mg of 5-methylthio-4-(4-phenylimidazol-2-yl)thiophene-2-carboxamidine trifluoroacetate as a colorless glass. 'H-NMR (300 MHz, CD 3 OD) 8 8.23 (s, 1H), 7.80 (s, 1H), 7.79 (d, 2H, J=7 Hz), 7.48 (min, 2H), 7.39 (min, 1H), and 2.78 (s, 3H). Mass spectrum (electrospray ionization) calcd. for C 15
H
14
N
4
S
2 : 315.1 (M + H). Found: 30 315.3. Also isolated was 4 mg of 5-methylthio-4-[N-(2-oxo-2 phenylethyl)carbamoyl]-thiophene-2-carboxamidine trifluoroacetate as a colorless glass. 'H-NMR (300 MHz, DMSO-d 6 ) 8 9.30 (broad s, 2H), 8.86 (broad s, 2H), 8.68 (t, 1H, J=5.4 Hz), 8.43 (s, 1H), 8.04 (d, 2H, J=7 Hz), 7.70 (t, 1H, J=7 Hz), 7.58 (t, 2H, WO 00/47578 -143- PCT/US99/18065 J=7 Hz), 4.78 (d, 2H, J=5.4 Hz), and 2.63 (s, 3H). Mass spectrum (electrospray ionization) calcd. for C 15
H
1 sN 3 0 2
S
2 : 334.1 (M + H). Found: 334.3. Example 82 5 4-(4-Phenyl-1,3-thiazol-2-yl)thiophene-2-carboxamidine hydrochloride a) 4-Bromothiophene-2-carboxylic acid: To a cooled (0 oC) solution of 10.0 g (47.1 mmol based on 90 % purity) of 4-bromothiophene-2-carbaldehyde (Aldrich Chemical Company, Milwaukee, WI) in 200 mL of t-butanol was added 100 mL of 20 10 % (w/v) NaH 2
PO
4 followed by 60 mL (0.566 mol) of 2-methyl-2-butene. Sodium chlorite (70.8 mmol based on 80 % purity) in 60 mL of water was added with stirring. After stirring the two-phase mixture vigorously for 16 h at room temperature, the pH of the aqueous layer was adjusted to 1-2 with 20 % HC1. The layers were separated and the aqueous layer extracted with EtOAc (2 x 120 mL). The combined organic 15 layers were dried (Na 2
SO
4 ) and concentrated in vacuo to afford 9.8 g of an off-white solid. Recrystallization from a minimum of MeCN (three crops) gave the title compound as a white solid (9.02 g, 93%). 'H-NMR (300 MHz, CDCl 3 ) 6 7.79 (d, 1H, J=1.5 Hz), and 7.55 (d, 1H, J=1.5 Hz). b) Methyl 4-bromothiophene-2-carboxylate: To a cooled (-20 0 C) solution of 20 6.02 g (29.1 mmol) of 4-bromothiophene-2-carboxylic acid (as prepared in the previous step) in 100 mL of anhyd MeOH under nitrogen was added 2.55 mL (34.9 mmol) of thionyl chloride dropwise at a rate to keep the temperature below -5oC (ca. 8-10 min). After stirring for 1 h at room temperature, the mixture was refluxed for 8 h, cooled, and concentrated in vacuo. The resulting 6.7 g of pale amber oil was 25 passed through a 150 g pad of silica gel with ca. 600 mL of CH 2 C1 2 (discarding the first 120 mL which contained a minor impurity) to afford, after concentration in vacuo, the title compound as a colorless oil (6.11 g, 95 %). 'H-NMR (300 MHz, CDCl 3 ) 8 7.69 (d, 1H, J=1.5 Hz), 7.45 (d, 1H, J=l.5 Hz), and 3.90 (s, 3H). c) Methyl 4-cyanothiophene-2-carboxylate: To a solution of 3.82 g (17.3 30 mmol) methyl 4-bromothiophene-2-carboxylate (as prepared in the previous step) in 10 mL of anhyd DMF was added 3.10 g (34.6 mmol) of copper (I) cyanide. The mixture was heated to reflux with stirring for 18 h, cooled and poured into 100 mL of 10 % (w/v) KCN. The mixture was extracted with EtOAc (3 x 60 mL) and the WO 00/47578 PCT/US99/18065 -144 combined extracts were washed with 150 mL each of water and brine. The dark solution was dried over Na 2
SO
4 , treated with decolorizing carbon, filtered and the resulting colorless solution concentrated in vacuo. The resulting light yellow solid was recrystallized from MeOH to afford the title compound as a cream-colored solid 5 (1.67 g, 58 %). 'H-NMR (300 MHz, CDCI 3 ) 8 8.09 (d, 1H, J=1.4 Hz), 7.93 (d, 1H, J=1.4 Hz), and 3.93 (s, 3H). IR (film): 2235 and 1712 cm-'. d) Methyl 4-(aminothioxomethyl)thiophene-2-carboxylate: A solution of 1.32 g (7.89 mmol) of methyl 4-cyanothiophene-2-carboxylate (as prepared in the previous step) in 200 mL of reagent grade MeOH was degassed with nitrogen through 10 a fritted gas dispersion tube for 10 min. Triethylamine (5.50 mL, 39.5 mmol) was added and hydrogen sulfide gas was bubbled into the solution at a vigorous rate for 5 min and then at a minimal rate (as measured through an outlet oil bubbler) for 5 h with stirring. The gas introduction was stopped and the mixture was capped and stirred for 19 h at room temperature. The mixture was concentrated in vacuo to a 15 yellow solid which was suspended in 10 mL of EtOH, cooled to -20 oC, and filtered washing with 5 mL of cold (-20 'C) EtOH. The resulting solid was dried under suction followed by high vacuum to afford the title compound as a beige solid (1.31 g, 82 %). 'H-NMR (300 MHz, DMSO-d 6 ) 8 9.85 (broad s, 1H), 9.51 (broad s, 1H), 8.50 (d, 1H, J=1.5 Hz), 8.28 (d, 1H, J=l.5 Hz), and 3.84 (s, 3H). 20 e) Methyl 4-(4-phenyl-1,3-thiazol-2-yl)thiophene-2-carboxylate: To a solution of 150 mg (0.745 mmol) of methyl 4-(aminothioxomethyl)-thiophene-2 carboxylate (as prepared in the previous step) in 6 mL of acetone was added 148 mg (0.745 mmol) of 2-bromoacetophenone. After refluxing for 2 h, the mixture was concentrated by boiling to a volume of ca. 2 mL. The resulting mixture was cooled ( 25 10 oC) and filtered washing with cold acetone (2 x 0.5 mL). A second crop was obtained from the mother liquors and the combined crops dried to afford the title compound as a beige solid (202 mg, 90 %). 'H-NMR (300 MHz, DMSO-d 6 ) 8 8.56 (d, 1H, J=l.5 Hz), 8.25 (d, 1H, J=l.5 Hz), 8.18 (s, 1H), 8.04 (d, 2H, J=7 Hz), 7.48 (t, 2H, J=7 Hz), 7.38 (t, 1H, J=7 Hz), and 3.89 (s, 3H). Mass spectrum (MALDI-TOF, a 30 cyano-4-hydroxycinnamic acid matrix) calcd. for CI 5
H
1
INO
2
S
2 : 302.0 (M + H). Found: 301.8.
WO 00/47578 -145- PCT/US99/18065 f) 4-(4-Phenyl-1,3-thiazol-2-yl)thiophene-2-carboxamidine hydrochloride: Methyl 4-(4-phenyl-l1,3-thiazol-2-yl)thiophene-2-carboxylate (160 mg, 0.531 mmol, as prepared in the previous step) was treated according to the procedure in Example 10, step (b) using 284 mg (5.31 mmol) of ammonium chloride in 2.6 mL of toluene 5 and 2.65 mL (5.30 mmol) of 2 M trimethylaluminum in toluene. The resulting light yellow solid was chromatographed on a 10 g silica SPE column (Waters Sep-Pak) with a gradient of 5-20% MeOH-CH 2 C2. The resulting pale amber glass was triturated with CH 2 C1 2 -MeCN and concentrated in vacuo to afford the title compound as a beige solid (68 mg, 45 %). 'H-NMR (300 MHz, DMSO-d 6 ) 8 9.51 (broad s, 2H), 10 9.09 (broad s, 2H), 8.71 (d, 1H, J=l.5 Hz), 8.61 (d, 1H, J=1.5 Hz), 8.21 (s, 1H), 8.05 (d, 2H, J=7 Hz), 7.50 (t, 2H, J=7 Hz), and 7.40 (t, 1H, J=7 Hz). Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) calcd. for C 14 H 1
N
3
S
2 : 286.0 (M + H). Found: 286.3. 15 Example 83 5-Methylthio-4-[4-benzyl(1,3-thiazol-2-yl)]thiophene-2-carboxamidine hydrochloride a) Bromo-3-phenylacetone: To a solution of 132 mL (1.00 mmol) of 20 phenylacetyl chloride in 1.0 mL of anhyd MeCN was added 1.05 mL (2.10 mmol) of a 2 M solution of trimethylsilyldiazomethane in hexane. After stirring 1 h at room temperature, the mixture was cooled (0oC) and 300 mL (1.50 mmol) of 30 wt % HBr in acetic acid was added dropwise (gas evolution). After stirring 15 min, the mixture was concentrated in vacuo and rapidly chromatographed on a 2 g silica SPE column 25 (Waters Sep-Pak) with 50 % CH 2 Cl 2 -hexane to afford the title compound as a pale yellow oil (201 mg, 94 %). 1 H-NMR (300 MHz, CDCl 3 ) 8 7.2-7.4 (m, 5H), 3.95 (s, 2H), 3.92 (s, 2H). b) Methyl 5-methylthio-4-[4-benzyl(1,3-thiazol-2-yl)]thiophene-2 carboxylate: Using a procedure similar to that of Example 10 with 171 mg (0.690 30 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (as prepared in Example 82, step (e)) in 4 mL of acetone and 147 mg (0.690 mmol) of 1 bromo-3-phenylacetone (as prepared in the previous step) afforded the title compound as a light tan powder (236 mg, 95 %). 'H-NMR (300 MHz, DMSO -d 6 ) 6 8.11 (S, WO 00/47578 -146- PCT/US99/18065 1H), 7.2-7.4 (m, 5H), 4.11 (s, 2H), 3.84 (s, 3H), and 2.72 (s, 3H). Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) calcd. for C 17
H
15
NO
2
S
3 : 362.0 (M + H). Found: 362.3. c) 5-Methylthio-4-[4-benzyl(1,3-thiazol-2-yl)]thiophene-2-carboxamidine 5 hydrochloride: Methyl 5-methylthio-4-[4-benzyl(1,3-thiazol-2-yl)]thiophene-2 carboxylate (60 mg, 0.166 mmol, as prepared in the previous step) was treated according to the procedure in Example 10, step (b) using 88.8 mg (1.66 mmol) of ammonium chloride in 0.5 mL of toluene and 0.830 mL (5.30 mmol) of 2 M trimethylaluminum in toluene to afford, after trituration from MeOH with Et 2 0, the 10 title compound as a yellow solid (38.2 mg, 60 %). IH-NMR (300 MHz, CD 3 OD) 8 8.43 (s, 1H), 7.16-7.33 (m, 5H), 4.15 (s, 2H), and 2.75 (s, 3H). Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) calcd. for C 16
H
15
N
3
S
3 : 346.0 (M + H). Found: 346.0. 15 Example 84 5-Methylthio-4-(4-phenyl(1,3-oxazol-2-yl))thiophene-2-carboxamidine hydrochloride a) Methyl 4-[N-(2-hydroxy-1-phenylethyl)carbamoyl]-5 20 methylthiothiophene-2-carboxylate: To a stirred suspension of 1.23 g (5.29 mmol) of 5-(methoxycarbonyl)-2-methylthiothiophene-3-carboxylic acid (as prepared in Example 79, step (a)) in 20 mL of anhyd CH 2
C
2 (under a CaSO 4 drying tube) was added 1.85 mL (21.2 mmol) of oxalyl chloride followed by 30 mL of anhyd DMF. After stirring for 2 h at room temperature, the mixture was concentrated in vacuo. 25 The resulting yellow solid was dissolved in 20 mL of anhyd CH 2 C1 2 , cooled (0oC) and 1.85 mL of N,N-diisopropylethylamine (10.6 mmol) and 1.02 g (7.41 mmol) of phenylglycinol was added and the mixture stirred for 1 h at room temperature. The mixture was concentrated to an oil and partitioned between 200 mL of EtOAc and 200 mL of saturated NaHCO 3 . The organic phase was washed with saturated NaHCO 3 30 (200 mL), 10 % (w/v) citric acid, and brine (200 mL), and dried over Na 2
SO
4 . After removing the solvent in vacuo, the residue was chromatographed on a 10 g silica SPE column (Waters Sep-Pak) with a gradient of 0-20 % EtOAc-CH 2
C
2 to afford the title compound as a light yellow solid (1.26 g, 68 %). 'H-NMR (300 MHz, CDCl 3 ) 8 8.00 WO 00/47578 -147- PCT/US99/18065 (s, 1H), 7.30-7.42 (min, 5H), 7.08 (d, 1H, J=7.2 Hz), 5.26 (min, 1H), 3.99 (t, 2H, J=5.4 Hz), 3.89 (s, 3H), 2.60 (s, 3H), and 2.33 (t, 1H, J=6.1 Hz). Mass spectrum (electrospray ionization) calcd. for C 16
H
1 7
NO
4
S
2 : 352.1 (M + H). Found: 352.0. b) Methyl 5-methylthio-4-[N-(2-oxo-l-phenylethyl)carbamoyl]thiophene-2 5 carboxylate: To a solution of 505 mg (1.44 mmol) methyl 4-[N-(2-hydroxy-1 phenylethyl)carbamoyl]-5-methylthiothiophene-2-carboxylate (as prepared in the previous step) in 20 mL of anhydrous CH 2 Cl 2 was added 856 mg (2.02 mmol) of Dess Martin reagent (Omega Chemical Company, Inc., Levis (Qc) Canada). After stirring in an open flask for 1.5 h at room temperature, the mixture was concentrated in vacuo. 10 to ca. 10 % volume and partitioned between 50 mL of EtOAc and 50 mL of saturated NaHCO 3 -brine (1:1). The organic phase were washed with brine (200 mL), dried over Na 2
SO
4 and concentrated in vacuo. Concentrated again from CH 2 C1 2 followed by high vacuum afforded the title compound as a light yellow foam (495 mg, 98 %) which was used in the next step without further purification. IH-NMR (300 MHz, 15 CDCl 3 ) 8 9.64 (s, 1H), 8.04 (s, 1H), 7.59 (d, 1H, J=5 Hz), 7.36-7.46 (mn, 5H), 5.76 (d, 1H, J=5 Hz), 3.90 (s, 3H), and 2.62 (s, 3H). c) Methyl 5-methylthio-4-(4-phenyl(1,3-oxazol-2-yl))thiophene-2 carboxylate: To a cooled (0OC) solution of 465 mg (1.33 mmol) methyl 5 methylthio-4-[N-(2-oxo-1 -phenylethyl)carbamoyl]thiophene-2-carboxylate (as 20 prepared in the previous step) in 6 mL of anhyd DMF was added 186 mL (2.00 mmol) of phosphorus oxychloride. After stirring for 14 h at room temperature, the mixture was treated with 10 mL of saturated NaHCO 3 and concentrated to dryness under high vacuum. The resulting residue was partitioned between 80 mL of EtOAc and 60 mL of water. The aqueous layer was extracted with EtOAc (2 x 10 mL) and the combined 25 organic phases washed with brine (60 mL), and dried over Na 2
SO
4 . The resulting 406 mg of amber-colored solid was recrystallized from CH 2 C1 2 -Et 2 O to remove the majority of a polar impurity as a cream-colored solid. The remaining mother liquors were chromatographed on a 10 g silica SPE column (Waters Sep-Pak) with a gradient of 40-100 % CH 2 C1 2 -hexane and the resulting residue triturated with Et 2 0-hexane 30 (2:1)) to afford the title compound as a light beige solid (114 mg, 26 %). 'H-NMR (300 MHz, CDCl 3 ) 8 8.24 (s, 1H), 7.93 (s, 1H), 7.83 (min, 2H), 7.43 (mn, 2H), 7.33 (inm, WO 00/47578 -148- PCT/US99/18065 1H), 3.91 (s, 3H), and 2.72 (s, 3H). Mass spectrum (ESI) calcd. for C, 6
H
1 3
NO
3
S
2 : 332.0 (M + H). Found: 332.2. d) 5-Methylthio-4-(4-phenyl(1,3-oxazol-2-yl))thiophene-2-carboxamidine hydrochloride: Methyl 5-methylthio-4-(4-phenyl(1,3-oxazol-2-yl))thiophene-2 5 carboxylate (80.3 mg, 0.242 mmol, as prepared in the previous step) was treated according to the procedure in Example 10, step (b) using 155 mg (2.90 mmol) of ammonium chloride in 1.45 mL of toluene and 1.45 mL (2.90 mmol) of 2 M trimethylaluminum in toluene. The resulting light yellow solid was chromatographed on a 5 g silica SPE column (Waters Sep-Pak) with 10% MeOH-CH 2 Cl 2 to give a light 10 yellow resin. Crystallization from MeOH- Et 2 0 (ca. 1:3) afforded the title compound as a yellow solid (62.2 mg, 82 %). H-NMR (300 MHz, DMSO-d 6 ) 8 9.39 (broad s, 2H), 8.97 (broad s, 2H), 8.78 (s, 1H), 8.60 (s, 1H), 7.89 (d, 2H, J=7 Hz), 7.49 (t, 2H, J=7 Hz), 7.38 (t, 1H, J=7 Hz), and 2.80 (s, 3H). Mass spectrum (ESI) calcd. for
C
15
H
13
N
3 0S 2 : 316.1 (M + H). Found: 316.2. 15 Example 85 4-[4-(4-hydroxy-3-methoxyphenyl) (1,3-thiazol-2-yl)]-5-methlylthiothiophene-2 carboxamidine hydrochloride 20 a) 4-(Chlorocarbonyl)-2-methoxyphenyl acetate: To a stirred suspension of 1.00 g (4.76 mmol) of 4-acetoxy-3-methoxybenzoic acid (Pfaltz and Bauer, Inc.) in 4 mL of anhyd CH 2 C1 2 (under a CaSO 4 drying tube) was added 4.15 mL (47.6 mmol) of oxalyl chloride followed by 25 mL of anhyd DMF. After stirring for 4 h at room temperature, the mixture was concentrated in vacuo to afford the title compound as 25 light yellow crystals (1.12 g, 103%). 'H-NMR (300 MHz, CDC13) 8 7.81 (dd, 1H, J=8.4, 2.1 Hz), 7.66 (d, 1H, 2.1 Hz), 7.19 (d, 1H, 8.4 Hz), 3.91 (s, 3H), and 2.35 (s, 3H). b) 4-(2-Bromoacetyl)-2-methoxyphenyl acetate: To a solution of 1.09 g (4.6 mmol) of 4-(chlorocarbonyl)-2-methoxyphenyl acetate (as prepared in the precious 30 step) in 10 mL of anhyd CH 2 C1 2 was added 10.0 mL (20.0 mmol) of a 2 M solution of trimethylsilyldiazomethane in hexane. After stirring 2 h at room temperature, the mixture was cooled (0 oC) and 3.20 mL (16.0 mmol) of 30 wt % HBr in acetic acid WO 00/47578 -149- PCT/US99/18065 was added dropwise (gas evolution). After stirring 5 min, the mixture was concentrated in vacuo and rapidly chromatographed on a 10-g silica SPE column (Waters Sep-Pak) with CH 2 C1 2 to afford the title compound as a light yellow crystalline solid (1.28 g, 97 %). 'H-NMR (300 MHz, CDCl 3 ) 6 7.63 (d, 1H, 1.9 Hz), 5 7.59 (dd, 1H, J=8.2, 1.9 Hz), 7.16 (d, 1H, 8.2 Hz), 4.43 (s, 2H), 3.91 (s, 3H), and 2.35 (s, 3H). c) 2-Methoxy-4-{2-[S-(methoxycarbonyl)-2-methylthio(3-thienyl)](1,3 thiazol-4-yl)}phenyl acetate: Using a procedure similar to that of Example 82, step (e) with 1.00 g (4.04 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene 10 2-carboxylate (Maybridge Chemical Company, Cornwall, UK) in 15 mL of reagent acetone and 1.16 g (4.04 mmol) of 4-(2-bromoacetyl)-2-methoxyphenyl acetate (as prepared in the previous step) afforded the title compound as 1.42 g of a yellow solid which, according to the 'H-NMR spectrum, consisted of a ca. 1:1 mixture of the title compound and the corresponding compound resulting from partial loss of the acetate. 15 'H-NMR (300 MHz, DMSO -d 6 ) 8 8.27 (s, 1H), 8.22 (s, 1H), 8.19 (s, 1H), 8.00 (s, 1H), 7.78 (d, 1H, 1.9 Hz), 7.67 (dd, 1H, J=8.2, 1.9 Hz), 7.61 (d, 1H, 1.9 Hz), 7.51 (dd, 1H, J=8.2, 1.9 Hz), 7.19 (d, 1H, 8.2 Hz), 6.86 (d, 1H, 8.2 Hz), 8.87 (m, 12H), 2.76 (s, 3H), 2.75 (s, 3H), and 2.28 (s, 3H). Mass spectrum (ESI) calcd. for C 19
H
17 NOsS 3 and
C
17
H
1 5
NO
3
S
3 436.0 (M + H) and 394.1 (M + H). Found: 436.1 and 394.2. The 20 mixture was used without further purification in the following step where formation of the amidine involves concomitant removal of the acetate. d) 4-[4-(4-hydroxy-3-methoxyphenyl) (1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxamidine hydrochloride: A portion of the mixture (500 mg, ca. 1.21 mmol as based on the IH-NMR integration) containing the 2-methoxy-4 25 {2-[5-(methoxycarbonyl)-2-methylthio(3-thienyl)](1,3-thiazol-4-yl)}phenyl acetate (as prepared in the previous step) was treated according to the procedure in Example 10, step (b) using 610 mg (11.4 mmol) of ammonium chloride in 5.7 mL of toluene and 5.70 mL (11.4 mmol) of 2 M trimethylaluminum in toluene. After chromatography of the resulting residue on a 10 g silica SPE column (Waters Sep 30 Pak) with a gradient of 5-20 % MeOH-CH 2 C1 2 to obtain a yellow glass which was recrystallized from MeOH-CH 2 C1 2 to afford the title compound as a pale yellow solid WO 00/47578 -150- PCT/US99/18065 (192 mg, 42 %). IH-NMR (300 MHz, DMSO-d 6 ) 6 9.35 (broad s, 2H), 9.27 (s, 1H), 8.97 (broad s, 2H), 8.62 (s, 1H), 8.04 (s, 1H), 7.62 (s, 1H), 7.54 (d, 1H J=8.2 Hz), 6.88 (d, 1H, J=8.2 Hz), 3.87 (s, 3H), and 2.79 (s, 3H). Mass spectrum (ESI) calcd. for
C
1 6
H
15
N
3 0 2
S
3 : 378.0(M + H). Found: 378.1. 5 Example 86 4-[4-(3-Hydroxy-4-methoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride 10 a) 3-Acetyloxy-4-methoxybenzoic acid: To a suspension of 600 mg (3.57 mmol) of 3-hydroxy-4-methoxybenzoic acid (Aldrich Chemical Company, Milwaukee, WI) in 5 mL of anhyd CH 2 C1 2 was added 1.31 mL (7.50 mmol) of N, N diisopropylethylamine and the mixture stirred until homogeneous (ca. 5 min). Acetyl chloride (305 mL, 4.28 mmol) was added dropwise over 2 min followed by 2.0 mg 15 ((0.016 mmol) of 4-dimethylaminopyridine. After stirring at room temperature for 1 h, the mixture was poured into 50 mL of EtOAc and washed with 1 M HCI (3 x 25 mL). The organic phase was extracted with saturated NaHCO 3 (6 x 15 mL) and the combined extracts saturated with solid NaCI and acidified to pH 2 with conc HCL. The resulting suspension was extracted with EtOAc (3 x 20mL) and the combined 20 extracts were dried over Na 2
SO
4 and concentrated in vacuo to afford the title compound as a light beige powder (463 mg, 62 %). IH-NMR (300 MHz, CDCl 3 ) 8 8.00 (dd, 1H, J=8.7, 2.0 Hz), 7.79 (d, 1H, 2.0 Hz), 7.00 (d, 1H, 8.7 Hz), 3.91 (s, 3H), and 2.34 (s, 3H). b) 3-(Chlorocarbonyl)-6-methoxyphenyl acetate: Using the procedure in 25 Example 85, step (a), 400 mg (1.90 mnimol) of 3-acetyloxy-4-methoxybenzoic acid (as prepared in the previous step) was treated with 663 mL (7.60 mmol) of oxalyl chloride and 25 mL of anhyd DMF for 2 h to afford, after workup, the title compound as a beige crystalline solid which was used in the following step without further purification. 30 c) 5-(2-Bromoacetyl)-2-methoxyphenyl acetate: Using the procedure in Example 85, step (b), the entire sample of 3-(chlorocarbonyl)-6-methoxyphenyl acetate (as prepared in the previous step) in 5 mL of anhyd CH 2
C
2 - was treated with 2.09 mL (4.18 mmol) of a 2 M solution of trimethylsilyldiazomethane in hexane and WO 00/47578 -151- PCT/US99/18065 456 mL (2.28 mmol) of 30 wt % HBr in acetic acid. Chromatography as in Example 85, step (b) followed by recrystallization from CH 2 C1 2 -hexane afforded the title compound as a faintly yellow solid (366 mg, 67 %). IH-NMR (300 MHz, CDCl 3 ) 6 7.79 (dd, 1H, J=8.6, 2.2 Hz), 7.70 (d, 1H, 2.2 Hz), 7.03 (d, 1H, 8.6 Hz), 4.38 (s, 5 2H), 3.92 (s, 3H), and 2.34 (s, 3H). d) 2-Methoxy-5-{2-[5JS-(methoxycarbonyl)-2-methylthio(3-thienyl)](1,3 thiazol-4-yl)}phenyl acetate: Using a procedure similar to that of Example 82, step (e) with 282 mg (1.14 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge Chemical Company, Cornwall, UK) 10 in 4 mL of acetone and 3.27 mg (1.14 mmol) of 5-(2-bromoacetyl)-2-methoxyphenyl acetate (as prepared in the previous step) afforded a yellow solid (374 mg) which, according to the 'H-NMR spectrum, consisted of a 3:7 mixture of the title compound and the corresponding compound resulting from partial loss of the acetate. Mass spectrum (ESI) calcd. for C q 9
H,
7 NOsS 3 and C 17
H,
5
NO
3
S
3 436.0 (M + H) and 394.1 (M 15 + H). Found: 436.0 and 394.0. The mixture was used without further purification in the following step where formation of the amidine involves concomitant removal of the acetate. e) 4-[4-(3-Hydroxy-4-methloxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothliophene-2-carboxamidine hydrochloride: A portion of the mixture (320 20 mg, ca. 0.788 mmol as based on the 'H-NMR spectrum) containing the 2-methoxy-5 {2-[5-(methoxycarbonyl)-2-methylthio(3-thienyl)](1,3-thiazol-4-yl) }phenyl acetate (as prepared in the previous step) was treated according to the procedure in Example 10, step (b), using 415 mg (7.76 mmol) of ammonium chloride in 3.5 mL of toluene and 3.88 mL (7.66 mmol) of 2 M trimethylaluminum in toluene. After 25 chromatography of the resulting residue on a 10 g silica SPE column (Waters Sep Pak) with 10-40 % MeOH-CH 2 C1 2 , a light yellow solid was obtained which was dissolved in 45 mL of DMF and filtered to remove silica gel. Concentration under high vacuum and recrystallization from MeOH-Et 2 0 afforded the title compound as a light tan solid (132 mg, 44 %). 'H-NMR (300 MHz, DMSO-d 6 ) 6 9.49 (broad s, 2H), 30 9.16 (broad s, 2H), 8.67 (s, 1H), 7.98 (s, 1H), 7.5 (obscured m, 3H), 7.00 (obscured d, 1H, J=8.3 Hz), 3.82 (s, 3H), and 2.79 (s, 3H). Mass spectrum (ESI) calcd. for
C,
6 H,sN 3 0 2
S
3 : 378.0(M + H). Found: 378.1.
WO 00/47578 PCT/US99/18065 -152 Example 87 5-Methylthio-4-(N-phenylcarbamoyl)thiophene-2-carboxamidine hydrochloride 5 a) Methyl 5-methylthio-4-(N-phenylcarbamoyl)thiophene-2-carboxylate: To 182 mg (0.785 mmol) of 5-(methoxycarbonyl)-2-methylthiothiophene-3 carboxylic acid (as prepared in Example 95) in 4 mL of anhyd CH 2 C1 2 was treated with 275 mL (3.15 mmol) of oxalyl chloride and 6 mL of anhyd DMF for 2 h similar to Example 79, step (a); followed by 206 mL (1.18 mmol) of NN 10 diisopropylethylamine and 85.9 mL (0.942 mmol) of aniline in 3 mL of anhyd CH 2 C1 2 for 20 min. The mixture was poured into 25 mL of EtOAc and washed with 1 M HCI (2 x 25 mL), saturated NaHCO 3 (2 x 25 mL), and brine (25 mL), and dried over Na 2
SO
4 . Removal of the solvent in vacuo, afforded the pure title compound as a light yellow solid (163 mg, 68 %). 'H-NMR (300 MHz, CDCl 3 ) 8 8.23 (broad s, 1H), 8.10 15 (s, lH), 7.63 (d, 2H, J=7 Hz), 7.36 (t, 2H, J=7 Hz), 7.15 (t, 2H, J=7 Hz), 3.90 (s, 3H), and 2.64 (s, 3H). b) 5-Methylthio-4-(N-phenylcarbamoyl)thiophene-2-carboxamidine hydrochloride: Methyl 5-methylthio-4-(N-phenylcarbamoyl)thiophene-2-carboxylate (60.0 mg, 0.195 mmol , as prepared in the previous step) was treated similarly to the 20 procedure in Example 10, step (b) using 310 mg (5.80 mmol) of ammonium chloride in 2 mL of toluene and 2.90 mL (5.80 mmol) of 2 M trimethylaluminum in toluene for 6 h. Chromatography of the resulting residue on a 2 g silica SPE column (Waters Sep-Pak) with a gradient of 5-20 % MeOH-CH 2
C
2 , followed by crystallization from MeOH-Et 2 0 afforded the title compound as a beige solid (40.3 mg, 71 %). IH-NMR 25 (300 MHz, DMSO-d 6 ) 6 10.24 (s, 1H), 9.34 (broad s, 2H), 9.05 (broad s, 2H), 8.75 (s, 1H), 7.73 (d, 2H, J=8 Hz), 7.36 (t, 2H, J=8 Hz), 7.11 (m, 1H), and 2.67 (s, 3H). Mass spectrum (ESI) calcd. for C 13
H
13 N3OS 2 : 292.1 (M + H). Found: 292.4.
WO 00/47578 PCT/US99/18065 -153 Example 88 and 89 5-Methylthio- 4 -JN-benzylcarbamoyl]thiophene-2-carboxamidinehydrochloride and 4-{Imino[benzylamino]methyl}-5-methylthiothiophene-2-carboxamidine hydrochloride 5 a) Methyl 5-methylthio-4-JN-benzylcarbamoyl]thiophene-2-carboxylate: The identical procedure of Example 87, step (a) was used with 103 mL (0.942 mmol) of benzylamine and the same amounts of all other reagents to afford the title compound as a light yellow solid (167 mg, 66 %). 1H-NMR (300 MHz, CDCI 3 ) 10 5 7.93 (s, 1H), 7.28-7.38 (m, 5H), 6.58 (broad s, 1H), 4.62 (s, 2H, J=5.7 Hz), 3.87 (s, 3H), and 2.60 (s, 3H). b) 5-Methylthio-4-[N-benzylcarbamoyl]thiophene-2 carboxamidinehydrochloride and 4-{Imino[benzylamino]methyl}-5 methylthiothiophene-2-carboxamidinehydrochloride: Methyl 5-methylthio-4-[N 15 benzylcarbamoyl]thiophene-2-carboxylate (62.7 mg, 0.195 mmol, as prepared in the previous step) was treated similarly to the procedure in Example 10, step (b) using 310 mg (5.80 mmol) of ammonium chloride in 2 mL of toluene and 2.90 mL (5.80 mmol) of 2 M trimethylaluminum in toluene for 6 h. Chromatography of the resulting residue on a 2 g silica SPE column (Waters 20 Sep-Pak) with a gradient of 5-20 % MeOH-CH 2 C1 2 , followed by crystallization from MeOH-Et 2 0 afforded 5-methylthio-4-[N-benzylcarbamoyl]thiophene-2 carboxamidinehydrochloride as a beige solid (21.1 mg, 35 %). IH-NMR (300 MHz, DMSO-d 6 ) 8 7.93 (s, 1H), 7.28-7.38 (m, 5H), 6.58 (broad s, 1H), 4.62 (s, 2H, J=5.7 Hz), 3.87 (s, 3H), and 2.60 (s, 3H). Mass spectrum (ESI) calcd. for Cl 4 HsN 3 0S 2 : 25 306.1 (M + H). Found: 306.6. Also isolated and crystallized from MeOH-Et 2 0 was the more polar 4 { imino[benzylamino]methyl } -5-methylthiothiophene-2-carboxamidinehydrochloride as a beige solid (32.0 mg, 54 %). 'H-NMR (300 MHz, DMSO-d 6 ) consistent with desired product as broad mixture of rotomers. Mass spectrum (ESI) calcd. for 30 C 14
H
16
N
4
S
2 : 305.1 (M + H). Found: 305.8.
WO 00/47578 -154- PCT/US99/18065 Example 90 and 91 4-JN-Methyl-N-benzylcarbamoyl]-5-methylthiothiophene-2-carboxamidine hydrochloride and 4-{Imino[methylbenzylamino]methyl}-5-methylthiothiophene-2 carboxamidinehydrochloride 5 a) Methyl 4-[N-methyl-N-benzylcarbamoylf-5-methylthiothiophene-2 carboxylate: The identical procedure of Example 87, step (a) was used with 122 mL (0.942 mmol) of N-benzylmethylamine and the same amounts of all other reagents to afford the title compound as a light yellow solid (169 mg, 64 %). IH-NMR (300 10 MHz, CDCl 3 ) 8 7.68 (s, 1H), 7.34 (m, 5H), 4.6 (broad m, 2H), 3.86 (s, 3H), 2.91 (inm, 3H), and 2.60 (s, 3H). b) 4-[N-Methyl-N-benzylcarbamoyl]-5-methylthiothiophene-2 carboxamidine hydrochloride and 4-{lmino[methylbenzylamino]methyl}-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-[N-methyl-N 15 benzylcarbamoyl]-5-methylthiothiophene-2-carboxylate (65.4 mg, 0.195 mmol, as prepared in the previous step) was treated similarly to the procedure in Example 10, step (a) using 310 ming (5.80 immol) of ammonium chloride in 2 mL of toluene and 2.90 mL (5.80 mmol) of 2 M trimethylaluminum in toluene for 6 h. Chromatography of the resulting residue on a 2 g silica SPE column (Waters 20 Sep-Pak) with a gradient of 5-20 % MeOH-CH 2
C
2 afforded 4-[N-methyl-N benzylcarbamoyl]-5-methylthiothiophene-2-carboxamidine hydrochloride as a amber colored glass (34.3 mg, 55 %). 'H-NMR (300 MHz, DMSO-d 6 ) 8 9.32 (broad s, 2H), 9.06 (broad s, 2H), 8.11 (s, 1H), 7.36 (mn, 5H), 4.66 (min, 2H), 2.88 (s, 3H) and 2.66 (s, 3H). Mass spectrum (ESI) calcd. for C 15
H,
7
N
3 0S 2 : 320.1 (M + H). Found: 320.4. 25 Also isolated and then crystallized from MeOH-Et 2 0 was the more polar 4- { imino [methylbenzylamino]methyl } -5-methylthiothiophene-2-carboxamidine hydrochloride as a beige solid (19.8 mg, 32 %). 1H-NMR (300 MHz, DMSO-d 6 ) consistent with desired product as broad mixture of rotomers. Mass spectrum (ESI) calcd. for C 15
H,
8
N
4
S
2 : 319.1 (M + H). Found: 319.6. 30 WO 00/47578 PCT/US99/18065 -155 Example 92 and 93 5-Methylthio-4-[N-(2-phenylethyl)carbamoyl]thiophene-2-carboxamidine hydrochloride and 4 -{Imino[(2-phenylethyl)amino]methyl}-5-methylthiothliiophene 2-carboxamidine hydrochloride 5 a) Methyl S-methylthio-4-JN-(2-phenylethyl)carbamoyl]thiophene-2 carboxylate: The identical procedure of Example 87, step (a) was used with 118 mL (0.942 mmol) of phenethylamine and the same amounts of all other reagents to afford the title compound as a light yellow solid (165 mg, 63 %). 'H-NMR (300 MHz, 10 CDC13) 8 7.86 (s, 1H), 7.30-7.35 (m, 5H), 6.44 (m, 1H), 3.87 (s, 3H), 3.70 (q, 2H, J=7 Hz), 2.93 (t, 2H, J=7 Hz), and 2.53 (s, 3H). b) 5-Methylthio-4-JN-(2-phenylethyl)carbamoyl]thiophene-2-carboxamidine hydrochloride and 4 -{ilmino[( 2 -phenylethyl)aminofmethyl}-5-methylthiothiophene 2-carboxamidine hydrochloride: Methyl 5-methylthio-4-[N-(2 15 phenylethyl)carbamoyl]thiophene-2-carboxylate (65.4 mg, 0.195 mmol, as prepared in the previous step) was treated similarly to the procedure in Example 10, step (a) using 310 mg (5.80 mmol) of ammonium chloride in 2 mL of toluene and 2.90 mL (5.80 mmol) of 2 M trimethylaluminum in toluene for 6 h. Chromatography of the resulting residue on a 2 g silica SPE column (Waters 20 Sep-Pak) with a gradient of 5-20 % MeOH-CH 2 C1 2 , followed by crystallization from MeOH-Et 2 0 afforded 5-methylthio-4-[N-(2-phenylethyl)carbamoyl]thiophene-2 carboxamidine hydrochloride as a beige solid (17.4 mg, 28 %). IH-NMR (300 MHz, DMSO-d 6 ) 6 8.8-9.3 (broad m, 4H), 8.48 (m, 1H), 8.35 (s, 1H), 7.26 (m, 5H), 3.44 (m, 2H), 2.82 (t, 3H, J=7.5 Hz), and 2.61 (s, 3H). Mass spectrum (ESI) calcd. for 25 C 15 Hi 7
N
3 0S 2 : 320.1 (M + H). Found: 320.4. Also isolated and crystallized from MeOH-Et20 was the more polar 4-{imino[(2-phenylethyl)amino]methyl}-5-methylthiothiophene-2-carboxamidine hydrochloride as a beige solid (19.1 mg, 31 %). IH-NMR (300 MHz, DMSO-d 6 ) 8 8.37 (s, 1H), 7.2-7.4 (m, 5H), 3.70 (t, 2H, J=7.6 Hz), 2.96 (t, 2H, J=7.6 Hz), and 30 2.71 (s, 3H). Mass spectrum (ESI) calcd. for C
I
sH 18
N
4
S
2 : 319.1 (M + H). Found: 319.5.
WO 00/47578 PCT/US99/18065 -156 Example 94 3-Amino-2-aza-3-[5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl))(2 thienyl)]prop-2-enenitrile 5 To 100 mg (0.302 mmol) of 5-methylthio-4-(4-phenyl(1,3-thiazol-2 yl))thiophene-2-carboxamidine (as prepared in Example 10, step b) in 3 mL of EtOH was added 29.6 mg (0.604 mmol) of cyanamide as a solution in 0.3 mL of water. The mixture was heated to reflux and 0.302 mL (0.302 mmol) of 1 M aqueous KOH was added. After 3 h, the mixture was cooled (0 0 C) and filtered washing with ice-cold 10 EtOH. The resulting solid was dried in vacuo to afford the title compound as a light yellow powder (78.4 mg, 73 %). 'H-NMR (300 MHz. DMSO-d 6 ) 8 9.31 (broad s, 1H), 8.70 (broad s, 1H), 8.63 (s, 1H), 8.19 (s, 1H), 8.09 (d, 2H, J=7 Hz), 7.49 (t, 2H, J=7 Hz), 7.39 (t, 1H, J=7 Hz), and 2.75 (s, 3H). Mass spectrum (MALDI-TOF, a cyano-4-hydroxycinnamic acid matrix) calcd. for C 16
H
12
N
4
S
3 : 357.0 (M + H). Found: 15 357.1. Example 95 5-(Methoxycarbonyl)-2-methylthiothiophene-3-carboxylic acid Methyl 4-cyano-5-methylthiothiophene-2-carboxylate (2.20 g, 10.3 mmol, 20 Maybridge Chemical Company, Cornwall, UK) and tetrafluorophthalic acid (2.45 g, 10.3 mmol) in an 8-mL sealable pressure tube (Ace Glass Company) with stir bar was heated to 160 oC. The molten mixture was stirred for 4 days, cooled and the resulting residue broken up and extracted by refluxing with 80 mL chloroform. The mixture was cooled, decolorizing carbon (ca. 0.5 g) was added and the mixture filtered 25 (Celite). The resulting solution was extracted with saturated NaHCO 3 (4 x 30 mL) and the combined aqueous extracts acidified to pH 1-2 with conc HCl and filtered to provide a light tan solid. After dissolving the solid in a minimum of 1 M K 2
CO
3 (35 40 mL) and filtering (washing with 10-20mL of water) to clarify the solution, it was slowly acidified to pH 6.5-7.0 with stirring and filtered (Celite) to remove a brown 30 precipitate. The pH adjustment and filtration was repeated and the resulting solution was saturated with solid NaCI and acidified to pH 1-2 with conc HC1. The precipitate was filtered, washed with water (3 x 10 mL) and dried over P 2 Os under high vacuum to afford the title compound as a cream-colored powder (1.24 g, 52 %). 'H-NMR WO 00/47578 -157- PCT/US99/18065 (300 MHz, DMSO-d 6 ) 8 13.14 (broad s, 1H), 7.89 (s, 1H), 3.82 (s, 3H) and 2.64 (s, 3H). Mass spectrum (ESI, negative mode) calcd. for C 8
HO
8 0 4
S
2 : 232.0 (M-). Found: 231.7. 5 Example 96 5-Ethylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine hydrochloride 5 a) Methyl 4-(4-phenyl(1,3-thiazol-2-yl))-5-(methylsulfonyl)thiophene-2 10 carboxylate: Using the procedure of Example 141, step (a) with 600 mg (1.73 mmol) of methyl 5-methylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate as prepared in Example 10, step (a) afforded 642 mg (98 %) of the title compound as a light yellow powder. IH-NMR (300 MHz, CDC13) 8 7.93 (s, 1H), 7.90 (m, 2H), 7.63 (s, 1H), 7.47 (m, 2H), 7.39 (m, 1H), 3.98 (s, 3H) and 3.73 (s, 3H). Mass spectrum 15 (ESI, m/z): calcd. for C 16
H
1 3
NO
4
S
3 380.0 (M+H), found 380.2. b) 4-(4-Phenyl)(1,3-thiazol-2-yl))-5-(methlylsulfonyl)thiophene-2 carboxamidine hydrochloride: Using the procedure of Example 141, step (b) with 560 mg (1.48 mmol) methyl 4-[4-(4-chlorophenyl)(1,3-thiazol-2-yl)]-5 (methylsulfonyl)thiophene-2-carboxylate as prepared in the previous step afforded 20 392 mg (66 %) of the title compound as a off-white solid. 'H-NMR (300 MHz, DMSO-d 6 ) 8 9.7 (broad s, 2H), 9.4 (broad s, 2H), 8.58 (s, 1H), 8.43 (s, 1H), 8.02 (d, 2H, J=7 Hz), 7.52 (t, 2H, J=7 Hz), 7.43 (t, 1H, J=7 Hz), and 3.90 (s, 3H). Mass spectrum (ESI, m/z): calcd. for C, 5
H,
3
N
3 0 2
S
3 364.0 (M+H), found 364.1. c) 5-Ethylthio-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine 25 hydrochloride: Using the procedure of Example 141, step (c) with 23.1 mg (0.0578 mmol) of the 4-(4-phenyl)(1,3-thiazol-2-yl))-5-(methylsulfonyl)thiophene-2 carboxamidine hydrochloride (as prepared in the previous step), 64.1 mL( 0.867 mmol) of ethanethiol (in 2 portions over 2 h) and 40.3 mL (0.231 mmol) of DIEA in 3 mL of methanol gave a yellow resin which was chromatographed on a 2 g silica SPE 30 column (Waters Sep-Pak) with a gradient of 0-15 % MeOH-CH 2 C1 2 , followed by trituration with CH 2 C1 2 to afford the title compound as an off-white solid (21.7 mg, 98 %). 'H-NMR (300 MHz, DMSO-d 6 ) 8 9.45 (broad s, 2H), 9.07 (broad s, 2H), 8.68 (s, 1H), 8.28 (s, 1H), 8.09 (d, 2H, J=7 Hz), 7.51 (t, 2H, J=7 Hz), 7.40 (t, 1H, J=7 Hz), WO 00/47578 -158- PCT/US99/18065 3.23 (q, 2H, J=7 Hz) and 1.42 (t, 3H, J=7 Hz). Mass spectrum (ESI) calcd. for
C,
6
H,
1
N
3
S
3 : 346.1 (M+ H). Found: 346.2. Example 97 5 5-Methylthio-4-[4-(phenoxymethyl)(1,3-thiazol-2-yl)]thiophene-2-carboxamidine hydrochloride a) 3-Bromo-1-phenoxyacetone: To a solution of 6.c (0.050 mmol) of phenoxyacetyl chloride in 250 mL of anhyd MeCN in a 1-dram short vial (Wheaton 10 Glass) was added 50 mL (0.100 mmol) of a 2 M solution of trimethylsilyldiazomethane in hexane and the vial capped with a PTFE-lined cap. After stirring 1 h at room temperature on a vortex shaker, the mixture was cooled (0 °C) and 21 mL (0.105 mmol) of 30 wt % HBr in acetic acid was added dropwise (gas evolution). After vortexing for 10 min, the mixture was concentrated in vacuo on a 15 vacuum centrifuge concentrator (Speed-Vac, Savant Instruments, Inc.) to provide an amber-colored oil which was used directly in the following step. b) Methyl 5-methylthio-4-[4-(phenoxymethyl)(1,3-thiazol-2-yl)]thiophene-2 carboxylate: To the 3-bromo-1-phenoxyacetone (as prepared in the previous step in a 1-dram vial) was added 14.8 mg (0.060 mmol) of methyl 4-(aminothioxomethyl)-5 20 methylthiothiophene-2-carboxylate (Maybridge Chemical Company, Cornwall, UK) as 1.48 mL of a 10 mg /mL solution in acetone. The vial was tightly capped and placed on a heated platform shaker (Innova model 4080, New Brunswick Scientific Co., Inc.) and vortexed at 55 *C and 250 rpm for 4 h. To the resulting mixture was added 50 mg (0.150 mmol) of diethylaminomethyl-polystyrene resin (Fluka Chemika 25 Biochemika, 3.0 mmol / g) as 0.50 mL of a 100 mg / mL suspension in acetone and the mixture vortexed briefly. Chloroacetylpolystyrene resin (30 mg, 0.150 mmol, Advanced ChemTech Inc., 5.0 mmol / g) was then added followed by (0.750 mg, 0.005 mmol) Nal as 100 mL of a 7.5 mg / mL solution in acetone. The mixture was again capped tightly and placed on a heated platform shaker and vortexed at 55 0 C and 30 250 rpm for 22 h. The mixture was filtered through a 2 mL fritted column (BioRad Biospin minicolumn) washing with acetone (2 x 0.5 mL) and MeOH (2 x 0.5 mL) into a 2 dram vial and concentrated on a vacuum centrifuge concentrator to afford 21.0 mg of the title compound as an off-white solid. 1H-NMR (300 MHz, DMSO-d 6 ) 6 8.17 WO 00/47578 -159- PCT/US99/18065 (s, 1H), 7.82 (s, 1H), 7.13 (min, 2H), 7.07 (m, 2H), 6.96 (min, 1H), 5.22 (s, 2H), 3.85 (s, 3H), and 2.74 (s, 3H). Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) calcd. for C 17
H
15
NO
3
S
3 :378.0 (M + H). Found: 378.3. c) 5-Methylthio-4-[4-(phenoxymethyl)(1,3-thiazol-2-yl)]thiophene-2 5 carboxamidine hydrochloride: The methyl 5-methylthio-4-[4-(phenoxymethyl)(1,3 thiazol-2-yl)]thiophene-2-carboxylate (as prepared in the previous step)under nitrogen in a 2 dram vial with a micro magnetic stir bar) was capped with an open-top phenolic cap containing a PTFE-backed silicone septum. A 1 M solution of the reagent freshly prepared from trimethylaluminum and ammonium chloride in toluene according to the 10 procedure in Example 10, step b (0.750 mL, 0.750 mmol) was added by syringe by puncturing the septum once with the needle to allow venting of gas followed by a second puncture to inject the reagent. The vial was placed in an aluminum heating block under nitrogen (Fisher Scientific Dry Bath Incubator fitted with a custom-made nitrogen manifold cover). The manifold was flushed with nitrogen and the reaction 15 stirred by means of a large magnetic stir motor placed inverted on top of the manifold. The reaction was heated to 100 oC for 4 h, and cooled to room temperature over ca. 2 h. The contents of the vial were quenched carefully into 0.5 g of silica gel in 2 mL of
CH
2 C1 2 , capped and shaken to homogeneity. The slurry was filtered through a 4-mL fritted column (Isolab microcolumn) into a 2-dram vial washing with CH 2 Cl 2 (2 x 1 20 mL), CH 2 C1 2 -MeOH (1:1, 1 x 1 mL) and MeOH(2 x I mL) and the filtrate concentrated on a vacuum centrifuge concentrator to a yellow solid. Filtration through a 500 mg silica SPE column (Supelco LC-Si) with 10 % MeOH -CH 2 C1 2 afforded the title compound as a yellow solid (14.8 mg). lH-NMR (300 MHz, DMSO-d 6 ) 8 9.45 (d, 2H, J=8.2 Hz), 9.11 (d, 2H, J=8.2 Hz), 8.97 (broad s, 2H), 8.65 25 (s, 1H), 7.90 (s, 1H), 7.0-7.5 (min, 5H), 5.25 (s, 2H), and 2.79 (s, 3H). Mass spectrum (MALDI-TOF, gentisic acid matrix) calcd. for C 17 HsNO 3
S
3 : 362.0 (M + H). Found: 361.7.
WO 00/47578 PCT/US99/18065 -160 Examples 98-126 Examples 98-104 were carried out using the procedure of Example 97, steps (b) and (c) using 0.050 mmol of the reagent specified in the table. Examples 105-126 5 were carried out using the procedure of Example 97, steps (a), (b) and (c) using 0.05 mmol of reagent. Mass Spectrum (ESI) Example Reagent Compound Formula Calcd Found (M+H) 98 1-bromo- 4-[4-(tert-butyl)(1,3- C13 H17 N3 S3 312.1 312.2 pinacolone thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 99 4-fluoro- 4-[4-(4- C15 H12 F N3 350.0 350.2 phenacyl fluorophenyl)(1,3- S3 bromide thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 100 4-cyano- 4-[4-(4- C16 H15 N5 S3 374.1 374.2 phenacyl amidinophenyl)(1,3 bromide thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 101 3-fluoro- 4-[4-(3- C15 H12 F N3 350.0 350.2 phenacyl fluorophenyl)(1,3- S3 bromide thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 102 4-(diethylamino)- 4-{4-[4-(diethylamino)- C19 H22 N4 S3 403.1 403.2 phenacyl phenyl](1,3-thiazol-2 bromide yl)}-5 methylthiothiophene-2 carboxamidine hydrochloride 103 3-chloro- 4-[4-(3- C15 H12 Cl N3 366.0 366.1 phenacyl chlorophenyl)(1,3- S3 bromide thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride WO 00/47578 PCT/US99/18065 -161 104 3,4-difluoro- 4-[4-(3,4- C15 H 11 F2 N3 368.0 368.2 phenacyl difluorophenyl)(1,3- S3 bromide thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 105 2,6-difluoro- 4-[4-(2,6- C15 H 11 F2 N3 368.0 368.2 benzoyl chloride difluorophenyl)(1,3- S3 thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 106 4-ethoxy-benzoyl 4-[4-(4- C17 H17 N3 0 376.1 376.2 chloride ethoxyphenyl)(1,3- S3 thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 107 4-chloro- 4-{4-[(4- C16 H14 Cl N3 396.0 396.1 phenoxyacetyl chlorophenoxy)- O S3 chloride methyl](1,3-thiazol-2 yl)}- 5 methylthiothiophene-2 carboxamidine hydrochloride 108 cyclopentane- 4-(4-cyclopentyl(1,3- C14 H17 N3 S3 324.1 324.2 carbonyl chloride thiazol-2-yl))-5 methylthiothiophene-2 carboxamidine hydrochloride 109 1-naphthoyl 5-methylthio-4-(4- C19 HS15 N3 S3 382.1 382.2 chloride naphthyl(1,3-thiazol-2 yl))thiophene-2 carboxamidine hydrochloride 110 3,5-dichloro- 4-[4-(3,5- C15 Hi 1 Cl2 N3 400.0 400.1 benzoyl chloride dichlorophenyl)(1,3- S3 thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 111 2,5- 4-[4-(2,5- C15 H 11 F2 N3 368.0 368.2 difluorobenzoyl difluorophenyl)(1,3- S3 chloride thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 112 9-fluorenone-4- 5-methylthio-4-[4-(9- C22 H15 N3 0 434.1 434.2 carbonyl chloride oxofluoren-4-yl)(1,3- S3 thiazol-2-yl)]thiophene 2-carboxamidine hydrochloride WO 00/47578 -162- PCT/US99/18065 113 3- 4-{4-[(3- C17 H17 N3 0 376.1 376.2 methoxyphenyl- methoxyphenyl)methyl]( S3 acetyl chloride 1,3-thiazol-2-yl)}-5 methylthiothiophene-2 carboxamidine hydrochloride 114 4-methyl valeroyl 4-[4-(3-methylbutyl)(1,3- C14 H19 N3 S3 326.1 326.2 chloride thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 115 3-(2- 4-{4-[3-(2- C19 H15 CI N4 447.0 447.1 chlorophenyl)-5- chlorophenyl)-5- O S3 methylisoxazole- methylisoxazol-4-yl](1,3 4-carbonyl thiazol-2-yl)}-5 chloride methylthiothiophene-2 carboxamidine hydrochloride 116 4-n-amyloxy- 5-methylthio-4-[4-(4- C20 H23 N3 0 418.1 418.2 benzoyl chloride pentyloxyphenyl)(1,3- S3 thiazol-2-yl)]thiophene 2-carboxamidine hydrochloride 117 1-(4- 4-{4-[(4-chlorophenyl)- C20 H20 Cl N3 434.1 434.3 chlorophenyl)- 1- cyclopentyl](1,3-thiazol- S3 cyclopentanecarb 2-yl)}-5 onyl-chloride methylthiothiophene-2 carboxamidine hydrochloride 118 4-(trifluoro- 5-methylthio-4-{4-[4- C16 H12 F3 N3 416.0 416.1 methoxy)benzoyl (trifluoromethoxy)phenyl O S3 chloride ](1,3-thiazol-2 yl)}thiophene-2 carboxamidine hydrochloride 119 3-chloro- 4-[4-(3- C17 H12 CI N3 422.0 422.1 benzo[b] chlorobenzo[b]thiophen- S4 thiophene-2- 2-yl)(1,3-thiazol-2-yl)] carbonyl chloride 5-methylthiothiophene-2 carboxamidine hydrochloride 120 3-(2-chloro-6- 4-{4-[3-(6-chloro-2- C19 H14 Cl F N4 465.0 465.1 fluorophenyl)-5- fluorophenyl)-5- O S3 methylisoxazole- methylisoxazol-4-yl](1,3 4-carbonyl thiazol-2-yl)}-5 chloride methylthiothiophene-2 carboxamidine hydrochloride 121 3-cyanobenzoyl 4-[4-(3- C16 H15 N5 S3 374.1 374.7 chloride amidinophenyl)(1,3 thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride WO 00/47578 PCT/US99/18065 -163 122 4- 4-{4-[(4- C17 H17 N3 0 376.1 376.2 methoxyphenyl- methoxyphenyl)- S3 acetyl chloride methyl](1,3-thiazol-2 yl)}- 5 methylthiothiophene-2 carboxamidine hydrochloride 123 3-(t-butyl)-1- 4-{4-[3-(tert- C16 H19 N5 S3 378.1 378.2 benzylpyrazole- butyl)pyrazol-5-yl](1,3 5- thiazol-2-yl)}-5 carbonyl methylthiothiophene-2 chloride carboxamidine hydrochloride 124 3-(4- 5-methylthio-4-[4-(1- C12 H13 N3 S3 296.0 296.2 chlorophenyl)- methylvinyl)(1,3-thiazol 2,2-dimethyl- 2-yl)]thiophene-2 propanoyl carboxamidine chloride hydrochloride 125 n-(1- 5-methylthio-4-(4-{ 1- C27 H24 N4 02 565.1 565.1 naphthalene- [(naphthylsulfonyl)amino S4 sulfonyl)-l- ]-2-phenylethyl}(1,3 phenylalanyl thiazol-2-yl))thiophene chloride 2-carboxamidine hydrochloride 126 2-bromo-5- , 4-[4-(2-bromo-5- C16 H14 BrN3 440.0 440.2 methoxybenzoyl methoxyphenyl)(1,3- O S3 chloride thiazol-2-yl)]-5 methylthiothiophene-2 carboxamidine hydrochloride 5 Example 127 a) 1-[3,5-Bis(trifluoromnethyl)phenyl]-2-bromoethan-1-one: A stirred suspension of 1 g (3.9 mmol) of 3,5-bis(trifluoromethyl)acetophenone (Lancaster, Windham, NH, USA) in dry methanol (20 mL) and 1 g (15 mmol, 2.6 eq) of poly(4 vinyl pyridinium tribromide) (Aldrich, Milwaukee, WI, USA) was protected from 10 moisture with dry nitrogen, and heated at reflux for 70 min. The polymer was filtered from the cooled solution and washed with methanol and twice with dichloromethane. The solvents were removed in vacuo to give 1-[3,5-bis(trifluoromethyl)phenyl]-2 bromoethan-1-one (1.2 g, 92 %). 'H-NMR (DMSO-d 6 ; 300 MHz) 5 8.43 (mn, 2H), 8.12 (min, 1H), 4.46 (s, 3H).
WO 00/47578 PCT/US99/18065 -164 b) Methyl 4-{4-[3,5-bis(trifluoromethyl)phenyl](1,3-thiazol-2-yl)}-5 methylthiothiophene-2-carboxylate: A solution of 75 mg (0.3 mmol) of methyl 4 (aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (Maybridge, Cornwall, UK) was reacted with 101 mg (0.3 mmol) of 1-[3,5-bis(trifluoromethyl)phenyl]-2 5 bromoethane-1-one in a manner similar Example 8, step (a)to give methyl 4-{4-[3,5 bis(trifluoromethyl)phenyl](1 ,3-thiazol-2-yl)}-5-methylthiothiophene-2-carboxylate (7 mg, 5 %) as a solid.
'
H-NMR (DMSO-d 6 ; 300 MHz) 8 8.75 (s, 1H), 8.73 (m, 2H), 8.29 (s, 1H), 8.13 (m, 1H), 3.87 (s, 3H), 2.79 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 18
H
1
INO
2
S
3
F
6 , 484.0 (M+H), found 484.0. 10 c) 4-{4-[3,5-Bis(trifluoromethyl)phenyl](1,3-thiazol-2-yl)}-5 methylthiothiophene-2-carboxamidine: Methyl 4-[4-3,5 bis(trifluoromethyl)phenyl](1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxylate (7 mg, 14.5 mmol) was treated in a manner similar to that for Example 10, step (b), to give 4-{4-[3,5-bis(trifluoromethyl)phenyl](1,3-thiazol-2-yl)} -5-methylthiothiophene 15 2-carboxamidine (6 mg, 89 %) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8.78 (s, 1H), 8.74 (s, 2H), 8.62 (s, 1H), 8.15 (s, 1H), 2.82 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C,, 7 H N3S3F6, 468.0 (M+H), found 468.0. 20 Example 128 a) 2-Bromo-1-[3-fluoro-5-(trifluoromethyl)phenyl]ethan-1-one: A stirred suspension of 1 g (4.5 mmol) of 3-fluoro-5-(trifluoromethyl)acetophenone (Lancaster, Windham, NH, USA) was treated in a manner similar to that for Example 127, step (a) to give of a 1:1 mixture of 2-bromo-1-[3-fluoro-5-(trifluoromethyl)phenyl]ethan 25 1-one and dibrominated product (1.6 g, 100%). 'H-NMR (DMSO-d 6 ; 300 MHz) 6 8.25-7.52 (m, 6H), 6.54 (s, 1H), 4.42 (s, 2H). b) Methyl 4-{4-[3-fluoro-5-(trifluoromethyl)phenyl](1,3-thiazol-2-yl)}-5 methylthiothiophene-2-carboxylate: A solution of 75 mg (0.3 mmol) of methyl 4 (aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (Maybridge, Cornwall, 30 UK) was reacted with of 86 mg (0.3 mmol) 2-bromo-1-[3-fluoro-5 (trifluoromethyl)phenyl]ethan-1-one in a manner similar to Example 8, step (a) to give, methyl 4-{4-[3-fluoro-5-(trifluoromethyl)phenyl](1,3-thiazol-2-yl)}-5- WO 00/47578 -165- PCT/US99/18065 -165 methylthiothiophene-2-carboxylate (41 mg, 31 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.59 (s, 1H), 8.29 (m, 1H), 8.27 (s, 1H), 8.25 and 8.21 (m, 1H, 1:1 ratio conformers), 7.73 and 7.70 (m, 1H, 1:1 ratio conformers). Mass spectrum (MALDI TOF, CHCA matrix, m/z): Calcd. for C 17
,H,
1
NO
2
S
3
F
4 , 434.0 (M+H), found 434.0. 5 c) 4-{4-[3-Fluoro-5-(trifluoromethyl)phenyl](1,3-thiazol-2-yl)}-5 methylthiothiophene-2-carboxamidine: Methyl 4-{4-[3-fluoro-5 (trifluoromethyl)phenyl](1,3-thiazol-2-yl)}-5-methylthiothiophene-2-carboxylate (40 mg, 0.92 mmol) was treated in a manner similar to that for Example 10, step (b), to give 4- {4-[3-fluoro-5-(trifluoromethyl)phenyl](1,3-thiazol-2-yl) } -5 10 methylthiothiophene-2-carboxamidine (31 mg, 81 %) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.36 (br s, 2H), 9.01 (br s, 2H), 8.68 (s, 1H), 8.63 (s, 1H), 8.30 (m, 1H), 8.25 and 8.22 (m, 1H, 1:1 ratio conformers), 7.75 and 7.73 (m, 1H, 1:1 ratio conformers), 2.82 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for Cl 6
HN
3
S
3
F
4 , 418.5 (M+H), found 418.0. 15 Example 129 a) 2 -Bromo-1-[3-fluoro-5-(trifluoromethyl)phenyl]propan-1-one: A stirred suspension of 1 g (4.5 mmol) of 1-[3-fluoro-5-(trifluoromethyl)phenyl]propan-1-one (Lancaster, Windham, NH, USA) was treated in a manner similar to that for Example 20 127, step (a) to give 2 -bromo-1-[3-fluoro-5-(trifluoromethyl)phenyl]propan-1-one (1.33 g, 99 %). 'H-NMR (DMSO-d 6 ; 300 MHz) 5 8.07 (m, 1H), 7.92 and 7.89 (m, 1H, 1:1 ratio conformers), 7.57 and 7.55 (m, 1H, 1:1 ratio conformers), 5.20 (q, 1H, J=6.6Hz), 1.93 (d, 3H, J=6.6 Hz). b) Methyl 4-{4-[3-fluoro-5-(trifluoromethyl)phenylf-5-methyl(1,3-thiazol-2 25 yl)}-5-mnethylthiothiophene-2-carboxylate: A solution of 75 mg (0.3 mmol) of methyl 4-(aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (Maybridge, Cornwall, UK) was reacted with 90 mg (0.3 mmol) of2-bromo-l-[3-fluoro-5 (trifluoromethyl)phenyl]propan-1-one in a manner similar to Example 8, step (a) to give, methyl 4- {(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-methyl(1,3-thiazol-2-yl) } -5 30 methylthiothiophene-2-carboxylate (31.9 mg, 24 %) as a solid.
'
H-NMR (DMSO-d 6 ; 300 MHz) 8 8.17 (s, 1H), 7.98 (m, 1H), 7.95 and 7.92 (m, 1H, 1:1 ratio conformers), 7.77 and 7.74 (m, 1H, 1:1 ratio conformers), 3.87 (s, 3H), 2.75 (s, 3H), 2.70 (s, 3H).
WO 00/47578 -166- PCT/US99/18065 Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C, 8
H
1 3
NO
2
S
3
F
4 , 448.0 (M+H), found 448.0. c) 4-{4-[3-Fluoro-5-(trifluoromethyl)phenylf-5-methyl(1,3-thiazol-2-yl)}-5 methylthiothiophene-2-carboxamidine: Methyl 4-{4-[3-fluoro-5 5 (trifluoromethyl)phenyl]-5-methyl(1,3-thiazol-2-yl) }-5-methylthiothiophene-2 carboxylate (30 mg, 0.067 mmol) was treated in a manner similar to that for Example 10, step (b), to give 4-{4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-methyl(1,3-thiazol-2 yl)}-5-methylthiothiophene-2-carboxamidine (32 mg, quantitive yield) as a yellow solid. 1 H-NMR (DMSO-d 6 ; 300 MHz) 8 9.42 (br s, 2H), 9.03 (br s, 2H), 8.60 (s, 1H), 10 7.98 (m, 1H), 7.95 and 7.92 (m, 1H, 1:1 ratio conformers), 7.79 and 7.76 (m, 1H, 1:1 ratio conformers), 2.78 (s, 3H), 2.71 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 17
H
1 3
N
3
S
3 F4, 432.0 (M+H), found 432.6. Example 130 15 a) 1-[3,5-Bis(trifluoromethyl)phenylf-2-bromopropan-1-one: A stirred suspension of 1 g (3.7 mmol) of 1-[3,5-bis(trifluoromethyl)phenyl]-propan-1-one (Lancaster, Windham, NJ, USA) treated in a manner similar to that for Example 127, step (a) to give 2-bromo-1-[3-fluoro-5-(trifluoromethyl)phenyl]propan-1-one (1.1 g, 86 %). 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.46 (m, 2H), 8.09 (m, 1), 5.26 (q, 1H, 20 J=6.6Hz), 1.96 (d, 3H, J=6.5 Hz). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 1
,H
7 OBrF 6 , 349.0 (M+H), found 348.9. b) Methyl 4-{4-J3,5-bis(trifluoromethyl)phenyl]-5-methliyl(1,3-thiazol-2-yl)} 5-methylthiothiophene-2-carboxylate: A solution of 75 mg (0.3 mmol) of methyl 4 (aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (Maybridge, Cornwall, 25 UK) was reacted with 105 mg 1-[3,5-Bis(trifluoromethyl)phenyl]-2-bromopropan-1 one in a manner similar to Example 8, step (a) to give, after preparative thin-layer chromatrography purification, methyl 4-{4-[3,5-bis(trifluoromethyl)phenyl]-5 methyl(1,3-thiazol-2-yl)}-5-methylthiothiophene-2-carboxylate (16.2 mg, 11 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.41 (m, 2H), 8.18 (m, 2H), 3.86 (s, 3H), 30 2.75 (s, 3H), 2.71 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for Cl 9
H
1 3
NO
2
S
3
F
6 , 498.0 (M+H), found 497.6.
WO 00/47578 PCT/US99/18065 -167 c) 4-{4-[3,5-Bis(trifluoromethyl)phenylf-5-methyl(1,3-thiazol- 2 -yl)}-5 methylthiothiophene-2-carboxamidine: Methyl 4-{4-[3,5 bis(trifluoromethyl)phenyl]-5-methyl( 1,3-thiazol-2-yl)} -5-methylthiothiophene-2 carboxylate (15 mg, 0.031 mmol) was treated in a manner similar to that for Example 5 10, step (b), to give 4-{4-[3,5-bis(trifluoromethyl)phenyl]-5-methyl(1,3-thiazol-2 yl)}-5-methylthiothiophene-2-carboxamidine (13 mg, 88 %) as a yellow solid. 'H NMR (DMSO-d 6 ; 300 MHz) 8 9.39 (br s, 2H), 8.94 (br s, 2H), 8.58 (s, 1H), 8.40 (inm, 2H), 8.19 (m, 1H), 2.79 (s, 3H), 2.73 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C, 8
H
13
N
3
S
3
F
6 , 482.0 (M+H), found 482.5. 10 Example 131 a) 2-Bromo-1,2-diphenylethan-l-one: A stirred suspension of 0.2 g (1 mmol) of deoxybenzoin was treated in a manner similar to that for Example 127, step (a) to give 2-bromo-1,2-diphenylethan-1-one (270 mg, 98 %). 'H-NMR (DMSO-d 6 ; 15 300 MHz) 8 8.10-8.06 (min, 2H), 7.95-7.31 (m, 8H), 7.21 (s, 1H). b) Methyl 4-(4,5-diphenyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate: A solution of 75 mg (0.3 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge, Cornwall, UK) was reacted with 92 mg, 0.3 mmol) of 2-bromo-1,2-diphenylethan-1-one in a manner similar to Example 20 8, step (a) to give, after preparative thin-layer chromatrography purification, methyl 4 (4,5-diphenyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxylate (9 mg, 7 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 8.94 (br s, 0.4H), 8.66 (s, 1H), 8.60 (br s, 0.3 H), 8.08 (s, 1H), 7.93 and 7.20 (AB quartet, 2H, J=8.7 Hz), 7.68 and 7.35 (AB quartet, 2H, J=8.2 Hz), 2.77 (s, 3H), ), 2.33 (s, 3H). Mass spectrum (MALDI-TOF, 25 CHCA matrix, m/z): Calcd. for C 22
HI
7
NO
2
S
3 , 424.0 (M+H), found 424.3. c) 4-(4,5-Diphenyl(1,3-thziazol-2-yl))-5-methylthiothiophene-2 carboxamidine: Methyl 4-(4,5-diphenyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (9 mg, 0.021 mmol) was treated in a manner similar to that for Example 10, step (b), to give 4-(4,5-diphenyl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 30 carboxamidine (3 mg, 35 %) as a brown oil. Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 21
H,
7
N
3
S
3 , 408.1 (M+H), found 408.0.
WO 00/47578 PCT/US99/18065 -168 Example 132 a) Methyl 4-(4-benzo[b]thiophen-2-yl(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate: A solution of 75 mg (0.3 mmol) of methyl 4 (aminothioxomethyl)-5-methylthiothiophene-2-carboxylate was reacted with 77 mg 5 (0.3 mmol) of 3-bromoacetylbenzo[b]thiophene (Maybridge, Cornwall, UK) in a manner similar to Example 8, step (a) to give, after preparative thin-layer chromatrography purification, methyl 4-(4-benzo[b]thiophen-2-yl(1,3-thiazol-2-yl)) 5-methylthiothiophene-2-carboxylate (28 mg, 23 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 5 8.63 (d, 1H, J=7.4 Hz), 8.30 (s, 1H), 8.25 (s, 1H), 8.22 (s, 1H), 7.53-7.46 10 (min, 2H), 3.87 (s, 3H), 2.78 (s, 3H). b) 4-(4-Benzo[b]thiophen-2-yl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxamidine: Methyl 4-(4-benzo[b]thiophen-2-yl(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate (28 mg, 0.69 mmol) was treated in a manner similar to that for Example 10, step (b), to give 4-(4-benzo[b]thiophen-2-yl(1,3 15 thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine (17 mg, 64 %) as a brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.22 (br s, 4H), 8.68 (s, 1H), 8.66 (d, 1H, J=7.6 Hz), 8.30 (s, 1H), 8.25 (s, 1H), 8.10 (d, 1H, J=7.3 Hz), 7.55-7.45 (inm, 2H), 2.81 (s, 3H). Mass spectrum (MALDI-TOF, GA matrix, m/z): Calcd. for
C,
7
H
13
N
3
S
4 , 388.0 (M+H), found 388.2. 20 Example 133 a) Methyl 4-(4-benzo[d]benzo[3,4-bffuran-3-yl(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate: A solution of 75 mg (0.3 mmol) of methyl 4 (aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (Maybridge, Cornwall, 25 UK) was reacted with 86 mg (0.3 mmol) of 2-(bromoacetyl)-dibenzofuran (Aldrich, Milwaukee, WI, USA) in a manner similar to Example 8, step (a) to give, after preparative thin-layer chromatrography purification, methyl 4-(4,5-diphenyl(1,3 thiazol-2-yl))-5-methylthiothiophene-2-carboxylate (45 mg, 36 %) as a solid. 'H NMR (DMSO-d 6 ; 300 MHz) 8 8.83-7.44 (min, 7H), 8.29 (s, 1H), 8.27 (s, 1 H), 3.88 (s, 30 3H), 2.80 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for
C
22
HI
5
NO
3
S
3 , 438.0 (M+H), found 438.5.
WO 00/47578 PCT/US99/18065 -169 b) 4-4-Benzo[d]benzo[3,4-bffuran-3-yl(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine: Methyl 4-(4-benzo[d]benzo[3,4-b]furan-3 yl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxylate (45 mg, 0.11 mmol) was treated in a manner similar to that for Example 10, step (b), to give 4-4 5 benzo[d]benzo[3,4-b]furan-3-yl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxamidine (16.8 mg, 36 %) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.72-9.10 (min, 3H), 8.84 - 7.31 (min, 9H), 2.84 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 2 1 HIsN 3
OS
3 , 422.0 (M+H), found 421.9. 10 Example 134 a) Methyl 4-(4-(4-nitrophenyl)(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate: A solution of 1 g (4 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiothiophene-2-carboxylate (Maybridge, Cornwall, UK) was reacted with 987 mg (4 mmol) of 2-bromo-4'-nitroacetophenone in a manner similar to Example 8, step 15 (a) to give methyl 4-(4-(4-nitrophenyl)(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (1.7 g, quantitive yield) as a brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 8.57 (s, 1H), 8.34 (s, 4H), 8.25 (s, 1H), 3.94 (s, 3H), 3.81 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 1 6
H
12
N
2 0 4
S
3 393.0 (M+H), found 392.8. 20 b) Methyl 4-(4-(4-aminophenyl)(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate: Methyl 4-(4-(4-nitrophenyl)(1,3-thiazol-2-yl))-5-methylthiothiophene 2-carboxylate (800 mg, 2 mmol) was dissolved in 150 mL tetrahydrofuran and treated with 20 % titanium chloride solution (Fisher Scientific, Pittsburgh, PA, USA) for 1 h. The mixture was poured into 2 M sodium hydroxide solution (100 mL), extracted 25 with dichloromethane (4 x 50 mL). The combined organic layers were washed with saturated brine solution and dried over anhydrous sodium sulfate. The solid was filtered off, and the solvent removed in vacuo. This material was purified by column chromatography on silica gel (30 g) eluting with dichloromethane:methanol 98/2 (v:v) to give methyl 4-(4-(4-aminophenyl)(1,3-thiazol-2-yl))-5-methylthiothiophene-2 30 carboxylate (500 mg, 69 %) as a solid. 'H-NMR (DMSO-d6; 300 MHz) 8 8.17 (s, 1H), 7.77 (s, 1H), 7.74 and 6.62 (AB quartet, 2H, J=8.6 Hz), 5.35 (s, 2H), 3.86 (s, WO 00/47578 PCT/US99/18065 -170 3H), 2.74 (s, 3H).). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for
C
16
H
I4
N
2 0 2
S
3 363.0 (M+H), found 362.4. c) Methyl 4
-(
4 -{4-[(methylsulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate: Methyl 4-(4-(4-aminophenyl)(1,3-thiazol-2 5 yl))-5-methylthiothiophene-2-carboxylate (200 mg, 0.55 mmol) was dissolved in dry dichloromethane (20 mL). To this, N-methyl morpholine (150 pIL, 1.38 mmol) and dimethylaminopyridine (6.1 mg, 0.055 mmol) were added, the mixture was cooled on an ice bath, and methanesulfonyl chloride (43 pL, 0.55 mmol) was added dropwise. The mixture was then stirred for 8 days at room temperature. The mixture was 10 partitioned between saturated sodium bicarbonate (50 mL) and dichloromethane (20 mL). The aqueous layer was extracted with dichloromethane (3 x 20 mL), and the combined organic layers were washed with saturated sodium bicarbonate (20 mL), brine (2 x 20 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo. Column chromatrography on silica gel (100 g) eluting with 15 dichloromethane:methanol 99/1 (v:v), gave methyl 4-(4-{4 [(methylsulfonyl)amino]phenyl} (1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (155 mg, 64 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.92 (s, 1H), 8.22 (s, 1H), 8.11 (s, 1H), 8.40 and 6.90 (AB quartet, 2H, J=8.7 Hz), 3.87 (s, 3H), 3.05 (s, 3H), 2.76 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix m/z): 20 Calcd. for C, 7
H
1 6
N
2 0 4
S
4 441.0 (M+H), found 441.2. d) 4
-(
4
-{
4 -[(Methylsulfonyl)amino]phenyl}(1,3-thiiazol-2-yl))-5 methylthiothiophene-2-carboxamidine: Methyl 4-(4-{4 [(methylsulfonyl)amino]phenyl} (1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (81 mg, 0.184 mmol) was treated in a manner similar to that for Example 25 10, step (b), to give 4
-(
4
-{
4 -[(methylsulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine (24.9 mg, 32 %) as a light brown solid. 'H NMR (DMSO-d 6 ; 300 MHz) 6 10.0 (br s, 1H), 9.3 (br s, 2H), 8.98 (s, 1H), 8.65 (s, 1H), 8.21 (s, 1H), 7.98 and 7.5 (AB quartet, 2H, J=8.6 Hz), 3.05 (s, 3H), 2.79 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C, 6
H,
6
N
4 0 2
S
4 425.0 30 (M+H), found 425.1.
WO 00/47578 PCT/US99/18065 -171 Example 135 a) Methyl 4
-(
4 -{4-J(phenylsulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 methylthiothliiophene-2-carboxylate: Methyl 4-(4-(4-aminophenyl)(1,3-thiazol-2 yl))-5-methylthiothiophene-2-carboxylate (100 mg, 0.28 mmol) was dissolved in dry 5 dichloromethane (10 mL). To this, N-methyl morpholine (46 tL, 0.42 mmol) and dimethylaminopyridine (3.4 mg, 0.028 mmol) were added, the mixture was cooled on an ice bath, and benzenesulfonyl chloride 35 tL, 0.28 mmol) was added dropwise. The mixture was then stirred for 24 h at room temperature. Workup was carried out as in Example 134, step (c). Trituration with dichloromethane and methanol gave 10 methyl 4-(4- {4-[(phenylsulfonyl)amino]phenyl } (1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate (44 mg, 31 %) as a crystalline solid.
'
H-NMR (DMSO-d 6 ; 300 MHz) 6 10.46 (s, 1H), 8.19 (s, 1H), 8.05 (s, 1H), 7.91 and 7.19 (AB quartet, 2H, J=8.7 Hz), 7.81 (m, 2H), 7.64-7.54 (m, 3H) 3.85 (s, 3H), 2.74 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 22
H
1 sN 2 0 4
S
4 504.2 15 (M+H), found 504.1 b) 4
-(
4 -{4-J(Phenylsulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine: Methyl 4-(4-{4 [(phenylsulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (30 mg, 0.060 mmol) was treated in a manner similar to that for Example 20 10, step (b), to give 4-(4-{4-[(phenylsulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine (12.6 mg, 43 %) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.13 (br s, 3H), 8.60 (s, 1H), 8.08 (s, 1H) 7.93 and 7.20 (AB quartet, 2H, J=8.7 Hz), 7.82-7.79 (m, 2H), 7.65-7.53 (m, 3H) 3.85 (s, 3H), 2.74 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 2 1 HlsN 4 0 2
S
4 , 25 87.0 (M+H), found 487.7. Example 136 a) Methyl 4 -(4-{4-[(trifluoromethylsulfonyl)amino]phenyl}(1,3-thiazol-2 yl))-5-methylthiothiophene-2-carboxylate: Methyl 4-(4-(4-aminophenyl)(1,3-thiazol 30 2 -yl))-5-methylthiothiophene-2-carboxylate (200 mg, 0.55 mmol) was dissolved in dry pyridine (20 mL). The mixture was cooled on an ice bath, and trifluoromethanesulfonic anhydride (0.5 mL, 3 mmol) was added. The mixture was WO 00/47578 -172- PCT/US99/18065 then stirred for 1.5 h at room temperature. Workup was carried out as in Example 134, step (c). Column chromatrography on silica gel (30 g) eluting with hexanes:ethyl acetate 7/3 (v:v), followed by preparative thin layer chromatography eluting with dichloromethane:methanol 99/1 (v:v) gave methyl 4-(4-{4 5 [(trifluoromethylsulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (160 mg, 59 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.48 and 7.87(s, 3/2 ratio conformers, 1H), 8.23 (s, 1H), 8.21 (s, 1H), 8.29 and 7.84 (AB quartet, 2H, 2/3 ratio conformers, J=8.7 Hz), 8.10 and 7.37 (AB quartet, 2H, J=8.7 Hz), 3.87 and 3.86 (s, 2/3 ratio conformers, 3H), 2.77 and 2.76 (s, 2/3 ratio 10 conformers, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for
C
17
H
13
N
2 0 4
S
4
F
4 495.0 (M+H), found 495.6 b) 4-(4-{4-[(Trifluoromethylsulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine: Methyl 4-(4-{4 [(trifluoromethylsulfonyl)amino]phenyl} (1,3-thiazol-2-yl))-5-methylthiothiophene-2 15 carboxylate (30 mg, 0.061 mmol) was treated in a manner similar to that for Example 10, step (b), to give of4-(4-{4-[(trifluoromethylsulfonyl)amino]phenyl}(1,3-thiazol-2 yl))-5-methylthiothiophene-2-carboxamidine (21.6 mg, 74 %) as a light brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.39 (br s, 2H), 8.97 (br s, 2H), 8.64 (s, 1H), 8.24 (s, 1H), 8.12 and 7.39 (AB quartet, 2H, J=8.7 Hz), 4.78 (br s, 1H), 2.79 (s, 3H). Mass 20 spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for Cl 6
H
13
N
4 0 2
S
4
F
3 , 479.0 (M+H), found 479.5. Example 137 a) Methyl 4-(4-{4-J(toluenesulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 25 methylthiothiophene-2-carboxylate: Methyl 4-(4-(4-aminophenyl)(1,3-thiazol-2 yl))-5-methylthiothiophene-2-carboxylate (33 mg, 0.09 mmol) was dissolved in dry dichloromethane (5 mL). To this, N-methyl morpholine (10 pL, 0.09 mmol) andp toluenesulfonyl chloride (17 mg, 0.09 mmol) was added and the mixture was stirred at room temperature for 5 days. Workup was carried out as in Example 134, step (c). 30 Trituration with dichloromethane and methanol gave methyl 4-(4-{4 [(toluenesulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (20 mg, 43 %) as a brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 10.39 WO 00/47578 -173- PCT/US99/18065 (s, 1H), 8.19 (s, 1H), 8.05 (s, 1H), 7.91 and 7.18 (AB quartet, 2H, J=8.7 Hz), 7.68 and 7.35 (AB quartet, 2H, J=8.2 Hz), 3.85 (s, 3H), 2.74 (s, 3H), 2.27 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C, 3
H
2 020 4
S
4 , 517.2 (M+H), found 517.0. 5 b) 4-(4-{4-[(Toluenesulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxamidine: Methyl 4-(4-{4 [(toluenesulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5-methylthiothiophene-2 carboxylate (15 mg, 0.029 mmol) was treated in a manner similar to that for Example 10, step (b), to give 4-(4-{4-[(toluenesulfonyl)amino]phenyl}(1,3-thiazol-2-yl))-5 10 methylthiothiophene-2-carboxamidine (17.9 mg, 81 %) as a light brown solid. 'H NMR (DMSO-d 6 ; 300 MHz) 8 8.94 (br s, 0.4H), 8.66 (s, 1H), 8.60 (br s, 0.3 H), 8.08 (s, 1H), 7.93 and 7.20 (AB quartet, 2H, J=8.7 Hz), 7.68 and 7.35 (AB quartet, 2H, J=8.2 Hz), 2.77 (s, 3H), 2.33 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 22
H
20
N
4 0 2
S
4 :501.1 (M+H), found 501.1. 15 Example 138 a) Methyl 4-[4-(4-chlorophenyl) (1,3-thiazol-2-yl)]-5 (methylsulfinyl)thiophene-2-carboxylate: To a stirred solution of 764 mg (2 mmol) of methyl 4-[4-(4-chlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 20 carboxylate (Maybridge, Cornwall, UK) dissolved in 1,1,1,3,3,3 hexafluoroisopropanol (2.5 mL) was added 30% hydrogen peroxide (0.45 mL, 4 mmol). This solution was stirred for 45 h at room temperature. Dichloromethane (10 mL) was added after 2 hours. Additional hydrogen peroxide (2 x 0.45 mL portions) was added after 4 hours and 24 hours. The mixture was quenched with 10% sodium 25 sulfite in brine (4 mL). The organic layer was separated, dried over anhydrous sodium sulfate, and the solvents removed in vacuum. Column chromatography on silica gel (45 g), eluting with dichloromethane:methanol 99/1 (v:v) gave methyl 4-[4 (4-chlorophenyl)(1,3-thiazol-2-yl)]-5-(methylsulfinyl)thiophene-2-carboxylate (720 mg, 90 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.37 (s, 1H), 8.30 (s, 1H), 30 8.05 and 7.52 (AB quartet, 2H, J=8.6 Hz), 3.91 (s, 3H), 3.16 (s, 3H). Mass spectrum (MALDI-TOF, GA matrix, m/z): Calcd. for C 16
H
1 2
NO
3
S
3 C1:398.0 (M+H), found 397.8.
WO 00/47578 -174- PCT/US99/18065 b) 4-[4-(4-Chlorophenyl) (1,3-thiazol-2-yl)]-5-(methylsulfinyl)thiophene-2 carboxamidine: Methyl 4-[4-(4-chlorophenyl)(1,3-thiazol-2-yl)]-5 (methylsulfinyl)thiophene-2-carboxylate (100 mg, 0.25 mmol) was treated in a manner similar to that for Example 10, step (b), to give, after preparative thin layer 5 chromatography purification eluting with dichloromethane:methanol:acetic acid 9/1/0.5 (v:v:v), 4-[4-(4-chlorophenyl)(1,3-thiazol-2-yl)]-5-(methylsulfinyl)thiophene 2-carboxamidine (18.2 mg, 19 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.33 (s, 1H), 8.22 (s, 1H), 8.05 and 7.57 (AB quartet, 2H, J=8.6 Hz), 3.12 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix m/z): Calcd. for C 15
H
1 2
N
3 0S 3 C1 382.0 (M+H), 10 found 382.1. Example 139 a) Methyl 4-cyano-5-(methylsulfonyl)thiophene-2-carboxylate: To a stirred solution of (4.5 g, 21 mmol) of methyl 4-cyano-5-methylthiothiophene-2-carboxylate 15 (Maybridge, Cornwall, UK) was dissolved in dichloromethane (250 mL) and treated with m-chloroperbenzoic acid (15.3 g, 90 mmol) at room temperature for 2.25 h. The mixture was filtered and the solid washed with dichloromethane (2 x 50 mL). The filtrate was washed with sodium bicarbonate (2 x 100 mL), sodium thiosulfate (100 mL), sodium bicarbonate (100 mL), water (100 mL), brine (100 mL), and dried over 20 anhydrous sodium sulfate. The solvent was removed in vacuo to give methyl 4 cyano-5-(methylsulfonyl)thiophene-2-carboxylate (4.91 g, 95%) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.44 (s, 1H), 3.91 (s, 3H), 3.58 (s, 3H). b) Methyl 4-cyano-5-methoxythiophene-2-carboxylate: Methyl 4-cyano-5 (methylsulfonyl)thiophene-2-carboxylate (2 g, 8 mmol) was refluxed with 0.5 M 25 sodium methoxide in methanol (16 mL) for 15 minutes. The solution was cooled, the crystallized solid collected on a Btichner funnel and washed with methanol (50 mL) to give methyl 4-cyano-5-methoxythiophene-2-carboxylate (1.145 g, 73%) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.87 (s, 1H) 4.19 (s, 3H), 3.82 (s, 3H). c) Methyl 4-(aminothioxomethyl)-5-methoxythiophene-2-carboxylate: 30 Methyl 4-cyano-5-methoxythiophene-2-carboxylate (1 g, 5 mmol) was dissolved in dry methanol (150 mL) and triethylamine (3.5 mL, 25.4 mmol) was added. After degassing the solution with argon for 10 minutes, hydrogen sulfide gas was bubbled WO 00/47578 PCT/US99/18065 -175 through the solution for 5 h. After stirring 18 h at room temperature, the solution was degassed by bubbling argon (6 h), concentrated to 20 mL and acetone (20 mL) was added. The dark solid was collected on a Btichner funnel and washed with acetone. Recrystallize solid from hot ethanol (15 mL) to give methyl 4-(aminothioxomethyl) 5 5-methoxythiophene-2-carboxylate (683 mg, 59 %) as a brown oil. Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for CH 9
NO
3
S
2 232.0 (M+H), found 232.4 d) Methyl 5-methoxy-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate: A solution of 400 mg (1.73 mmol) of methyl 4-(aminothioxomethyl)-5 10 methoxythiophene-2-carboxylate was reacted with 345 mg (1.73 mmol) of 2 bromoacetophenone (Aldrich, Milwaukee, WI, USA) in a manner similar to Example 8, step (a) to give methyl 5-methoxy-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2 carboxylate (56 mg, 10 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 8.22 (s, 1H), 8.14 (s, 1H), 8.05 (m, 2H), 7.47 (m, 2H), 7.36 (m, 1H), 4.26 (s, 3H), 3.85 (s, 3H). 15 e) S-Methoxy-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine: Methyl 5-methoxy-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate (55 mg, 0.16 mmol) was treated in a manner similar to that for Example 10, step (b), to give 5 methoxy-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine (36 mg, 69 %) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.34 (br s, 2H), 8.94 (br s, 2H), 8.70 20 (s, 1H), 8.20 (s, 1H), 8.07 (m, 2H), 7.49 (m, 2H), 7.38 (m, 1H), 4.32 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 15
H,
3 N3OS 2 316.5 (M+H), found 316.1 Example 140 a) Methyl 4-cyano-5-[(4-methoxyphenyl)methylthio]thiophene-2 25 carboxylate: To a stirred solution of 2.5 g (10 mmol) of methyl 4-cyano-5 (methylsulfonyl)thiophene-2-carboxylate (Example 139, step (a)) in dry methanol (15 mL) was added p-methoxybenzylmercaptan (3.8 mL, 28 mmol) and triethylamine (1.4 mL, 10 mmol). This solution was refluxed for 15 min and cooled. The resulting solid was collected on a biichner funnel and washed with methanol (2 x 25 mL) to methyl 30 4 -cyano-5-[(4-methoxyphenyl)methylthio]thiophene-2-carboxylate (2.84 g, 89 %) as a solid.
WO 00/47578 -176- PCT/US99/18065 b) Methyl 4-(aminothioxomethyl)-S-J(4 methoxyphenyl)methylthio]thiophene-2-carboxylate: Methyl 4-cyano-5-[(4 methoxyphenyl)methylthio]thiophene-2-carboxylate (2.5 g, 7.8 mmol) was treated as in Example 139, step (c) to give methyl 4-(aminothioxomethyl)-5-[(4 5 methoxyphenyl)methylthio]thiophene-2-carboxylate (1.32 g, 48 %) as a solid. 'H NMR (DMSO-d 6 ; 300 MHz) 5 9.64 (s, 1H), 9.28 (s, 1H), 8.08 (s, 1H), 7.35 and 6.92 (AB quartet, 2H, J=8.7 Hz), 4.27 (s, 2H), 3.82 (s, 3H), 3.75 (s, 3H). c) Methyl S-(methoxyphenylthio)-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2 carboxylate: A solution of 1.2 g (3.4 mmol) of methyl 4-(aminothioxomethyl)-5-[(4 10 methoxyphenyl)methylthio]thiophene-2-carboxylate was reacted with 676 mg (3.4 mmol) of 2-bromoacetophenone (Aldrich, Milwaukee, WI, USA) in a manner similar to'Example 8, step (a) to give methyl 5-(methoxyphenylthio)-4-(4-phenyl(1,3-thiazol 2-yl))thiophene-2-carboxylate (755 mg, 49 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.26 (s, 1H), 8.22 (s, 1H), 8.04 (m, 2H), 7.48 (m, 2H), 7.38 (m, 1H), 7.33 and 15 6.89 (AB quartet, 2H, J=8.7 Hz), 4.40 (s, 2H), 3.86 (s, 3H), 3.72 (s, 3H). d) 5-(Methoxyphenylthio)-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2 carboxamidine: Methyl 5-(methoxyphenylthio)-4-(4-phenyl(1,3-thiazol-2 yl))thiophene-2-carboxylate (100 mg, 0.22 mmol) was treated in a manner similar to that for Example 10, step (b), to give 5-methoxy-4-(4-phenyl(1,3-thiazol-2 20 yl))thiophene-2-carboxamidine (94 mg, 91 %) as an orange solid. 'H-NMR (DMSO d 6 ; 300 MHz) 8 9.49 (br s, 2H), 9.15 (br s, 2H), 8.70 (s, 1H), 8.26 (s, 1H), 8.07 (m, 2H), 7.49 (m, 2H), 7.40 (m, 1H), 7.35 and 6.90 (AB quartet, 2H, J=8.7 Hz), 4.41 (s, 2H), 3.73 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for
C
22
H,
9
N
3 0S 3 438.5 (M+H), found 438.1. 25 Example 141 a) Methyl 4-[4-(4-chlorophenyl) (1,3-thiazol-2-yl)]-5 (methylsulfonyl)thiophene-2-carboxylate: To a stirred solution of 1 g (2.6 mmol) of methyl 4-[4-(4-chlorophenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2-carboxylate 30 (Maybridge, Cornwall, UK) was dissolved in dry dichloromethane (50 mL) and treated with m-chloroperbenzoic acid (1.94 g, 11.3 mmol) at room temperature for 1.5 h. The solution was filtered and the solid washed with dichloromethane. The WO 00/47578 PCT/US99/18065 -177 filtrate was washed with sodium bicarbonate solution (2 x 20 mL), sodium thiosulfate solution (20 mL), sodium bicarbonate solution (20 mL), brine (20 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo to give methyl 4-[4-(4 chlorophenyl)(1,3-thiazol-2-yl)]-5-(methylsulfonyl)thiophene-2-carboxylate (826 mg, 5 77 %) as a tan solid. Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for
C
1 6
H
1 2
NO
4
S
3 C1 414.0 (M+H), found 414.8. b) 4-[4-(4-Chlorophenyl) (1,3-thiazol-2-yl)]-5-(methylsulfonyl)thiophene-2 carboxamidine: Methyl 4-[4-(4-chlorophenyl)(1,3-thiazol-2-yl)]-5 (methylsulfonyl)thiophene-2-carboxylate (200 mg, 0.4 mmol) was treated in a manner 10 similar to that for Example 10, step (b), to give 4-[4-(4-chlorophenyl)(1,3-thiazol-2 yl)]-5-(methylsulfonyl)thiophene-2-carboxamidine (85 mg, 53 %) as a yellow solid. c) 4-[4-(4-Chlorophenyl)(1,3-thiazol-2-yl)]-5-(phenylmethylthio)thiophene 2-carboxamidine: A stirred solution of 80 mg (0.2 mmol) of 4-[4-(4 chlorophenyl)(1, 3 -thiazol- 2 -yl)]-5-(methylsulfonyl)thiophene-2-carboxamidine 15 benzyl mercaptan (115 gL, 0.980 imol) was treated in a manner similar to that for Example 140, step (a) to give, after silica gel column chromatography (20 g) eluting with dichloromethane:methanol:acetic acid 9/1/0.5 (v:v:v), 4-[4-(4-chlorophenyl)(1,3 thiazol-2-yl)]-5-(phenylmethylthio)thiophene-2-carboxamidine (75 mg, 85 %) as a pale orange solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 9.44 (br s, 2H), 9.03 (br s, 2H), 20 8.67 (s, 1H), 8.33 (s, 1H), 8.08 and 7.56 (AB quartet, 2H, J=8.7 Hz), 7.54-7.17 (m. 5H), 4.45 (s, 2H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for
C
21
H,
6
N
3
S
3 C1 442.0 (M+H), found 442.7. Example 142 25 a) 1-S[5-(tert-Butyl)-2-methyl(3-furyl)]-2-bromoethan-l-one: A solution of 1 g (5 mmol) of 5-(tert-butyl)-2-methylfuran-3-carbonyl chloride (Maybridge, Cornwall, UK) dissolved in dry acetonitrile (4 mL) and 6.25 mL (12.5 mmol) of 2 M trimethylsilyldiazomethane in hexanes (Aldrich, Milwaukee, WI) was stirred 1.75 h at room temperature and the mixture was cooled on an ice bath for 5 min. To this, 30% 30 hydrogen bromide in acetic acid (2 mL, 10 mmol) was added dropwise over 10 min. This was stirred an additional 20 minutes on an ice bath. Evaporation of the solvents gave 1-[5-(tert-butyl)-2-methyl(3-furyl)]-2-bromoethan-1-one (1 g, 77 %) as a brown WO 00/47578 -178- PCT/US99/18065 oil. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 6.50 (s, 1H), 4.57(s, 2H), 2.52 (s, 1H), 1.24 (s, 9H). Mass spectrum (LCA, m/z): Calcd. for CizHi 5
O
2 Br, 259.1 and 261.1 (M+H), found 259.1 and 261.1. b) Methyl 4-{4-[5-(tert-butyl)-2-methyl(3-furyl)](1,3-thiazol-2-yl)}-5 5 methylthiothiophene-2-carboxylate: A solution of 955 mg (3.86 mmol) of methyl 4 (aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (Maybridge, Cornwall, UK) was reacted with 1 g (3.86 mmol) of 1-[5-(tert-butyl)-2-methyl(3-furyl)]-2 bromoethan-1-one (1 g) in a manner similar to Example 8, step (a) to give methyl 4 {4-[5-(tert-butyl)-2-methyl(3-fuiryl)](1,3-thiazol-2-yl) }-5-methylthiothiophene-2 10 carboxylate (999 mg, 64 %) as a red-brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.14 (s, 1H), 7.74 (s, 1H), 6.46 (s, 1H), 3.86 (s, 3H), 2.74 (s, 3H), 2.66 (s, 3H), 1.27 (s, 9H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for Cl 9
H
2 1 NO3S 3 , 408.1 (M+H), found 408.0. c) 4-{4-[5-(tert-Butyl)-2-methyl(3-furyl)](1,3-thiazol-2-yl)}-5 15 methylthiothiophene-2-carboxamidine: Methyl 4-{4-[5-(tert-butyl)-2-methyl(3 furyl)](1,3-thiazol-2-yl)}-5-methylthiothiophene-2-carboxylate (940 mg, 2.3 mmol) was treated in a manner similar to that for Example 10, step (b) to give 4-{4-[5-(tert butyl)-2-methyl(3-furyl)](1,3-thiazol-2-yl) }-5-methylthiothiophene-2-carboxamidine (930 mg, quantitive yield) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.42 20 (br s, 2H), 9.03 (br s, 2H), 8.59 (s, 1H), 7.77 (s, 1H), 6.47 (s, 1H), 2.78 (s, 3H), 2.68 (s, 3H), 1.27 (s, 9H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for CsH 2 1
N
3 0S 3 , 392.1 (M+H), found 392.1. Example 143 25 a) 1-[3-(tert-Butyl)-1-benzylpyrazol-5-yl]-2-bromoethan-1-one: A solution of 1 g (3.6 mmol) of 3-(tert-butyl)-1-benzylpyrazole-5-carbonyl chloride (Maybridge, Cornwall, UK) was dissolved in dry acetonitrile (4 mL) and 4.5 mL (9 mmol) of 2 M trimethylsilyldiazomethane in hexanes (Aldrich, Milwaukee, WI, USA) was added. After stirring 1 h 20 min at room temperature, the mixture was cooled on an ice bath 30 for 5 min. To this, 30% hydrogen bromide in acetic acid (2 mL, 10 mmol) was added dropwise over 15 min. This was stirred an additional 15 minutes on an ice bath. Filtration of the precipitated solid and evaporation of the solvents gave 1-[3-(tert- WO 00/47578 -179- PCT/US99/18065 butyl)-1-benzylpyrazol-5-yl]-2-bromoethan-1-one (1.47 g, quantitive yield) as an orange solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 7.33-7.06 (m, 5H), 7.08 (s, 1H), 5.64 (s, 2H), 4.57 (s, 2H), 1.28 (s, 9H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 16 Hi 9 N2OBr, 335.1 and 337.1 (M+H), found 335.6 and 337.6. 5 b) Methyl 4-{4-[3-(tert-Butyl)-1-benzylpyrazol-5-yl](1,3-thiazol-2-yl)}-5 methylthiothiophene-2-carboxylate: A solution of 823 mg (3.3 mmol of methyl 4 (aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (Maybridge, Cornwall, UK) was reacted with 1.36 g (3.3 mmol) of 1-[3-(tert-butyl)-1-benzylpyrazol-5-yl]-2 bromoethan-1-one in a manner similar to Example 8, step (a) to give methyl 4-{4-[3 10 (tert-butyl)- 1-benzylpyrazol-5-yl](1,3-thiazol-2-yl) }-5-methylthiothiophene-2 carboxylate (1.25 g, 79 %) as a crystalline solid. 1 H-NMR (DMSO-d 6 ; 300 MHz) 8 8.11 (s, 1H), 8.05 (s, 1H), 7.28-6.99 (m, 5H), 6.70 (s, 1H), 5.88 (s, 2H), 3.86 (s, 3H), 2.70 (s, 3H), 1.30 (s, 9H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 24
H
25
N
3 0 2
S
3 , 484.1 (M+H), found 483.9. 15 c) 4-{4-[3-(tert-Butyl)-1-benzylpyrazol-5-yl](1,3-thiazol-2-yl)}-5 methylthiothiophene-2-carboxamidine: Methyl 4-{4-[3-(tert-butyl)-1 benzylpyrazol-5-yl](1,3-thiazol-2-yl) } -5-methylthiothiophene-2-carboxylate (1.2 mg, 2.6 mmol) was treated in a manner similar to that for Example 10, step (b) to give 4 {4- [3-(tert-butyl)- 1-benzylpyrazol-5-yl](1,3-thiazol-2-yl) } -5-methylthiothiophene-2 20 carboxamidine (1.21 g, quantitive yield) as a yellow solid. 1 H-NMR (DMSO-d 6 ; 300 MHz) 8 9.43 (br s, 1H), 9.07 (br s, 1H), 8.60 (s, 1H), 8.04 (s, 1H), 7.37-6.97 (m, 5H), 6.70 (s, 1H), 5.92 (s, 2H), 2.73 (s, 3H), 1.30 (s, 9H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C 23
H
2 5
N
5
S
3 , 468.1 (M+H), found 468.1. 25 Example 144 a) 4-Bromo-5-methylthiophene-2-carboxylic acid: A stirred solution of 1 g (3.9 mmol) of 2-methyl-3,5-dibromothiophene (prepared by the method of Kano, S.et al., Heterocycles 20(10):2035, 1983) in dry tetrahydrofuran (10 mL) was cooled to 78 'C and 2 M n-butyllithium in cyclohexane (1.93 mL, 3.87 mmol) was added over 3 30 min. After stirring 3 min at -78 oC, the mixture was added to tetrahydrofuran (100 mL) with dry ice suspended. This mixture was allowed to stir and warm to room temperature. To this, 6 N hydrochloric acid (50 mL) was added carefully. Then, WO 00/47578 -180- PCT/US99/18065 water (50 mL) was added and the layers were separated. The aqueous layer was extracted with diethyl ether (4 x 30 mL). The combined organic layers were washed with water, brine, and dried over anhydrous sodium sulfate. The solvents were removed in vacuo to give an 85/15 mixture of 4-bromo-5-methylthiophene-2 5 carboxylic acid and 5-bromothiophene-2-carboxylic acid (780 mg, 90 %) as a tan solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 13.33 (br s, 1H), 7.62 (s, 1H), 7.56 and 7.34 (AB quartet, 0.35H, J=3.9 Hz), 2.41 (s, 3H). Gas Chromotography/Mass spectroscopy (m/z): Calcd. for C 6
H
5
O
2 SBr, 220.9 and 222.9 (M+H), found 221.3 and 223.3. Calcd. for CH 3 0O 2 SBr, 206.9 and 208.9 (M+H), found 207.3 and 209.3. 10 b) Methyl 4-bromo-5-methylthiophene-2-carboxylate: A solution of 780 mg (3.5 mmol) of an 85/15 mixture of 4-bromo-5-methylthiophene-2-carboxylic acid and 5-bromothiophene-2-carboxylic acid was dissolved in methanol (50 mL) and treated with 9 ml (18 mmol) 2 M trimethylsilyldiazomethane in hexanes (Aldrich, Milwaukee, WI, USA). Evaporation of the solvents gave an 8/2 mixture of methyl 4 15 bromo-5-methylthiophene-2-carboxylate and methyl 5-bromothiophene-2-carboxylate (858 mg, quantitive yield) as a brown oil. Gas Chromotography/Mass spectroscopy (m/z): Calcd. for C 7
H
7
O
2 SBr, 234.9 and 236.9 (M+H), found 235.3 and 237.3. Calcd. for C 6
H
4 0 2 SBr, 220.9 and 222.9 (M+H), found 221.3 and 223.3. c) Methyl 4 -cyano-5-methylthiophene-2-carboxylate: A solution of an 8/2 20 mixture of 823 mg (3.5 mmol) of methyl 4-bromo-5-methylthiophene-2-carboxylate and methyl 5-bromothiophene-2-carboxylate was dissolved in dry dimethylformamide (5 mL) and refluxed with copper cyanide (345 mg, 3.9 mmol) for 7 hours. The cooled solution was poured into 0.1 M aqueous sodium cyanide solution (200 mL) and extracted with diethyl ether (5 x 30 mL). The organic layers were washed with brine 25 (2 x 30 mL), dried over anhydrous sodium sulfate, and the solvents removed in vacuo. The resulting brown solid was purified by column chromatography on silica gel eluting with hexanes:ethyl acetate 9/1 (v:v) to give a 95/5 mixture of methyl 4-cyano 5-methylthiophene-2-carboxylate and methyl 5-methylthiophene-2-carboxylate (369 mg, 68 %) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.06 (s, 1H), 8.05 and 30 7.90 (2H, 0.1 H, J=4.0 Hz, minor component), 3.87 (s, 3H, minor component), 3.84 (s, 3H) 2.68 (s, 3H).
WO 00/47578 -181- PCT/US99/18065 d) Methyl 4 -(aminothioxomethyl)-S-methylthiophene-2-carboxylate: A stirred solution of 804 mg (4.4 mmol) of methyl 4-cyano-5-methylthiophene-2 carboxylate was treated in a manner similar to Example 139, step (c) to give, after fractional crystallization ethanol of the unreacted starting nitrile, a 2:3 ratio of methyl 5 4-(aminothioxomethyl)-5-methylthiophene-2-carboxylate and methyl 4-cyano-5 methylthiophene-2-carboxylate (457 mg, 48 %) as a light brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 9.93 (br s, 1H, minor), 9.34 (br s, 1H, minor), 8.06 (s, 1H, major), 7.77 (s, 1H, minor component), 3.84 (s, 3H, minor), 3.81 (s, 3H, major), 2.68 (s, 3H, major), 2.61 (s, 2H, minor). Mass spectrum (MALDI-TOF, CHCA matrix, 10 m/z): Calcd. for CsH 9
NO
2
S
2 216.0 (M+H), found 216.4. e) Methyl S-methyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate: A solution of 200 mg (0.93 mmol) of methyl 4-(aminothioxomethyl)-5 methylthiophene-2-carboxylate was reacted with 185 mg (0.93 mmol) of 2 bromoacetophenone in a manner similar to Example 8, step (a) to give, after 15 purification by preparative thin layer chromatography eluting with hexanes:ethyl acetate 7/3 (v:v), a mixture of methyl 5-methyl-4-(4-phenyl(1,3-thiazol-2 yl))thiophene-2-carboxylate and methyl 4-cyano-5-methylthiophene-2-carboxylate (96 mg, 36 %) as a solid. f) 5-Methyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine: 20 Methyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate (64 mg, 0.23 mmol) was treated in a manner similar to Example 10, step (b) to give, after preparative high pressure liquid chromatography (Dynamax C 18 column, 300 A pore size, 10 ptm particle size, 40% to 100% acetonitrile over 30 minutes in 0.1% aqueous trifluoroacetic acid) 5-methyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2 25 carboxamidine (0.6 mg, 0.9 %) as an an off-white solid. 'H-NMR (CD 3 OD, 300 MHz) 8 8.44 (s, 1H), 8.02 (m, 2H), 7.92 (s, 1H), 7.45 (m, 2H), 7.36 (m, 1H), 2.96 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for C, 5 Hi 3
N
3
S
2 300.1 (M+H), found 300.6. g) 5-(4-Phenyl-1,3-thiazol-2-yl)thiophene-2-carboxamide: From the HPLC 30 purified mixture in the previous step was isolated 5-(4-phenyl-1,3-thiazol-2 yl)thiophene-2-carboxamide as an off-white solid (2 mg). 'H-NMR (Methanol-d 4 ; 300 MHz) 8 7.99 (m, 2H), 7.97 (s, 1H), 7.95 and 7.78 (AB quartet, 2H, J=4.2 Hz), WO 00/47578 -182- PCT/US99/18065 7.48-7.35 (m, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for Cl 4
HIIN
3
S
2 286.0 (M+H), found 286.2. Example 145 5 a) Methyl 4-[4-(3,4-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiophene 2-carboxylate: A solution of 257 mg (0.48 mmol, based on a mixture containing 60% nitrile) of methyl 4-(aminothioxomethyl)-5-methylthiophene-2-carboxylate was reacted with 124 mg (0.48 mmol) of 2-bromo-(3',4'-dimethoxy)-acetophenone (Example 31, step (a)) was reacted in a manner similar to Example 8, step (a) to give 10 methyl 4-[4-(3,4-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiophene-2 carboxylate (95 mg, 53 %) as a solid. Mass spectrum (MALDI-TOF, CHCA matrix, m/z): Calcd. for CH 17
,NO
4
S
2 376.1 (M+H), found 376.3. b) 4-[4-(3,4-Dimethoxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiophene-2 carboxamide: Methyl 4-[4-(3,4-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5 15 methylthiophene-2-carboxylate (95 mg, 0.25 mmol) was treated in a manner similar to Example 10, step (b) to give 4-[4-(3,4-dimethoxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiophene-2-carboxamide (8 mg, 9 %) as a yellow solid. 'H-NMR (Methanol d 4 ; 300 MHz) 8 8.42 (s, 1H), 7.81 (s, 1H), 7.61 (m, 2H), 7.03 (m, 1H), 3.92 (s, 3H), 3.88 (s, 3H), 2.95 (s, 3H). Mass spectrum (MALDI-TOF, CHCA matrix, m/z): 20 Calcd. for C, 7
H,
7
N
3 0 2
S
2 360.1 (M+H), found 360.2. Example 146 a) 4-Bromo-5-methylthiophene-2-carboxylic acid: A solution of 27.65 g (108 mmol) of 2-methyl-3,5-dibromothiophene (prepared by the method of Kano, S.et 25 al., Heterocycles 20(10):2035, 1983) was dissolved in dry tetrahydrofuran (280 mL), cooled to -78 'C and 2 M n-butyl lithium in cyclohexane (54 mL, 108 mmol) was added over 10 min. After stirring 20 min at -78 oC, dry carbon dioxide gas was bubbled through the solution for 1.5 h as the mixture was allowed to warm to room temperature. To this 6 N hydrochloric acid (100 mL) was added carefully. The layers 30 were separated and the aqueous layer was extracted with diethyl ether (4 x 50 mL). The combined organic layers were washed with brine, and dried over anhydrous sodium sulfate. The solvents were removed in vacuo to give 4-bromo-5- WO 00/47578 -183- PCT/US99/18065 methylthiophene-2-carboxylic acid (22.4 g, 94 %) as an off-white solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 13.34 (br s, 1H), 7.61 (s, 1H), 2.41 (s, 3H). b) Isopropyl 4-bromo-S-methylthiophene-2-carboxylate: A solution of 5 g (22.6 mmol) of 4-bromo-5-methylthiophene-2-carboxylic acid was dissolved in dry 5 dichloromethane (200 mL) and reacted with oxalyl chloride (2 mL, 22.6 mmol) and dimethylformamide (100 ptL) stirring on an ice bath for 30 min and then at room temperature for 2.5 h. The solvents were removed in vacuo and the residue was passed through silica gel, eluting off with hexanes:ethyl acetate 7/3 (v:v), ethyl acetate, and dichloromethane. The solvents were removed in vacuo and the resulting 10 oil dissolved in dry dichloromethane (100 mL). This solution was reacted with dry pyridine (9 mL, 113 mmol) and dry isopropanol (40 mL, 522 mmol) for 88 h. The solvents were removed in vacuo and the residue partitioned between sodium bicarbonate (150 mL) and dichloromethane (75 mL). The aqueous layers were extracted with dichloromethane (2 x 20 mL), and the combined organic layers were 15 washed with sodium bicarbonate (30 mL), brine (30 mL), and dried over anhydrous sodium sulfate. The solvents were removed in vacuo. The residue was purified by column chromatography eluting with hexanes:ethyl acetate 9/1 (v:v) to give isopropyl 4-bromo-5-methylthiophene-2-carboxylate (1.91 g, 32 %) as a pale yellow oil. 'H NMR (DMSO-d 6 ; 300 MHz) 8 7.66 (s, 1H), 5.07 (septet, 1H, J=6.2 Hz), 2.42 (s, 3H), 20 1.29 (d, 6H, J=6.2 Hz). Mass spectrum (ESI, m/z): Calcd. for C 9 HizO 2 SBr 264.2 (M+H), found 264.8. c) Isopropyl 4-cyano-5-methylthiophene-2-carboxylate: A stirred solution of 1.9 g (7.3 mmol) of isopropyl 4-bromo-5-methylthiophene-2-carboxylate was dissolved in dry dimethylformamide (30 mL) and refluxed with copper cyanide (785 25 mg, 8.8 mmol) for 16 hours. The cooled solution was poured into 0.1 M aqueous sodium cyanide solution (300 mL) and extracted with diethyl ether (4 x 40 mL). The organic layers were washed with brine (2 x 40 mL), dried over anhydrous sodium sulfate, and the solvents removed in vacuo. Column chromatography on silica gel eluting with hexanes:ethyl acetate 9/1 (v:v), gave isopropyl 4-cyano-5 30 methylthiophene-2-carboxylate (960 mg, 63 %) as a yellow crystalline solid 1 H-NMR (DMSO-d 6 ; 300 MHz) 8 8.01 (s, 1H), 5.09 (septet, 1H, J=6.2 Hz), 2.67 (s, 3H), 1.29 (d, 6H, J=6.2 Hz).
WO 00/47578 PCT/US99/18065 -184 d) Isopropyl 4-(aminothioxomethyl)-5-methylthiophene-2-carboxylate: A stirred solution of 960 mg (4.59 mmol) of isopropyl 4-cyano-5-methylthiophene-2 carboxylate was treated in a manner similar to Example 139, step (c) to give, after crystallization from diethyl ether, isopropyl 4-(aminothioxomethyl)-5 5 methylthiophene-2-carboxylate (623 mg, 56 %) as a solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 9.93 (br s, 1H), 9.34 (br s, 1H), 7.54 (s, 1H), 5.07 (septet, 1H, J=6.2 Hz), 2.60 (s, 3H), 1.29 (d, 6H, J=6.2 Hz). Mass spectrum (MALDI-TOF, GA matrix, m/z): Calcd. for Co 10
H
1 3
NO
2
S
2 244.0 (M+H), found 243.8. e) Isopropyl 5-methyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2 10 carboxylate: A solution of 375 mg (1.54 mmol) of isopropyl 4-(aminothioxomethyl) 5-methylthiophene-2-carboxylate was reacted with 307 mg (1.54 mmol) of 2 bromoacetophenone (Aldrich, Milwaukee, WI, USA) in a manner similar to Example 8, step (a) to give, after crystallization from methanol, isopropyl 5-methyl-4-(4 phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate (347 mg, 66%) as light brown 15 needles. 1 H-NMR (DMSO-d 6 ; 300 MHz) 8 8.23 (s, 1H), 8.09 (s, 1H), 8.05 (m, 2H), 7.49 (m, 2H), 7.38 (m, 1H), 5.13 (septet, 1H, J=6.2 Hz), 2.86 (s, 3H), 1.33 (d, 6H, J=6.2 Hz). Mass spectrum (ESI, m/z): Calcd. for C 18
H
1 7
NO
2
S
2 344.1 (M+H), found 344.1. J) 5-Methyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine: 20 Isopropyl 5-methyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate (340 mg, 0.99 mmol) was treated in a manner similar to Example 10, step (b) to give 5-methyl 4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine (360 mg, quantitive yield) as a yellow solid. This material was dissolved in dry methanol (20 mL) and treated with 1 M HCI (g) in diethyl ether. Evaporation of the solvents in vacuo and 25 recrystallization from methanol gave the hydrochloride salt of 5-methyl-4-(4 phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine (252 mg, 76 %) as a light brown crystalline solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 9.45 (br s, 2H), 9.10 (br s, 2H), 8.56 (s, 1H), 8.27 (s, 1H), 8.06 (m, 2H), 7.50 (m, 2H), 7.40 (m, 1H), 2.93 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 15
H
1 3
N
3
S
2 300.1 (M+H), found 300.2. 30 WO 00/47578 -185- PCT/US99/18065 Example 147 a) 2-Methyl-5-[(methylethyl)oxycarbonyl]thiophene-3-carboxylic acid: A stirred mixture of 500 mg (2.39 mmol) of isopropyl 2-methyl-3-cyanothiophene-5 carboxylate and tetrafluorophthalic acid (570 mg, 2.39 mmol) was heated in a glass 5 bomb at 160 oC for 66 hours. The cooled residue was digested in hot chloroform (30 mL), treated with norite, and filtered through celite. The celite was washed with hot chloroform (30 mL). The cooled chloroform extracts were filtered and extracted with saturated sodium bicarbonate (4 x 10 mL). The basic extracts were washed with chloroform, filtered through celite, and acidified to pH 1 with concentrated 10 hydrochloric acid. The solid was collected by filtration and washed with water (3 x 10 mL) to give 2 -methyl-5-[(methylethyl)oxycarbonyl]thiophene-3-carboxylic acid (288 mg, 53 %) as a light brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 13.03 (br s, 1H), 7.85 (s, 1H), 5.08 (septet, 1H, J=6.2 Hz), 2.71 (s, 3H), 1.29 (d, 6H, J=6.2 Hz). Mass spectrum (ESI, m/z): Calcd. for C 1 oH 1 2 0 4 S 229.1 (M+H), found 228.8 15 b) Isopropyl 4-(2-bromoacetyl)-5-methylthiophene-2-carboxylate: A stirred solution of 300 mg (1.3 mmol) of 2-methyl-5-[(methylethyl)oxycarbonyl]thiophene 3-carboxylic acid was dissolved in dry dichloromethane (10 mL) and treated with oxalyl chloride (174 pL, 2 mmol) and dimethylformamide (50 pL). The mixture was stirred at room temperature for 1.25 h, the solvents removed in vacuo, and the residue 20 passed through silica gel (1 inch in a 60 mL sintered-glass Btichner funnel) and eluted off with dichloromethane (150 mL). This material was treated in a manner similar to Example 142, step (a) to give isopropyl 4-(2-bromoacetyl)-5-methylthiophene-2 carboxylate (266 mg, 67 %) as a solid. c) Isopropyl 4
-(
2 -amino(1,3-thiazol-4-yl))-5-methylthliiophene-2-carboxylate: 25 A solution of 260 mg (0.85 mmol) of isopropyl 4-(2-bromoacetyl)-5 methylthiophene-2-carboxylate was reacted with 65 mg (0.85 mmol) of thiourea in a manner similar to Example 8, step (a) to give isopropyl 4-(2-amino(1,3-thiazol-4-yl)) 5-methylthiophene-2-carboxylate (257 mg, quantitive yield) as a white solid. 'H-NMR (DMSO-d, 6 ; 300 MHz) 5 7.90 (s, 1H), 6.93 (s, 1H), 5.09 (septet, 1H, J=6.2 Hz), 2.61 30 (s, 3H), 1.29 (d, 6H, J=6.2 Hz). Mass spectrum (ESI, m/z): Calcd. for C,2HI 4
N
2 0 2
S
2 283.1 (M+H), found 283.1 WO 00/47578 -186- PCT/US99/18065 d) 4
-(
2 -Amino(1,3-thiazol-4-yl))-5-methylthiophene-2-carboxamidine: Isopropyl 4-(2-amino(1,3-thiazol-4-yl))-5-methylthiophene-2-carboxylate (240 mg, 0.85 mmol) was treated in a manner similar to Example 10, step (b) to give 4-(2 amino(1, 3 -thiazol-4-yl))-5-methylthiophene-2-carboxamidine (20 mg, 10 %) as a 5 solid. 'H NMR (DMSO-d 6 , 300 MHz): 6 9.30 (br s, 2H), 8.99 (bs, 2H), 8.28 (s, 1H), 6.78 (s, 1H), 2.71 (s, 3H); Mass Spectrum (ESI, m/z) calcd. for C 9
H
10
N
4
S
2 , 238.8 (M+H), found 239.2. Example 148 10 a) 4-Bromo-5-ethylthiophene-2-carboxylic acid: A stirred solution of 10 g (35 mmol) of 4,5-dibromothiophene-2-carboxylic acid (Lancaster, Windham, NH, USA) in dry THF (100 mL) was cooled to -78 oC. To this, 35 mL (70 mmol) of 2.0 M n-butyllithium in cyclohexane (Aldrich, Milwaukee, WI, USA) was added dropwise over 15 min, and the reaction was allowed to stir for 15 min at -78 oC. The 15 mixture was quenched with ethyl iodide (2.8 mL, 35 mmol) and allowed to warm to room temperature. The mixture was carefully poured into 6N hydrochloric acid (100 mL) and extracted with diethyl ether (4 x 50 mL). The organic layers were washed with water (2 x 50 mL), brine (50 mL), and dried over anhydrous sodium sulfate. The solvents were removed in vacuo to give 2-ethyl-3-bromo-thiophene-5-carboxylate (7 20 g, 85 %) as a dark solid. 1 H-NMR (DMSO-d6; 300 MHz) 8 13.25 (br s, 1H), 7.62 (s, 1H), 2.80 (q, 2H, J=7.5 Hz), 1.23 (t, 3H, J=7.5 Hz). b) Isopropyl 4-bromo-5-ethylthiophene-2-carboxylate: A solution of 7 g (30 mmol) of 4-bromo-5-ethylthiophene-2-carboxylic acid was dissolved in dry dichloromethane (200 mL) and treated with oxalyl chloride (3.2 mL, 36 mmol) and 25 dimethylformamide (0.5 mL) for 18.5 h. The solvents were removed in vacuo and the residual brown oil was passed through silica gel (2 inches in a 350 mL scintered-glass Btichner funnel) and eluted with 700 mL of hexanes:ethyl acetate 9/1 (v:v). The elutate was concentrated in vacuo and the oil dissolved in dry dichloromethane (200 mL). This solution was treated with pyridine (12 mL, 150 mmol) and dry isopropanol 30 (60 mL, 750 mmol) for 4 h at room temperature. The solvents were removed in vacuo and the residue partioned between dichloromethane (100 mL) and water (200 mL). The aqueous layers were extracted with dichloromethane (2 x 30 mL). The combined WO 00/47578 -1 87- PCT/US99/18065 organic layers were extracted with sodium bicarbonate (2 x 30 mL), brine (30 mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo. Purification by column chromatography on silica gel (250 g) eluting with hexanes:ethyl acetate 95/5 (v:v) gave isopropyl 2-ethyl-3-bromo-thiophene-5 5 carboxylate (4 g, 48 %) as a yellow oil. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 7.66 (s, 1H), 5.89 (septet, 1H, J=6.2 Hz), 2.80 (q, 2H, J=7.5 Hz), 1.29 (d, 6H, J=6.0 Hz), 1.24 (t, 3H, J=7.5 Hz). c) Isopropyl 4-cyano-5-ethylthiophene-2-carboxylate: A stirred solution of 4 g (14.4 mmol) of isopropyl 4-bromo-5-ethylthiophene-2-carboxylate was refluxed in 10 dry dimethylformamide (50 mL) with copper cyanide (1.94 g, 22 mmol) for 8 hours. The cooled mixture was poured into 0.1 M sodium cyanide (500 mL) and extracted with diethyl ether (4 x 50 mL). The organic layers were washed twice with brine (50 mL) and dried over anhydrous sodium sulfate. The solvents were removed in vacuo. Column chromatography on silica gel (400 g), eluting with hexanes:ethyl acetate 9/1 15 (v:v) gave isopropyl 2-ethyl-3-cyano-thiophene-5-carboxylate (1.7 g, 53 %) as a pale yellow oil. 'H-NMR (DMSO-d 6 ; 300 MHz) a 8.03 (s, 1H), 5.10 (septet, 1H, J=6.2 Hz), 3.04 (q, 2H, J=7.5 Hz), 1.31 (t, 3H, J=7.5 Hz), 1.30 (d, 6H, J=6.2 Hz). Mass spectrum (ESI m/z): Calcd. for CHI 3
NO
2 S 224.1 (M+H), found 224.0. d) Isopropyl 4-(aminothioxomethyl)-5-ethylthiophene-2-carboxylate: A 20 stirred solution of 1.7 g (7.6 mmol) of isopropyl 4-cyano-5-ethylthiophene-2 carboxylate was treated as in Example 139, step (c) to give isopropyl 5-ethyl-4 (aminothioxomethyl)-5-ethylthiophene-2-carboxylate (1.45 g, 74 %) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 9.93 (br s, 1H), 9.39 (br s, 1H), 8.04 (s, 1H), 5.08 (septet, 1H, J=6.2 Hz), 3.08 (q, 2H, J=7.5 Hz), 1.29 (d, 6H, J=6.2 Hz), 1.24 (t, 3H, 25 J=7.5 Hz). e) Isopropyl S-ethyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate: A solution of 450 mg (1.75 mmol) of isopropyl 5-ethyl-4-(aminothioxomethyl)-5 ethylthiophene-2-carboxylate was reacted with 348 mg (1.75 mmol) of 2 bromoacetophenone (Aldrich, Milaukee, WI, USA) in a manner similar to Example 8, 30 step (a) to give isopropyl 5-ethyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2 carboxylate (303 mg, 49%) as an off-white solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 8.22 (s, 1H), 8.07 (s, 1H), 8.03 (m, 2H), 7.49 (m, 2H), 7.38 (m, 1H), 5.13 (septet, WO 00/47578 -188- PCT/US99/18065 1H, J=6.2 Hz), 3.34 (q, 2H, J=7.4 Hz), 1.39 (t, 3H, J=7.4 Hz), 1.33 (d, 6H, J=6.2 Hz). Mass spectrum (ESI, m/z): Calcd. for C 19
HI
9
NO
2
S
2 358.1 (M+H), found 358.1. J) 5-Ethyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine: Isopropyl 5-ethyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxylate (250 mg, 5 0.70 mmol) was treated in a manner similar to that for Example 10, step (b), to give 5 ethyl-4-(4-phenyl(1,3-thiazol-2-yl))thiophene-2-carboxamidine (148 mg, 67 %) as a yellow solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 9.44 (br s, 2H), 9.07 (br s, 2H), 8.54 (s, 1H), 8.26 (s, 1H), 8.05 (m, 2H), 7.50 (m, 2H), 8.70 (s, 1H), 7.40 (m, 1H), 3.44 (q, 2H, J=7.4 Hz), 1.42 (t, 3H, J=7.4 Hz). Mass spectrum (ESI, m/z): Calcd. for 10 C 16
H
15
N
3
S
2 314.1 (M+H), found 314.2. Example 149 a) Isopropyl 4-[4-(3-hydroxyphenyl) (1,3-thiazol-2-yl)]-5-methylthliophene 2-carboxylate: A solution of 1.97 g (8.1 mmol) of isopropyl 4-(aminothioxomethyl) 15 5-methylthiophene-2-carboxylate was reacted with 1.74 g (8.1 mmol) of 3'-hydroxy 2-bromoacetophenone (Example 40, step (a)) were reacted in a manner similar to Example 8, step (a) to give, after column chromatography on silica gel eluting with hexane:ethyl acetate 7/3 (v:v), crystallization from acetonitrile, and recrystallization from hexanes, isopropyl 4-[4-(3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5 20 methylthiophene-2-carboxylate (1.4 g, 48%) as brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 6 9.57 (br s, 1H), 8.14 (s, 1H), 8.08 (s, 1H), 7.46 (m, 2H), 7.26 (m, 1H),), 6.78 (m, 1H), 5.12 (septet, 1H, J=6.2 Hz), 2.85 (s, 3H), 1.33 (d, 6H, J=6.2 Hz). Mass spectrum (ESI, m/z): Calcd. for C 18
H
17
NO
3
S
2 360.1 (M+H), found 360.1. b) 4-[4-(3-Hydroxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiophene-2 25 carboxamide: Isopropyl 4-[4-(3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiophene-2-carboxylate (1.4 g, 3.89 mmol) was treated in a manner similar to Example 10, step (b) to give 4-[4-(3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiophene-2-carboxamide (360 mg, 31 %) as a brown solid. 1 H-NMR (DMSO d 6 ; 300 MHz) 5 9.62 (br s, 1H), 9.45 (br s, 2H), 9.09 (br s, 2H), 8.53 (s, 1H), 8.16 (s, 30 1H), 7.47 (m, 2H), 7.27 (m, 1H), 6.80 (m, 1H), 2.93 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C, 5 sH, 3
N
3
OS
2 316.1 (M+H), found 316.2.
WO 00/47578 -1 89- PCT/US99/18065 Example 150 a) (tert-Butoxy)-N-({4-[4-(3-hydroxyphenyl) (1,3-thiazol-2-yl)]-5-methyl(2 thienyl)}iminomethyl)carboxamide: A stirred solution of 320 mg (1 mmol) of 4-[4 (3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiophene-2-carboxamide was 5 dissolved in dry dimethylformamide (50 mL) and treated with 262 mg (1.2 mmol) of di-tert-butyl-dicarbonate (Acros, Pittsburgh, PA, USA) and diisopropylethylamine (261 ptL, 1.5 mmol) for 64 hours at room temperature. The mixture was poured into sodium bicarbonate solution (200 mL) and extracted with dichloromethane (6 x 30 mL). The organic extracts were washed twice with brine (50 mL) and dried over 10 anhydrous sodium sulfate. The solvents were in vacuo and column chromatography on silica gel (100 g) eluting with dichloromethane:methanol 95/5 (v:v) gave (tert butoxy)-N-({4-[4-(3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methyl(2 thienyl)}iminomethyl)carboxamide (247 mg, 59 %) as a yellow oil. 'H-NMR (DMSO-d 6 ; 300 MHz) 5 9.56 (s, 1H), 9.12 (br s, 2H), 8.47 (s, 1H), 8.09 (s, 1H), 7.46 15 (m, 2H), 7.26 (m, 1H), 6.78 (m, 1H), 2.83 (s, 3H), 1.45 (s, 9H). Mass spectrum (ESI, m/z): Calcd. for C 2 0
H
21
N
3 0 3
S
2 416.1 (M+H), found 415.7 b) Methyl 2
-{
3
-[
2 -(5-{[(tert-butoxy)carbonylamino]iminomethyl}-2-methyl 3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetate: A stirred solution of 247 mg (0.595 mmol) of (tert-butoxy)-N-({4-[4-(3-hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methyl(2 20 thienyl)}iminomethyl)carboxamide was dissolved in dry dimethylformamide (4 mL) and treated with cesium carbonate (291 mg, 0.89 mmol) and methyl bromoacetate (136 mg, 0.89 mmol) for 3 h at 60 oC. The mixture was poured into water (50 mL) and extracted with dichloromethane (9 x 10 mL). The organic extracts were washed with brine (10 mL) and dried over anhydrous sodium sulfate. The solvents were 25 removed in vacuo and column chromatography on silica gel (50 g) eluting with dichloromethane:methanol 98/2 (v:v) gave methyl 2-{3-[2-(5- { [(tert butoxy)carbonylamino]iminomethyl}-2-methyl-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetate (178 mg, 61 %) as an oil. Mass spectrum (ESI, m/z): Calcd. for
C
23
H
25
N
3 0 5
S
2 488.1 (M+H), 388.1 ((M-BOC)+H), found 487.8, 388.2. 30 c) Methyl 2-{3-[2-(S-amidino-2-methyl-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetate: Methyl 2- {3-[2-(5- { [(tert-butoxy)carbonylamino]iminomethyl} 2 -methyl-3-thienyl)-1,3-thiazol-4-yl]phenoxy} acetate (15 mg, 0.031 mmol) treated WO 00/47578 -190- PCT/US99/18065 with dichloromethane:trifluoroacetic acid 1/1 (v:v) with 2.5% water added at room temperature for 1.5 h. Removal of the solvents in vacuo gave methyl 2-{3-[2-(5 amidino-2-methyl-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetate (8.1 mg, 52 %) as a brown solid. 'H-NMR (DMSO-d 6 ; 300 MHz) 8 9.38 (br s, 2H), 8.94 (br s, 2H), 8.51 5 (s, 1H), 8.31 (s, 1H), 7.62 (m, 2H), 7.41 (m, 1H), 6.96 (m, 1H), 4.89 (s, 2H), 3.72 (s, 3H), 2.92 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 18
H
17
N
3 0 3
S
2 388.1 (M+H), found 388.3. Example 151 10 a) 2-{3-[2-(5-{[(tert-Butoxy)carbonylamino]iminomethyl}-2-methyl-3 thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid: A stirred solution of 50 mg (0.11 mmol) of methyl 2- { 3- [2-(5- { [(tert-butoxy)carbonylamino]iminomethyl } -2-methyl-3 thienyl)-1,3-thiazol-4-yl]phenoxy}acetate was dissolved in tetrahydrofuran (10 mL) and treated 2M aqueous sodium hydroxide solution (2 mL) at room temperature for 1 15 h 10 min. The solvents were removed in vacuo. Purification by passing the solid through silica gel (1 inch in a 60 mL sintered-glass Bichner funnel) eluting with dichloromethane:methanol 8/2 (v:v) gave 2- {3-[2-(5-{ [(tert butoxy)carbonylamino]iminomethyl}-2-methyl-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetic acid (44 mg, 88 %) as a yellow solid. 1 H-NMR (DMSO-d 6 ; 300 20 MHz) 8 9.38 (br s, 2H), 8.94 (br s, 2H), 8.51 (s, 1H), 8.31 (s, 1H), 7.62 (m, 2H), 7.41 (m, 1H), 6.96 (m, 1H), 4.89 (s, 2H), 3.72 (s, 3H), 2.92 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 2 2
H
2 3
N
3 0 5
S
2 474.1 (M+H), 374.1 ((M-BOC)+H) found 374.2, 473.7. b) 2-{3-[2-(5-Amidino-2-methyl-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic 25 acid: Methyl 2- {3-[2-(5- { [(tert-butoxy)carbonylamino]iminomethyl}-2-methyl-3 thienyl)-l,3-thiazol-4-yl]phenoxy}acetate (4 mg, 0.0084 mmol) was treated with dichloromethane:trifluoroacetic acid 1/1 (v:v) with 2.5% water added at room temperature for 2 h 35 min. Removal of the solvents in vacuo gave 2-{3-[2-(5 amidino-2-methyl-3-thienyl)-1,3-thiazol-4-yl]phenoxy}acetic acid (2.9 mg, 71 %) as a 30 solid. Mass spectrum (ESI, m/z): Calcd. for CiHlsN 3 0 3
S
2 373.1 (M+H), found 374.2.
WO 00/47578 PCT/US99/18065 -191 c) tert-Butyl 4
-(
2
-{
3
-[
2 -(S-{[(tert-butoxy)carbonylamino]iminomethyl}-2 methyl- 3 -thienyl)-1, 3 -thiazol-4-yl]phenoxy}acetyl)piperazinecarboxylate: A stirred solution of 40 mg (0.084 mmol) of 2-{3-[2-(5-{[(tert butoxy)carbonylamino]iminomethyl } -2-methyl-3-thienyl)- 1,3-thiazol-4 5 yl]phenoxy}acetic acid dissolved in dry dimethylformamide (5 mL) was treated with hydroxybenzotriazole (23 mg, 0.17 mmol), 32 mg (0.17 mmol) of N-tert butoxycarbonyl-piperazine (Lancaster, Wiridham, NH, USA), 65 mg (0.17 mmol) of O-(7-azabenzotriazol- 1-yl)-N,N,NAN'-tetramethyluronium hexafluorophosphate (HATU) at room temperature for 20 h. The mixture was partitioned between 10 dichloromethane (50 mL) and brine (50 mL). The aqueous layers were extracted twice with dichloromethane (50 mL) and the combined organic layers were washed with brine (50 mL) and dried over anhydrous sodium sulfate. The solvents were removed in vacuo. Purification preparative thin layer chromatography eluting with dichloromethane:methanol 95/5 (v:v) gave tert-butyl 4-(2- {3-[2-(5- { [(tert 15 butoxy)carbonylamino]iminomethyl}-2-methyl-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetyl)piperazinecarboxylate (25 mg, 46%) as a white solid. iH-NMR (DMSO-d 6 ; 300 MHz) 5 9.13 (br s, 2H), 8.50 (s, 1H), 8.20 (s, 1H), 7.63 (m, 2H), 7.39 (m, 1H), 6.95 (m, 1H), 4.93 (s, 2H), 3.47-3.34 (m, 8H), 2.82 (s, 3H), 1.45 (s, 9H), 1.42 (s, 9H). Mass spectrum (ESI, m/z): Calcd. for C 3 1
H
39
N
5 0 6
S
2 642.3 (M+H), 542.3 20 ((M-BOC)+H), 442.3 ((M-2 BOC)+H), found 642.0, 542.2, 442.3. d) 5-Methliyl- 4
-{
4 -[3-(2-oxo-2-piperazinylethoxy)phenyl](1,3-thiazo-2 yl)}thiophene-2-carboxamidine: tert-Butyl 4-(2-{3-[2-(5-{ [(tert butoxy)carbonylamino]iminomethyl}-2-methyl-3-thienyl)-1,3-thiazol-4 yl]phenoxy}acetyl)piperazinecarboxylate (25 mg, 0.039 mmol) treated with 25 dichloromethane:trifluoroacetic acid 1/1 (v:v) with 2.5% water added at room temperature for 2 h. Removal of the solvents in vacuo gave 5-methyl-4-{4-[3-(2-oxo 2 -piperazinylethoxy)phenyl](1,3-thiazol-2-yl) } thiophene-2-carboxamidine (27.4 mg, quantitive yield) as an off-white solid. 1 H-NMR (Methanol-d 4 ; 300 MHz) 8 8.41 (s, 1H), 7.94 (s, 1H), 7.67 (m, 2H), 7.39 (m, 1H), 7.00 (m, 1H), 4.96 (s, 2H), 3.88 (m, 30 4H), 3.25 (m, 4H), 2.95 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 21
H
23
N
5 0 2
S
2 442.1 (M+H), found 442.4.
WO 00/47578 -192- PCT/US99/18065 Example 152 Methyl 4-(2-bromoacetyl)-5-methylthiothiophene-2-carboxylate To a stirring slurry of 2-methylthio-(5-carbomethoxy)-thiophene-3-carboxylic 5 acid (2.0 g, 8.61 mmol) in 28 mL of CH 2
C
2 under N 2 containing 0.8 mL DMF at 0 oC was added oxalyl chloride (1.9 equiv, 16.3 mmol) slowly via syringe. The reaction was allowed to warm to ambient temperature after 1 h, and then stirred an additional 1 h. The reaction mixture was filtered through a 20 cm pad of silica gel in a 30 mL sintered glass funnel wetted with 50% ethyl acetate-hexanes and further eluted with 10 the same solvent system until the eluent showed no product by UV visualization. The solvent was concentrated in vacuo, azeotroped with toluene (lx), and dried under vacuum to afford the acid chloride (1.52 g) as a light yellow solid. The acid chloride was dissolved in 20 mL of CH 3 CN, cooled to 0 oC, and treated with TMSCHN 2 (2.1 equiv, 6.3 mL, 2 M in hexanes) dropwise via syringe. The reaction was allowed to 15 warm to ambient temperature (0.5 h), cooled back to 5oC and immediately treated with 30% HBr-acetic acid (0.66 mL) dropwise via an addition funnel. After 15 min. at 0 oC, the reaction diluted with 20 mL of ether, filtered and thoroughly washed with ether (3x20 mL). The yellow solids were dried under vacuum to afford methyl 4-(2 bromoacetyl)-5-methylthiothiophene-2-carboxylate (1.0 g, 37% yield) as a yellow 20 powder.' H NMR (DMSO-d 6 , 300 MHz) 8 2.66 (s, 3H), 3.84 (s, 3H), 5.03 (s, 2H), 8.29 (s, 1H). Example 153 Isopropyl- 4-(2-bromoacetyl)-5-methylthiophene-2-carboxylate 25 To a stirring slurry of 2 -methyl-(5-carboisopropoxy)-thiophene-3-carboxylic acid (0.40 g, 1.75 mmol) in 15 mL of CH 2
C
2 under N 2 containing 0.8 mL DMF at 0 'C was added oxalyl chloride (1.9 equiv, 3.32 mmol,) slowly via syringe. The reaction was allowed to warm to ambient temperature after 1 h, and then stirred an additional 1 30 h. The solvent was concentrated in vacuo, azeotroped with toluene (lx), and dried under vacuum to afford the acid chloride (0.397 g, 1.60 mmol) as a light yellow solid. The acid chloride was dissolved in 7 mL of CH 3 CN, cooled to 0 'C, and treated with
TMSCHN
2 (2.1 equiv, 1.68 mL, 2 M in hexanes) dropwise via syringe. The reaction WO 00/47578 -1 93- PCT/US99/18065 was allowed to warm to ambient temperature (0.5 h), cooled back to 5 0 C and immediately treated with 30% HBr-acetic acid (0.5 mL) dropwise via an addition funnel. After 15 min. at 0 oC, the reaction mixture was filtered through a 10 cm pad of silica gel in a 15 mL sintered glass funnel wetted with 50% ethyl acetate-hexanes and 5 further eluted with the same solvent system until the eluent showed no product by UV visualization. The solvent was concentrated in vacuo dried under vacuum to afford isopropyl- 4-(2-bromoacetyl)-5-methylthiophene-2-carboxylate (0.329 g, 61% yield) as an oil which solidified upon standing to a tan solid. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.31 (d, 6H, J=6.3 Hz), 2.71 (s, 3H), 4.60 (s, 2H), 5.09 (min, 1H), 8.08 (s, 1H). 10 Example 154 a) Methyl 5-methylthio-4-[2-(phenylamino)-(1,3-thiazol-4-yl)]-thiophene-2 carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5-methylthiothiophene-2 carboxylate (60.5 mg, 0.19 mmol) was slurried in 4 mL of acetone with phenyl 15 thiourea (1 equiv, 30 mg) and heated to 70 oC . After 3 h the reaction was allowed to cool to room temperature, filtered, and dried in vacuo to give 62.5 mg (69% yield) of methyl 5-methylthio-4-[2-(phenylamino)-(1,3-thiazol-4-yl)]-thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.65 (s, 3H), 3.83 (s, 3H), 6.95-6.99 (min, 1H), 7.28-7.35 (min, 4H), 7.67 (d, 1H, J= 1.4, 7.7 Hz), 8.06 (s, 1H), 10.54 (s, 1H); 20 Mass Spectrum (ESI) m/z calcd. for C, 6
H,
4
N
2 0 2
S
3 , 362.49 (M+H), found 363.7. b) S-Methylthio-4-[2-(phenylamino)(1,3-thiazol-4-yl)]thiophene-2 carboxamidine hydrochloride: To a flame-dried flask containing 57.8 mg (8 equiv, 1.08 mmol) of NH 4 Cl under N 2 was charged 1.3 mL of toluene. AlMe 3 (8 equiv, 2M/hexanes, 0.54 mL) was added dropwise to the stirred slurry over a 3 min. period, 25 and allowed to stir another 5 min. At this time methyl 5-methylthio-4-[2 (phenylamino)-(1,3-thiazol-4-yl)]-thiophene-2-carboxylate hydrobromide (1 equiv, 60 mg, 0.135 mmol) was quickly added in one portion and the resultant mixture was immersed in a 120 oC oil bath. After 2 h 10 min. at this temperature, TLC (silica gel 60 F 25 4 , Merck KGaA, Darmstadt, Germany, 9:1:0.5 CH 2
C
2 -MeOH-AcOH eluent) 30 indicated the reaction to be complete by disappearance of the starting material. The reaction was allowed to cool to ambient temperature, then added via pipette to a stirred slurry of 1.3 g of SiO 2 in 20 mL of CHC1 3 . The residual residue in the flask was WO 00/47578 PCT/US99/18065 -194 rinsed with 4 mL of MeOH, briefly sonicated and added to the SiO 2 slurry. The slurry was stirred for 10 min. and then filtered through a 15-mL sintered glass funnel containing 20 cm of SiO 2 with 50% CHCl 3 -MeOH. The yellow fraction was collected, discarding the forerun. TLC indicated the product was essentially pure. The solvent 5 was removed in vacuo, and the residue triturated with 10% MeOH-CH 2 C1 2 . The solids were removed by filtration. The solvent was concentrated in vacuo to give 30.1 mg (66% yield) of 5-methylthio-4-[2-(phenylamino)-(1,3-thiazol-4-yl)]thiophene-2 carboxamidine hydrochloride as a red-brown powder. 'H NMR (DMSO-d 6 , 300 MHz) 5 2.73 (s, 3H), 6.94- 7.00 (min, 1H), 7.15 (s, 1H), 7.30-7.35 (min, 1H), 7.78 (d, 1H, J=8.7 10 Hz), 8.49 (s, 1H), 8.87 (bs, 2H), 9.31 (bs, 2H), 10.38 (s, 1H); Mass Spectrum (ESI) m/z calcd. for Ci 5
HI
4
N
4
S
3 , 346.50 (M+H), found 347.2. Example 155 a) Methyl 4-{2-[(2-chlorophenyl)amino](1,3-thiazol-4-yl)}-5 15 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (50 mg) was allowed to react with 2-chlorophenyl thiourea (26.7 mg) as described in Example 154, step (a), to give 58 mg (75%) of methyl 4- {2-[(2-chlorophenyl)amino]-(1,3-thiazol-4-yl) }-5-methylthiothiophene-2 carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.66 (s, 3H), 3.82 (s, 20 3H), 7.04 (min, 1H), 7.32-7.38 (min, 2H), 7.47 (dd, 1H, J= 1.4, 8.7 Hz), 8.12 (s, 1H), 8.56 (dd, 1H, J=l.4, 8.3 Hz), 9.75 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C1 6
H,
3
CIN
2 0 2
S
3 , 396.94 (M+H), found 397.1. b) 4-{2-[(2-Chlorophenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene 2-carboxamidine hydrochloride: Methyl 4- {2-[(2-chlorophenyl)amino]-(1,3-thiazol 25 4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (40 mg, 0.08 mmol) was treated as described in Example 154, step (b) to give 24 mg (71.8%)of4-{2-[(2 chlorophenyl)amino]-(1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.71 (s, 3H), 7.04 (td, 1H, J=1.4, 7.8 Hz), 7.21 (s, 1H), 7.35 (t, 1H, J=8.5 Hz), 8.42 (s, 1H), 8.57 (dd, 1H, J=1.3, 8.3 30 Hz), 8.80 (bs, 2H), 9.26 (bs, 2H), 9.79 (s, 1H); Mass Spectrum (ESI) m/z calcd. for
C,
5 sHI 4
N
4
S
3 C1, 380.94 (M+H), found 381.1.
WO 00/47578 -195- PCT/US99/18065 Example 156 a) Methyl 4-(2-amino(1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5-methylthiothiophene-2 carboxylate (50 mg, 0.16 mmol) was allowed to react with thiourea (12 mg) as 5 described in Example 154, step (a), to give 54 mg (70% yield) of methyl 4-(2-amino (1,3-thiazol-4-yl))-5-methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.69 (s, 3H), 3.83 (s, 3H), 7.00 (s, 1H), 8.05 (s, 1H); Mass Spectrum (ESI) m/z calcd. for Co 10
H
10 0 2
S
3
N
2 , 286.41 (M+H), found 287.1; b) 4-(2-Amino-(1,3-thiazol-4-yl))-5-methylthiothiophene-2-carboxamidine 10 hydrochloride: Methyl 4-(2-amino-(1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxylate hydrobromide (110 mg, 0.29 mmol) was treated as described in Example 154, step (b). The resultant amidine (74 mg) was stirred in 3 mL of dry methanol under N 2 and treated with ca. lmL of ether saturated with dry HCI gas. Dry ether (1.5 mL) was then added and the result was allowed to sit for 2 h at ambient temperature 15 and then filtered to give 40 mg (45% yield) of 4-(2-amino-(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.69 (s, 3H), 6.90 (s, 1H), 8.44 (s, 1H), 9.20, 9.42 (s, 4H, NH); Mass Spectrum (ESI) m/z calcd.C 9 HioN 4
S
3 , 270.4 (M+H), found 271.2. 20 Example 157 a) Methyl 4-{2-[(2,5-dimethoxyphenyl)amino]j(1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (49.4 mg, 0.15 mmol) was allowed to react with 2,5-dimethoxy phenyl thiourea (37.2 mg) as described in Example 154, step (a), to 25 give 65.5 mg (87% yield) of methyl 4-({2-[(2,5-dimethoxyphenyl)amino](1,3-thiazol 4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.66 (s, 3H), 3.76 (s, 3H), 3.81 (s, 3H), 3.83 (s, 3H), 6.49 (dd, 1H, J=3.0, 8.8 Hz), 6.92 (d, 1H, J=8.9 Hz), 7.26 (s, 1H), 8.17 (s, 1H), 8.37 (d, 1H. J=3.1 Hz), 9.70 (s, 1H); Mass Spectrum (ESI) m/z calcd. for CIHisN 2 0 4
S
3 , 422.54 (M+H), found 423.1. 30 b) 4-{2-[(2,5-Dimethoxyphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine: Methyl 4- { 2-[(2,5 dimethoxyphenyl)amino]-(1,3-thiazol-4-yl) } -5-methylthiothiophene-2-carboxylate WO 00/47578 PCT/US99/18065 -196 hydrobromide (45.5 mg, 0.09 mmol) was treated as described in Example 154, step (b), followed by preparative thin layer chromatography (500 mm silica gel plate, J.T. Baker, Phillipsburg, NJ, 10%-methanol-CH,Cl 2 -sat'd. NH 3 eluent) to give 9.9 mg (27% yield of 4- {2-[(2,5-dimethoxyphenyl)amino](1,3-thiazol-4-yl)}-5 5 methylthiothiophene-2-carboxamidine. 'H NMR (DMSO-d 6 , 300 MHz) 6 2.60 (s, 3H), 3.73 (s, 3H), 3.81 (s, 3H), 6.48 (dd, 1H, J=3.1, 8.8 Hz), 6.92 (d, 1H, J=7.9 Hz), 7.05 (s, 1H), 7.5 (bs, 2H), 8.04 (s, 1H), 8.34 (d, 1H, J=1.0 Hz), 9.6 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C, 7
H,
8
N
4 0 2
S
3 , 406.55 (M+H), found 407.1. 10 Example 158 a) Methyl 4-{2-[(3-methoxyphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (53.3 mg, 0.17 mmol) was allowed to react with 2-methoxy phenyl thiourea (34.5 mg) as described in Example 154, step (a), to give 15 61 mg (76% yield) of methyl 4-{2-[(3-methoxyphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 5 2.67 (s, 3H), 3.78 (s, 3H), 3.83 (s, 3H), 6.53 (d, 1H, J=6.8 Hz), 7.13-7.24 (m, 2H), 7.29 (s, 3H), 7.59 (m, 1H), 8.16 (s, 3H), 10.32 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C,
,
H,
6
N
2 0 3
S
3 , 392.52 (M+H), found 393.2. 20 b) 4-{2-J(3-Methlioxyphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4- {2-[(3 methoxyphenyl)amino]-(1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (54.6 mg, 0.11 mmol) was treated as described in Example 154, step (b) to give 25.2 mg (56%) of 4-{2-[(3-methoxyphenyl)amino](1,3-thiazol-4-yl)}-5 25 methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.71 (s, 3H), 3.77 (s, 3H), 6.54 (m, 1H), 7.15 (s, 3H), 7.19-7.28 (m, 2H), 7.47 (m, 1H), 8.46 (s, 1H), 8.86 (bs, 2H), 9.28 (bs, 2H), 10.36 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C, 6
H,
6
N
4 0S 3 , 376.52 (M+H), found 377.2. 30 Example 159 a) Methyl 4-[2-[(4-methoxyphenyl)amino](1,3-thiazol-4-yl)}-S methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5- WO 00/47578 PCT/US99/18065 -197 methylthiothiophene-2-carboxylate (41.3 mg, 0.13 mmol) was allowed to react with 5-methoxy phenyl thiourea (26.8 mg) as described in Example 154, step (a) to give 25 mg (41% yield) of methyl 4-{2-[(4-methoxyphenyl)aminoj(1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 5 2.64, 2.68 (s, 3H rotomer) ), 3.72, 3.73 (s, 3H rotomer), 3.83 (s, 3H), 6.91 (dd, 2H, J=6.7, 8.8 Hz), 7.21 (s, 1H), 7.59 (d, 1H, J=9.0 Hz), 7.67 (d, 1H, J=9.0 Hz), 8.05, 8.13 (s, 1H rotomer), 10.16, 10.34 (bs, 1H, rotomer); Mass Spectrum (ESI) m/z calcd. for
C,
7 Hj 6
N
2 0 2
S
3 , 392.52 (M+H), found 393.1. b) 4-{2-[(4-Methoxyphenyl)amino](1,3-thiazol-4-yl)}-5 10 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-{2-[(4 methoxyphenyl)amino]-(1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (22 mg, 0.046 mmol) was treated as described in Example 154, step (b) to give 11.5 mg (61% yield) of 4-{2-[(4-methoxyphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 15 8 2.72 (s, 3H), 3.73 (s, 3H), 6.91 (d, 2H, J=9.0 Hz), 7.08 (s, 1H), 7.69 (d, 2H, J=9.1 Hz), 8.44 (s, 1H), 8.83 (bs, 2H), 9.28 (bs, 2H), 10.15 (s, 1H);Mass Spectrum (ESI) m/z calcd. for C, 6 H16N 4 0S 3 , 376.52 (M+H), found 377.1. Example 160 20 a) Methyl 4-(2-{[4-(dimethylamino)phenyl]aminoj(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (50 mg, 0.16 mmol) was allowed to react with 4 N,N-dimethylaminophenyl thiourea (31.5 mg) as described in Example 154, step (a), to give 53.2 mg (75% yield) of methyl 4-(2-{ [4-(dimethylamino)phenyl]amino} (1,3 25 thiazol-4-yl))-5-methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO d 6 , 300 MHz) 8 2.69 (s, 3H), 3.15 (s, 6H), 3.83 (s, 3H), 7.36 (s, 1H), 7.55 (bs, 2H), 7.88 (d, 2H, J=8.3 Hz), 8.16 (s, 1H), 10.56 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C, 8
HI
9
N
3 0 2
S
3 , 405.56 (M+H), found 406.1. b) 4-(2-{[14-(Dimethylamino)phenylfaminoj(1,3-thiazol-4-yl))-5 30 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-(2-{[4 (dimethylamino)phenyl]amino}(1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxylate hydrobromide (50 mg, 0.10 mmol) was treated as described in Example WO 00/47578 PCT/US99/18065 -198 154, step (b) to give 9.4 mg (22% yield) of 4-{2-[(4-methoxyphenyl)amino](1,3 thiazol-4-yl)} -5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 2.70 (s, 3H), 2.84 (s, 6H), 6.75 (d, 2H, J=9.2 Hz), 7.00 (s, 1H), 7.56 (d, 2H, J=9.1 Hz), 8.31 (s, 1H), 8.68 (bs, 3H), 9.92 (bs, 1H). 5 Example 161 a) Methyl 4-{2-[(4-chloro-2-methylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (50 mg, 0.16 mmol) was allowed to react with 2 10 methyl-4-chlorophenyl thiourea (32.1 mg) as described in Example 154, step (a), to give 62.2 mg (79% yield) of methyl 4-{2-[(4-chloro-2-methylphenyl)amino](1,3 thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO d 6 , 300 MHz) 8 2.28, 2.29 (s, 3H rotomer), 2.62, 2.66 (s, 3H rotomer), 3.82 (s, 3H), 7.21-7.29 (m, 3H), 8.04, 8.11 (s, 1H rotomer), 8.17 (d, 1H, J=8.8 Hz), 8.30 (d, 1H, 15 J=8.4 Hz), 9.44 (s, 1H), 9.59 (s, 1H); Mass Spectrum (ESI) m/z calcd. for
C,
7
H,
5 C1N 2 0 2
S
3 , 410.96 (M+H), found 411.1. b) 4-{2-J(4-Chloro-2-methylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-{2-[(4-chloro-2 methylphenyl)amino]-(1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate 20 hydrobromide (55 mg, 0.17 mmol) was treated as described in Example 154, step (b) to give 16 mg (22% yield) of 4-{2-[(4-chloro-2-methylphenyl)amino](1,3-thiazol-4 yl)}-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 5 2.30 (s, 3H), 2.70 (s, 3H), 7.15 (s, 1H), 7.23 -7.29 (mn, 2H), 8.34 (d, 1H, J=8.6 Hz), 8.44 (s, 1H), 8.86 (bs, 2H), 9.29 (bs, 2H), 9.47 (s, 1H); Mass Spectrum (ESI) m/z 25 calcd. for C, 6
H,
5 C1N 4
S
3 , 394.97 (M+H), found 395.1. Example 162 a) Methyl 4-{2-[(diphenylmethyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 30 methylthiothiophene-2-carboxylate (50 mg, 0.16 mmol) was allowed to react with diphenylmethane thiourea (38 mg) as described in Example 154, step (a), to give 145 mg (100% yield) of methyl 4-{2-[(diphenylmethyl)amino](1,3-thiazol-4-yl)}-5- WO 00/47578 -199- PCT/US99/18065 methylthiothiophene-2-carboxylate hydrobromide after removal of solvent in vacuo. 'H NMR (DMSO-d 6 , 300 MHz) 5 2.50 (s, 3H), 2.80 (s, 3H), 6.13, 6.18 (d, 1H rotomer, J=7.9 Hz), 7.23-7.41 (m, 11H), 8.00, 8.02 (s, 1H rotomer), 8.73, 8.86 (d, 1H, rotomer, J=8.0 Hz); Mass Spectrum (ESI) m/z calcd. for C 23
H
20
N
2 0 2
S
3 , 452.62 5 (M+H), found 453.0. b) 4-{2-J(Diphenylmethyl)aminoj(1,3-thiazol-4-yl)}-5-methylthiothiophene 2-carboxamidine: Methyl 4-{2-[(diphenylmethyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide. (96.3 mg. 0.18 mmol) was treated as described in Example 154, step (b) to give 16 mg (20% yield) of 4-{2 10 [(diphenylmethyl)amino](1,3-thiazol-4-yl) }-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 2.59 (s, 3H), 6.23 (d, 1H, J=7.9 Hz), 6.84 (s, 1H), 7.22-7.40 (m, 10 H), 8.09 (bs, 3H), 8.12 (s, 1H), 8.68 (d, 1H, J=8.4 Hz); Mass Spectrum (ESI) m/z calcd. for C 22
H
20
N
4
S
3 , 436.62 (M+H), found 437.1. 15 Example 163 a) Methyl 5-methylthio-4-{2-[(3-phenylpropyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (131 mg, 0.42 mmol) was allowed to react with propylphenyl thiourea (82.3 mg) in DMF as described in Example 154, step (a), then 20 filtered through a 5 cm pad of silica gel in a 15 mL glass fritted funnel with 10% methanol-CHCl 3 . Concentration of the solvent in vacuo gave 203 mg (100% yield) of methyl 5-methylthio-4- { 2-[(3-phenylpropyl)amino](1,3-thiazol-4-yl) }thiophene-2 carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.89 (m, 2H), 2.62 (s, 3H), 2.63-2.71 (m, 2H), 3.27-3.39 (m, 2H), 3.82 (s, 3H), 6.97 (s, 1H), 7.15-7.31 (m, 25 5H), 8.06 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C1 9
H
2 0
N
2 0 2
S
3 , 404.57 (M+H), found 405.1. b) 5-Methylthio-4-{2-[(3-phenylpropyl)amino](1,3-thiazol-4-yl)}thiophene 2-carboxamidine hydrochloride: Methyl -5-methylthio-4-{2- [(3 phenylpropyl)amino](1,3-thiazol-4-yl)}thiophene-2-carboxylate hydrobromide (112 30 mg, 0.23 mmol) was treated as described in Example 154, step (b) to give 16 mg (16% yield) of 4- { 2-[(diphenylmethyl)amino](1,3-thiazol-4-yl) }-5 methylthiothiophene-2-carboxamidine hydrochloride, which was further purified by WO 00/47578 -200- PCT/US99/18065 preparative thin layer chromatography using 20%-methanol-CH 2 Cl 2 -sat'd. NH 3 as eluent. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.89 (m, 2H), 2.54 (s, 1H), 2.66 (at, 2H, J=7.3 Hz), 3.31 (m, 2H), 6.69 (bs, 3H), 6.76 (s, 1H), 7.15-7.31 (m, 5H), 7.69 (m, 1H), 7.84 (s, 1H); Mass Spectrum (ESI) m/z calcd. for Cj 8
H
2 0
N
4
S
3 , 388.58 (M+H), found 5 389.2. Example 164 a) Methyl 5-methylthio-4-{2-J(2,4,5-trimethylphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 10 methylthiothiophene-2-carboxylate (60 mg, 0.21 mmol) was allowed to react with 2,4,5-trimethylphenyl thiourea as described in Example 154, step (a) to give 42.3 mg (41% yield) of methyl 5-methylthio-4- {2-[(2,4,5-trimethylphenyl)amino](1,3-thiazol 4-yl)}thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.16 (s, 3H), 2.18 (s, 3H), 2.19 (s, 3H), 2.64 (s, 3H), 3.82 (s, 3H), 6.97 (s, 1H), 7.18 (s, 15 1H), 7.86 (s, 1H), 8.12 (s, 1H), 9.29 (s, 1H); Mass Spectrum (ESI) m/z calcd. for Cj 9
H
2 0
N
2 0 2
S
3 , 404.57 (M+H), found 405.1. b) 5-Methylthio-4-{2-[(2,4,5-trimethylphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxamidine hydrochloride: Methyl -5-methylthio-4-{ 2-[(2,4,5 trimethylphenyl)amino](1,3-thiazol-4-yl)}thiophene-2-carboxylate hydrobromide 20 (37.3 mg, 0.07 mmol) was treated as described in Example 154, step (b) to give 28.3 mg (95% yield) of 5-methylthio-4- {2-[(2,4,5-trimethylphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 6 2.16 (s, 3H), 2.19 (s, 3H), 2.20 (s, 3H), 2.68 (s, 3H), 6.97 (s, 1H), 7.03 (s, 1H), 7.84 (s, 1H), 8.41 (s, 1H), 8.84 (bs, 2H), 9.26 (bs, 3H); Mass Spectrum (ESI) m/z calcd. for 25 C18H 20
N
4
S
3 , 388.58 (M+H), found 389.2. Example 165 a) Methyl 4-{2-J(2-fluorophenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothliophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 30 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with 2 fluorophenyl thiourea as described in Example 154, step (a) to give 55.6 mg (70% yield) of methyl 4- {2-[(2-fluorophenyl)amino](1,3-thiazol-4-yl) }-5- WO 00/47578 -201- PCT/US99/18065 methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.68 (s, 3H), 3.83 (s, 3H), 6.96-7.04 (m, 1H), 7.14-7.29 (m, 3H), 7.35 (s, 1H), 8.06, 8.14 (s, 1H rotomer), 8.61 (td, 1H rotomer, J=l.5, 8.5 Hz), 10.14, 10.30 (s, 1H rotomer); Mass Spectrum (ESI) m/z calcd. for C, 6
H
13
FN
2 0 2
S
3 , 380.48 (M+H), found 5 381.1. b) 4-{2-J(2-Fluorophenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene 2-carboxamidine hydrochloride: Methyl 4-{2-[(2-fluorophenyl)amino](1,3-thiazol 4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (55.6 mg, 0.13 mmol)) was treated as described in Example 154, step (b) to give 12.4 mg (24 %) of 4-{2-[(2 10 fluorophenyl)amino](1,3-thiazol-4-yl) }-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz); 8 2.72 (s, 3H), 3.16 (s, 3H), 6.97 7.08 (m, 1H), 7.18-7.36 (m, 4H), 8.49 (s, 1H), 8.70 (td, 1H, 1.4, 8.4 Hz), 8.92 (bs, 2H), 9.32 (bs, 2H), 10.18 (d, 1H, J=1.6 Hz); Mass Spectrum (ESI) m/z calcd. for C1 5
H
13
FN
4
S
3 , 364.49 (M+H), found 365.1. 15 Example 166 a) Methyl 4-{2-J(3-chloro-2-methylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with 2 20 methyl-3-chlorophenyl thiourea (39 mg) as described in Example 154, step (a) to give 61.8 mg (66% yield) of methyl 4- {2-[(3-chloro-2-methylphenyl)amino](1,3-thiazol-4 yl)}-5-methylthiothiophene-2-carboxylate hydrobromide. Mass Spectrum (ESI) m/z calcd. for C,, 7
H,
5 C1N 2 0 2
S
3 , 410.96 (M+H), found 411.1. b) 4-{2-[(3-Chloro-2-methylphenyl)amino](1,3-thiazol-4-yl)}-5 25 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-{2-[(3-chloro-2 methylphenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (61.8 mg, 0.12 mmol) was treated as described in Example 154, step (b) to give 46.7 mg (90%/o yield) of 4-{2-[(3-chloro-2-methylphenyl)amino](1,3 thiazol-4-yl) } -5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR 30 (DMSO-d 6 , 300 MHz) 8 2.34 (s, 3H), 2.69 (s, 3H), 7.15 (s, 1H), 7.18-7.26 (m, 2H), 8.12 (d, 1H, J=7.9 Hz), 8.41 (s, 1H), 8.84 (bs, 2H), 9.27 (bs, 2H), 9.61 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C, 6
H,
5
CIN
4
S
3 , 394.97 (M+H), found 395.1.
WO 00/47578 PCT/US99/18065 -202 Example 167 a) Methyl 4-(2-{[J2-(methylethyl)phenylfaminoj(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 5 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with 2 isopropyl phenyl thiourea (40 mg) as described in Example 154, step (a) to give 33.1 mg (36% yield) of methyl 4-(2- { [2-(methylethyl)phenyl]amino} (1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide. 1H NMR (DMSO-d 6 , 300 MHz) 6 1.17 (d, 6H, J=6.7 Hz), 2.60, 2.65 (s, 3H rotomer), 3.27 (s, 1H), 3.82 (s, 3H), 7.13 10 (s, 1H), 7.14-7.25 (m, 2H), 7.34-7.37 (m, 1H), 7.78 (m, 1H), 7.99, 8.08 (s, 1H rotomer), 9.52, 9.61 (bs, 1H rotomer); Mass Spectrum (ESI) m/z calcd. for
C
1 9
H
2 0
N
2 0 2
S
3 , 404.57 (M+H), found 405.1. b) 4-(2-{[2-(Methylethyl)phenyl]amino}(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-(2- { [2 15 (methylethyl)phenyl]amino } (1,3-thiazol-4-yl))-5-methylthiothiophene-2-carboxylate hydrobromide (33.1 mg, 0.06 mmol) was treated as described in Example 154, step (b) to give 22.4 mg (88%) of 4-(2- { [2-(methylethyl)phenyl]amino } (1,3-thiazol-4-yl)) 5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.19 (d, 6H, J=6.8 Hz), 2.70 (s, 3H), 3.32 (m, 1H), 7.04 (s, 1H), 7.14-7.25 (m, 20 2H), 7.35 (dd, 1H, J=1.4, 7.5 Hz), 7.86 (dd, 1H, J=1.4, 7.9 Hz), 8.37 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C, 8
H
20
N
4
S
3 , 388.58 (M+H), found 389.2. Example 168 a) Methyl 5-methylthio-4-(2-{[4-(phenylmethoxy)phenyl]amino](1,3-thiazol 25 4-yl))thiophene-2-carboxylate: Methyl 4-(2-bromoacetyl)-5-methylthiothiophene-2 carboxylate (336.3 mg, 1.08 mmol) was allowed to react with 4-benzyloxyphenyl thiourea (279 mg) as described in Example 154, step (a) to give 450 mg (76% yield) of methyl 4-(2- { [4-phenylmethoxyphenyl]amino} (1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide. Mass Spectrum (ESI) m/z calcd. 30 for C 2 3
H
2 0N 2 0 3
S
3 , 468.61 (M+H), found 469.2. b) 5-Methylthio-4-(2-{[4-(phenylmethoxy)phenyl]amino}(1,3-thiazol-4 yl))thiophene-2-carboxamidine hydrochloride: Methyl 4-(2-{ [4- WO 00/47578 PCT/US99/18065 -203 phenylmethoxyphenyl]amino}(1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxylate hydrobromide (100 mg, 0.18 mmol) was treated as described in Example 154, step (b) to give 23.9 mg (27% yield) 5-methylthio-4-(2-{ [4 (phenylmethoxy)phenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxamidine 5 hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.73 (s, 3H), 5.08 (s, 2H), 7.00 (d, 2H, J=8.2 Hz), 7.09 (s, 1H), 7.31-7.47 (m, 5H), 7.70 (d, 2H, J=8.0 Hz), 8.47 (s, 1H), 8.88 (bs, 2H), 9.30 (bs, 2H), 10.20 (s, 1H); Mass Spectrum (ESI) m/z calcd. for
C
22
H
2 0N 4 0S 3 , 452.62 (M+H), found 453.1. 10 Example 169 a) Methyl 4-{2-[(2-bromophenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with 2 bromophenyl thiourea (44 mg) as described in Example 154, step (a) to give 63.1 mg 15 (64% yield) of methyl 4-{2-[(2-bromophenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.65 (s, 3H), 3.82 (s, 3H), 7.00 (m, 1H), 7.33 (s, 1H), 7.40 (m, 1H), 7.64 (dd, 1H, J=1.4, 7.9 Hz), 8.04, 8.11 (s, 1H rotomer), 8.27, 8.37 (dd, 1H 9.60, 9.80 (bs, 1H rotomer, J=1.5, 8.2 Hz), Mass Spectrum (ESI) m/z calcd. for C 16
H,
3 BrN,02S 3 , 441.39 20 (M+H), found 441.1. b) 4-{2-[(2-Bromophenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene 2-carboxamidine hydrochloride: Methyl 4- { 2-[(2-bromophenyl)amino](1,3-thiazol 4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (63.1mg, 0.12 mmol) was treated as described in Example 154, step (b) to give 47.9 mg (86% yield) of 4-{2-[(2 25 bromophenyl)amino](1,3-thiazol-4-yl) }-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.70 (s, 3H), 7.01 (m 1H), 7.20 (s, 1H), 7.40 (m, 1H), 7.65 (dd, 1H, J=1.5, 8.0), 8.38 (dd, 1H, J=1.5, 8.3 Hz), 8.44 (s, 1H), 8.89 (bs, 2H), 9.30 (bs, 2H), 9.62 (s, 1H); Mass Spectrum (ESI) m/z calcd. for
C,
5
H,
3 BrN 4
S
3 , 425.39 (M+H), found 425.1. 30 WO 00/47578 PCT/US99/18065 -204 Example 170 a) Methyl 4-{2-J(2,6-dichlorophenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with 5 2,6-dichlorophenyl thiourea (42 mg) as described in Example 154, step (a) to give 63.1 mg (65% yield) of methyl 4-{2-[(2,6-dichlorophenyl)amino](1,3-thiazol-4-yl)} 5-methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.59 (s, 3H), 3.8 (s, 3H), 7.15 (s, 1H), 7.36 (m, 1H), 7.61 (m, 2H), 7.97 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C! 6
HI
2 C1 2
N
2 0 2
S
3 , 431.38 (M+H), found 431.0. 10 b) 4-{2-[(2,6-Dichlorophenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4- { 2-[(2,6 dichlorophenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (43 mg, 0.08 mmol) was treated as described in Example 154, step (b) to give 14.5 mg (40% yield) of 4- { 2-[(2,6-dichlorophenyl)amino](1,3-thiazol-4-yl)} 15 5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.69 (s, 3H), 7.15 (s, 1H), 7.18-7.26 (m, 2H), 8.13 (d, 1H, J=7.5 Hz), 8.41 (s, 1H), 8.84 (bs, 2H), 9.27 (bs, 2H), 9.61 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for
C
15
H
12
CI
2
N
4
S
3 , 415.39 (M+H), found 415.1. 20 Example 171 a) Methyl 4-{2-[(2-bromo-4-methylphenyl)aminoj(1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with 2 25 bromo-4-methylphenyl thiourea (47 mg) as described in Example 154, step (a) to give 62 mg (61% yield) of methyl 4- {2-[(2-bromo-4-methylphenyl)amino](1,3-thiazol-4 yl)}-5-methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 5 2.28 (s, 3H), 3.82 (s, 3H), 7.21 (m, 1H), 7.27 (s, 1H), 7.48 (m, 1H), 8.14, 8.17 (s, 1H rotomer), 9.52, 9.72 (bs, 1H rotomer); Mass Spectrum (ESI) m/z calcd. for 30 C, 7 HBrN202S3, 455.42 (M+H), found 455.0. b) 4-{2-[f(2-Bromo-4-methylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-{2-[(2-bromo-4- WO 00/47578 PCT/US99/18065 -205 methylpheny)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (62 mg, 0.11 mmol) was treated as described in Example 154, step (b) to give 26 mg (50% yield) of 4-{2-[(2-bromo-4-methylphenyl)amino](1,3-thiazol-4 yl) }-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 5 300 MHz) 8 2.28 (s, 3H), 2.70 (s, 3H), 7.14 (s, 1H), 7.21 (dd, 1H, J=1.6, 8.5 Hz), 7.49 (d, 1H, J=1.5 Hz), 8.16 (d, 1H, 8.3 Hz), 8.41 (s, 1H), 8.85 (bs, 2H), 9.28 (bs, 2H), 9.53 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 16 HsBrN 4
S
3 , 439.42 (M+H), found 439.1. 10 Example 172 a) Methyl 5-methylthio-4-{2-[(2-morpholin-4-ylethyl)amino]j(1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (100 mg, 0.32 mmol), was allowed to react with 1-ethylmorpholinothiourea (61.2 mg) as described in Example 154, step (a) to give 15 120.8 mg (79% yield) methyl 5-methylthio-4- {2-[(2-morpholin-4-ylethyl)amino](1,3 thiazol-4-yl)}thiophene-2-carboxylate hydrobromide. 1 H NMR (CD 3 OD, 300 MHz) 8 2.64 (s, 3H), 3.43-3.52 (m, 5H), 3.83-3.86 (m, 10H), 6.95 (s, 1H), 8.04 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 16
H
2 1
N
3 0 3
S
3 , 399.55 (M+H), found 400.1. b) 5-Methylthio-4-{2-[(2-morpholin-4-ylethyl)amino](1,3-thiazol-4 20 yl)}thiophene-2-carboxylate hydrochloride: Methyl- 5-methylthio-4-{2-[(2 morpholin-4-ylethyl)amino](1,3-thiazol-4-yl)}thiophene-2-carboxylate hydrobromide (62 mg, 0.12 mmol) was treated as described in Example 154, step (b) to give 26 mg (52% yield) of 5-methylthio-4-{2-[(2-morpholin-4-ylethyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.69 25 (s, 3H), 3.16-3.95 (m, 15H), 6.96 (s, 1H), 8.01 (bs, 1H), 8.49 (s, 1H), 8.84 (bs, 2H), 9.28 (bs, 2H), 10.49 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C,sH 21
N
5 0S 3 , 383.56 (M+H), found 384.2. Example 173 30 a) Methyl 4-{2-[(2,3-dichlorophenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with WO 00/47578 -206- PCT/US99/18065 2,3-dichlorophenylthiourea (42 mg) as described in Example 154, step (a) to give 60.5 mg (62% yield) methyl 4-{2-[(2,3-dichlorophenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.66 (s, 3H), 3.82 (s, 3H), 7.27 (dd, 1H, J=1.5, 6.5 Hz), 7.36 (d, 1H, J=8.2 Hz), 7.43 5 (s, 1H), 8.14 (s, 1H), 8.62 (dd, 1H, J=1.5, 8.4 Hz), 9.95 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C, 6
H
12 C1 2
N
2 0 2
S
3 , 431.38 (M+H), found 431.1. b) 4-2-[(2,3-Dichlorophenyl)amino](1,3-thiazol-4-yl)}-S methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-{2-[(2,3 dichlorophenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate 10 hydrobromide (60.5 mg, 0.11 mmol) was treated as described in Example 154, step (b) to give 15 mg (30% yield) of 4-{2-[(2,3-dichlorophenyl)amino](1,3-thiazol-4-yl)} 5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 5 2.71 (s, 3H), 7.27-7.28-7.41 (m, 2H), 8.45 (s, 1H), 8.63 (dd, 1H, J=1.5, 8.4 Hz), 8.84 (bs, 2H), 9.29 (bs, 2H), 9.99 (s, 1H); Mass Spectrum (ESI) m/z calcd. for 15 CisHI 2 C1 2
N
4
S
3 , 415.34 (M+H), found 415.1. Example 174 a) Methyl 5-methylthio-4-{2-J(3,4,5-trimethoxyphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 20 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with 2,3,4-trimethoxyphenylthiourea (46 mg) as described in Example 154, step (a) to give 61.8 mg (63% yield) of methyl 5-methylthio-4-{2-[(3,4,5 trimethoxyphenyl)amino](1,3-thiazol-4-yl)}thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.67 (s, 3H), 3.81 (s, 6H), 3.82 (s, 3H), 7.11 (s, 2H), 25 7.25 (s, 1H), 8.19 (s, 1H), 10.25 (s, 1H); Mass Spectrum (ESI) m/z calcd. for
C,
8
H
2 0
N
4 0 3
S
3 , 436.56 (M+H), found 437.1. b) 5-Methylthio-4-(2-[(3,4,5-trimethoxyphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxamidine hydrochloride: Methyl 5-methylthio-4-{2-[(3,4,5 trimethoxyphenyl)amino](1,3-thiazol-4-yl)}thiophene-2-carboxylate hydrobromide 30 (61.8 mg, 0.11 mmol) was treated as described in Example 154, step (b) to give 14 mg (27% yield) of 5-methylthio-4-{2-[(3,4,5-trimethoxyphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 6 WO 00/47578 -207- PCT/US99/18065 2.70 (s, 3H), 3.61 (s, 3H), 3.80 (s, 6H), 7.08 (s, 2H), 7.14 (s, 1H), 8.44 (s, 1H), 8.84 (bs, 2H), 9.26 (bs, 2H), 10.29 (s, 1H); Mass Spectrum (ESI) m/z calcd. for
C
18 sH 2 0
N
4 0 3
S
3 , 436.56 (M+H), found 437.1. 5 Example 175 a) Methyl 5-methylthio-4-{2-J(2-piperidylethyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (100 mg, 0.32 mmol) was allowed to react with N ethylpiperidylthiourea (60.6 mg) as described in Example 154, step (a) to give 90 mg 10 (59% yield) of methyl 5-methylthio-4- {2-[(2-piperidylethyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrobromide. 1 H NMR (DMSO-d 6 , 300 MHz) 6 1.41 (m, 2H), 1.70-1.79 (m, 6H), 2.65 (s, 3H), 2.95 (m, 2H), 3.52 (m, 2H), 3.73 (m, 2H), 3.82 (s, 3H), 7.08 (s, 1H), 7.96 (at, 1H, J=5.3 Hz), 8.09 (s, 1H), 9.40 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C 17
H
23
N
3 0 2
S
3 , 397.6 (M+H), found 398.1. 15 b) 5-Methylthio-4-{2-[(2-piperidylethyl)amino](1,3-thiazol-4-yl)}thiophene 2-carboxamidine hydrochloride: Methyl 5-methylthio-4- { 2-[(2 piperidylethyl)amino](1,3-thiazol-4-yl) } thiophene-2-carboxylate hydrobromide (72 mg, 0.15 mmol) was treated as described in Example 154, step (b) to give 26.8 mg (43% yield) of 5-methylthio-4- {2-[(2-piperidylethyl)amino](1,3-thiazol-4 20 yl)}thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.40 (m, 2H), 1.72-1.79 (m, 6H), 2.69 (s, 3H), 2.96 (m, 2H), 3.51 (m, 2H), 3.76 (m, 2H), 6.97 (s, 1H), 8.08 (t, 1H, J=5.5 Hz), 8.60 (s, 1H), 8.95 (bs, 1H), 9.35 (bs, 2H), 10.25 (s, 1H); Mass Spectrum (ESI) m/z calcd. for Ci 6
H
2 3
N
5
S
3 , 381.1 (M+H), found 382.2. 25 Example 176 a) Methyl 4-(2-{[(4-methylphenyl)methyl]amino}(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (111 mg, 0.35 mmol) was allowed to react with 4 methylphenylmethylthiourea as described in Example 154, step (a) to give 125 mg 30 (81% yield) of methyl 4-(2- { [(4-methylphenyl)methyl]amino } (1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide. Mass Spectrum (ESI) m/z calcd. for C, 1 8
H
18
N
2 0 2
S
2 , 358.5 (M+H), found 359.1.
WO 00/47578 -208- PCT/US99/18065 b) 4-(2-{[(4-Methylphenyl)methylfamino} (1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-(2- { [(4 methylphenyl)methyl]amino } (1,3-thiazol-4-yl))-5-methylthiothiophene-2-carboxylate hydrobromide (118 mg, 0.26 mmol) was treated as described in Example 154, step (b) 5 to give 58.2 mg (54% yield) of 4-(2-{ [(4-methylphenyl)methyl]amino}(1,3-thiazol-4 yl))-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.27 (s, 3H), 2.66 (s, 3H), 4.49 (d, 2H, J=5.7 Hz), 6.88 (s, 1H), 7.13 (d, 2H, J=7.8 Hz), 7.27 (d, 2H, J=8.0 Hz), 8.20 (t, 1H, J=5.8 Hz), 8.42 (s, 1H), 8.90 (bs, 2H), 9.27 (bs, 2H); Mass Spectrum (ESI) m/z calcd. for C, 7
H,
8
N
4
S
3 , 374.55 (M+H), 10 found 375.2. Example 177 a) Amino{[4-(4-chlorophenoxy)phenyl]amino}methane-1-thione: Unless otherwise indicated, all thioureas, isothiocyanates, thioamides and amines were 15 purchased from Maybridge Chemical Co. Ltd.(Cornwall, U.K.), Transworld Chemical Co. (Rockville, MD), or Aldrich Chemical Co., (Milwaukee, WI). (i) 4-Amino 4'-chlorodiphenylether (TCI America, Portland OR, 520 mg, 2.03 mmol) was slurried in 10 mL of ether and treated with ca. 1 mL of ether saturated with HCJ gas. After 5 min. the solvent was removed in vacuo. To a stirring biphasic solution amine-HCI 20 salt in 20 mL CHC1 3 -sat'd NaHCO 3 (1:1, v/v) at ambient temperature was added thiophosgene (1.2 equiv, 2.4 mmol) in 5 mL of CHC1 3 dropwise via an addition funnel. The reaction was vigorously stirred for 1 h (TLC, 50% ethyl acetate-hexanes indicates clean conversion to a higher Rf spot), at which time the layers were separated, the aqueous layer extracted with CHC1 3 (1x20 mL), and the combined 25 organic layers washed with brine (1x20 mL) and dried (Na 2
SO
4 ). Concentration of the solvent in vacuo yielded the crude 4-(4-chlorophenoxy)-phenylisothiocyanate (414 mg). (ii) The 4-(4-chlorophenoxy)-phenylisothiocyanate was transferred to an Ace Glass pressure tube equipped with a Teflon coated stir bar and treated with a 2.0 M solution of NH 3 in 5 ml methanol (Aldrich Chemical Co., Milwaukee, WI)). The tube 30 was sealed and immersed in a 80 oC oil bath. After 2 h, the reaction was cooled to 0 'C in an ice bath. The precipitates were filtered and dried under vacuum to yield amino { [4-(4-chlorophenoxy)phenyl]amino }methane- 1-thione (328 mg, 79%). 'H WO 00/47578 PCT/US99/18065 -209 NMR (DMSO-d 6 , 300 MHz) 8 7.02 (m, 4H), 7.41 (m, 4H), 9.65 (s, 1H); Mass Spectrum (ESI) m/z called. for C, 3
H,,CIN
2 OS, 278.8 (M+H), found 279.4. b) Methyl 4 -(2-{[4-(4-chlorophenoxy)phenyl]amino(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 5 methylthiothiophene-2-carboxylate (309 mg, 1.0 mmol) was allowed to react with amino { [4-(4-chlorophenoxy)phenyl]amino}methane-1 -thione (297 mg) as described in Example 154, step (a) to give 410 mg (72% yield) of methyl 4-(2- { [4-(4 chlorophenoxy)phenyl]amino} (1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxylate hydrobromide. Mass Spectrum (ESI) m/z calcd. for C 22
H,
7 C1N 2 0 3
S
3 , 10 489.1 (M+H), found 489.1. c) 4-(2-{[4-(4-Chlorophenoxy)phenyl]amino}(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-(2- {[4-(4 chlorophenoxy)phenyl]amino}(1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxylate hydrobromide (300 mg, 0.52 mmol) was treated as described in Example 15 154, step (b) to give 129.9 mg (49% yield) of 4-(2-{[4-(4 chlorophenoxy)phenyl]amino}(1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.72 (s, 3H), 6.97 (m, 2H), 7.07 (m, 2H), 7.15 (s, 1H), 7.40 (m, 2H), 7.85 (m, 2H), 8.46 (s, 1H), 8.82 (bs, 2H), 9.27 (bs, 2H), 10.43 (bss, 1H); Mass Spectrum (ESI) m/z calcd. for 20 C 21
H,
7
CIN
4 0S 3 , 473.1 (M+H), found 473.2, 475.1. Example 178 a) Methyl 5-methylthio-4-[2-({4-[5-(trifluoromethyl) (2 pyridyloxy)]phenyl}amino)(1,3-thiazol-4-yl)]thiophene-2-carboxylate: Methyl 4-(2 25 bromoacetyl)-5-methylthiothiophene-2-carboxylate (70 mg, 0.23 mmol) was allowed to react with 4 -[5-(trifluoromethyl)pyrid-2-yloxy]thiobenzamide (50 mg) as described in Example 154, step (a) to give 115 mg (98% yield) of methyl 5-methylthio-4-[2-({4 [5-(trifluoromethyl)(2-pyridyloxy)]phenyl}amino)(1,3-thiazol-4-yl)]thiophene-2 carboxylate. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.70 (s, 3H), 3.85 (s, 3H), 7.38 (m, 30 3H), 8.10 (m, 1H), 8.18 (s, 1H), 8.28 (dd, 1H, J=2.7, 8.8 Hz), 8.32 (s, 1H), 8.60 (m, 1H); Mass Spectrum (ESI) m/z calcd. for C 22
H
1 5
F
3
N
2 0 3
S
3 , 508.56 (M+H), found 509.2.
WO 00/47578 -210- PCTIUS99/18065 b) 5-Methylthio-4-[2-({4-[5JS-(trifluoromethyl) (2 pyridyloxy)]phenyl}amino)(1,3-thiazol-4-yl)]thiophene-2-carboxamidine hydrochloride: Methyl 5-methylthio-4-[2-({4-[5-(trifluoromethyl)(2 pyridyloxy)]phenyl } amino)(1,3-thiazol-4-yl)]thiophene-2-carboxylate (95 mg, 0.18 5 mmol) was treated as described in Example 154, step (b) to give 30.3 mg (32% yield) of 5-methylthio-4-[2-({4-[5-(trifluoromethyl)(2-pyridyloxy)]phenyl } amino)(1,3 thiazol-4-yl)]thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.75 (s, 3H), 7.34 (d, 1H, J=8.7 Hz), 7.41 (m, 2H), 8.01 (s, 1H), 8.10-8.14 (m, 2H), 8.29 (dd, 1H, J=2.5, 8.4 Hz), 8.60 (m, 1H), 8.63 (s, 1H), 8.91 (bs, 2H), 9.31 (bs, 10 2H); Mass Spectrum (ESI) m/z calcd. for C 21
H,
5
F
3
N
4 0S 3 , 492.6 (M+H), found 493.1. Example 179 a) Methyl 4-(2-{[4-phenoxyphenyl]amino}(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 15 methylthiothiophene-2-carboxylate (200 mg, 0.64 mmol) was allowed to react with 4 phenoxyphenylthiourea (158 mg) as described in Example 154, step (a) to give 300 mg (88% yield) of methyl 4-(2-{ [4-(phenoxy)phenyl]amino} (1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxylate hydrobromide. Mass Spectrum (ESI) m/z calcd. for C 22 HsN 2 0 3
S
3 , 454.6 (M+H), found 455.2. 20 b) 4-(2-{[4-Phenoxyphenyl]aminoj(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-(2- { [4 (phenoxy)phenyl]amino}(1,3-thiazol-4-yl))-5-methylthiothiophene-2-carboxylate hydrobromide (230 mg, 0.42 mmol) was treated as described in Example 154, step (b) and purified by preparative thin layer chromatography (20% methanol-CH 2 Cl 2 -sat'd. 25 NH 3 , 500 mm silica gel plate, J.T. Baker, Phillipsburg, NJ) to give 86 mg (47% yield) of the product. A 46 mg aliquot was dissolved in 1 mL of methanol, treated with 3 drops of ether saturated with HCI gas, and concentrated in vacuo with toluene (2x5mL) to give 42.3 mg (21% yield) of 4-(2-{ [4-phenoxyphenyl]amino}(1,3-thiazol 4-yl))-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 30 300 MHz) 6 2.71 (s, 3H), 6.97-7.11 (m, 4H), 7.15 (s, 1H), 7.36 (m, 2H), 7.72, 7.85 (d, 2H rotomer, J=8.7 Hz), 8.36, 8.55 (s, 1H rotomer), 9.00 (bs, 2H), 9.35 (bs, 2H), 10.49 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 21
H,
8
N
4 0S 3 , 438.6 (M+H), found 439.2.
WO 00/47578 PCT/US99/18065 -211 Example 180 a) Amino([4-(phenylamino)phenylfamino}methane-1-thione: 4 Aminodiphenylamine (500 mg, 2.71 mmol) was treated as described in Example 177, 5 step (a) and recrystallized from toluene to give 350 mg (53% yield) of amino { [4 (phenylamino)phenyl]amino}methane-1-thione. 'H NMR (DMSO-d 6 , 300 MHz) 6 6.80 (m, 1H), 7.01-7.24 (m, 8H), 8.15 (s, 1H), 9.45 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 13
H,
3
N
3 S, 243.33 (M+H), found 244.2. b) Methyl 5-methylthio-4-(2-([4-(phenylamino)phenyl]amino}(1,3-thiazol-4 10 yl))thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (90 mg, 0.28 mmol) was allowed to react with amino { [4-(phenylamino)phenyl]amino}methane-1-thione (70.8 mg) as described in Example 154, step (a) to give 71 mg (47% yield) of methyl 5-methylthio-4-(2-{ [4 (phenylamino)phenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxylate 15 hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.66 (s, 3H), 3.82 (s, 3H), 6.73 (m, 1H), 6.96-7.24 (m, 9H), 7.63 (d, 1H, J=8.6 Hz), 8.12 (s, 1H), 10.13 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C 2 2
H,
9
N
3 0 2
S
3 , 453.60 (M+H), found 454.2. c) S-Methylthio-4-(2-{[4-(phenylamino)phenyl]aminoj(1,3-thiazol-4 yl))thiophene-2-carboxamidine hydrochloride: Methyl 5-methylthio-4-(2- { [4 20 (phenylamino)phenyl]amino } (1,3-thiazol-4-yl))thiophene-2-carboxylate hydrobromide (71 mg, 0.13 mmol) was treated as described in Example 154, step (b) to give 23.3 mg (38% yield) of 5-methylthio-4-(2-{ [4 (phenylamino)phenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.72 (s, 3H), 6.74 (t, 1H, J=7.3 Hz), 25 6.98 (d, 1H, J=7.6 Hz), 7.08 (m, 2H), 7,18 (m, 2H), 7.66 (d, 2H, J=8.9 Hz), 7.99 (s, 1H), 8.45 (s, 1H), 9.03 (bs, 4H), 10.17 (s, 1H); Mass Spectrum (ESI) m/z calcd. for
C
2 1H, 9 NsS 3 , 437.59 (M+H), found 438.2. Example 181 30 a) Amino{[4-benzylphenylfamino}methane-1-thione: 4-Benzylphenylamine (500 mg, 2.73 mmol) was treated as described in Example 177, step (a) to give 410 mg (62% yield) of amino {[4-benzylphenyl]amino}methane-1-thione. 'H NMR WO 00/47578 -212- PCT/US99/18065 (DMSO-d 6 , 300 MHz) 8 3.89 (s, 2H), 7.14-7.28 (m, 9H), 9.59 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C,4HI 4
N
2
S
3 , 242.1 (M+H), found 243.2. b) Methyl 5-methylthio-4-(2-{[4-benzylphenyl]amino}(1,3-thiazol-4 yl))thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 5 methylthiothiophene-2-carboxylate (90 mg, 0.28 mmol) was allowed to react with amino { [4-benzylphenyl]amino }methane- 1-thione (70.5 mg) as described in Example 154, step (a) to give 70.1 (47% yield) of methyl 5-methylthio-4-(2-{ [4 benzylphenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.66 (s, 3H), 3.82 (s, 3H), 3.87 (s, 2H), 7.14 -7.30 (m, 10 8H), 7.66 (d, 2H, J=8.5 Hz), 8.12 (s, 1H), 10.23 (s, 1H); (Mass Spectrum (ESI) m/z calcd. for C 2 2
H,
9
N
3 0 2
S
3 , 453.6 (M+H), found 454.2. c) 5-Methylthio-4-(2-{[4-benzylphenyl]amino}(1,3-thiazol-4-yl))thiophene 2-carboxamidine hydrochloride: Methyl 5-methylthio-4-(2-{ [4 benzylphenyl]amino} (1,3-thiazol-4-yl))thiophene-2-carboxylate hydrobromide (82.2 15 mg, 0.15 mmol) was treated as described in Example 154, step (b) to give 33.4 mg (47% yield) of 5-methylthio-4-(2-{ [4-benzylphenyl]amino} (1,3-thiazol-4 yl))thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.72 (s, 3H), 3.89 (s, 2H), 7.12 (s, 1H), 7.16-7.29 (m, 7H), 7.69 (d, 2H, J=8.6 Hz), 8.43 (s, 1H), 9.02 (bs, 4H), 10.28 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 22
H
20 N4S 3 , 20 436.6 (M+H), found 437.2. Example 182 a) ({4-[(Aminothioxomethyl)amino]phenyl}sulfonyl)piperidine: 4 Aminophenylsulphonylpiperidine (500 mg, 2.08 mol) was treated as described in 25 Example 177, step (a) to give 382 mg (61% yield) of({4 [(aminothioxomethyl)amino]phenyl}sulfonyl)piperidine. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.34 (m, 2H), 1.53 (m, 4H), 2.85 (m, 4H), 7.62 (m, 2H), 7.78 (m, 2H), 10.10 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C, 2
H,
7
N
3 0 2
S
2 , 299.4 (M+H), found 300.2. 30 b) Methyl 5-methylthio-4-(2-{[4-(piperidylsulfonyl)phenyl]aminoj(1,3 thiazol-4-yl))thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (90 mg, 0.28 nmmol) was allowed to react with WO 00/47578 -213- PCT/US99/18065
({
4 -[(aminothioxomethyl)amino]phenyl}sulfonyl)piperidine (87.1 mg) as described in Example 154, step (a) to give 105 mg (63% yield) of methyl 5-methylthio-4-(2-{[4 (piperidylsulfonyl)phenyl]amino } (1,3-thiazol-4-yl))thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.33 (m, 2H), 1.52 (m, 4H), 2.69 (s, 5 3H), 2.84 (m, 4H), 3.82 (s, 3H), 7.43 (s, 1H), 7.66 (m, 2H), 7.98 (m, 2H), 8.16 (s, 1H), 10.85 (s, 1H); (Mass Spectrum (ESI) m/z calcd. for C 21
H
23
N
3 0 4
S
4 , 509.69 (M+H), found 510.2. c) 5-Methylthio-4-(2-{[4-(piperidylsulfonyl)phenyl]amino}(1,3-thiazol-4 yl))thiophene-2-carboxamidine hydrochloride: Methyl 5-methylthio-4-(2- { [4 10 (piperidylsulfonyl)phenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxylate hydrobromide (105 mg, 0.17 mmol) was treated as described in Example 154, step (b) to give 30.3 mg (34% yield) of 5-methylthio-4-(2-{[4 (piperidylsulfonyl)phenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.36 (m, 2H), 1.54 (m, 4H), 2.76 (s, 15 3H), 2.86 (m, 4H), 7.30 (s, 1H), 7.68 (d, 2H, J=8.8 Hz), 8.03 (d, 2H, J=8.8 Hz), 8.51 (s, 1H), 8.84 (bs, 2H), 9.28 (bs, 2H), 10.94 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 20
H
23
NSO
2 S,, 493.69 (M+H), found 494.2. Example 183 20 a) Amino(3-quinolylamino)methane-1-thione: 3-Aminoquinoline (500 mg, 3.46 mmol) was treated as described in Example 177, step (a) to give 285 mg (41% yield) of amino(3-quinolylamino)methane-1-thione. 'H NMR (DMSO-d 6 , 300 MHz) 6 7.57 (m, 1H), 7.67 (m, 1H), 7.94 (m, 2H), 8.41 (d, 1H, J=2.4 Hz), 8.85 (d, 1H, J=2.5 Hz), 10.03 (s, 1H); Mass Spectrum (ESI) m/z calcd. for CoH 9
N
3 S, 203.3 (M+H), 25 found 204.1. b) Methyl 5-methylthio-4-[2-(3-quinolylamino)(1,3-thiazol-4-yl)]thiophene 2-carboxylate: Methyl 4-(2-bromoacetyl)-5-methylthiothiophene-2-carboxylate (90 mg, 0.28 mmol) was allowed to react with amino(3-quinolylamino)methane-1-thione (59.1 mg) as described in Example 154, step (a) to give 107.5 mg (78% yield) of 30 methyl 5-methylthio-4-[2-(3-quinolylamino)(1,3-thiazol-4-yl)]thiophene-2 carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.75 (s, 3H), 3.84 (s, WO 00/47578 PCT/US99/18065 -214 3H), 7.52 (s, 1H), 7.92-8.05 (m, 2H), 8.22 (s, 1H), 9.22 (m, 2H); Mass Spectrum (ESI) m/z calcd. for C 19
H,,N
3 0 2
S
3 , 413.54 (M+H), found 414.1. c) 5-Methylthio-4-[2-(3-quinolylamino)(1,3-thiazol-4-yl)]thiophene-2 carboxamidine hydrochloride: Methyl 5-methylthio-4-[2-(3-quinolylamino)(1,3 5 thiazol-4-yl)]thiophene-2-carboxylate hydrobromide (107.5 mg, 0.21 mmol) was treated as described in Example 154, step (b) to give 4.5 mg (4.9% yield) of 5 methylthio-4-[2-(3-quinolylamino)(1,3-thiazol-4-yl)]thiophene-2-carboxamidine hydrochloride. 1 H NMR (DMSO-d 6 , 300 MHz) 8 2.80 (s, 3H), 7.29 (s, 1H), 7.59 (m, 2H), 7.93 (m, 2H), 8.54 (s, 1H), 8.89 (bs, 2H), 8.91 (m, 1H), 9.16 (m, 1H), 9.29 (bs, 10 2H), 10.97 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 8
H
15
NS
3 , 397.5 (M+H), found 398.1. Example 184 a) Methyl 5-methylthio-4-J2-(2-naphthylamino) (1,3-thiazol-4-yl)]thiophene 15 2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5-methylthiothiophene-2 carboxylate (65 mg, 0.21 mmol) was allowed react with 2-napthylthiourea (42.4 mg) as described in Example 154, step (a) to give 82.5 mg (80% yield)of methyl 5 methylthio-4-[2-(2-naphthylamino)(1,3-thiazol-4-yl)]thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.67 (s, 3H), 3.83 (s, 3H), 7.31 (s, 20 1H), 7.50-7.67 (m, 4H), 7.93 (m, 1H), 8.15 (s, 1H), 8.31-8.35 (m, 1H), 8.46 (d, 1H, J=7.6), 10.22 (s, 1H)); Mass Spectrum (ESI) m/z calcd. for C 2 0
H
1 6
N
2 0 2
S
3 , 412.6 (M+H), found 413.1. c) 5-Methylthio-4-J2-(2-naphthylamino) (1,3-thiazol-4-yl)]thiophene-2 carboxamidine hydrochloride: Methyl 5-methylthio-4-[2-(2-naphthylamino)(1,3 25 thiazol-4-yl)]thiophene-2-carboxylate hydrobromide (42.7 mg, 0.086 mmol) was treated as described in Example 154, step (b) to give 5.8 mg (16% yield) of 5 methylthio-4-[2-(2-naphthylamino)(1,3-thiazol-4-yl)]thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.72 (s, 3H), 7.12-7.27 (m, 3H), 7.50-7.68 (m, 3H), 7.94 (m, 1H), 8.32-8.35 (m, m, 1H), 8.51 (s, 1H), 8.97 (bs, 2H), 30 9.34 (bs, 2H), 10.26 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C, 19
H
1 6
N
4
S
3 , 396.6 (M+H), found 397.2.
WO 00/47578 PCT/US99/18065 -215 Example 185 a) Methyl 4-[2-(2H-benzof3,4-d]l,3-dioxolan-5-ylamino) (1,3-thiazol-4-yl)] 5-methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (65 mg, 0.21 mmol) was allowed to react with 5 2,3-methylenedioxyphenylthiourea (41.2 mg) as described in Example 154, step (a) to give 51 mg (50% yield) of methyl 4-[2-(2H-benzo[3,4-d]1,3-dioxolan-5-ylamino)(1,3 thiazol-4-yl)]-5-methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO d 6 , 300 MHz) 8 2.66 (s, 3H), 3.83 (s, 3H), 5.98 (s, 2H), 6.84-6.89 (m, 1H), 6.96, 7.04 (dd, 1H rotomer, J=2.2, 8.5 Hz), 7.25 (s, 1H), 7.46, 7.60 (d, 1H rotomer, J=2.1 Hz), 10 8.05, 8.13 (s, 1H rotomer), 10.19, 10.34 (s, 1H, rotomer); Mass Spectrum (ESI) m/z calcd. for Cl 7
H,
4
N
2 0 4
S
3 , 406.5 (M+H), found 407.1. b) 4-[2-(2H-Benzo[3,4-d]l,3-dioxolan-5-ylamino)(1,3-thiazol-4-yl)]-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-[2-(2H-benzo[3,4 d] 1,3-dioxolan-5-ylamino)(1, 3 -thiazol-4-yl)]-5-methylthiothiophene-2-carboxylate 15 hydrobromide (51 mg, 0.10 mmol) was treated as described in Example 154, step (b) to give 16.6 mg (39% yield) of 4-[2-(2H-benzo[3,4-d] 1,3-dioxolan-5-ylamino)(1,3 thiazol-4-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 6 2.71(s, 3H), 5.98 (s, 2H), 6.87 (d, 1H, J=8.2 Hz), 7.09-7.13 (m, 2H), 7.67 (d, 1H, J=2.4 Hz), 8.50 (s, 1H), 8.95 (bs, 2H), 9.33 (bs, 2H), 10.30 (s, 20 1H); Mass Spectrum (ESI) m/z calcd. for C, 6
H
14
N
4 0 2 S3, 390.51 (M+H), found 391.2; Example 186 a) Amino[(7-bromofluoren-2-yl)amino]methane--thione: 2-Amino-7 bromofluorene (500 mg, 1.90 mmol) was treated as described in Example 177, step 25 (a) to give 128 mg (21% yield) ofamino[(7-bromofluoren-2-yl)amino]methane-1 thione. 'H NMR (DMSO-d 6 , 300 MHz) 8 3.35 (s, 2H), 7.35 (d, 1H, J=8.3 Hz), 7.54 (d, 1H, J=8.0 Hz), 7.66 (s, 1H), 7.77-7.87 (m, 3H), 9.80 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C, 4 H,,BrN 2 S, 319.2 (M+H), found 320.1, 321.1. b) Methyl 4-{2-[(7-bromofluoren-2-yl)amino](1,3-thiazol-4-yl)}-5 30 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (90 mg, 0.28 mmol) was allowed to react with amino[(7-bromofluoren-2-yl)amino]methane-1-thione (92.8 mg) as described in WO 00/47578 -216- PCT/US99/18065 Example 154, step (a) to give 141 mg (82% yield) of methyl 4-{2-[(7-bromofluoren 2-yl)amino](1,3-thiazol-4-yl) }-5-methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.70 (s, 3H), 3.83 (s, 3H), 3.93 (s, 2H), 7.33 (s, 1H), 7.51 (dd, 1H, J=1.9, 8.0 Hz), 7.65 (dd, 1H, J=2.0, 8.4 Hz), 7.74 (ad, 2H, J=8.3 Hz), 5 7.83 (ad, 1H, J=8.4 Hz), 8.18 (s, 1H), 8.23 (d, 1H, J=1.4 Hz), 10.47 (s, 1H). c) 4-{2-[(7-Bromofluoren-2-yl)amino](1,3-thiazol-4-yl))}-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4-{2-[(7 bromofluoren-2-yl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (100 mg, 0.15 mmol) was treated as described in Example 154, step (b) 10 to give 3.3 mg (4% yield) of 4-{2-[(7-bromofluoren-2-yl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine hydrochloride. 1 H NMR (DMSO-d 6 , 300 MHz) 8 2.76 (s, 3H), 3.95 (s, 2H), 7.18 (s, 1H), 7.54 (dd, 1H, J=1.8, 10.0 Hz), 7.67-7.76 (m, 3H), 7.85 (d, 1H, J=8.2 Hz), 8.23 (s, 1H), 8.50 (s, 1H), 10.53 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 2 2
H,
7 BrN 4
S
3 , 513.5 (M+H), found 513.1, 515.1. 15 Example 187 a) Methyl 4-{2-[(4-cyclohexylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (65 mg, 0.21 mmol) was allowed to react with 4 20 cyclohexylphenylthiourea (49.2 mg) as described in Example 154, step (a) to give 45 mg (41% yield) of methyl 4- { 2-[(4-cyclohexylphenyl)amino]( 1,3-thiazol-4-yl) }-5 methylthiothiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.23-1.39 (m, 5H), 1.71-1.79 (m, 5H), 2.68 (s, 3H), 3.83 (s, 3H), 7.16 (d, 2H, J=8.6 Hz), 7.26 (s, 1H), 7.65 (d, 2H, J=8.7 Hz), 8.14 (s, 1H), 10.19 (s, 1H); Mass Spectrum 25 (ESI) m/z calcd. for C 2 2
H
2 4
N
2 0 2
S
3 , 444.64 (M+H), found 445.2. b) 4-{2-[(4-Cyclohexylphenyl)amino](1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl 4- { 2-[(4 cyclohexylphenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxylate hydrobromide (31.1 mg, 0.059 mmol) was treated as described in Example 154, step 30 (b) to give 12.8 mg (47% yield) of 4-{2-[(4-cyclohexylphenyl)amino](1,3-thiazol-4 yl)}-5-methylthiothiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.33-1.40 (m, 5H), 1.68-1.79 (m, 5H), 2.44 (m, 1H), 2.73 (s, 3H), 7.12 (s, WO 00/47578 -217- PCT/US99/18065 1H), 7.18 (d, 2H, J=8.7 Hz), 7.68 (d, 2H, J=8.7 Hz), 8.47 (s, 1H), 8.85 (bs, 2H), 9.32 (bs, 2H), 10.28 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 2 1H 24
N
4
S
3 , 428.64 (M+H), found 429.2. 5 Example 188 a) Amino{J4-(phenyldiazenyl)phenyl]amino}methane-1-thione: 4 Phenylazophenylisothiocyanate (314 mg, 1.30 mmol) was treated as described in Example 177, step (a), part (ii), to give 295 mg (88% yield) of amino { [4 (phenyldiazenyl)phenyl]amino}methane- -thione. 'H NMR (DMSO-d 6 , 300 MHz) 10 6 6.84 (m, 1H), 7.57 (m, 2H), 7.73 (m, 2H), 7.85-7.89 (m, 4H), 10.04 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 13
HI
2
N
4 S, 256.3 (M+H), found 257.2. b) Methyl 5-methylthio-4-(2-{[4-(phenyldiazenyl)phenyl]amino(1,3-thiazol 4-yl))thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (65 mg, 0.21 mmol) was allowed to react with 15 amino { [ 4 -(phenyldiazenyl)phenyl]amino}methane-1 -thione (53.8 mg) as described in Example 154, step (a) to give 80.6 mg (70% yield) of methyl 5-methylthio-4-(2-{[4 (phenyldiazenyl)phenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 6 2.72 (s, 3H), 3.84 (s, 3H), 7.46 (s, 1H), 7.49-7.61 (m, 3H), 7.84 (m, 2H), 7.91-8.02 (m, 4H), 8.20 (s, 1H), 10.83 (s, 1H); 20 Mass Spectrum (ESI) m/z calcd. for C 22
H,
8
N
4 0 2
S
3 , 466.6 (M+H), found 467.1. c) S-Methylthio-4-(2-{[4-(phenyldiazenyl)phenyl]aminoj(1,3-thiazol-4 yl))thiophene-2-carboxamidine hydrochloride: Methyl 5-methylthio-4-(2-{ [4 (phenyldiazenyl)phenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxylate hydrobromide (47.7 mg, 0.087 mmol) was treated as described in Example 154, step 25 (b) to give 32.8 mg (77% yield) of 5-methylthio-4-(2-{[4 (phenyldiazenyl)phenyl]amino}(1,3-thiazol-4-yl))thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.78 (s, 3H), 7.26 (s, 1H), 7.49 7.63 (m, 3H), 7.66-7.74 (m, 3H), 7.84-8.08 (m, 3H), 8.60 (s, 1H), 11.02 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C 21 Hs, 8
N
6
S
3 , 450.6 (M+H), found 451.1. 30 WO 00/47578 -218- PCT/US99/18065 Example 189 a) {3-[(Aminothioxomethyl)amino]phenyl}methan-1-ol: 3-Aminobenzyl alcohol (550 mg, 4.46 mmol) was treated as described in Example 177, step (a) to give 618 mg (76% yield) of {3-[(aminothioxomethyl)amino]phenyl}methan-1-ol. 'H 5 NMR (DMSO-d 6 , 300 MHz) 8 4.47 (d, 2H, J=5.6 Hz), 5.19 (t, 1H, J=5.7 Hz), 7.06 (d, 1H, J=6.2 Hz), 7.18-7.30 (m, 3H), 9.73 (s, 1H). b) Methyl-5-methylthio4-(2-{[3-(hydroxymethyl)phenyl]amino}(1,3-thiazol 4-yl))-thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (1.01 g, 3.26 mmol) was allowed to react with of 10 {3-[(aminothioxomethyl)amino]phenyl}methan-l1-ol as described in Example 154, step (a) to give 1.42 g (92% yield) of methyl-5-methylthio4-(2-{[3 (hydroxymethyl)phenyl]amino } (1,3-thiazol-4-yl))-thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 5 2.67 (s, 3H), 3.83 (s, 3H), 4.49 (s, 2H), 6.92 (m, 1H), 7.23-7.31 (m, 2H), 7.60 (m, 1H), 7.81 (bs, 1H), 8.17 (s, 1H), 10.29 15 (bs, 1H). c) 5-Methylthio 4-(2-{[3-(hydroxymethyl)phenyl]aminoj(1,3-thiazol-4-yl)) thiophene-2-carboxamidine hydrochloride: Methyl-5-methylthio4-(2-{[3 (hydroxymethyl)phenyl]amino}(1,3-thiazol-4-yl))-thiophene-2-carboxylate hydrobromide (700 mg, 1.47 mmol) was treated as described in Example 154, step (b) 20 using 1:9:1 methanol-CH 2
C
2 -DMF as eluent to give 195 mg (32% yield) of 5 methylthio 4-(2- { [3-(hydroxymethyl)phenyl]amino} (1,3-thiazol-4-yl))-thiophene-2 carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.71 (s, 3H), 4.50 (s, 2H), 6.93 (d, 1H, J=7.6 Hz), 7.15 (s, 1H), 7.21-7.27 (m, 1H), 7.38 (bs, 1H), 7.65 (d, 1H, J=8.1 Hz), 7.80 (s, 1H), 8.53 (s, 1H), 8.94 (bs, 2H), 9.32 (bs, 2H), 10.37 (s, 25 1H); Mass Spectrum (ESI) m/z calcd. for C 1 6
H,
6
N
4 0S 3 , 376.5 (M+H), found 377.2. Example 190 a) (tert-Butoxy)-N-[(4-{2-[(3-hydroxymethylphenyl)amino](1,3-thiazol-4 yl)}-5-methylthio(2-thienyl))iminomethyl]-carboxamide: 5-Methylthio 4-(2-{([3 30 (hydroxymethyl)phenyl]amino} (1,3-thiazol-4-yl))-thiophene-2-carboxamidine (103 mg, 0.27 mmol) was slurried in THF (4 mL) and treated with 0.5 mL of 0.5 N NaOH. tert-Butyldicarbonate (0.40 mmol) was added in one portion and the resulting mixture WO 00/47578 -219- PCT/US99/18065 was stirred overnight. The reaction was partitioned in CH 2 Cl 2 and water. The organic layer was separated and washed with brine (1x20 mL) and dried (Na 2
SO
4 ). Removal of the solvent in vacuo, followed by purification on preparative thin layer chromatography (500 mm silica gel plate, J.T.Baker, Phillipsburg, NJ, 1% methanol 5 CH 2 Cl 2 ), gave 45 mg (35% yield) of ((tert-Butoxy)-N-[(4-{2-[(3 hydroxymethylphenyl)amino](1,3-thiazol-4-yl)}-5-methylthio(2 thienyl))iminomethyl]-carboxamide. ' H NMR (DMSO-d 6 , 300 MHz) 6 1.44 (s, 9H), 2.66 (s, 3H), 4.49 (d, 2H, J=5.7 Hz), 5.15 (t, 1H, J1=5.5 Hz), 6.92 (d, 1H, J=7.5 Hz), 6.96 (s, 1H), 7.26 (min, 1H), 7.66 -7.75 (min, 2H), 8.38 (s, 1H), 8.98 (bs, 2H), 10.24 (s, 10 1H). b) (tert-Butoxy)-N-(imino[4-[2-([3-[(3 methylpiperidyl)methyl]phenyl}amino)(1,3-thiazol-4-yl)]-5-methylthio(2 thienyl)}methyl)carboxamide: To a stirring solution of ((tert-butoxy)-N-[(4- {2-[(3 hydroxymethylphenyl)amino](1,3-thiazol-4-yl)}-5-methylthio(2 15 thienyl))iminomethyl]-carboxamide (45 mg, 0.094 mmol) under N 2 was added triethylamine (2 equiv, 26.3 pl), followed by methansulfonyl chloride (Aldrich Chemical Co., Milwaukee, WI, 0.13 mmol, 10.2 l ). The reaction was stirred for 1 h, at which time the reaction was partitioned in CH 2 Cl 2 -water. The organic layer was washed with brine (20 mL), filtered through a 5-cm pad of silica gel in a 15-mL fritted 20 glass funnel and dried (Na 2
SO
4 ). Removal of the solvent in vacuo afforded the crude mesylate (44 mg) which was used immediately without further purification. To 25.3 mg (0.045 mmol) of the mesylate in 0.5 mL of DMF was added 3-methyl piperidine (0.18 mmol, 21.4 tL ) and the result was heated to 65oC in an oil bath for 4 h. The reaction was concentrated in vacuo and purified by preparative thin layer 25 chromatography (250 mm silica gel plate, 10% methanol-CH 2 C1 2 , J.T.Baker, Phillipsburg, NJ) to give 8.2 mg (32% yield) of (tert-butoxy)-N-(imino{4-[2-({3-[(3 methylpiperidyl)methyl]phenyl}amino)(1,3-thiazol-4-yl)]-5-methylthio(2 thienyl)}methyl)carboxamide. Mass Spectrum (ESI) m/z calcd. for C 27
H
3 5 NsO 2
S
3 , 557.8 (M+H), found 557.9, 458.2 (-C(O)OC(CH 3
)
3 . 30 c) 4-[2-([3-[(3-Methylpiperidyl)methyl]phenyl}amino)(1,3-thiazol-4-yl)]-5 methylthiothiophene-2-carboxamidine hydrochloride: (tert-Butoxy)-N-(imino{4-[2 ({3-[(3-methylpiperidyl)methyl]phenyl}amino)(1,3-thiazol-4-yl)]-5-methylthio(2- WO 00/47578 -220- PCT/US99/18065 thienyl)}methyl)carboxamide (8.2 mg, 0.014 mmol) was stirred 2 mL of a 10% 3N HCl-ethyl acetate solution at 0 'C for 30 min., at which time the solvent was removed in vacuo to give 8 mg (100% yield) of the 4-[2-({3-[(3 methylpiperidyl)methyl]phenyl}amino)(1,3-thiazol-4-yl)]-5-methylthiothiophene-2 5 carboxamidine hydrochloride. ' H NMR (DMSO-d 6 , 300 MHz) 80.83 (d, 3H, J=5.6 Hz), 1.54-2.48 (m, 5H), 2.52-2.63 (m, 4H), 2.66 (s, 3H), 4.23 (d, 2H, J=4.8 Hz), 7.15 7.23 (m, 2H), 7.41 (t, 1H, J=7.8 Hz), 7.86-7.92 (m, 2H), 8.63 (s, 1H), 9.01 (bs, 2H), 9.42 (bs, 2H), 10.63 (s, 1H); (Mass Spectrum (ESI) m/z calcd. for C 2 2
H
27
NSS
3 , 457.7 (M+H), found 458.2. 10 Example 191 a) Methyl-5-methylthio-4-{2-J(3-hydroxyphenyl)amino](1,3-thiazol-4-yl)} thiophene-2-carboxylate hydrobromide: Methyl 4-(2-bromoacetyl)-5 methylthiothiophene-2-carboxylate (60 mg, 0.19 mmol) was allowed to react with 3 15 hydroxyphenylthiourea (32.6 mg) as described in Example 154, step (a) to give 80.2 mg (92% yield) of methyl-5-methylthio-4- {2-[(3-hydroxyphenyl)amino](1,3-thiazol 4-yl)}-thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.67 (s, 3H), 3.83 (s, 3H), 6.38 (d, 1H, J=7.6 Hz), 7.06-7.12 (m, 2H), 7.20-7.29 (m, 2H), 8.14 (s, 1H), 10.17 (s, 1H). 20 b) 4-{2-[(3-Hydroxyphenyl)aminoj(1,3-thiazol-4-yl)}-5 methylthiothiophene-2-carboxamidine hydrochloride: Methyl-5-methylthio-4-{2 [(3-hydroxyphenyl)amino](1,3-thiazol-4-yl)}-thiophene-2-carboxylate hydrobromide (460 mg, 1.0 mmol) was treated as described in Example 154, step (b) to give 215 mg (54% yield) of 4-{2-[(3-hydroxyphenyl)amino](1,3-thiazol-4-yl)}-5 25 methylthiothiophene-2-carboxamidine hydrochloride. (Mass Spectrum (ESI) m/z calcd. for C,1 5
H,
4
N
4 0S 3 , 362.5 (M+H), found 363.2. c) (tert-Butoxy)-N-J(4-{2-[(4-hydroxyphenyl)amino](1,3-thziazol-4-yl)}-5 methylthio(2-thienyl))iminomethyl]carboxamide: To a stirring solution of 4-{2-[(3 hydroxyphenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2-carboxamidine 30 hydrochloride (215 mg, 0.48 mmol) in 4 mL of CH 2 Cl 2 -DMF (3:1, v/v) was added di isopropylethylamine (1.2 equiv). Di-tert-butyl dicarbonate (1.2 equiv, 127 mg, Aldrich Chemicals, Milwaukee, WI) was then added dropwise in 1 mL CH 2 C1 2 via an WO 00/47578 -221- PCT/US99/18065 addition funnel. The reaction was allowed to stir overnight, partitioned in CH 2 Cl 2 H 2 0, and the layers separated. The organic layer was dried (Na 2
SO
4 ) and concentrated in vacuo. The residue was purified by flash chromatography (1% methanol-CH 2
C
2 ) to give 60 mg (27% yield) of (tert-butoxy)-N-[(4-{2-[(4-hydroxyphenyl)amino](1,3 5 thiazol-4-yl)}-5-methylthio(2-thienyl))iminomethyl]carboxamide. 'H NMR (DMSO d 6 , 300 MHz) 8 1.44 (s, 9H), 2.72 (s, 3H), 6.38 (m, 1H), 6.96 (s, 1H), 7.06-7.12 (m, 2H), 7.28 (m, 1H), 8.35 (s, 1H), 9.00 (bs, 2H), 9.28 (s, 1H), 10.11 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 20
H
22
N
4 0 3
S
3 , 462.6 (M+H), found 462.7, 363.2 [ C(0)OC(CH 3 )z]. 10 d) (tert-Butoxy)-N-[[4-(2-[[3-(carbamoylmethoxy)phenyl]amino)(1,3 thiazol-4-yl))-5-methylthio(2-thienyl)]iminomethyl}carboxamide: To stirring solution of (tert-butoxy)-N-[(4- {2-[(4-hydroxyphenyl)amino](1,3-thiazol-4-yl) } -5 methylthio(2-thienyl))iminomethyl]carboxamide (65 mg, 0.14 mmol) in 1.5 mL of DMF was added sequentially Cs 2
CO
3 (1.5 equiv, 60.1 mg, Aldrich Chemicals, 15 Milwaukee, WI), bromoacetamide (1.2 equiv, 20.4 mg, Aldrich Chemicals, Milwaukee, WI), and a catalytic amount of KI. The reaction was warmed to 58 'C in an oil bath, stirred for 48 h, at which time another 0.6 equiv of bromoacetamide was added. Stirring was continued for another 24 h, at which time the reaction was filtered and concentrated in vacuo. The residue was purified by preparative thin layer 20 chromatography (50% ethyl acetate-hexanes) to give 9 mg (12% yield) of (tert butoxy)-N- { [4-(2- { [3 -(carbamoylmethoxy)phenyl]amino } (1,3 -thiazol-4-yl))-5 methylthio(2-thienyl)]iminomethyl}carboxamide. Mass Spectrum (ESI) m/z calcd. for
C
22
H
2 5
N
5 04S 3 , 519.7 (M+H), found 519.7, 420.7 [-C(0)OC(CH3)3]. e) 4-(2-{[4-(Carbamoylmethoxy)phenyl]amino](1,3-thiazol-4-yl))-5 25 methylthiothiophene-2-carboxamidine trifluoroacetate: To a stirring suspension of(tert-butoxy)-N- { [4-(2- { [3-(carbamoylmethoxy)phenyl] amino } (1,3-thiazol-4-yl))-5 methylthio(2-thienyl)]iminomethyl}carboxamide (ca. 4 mg, 0.007 mmol) in CH 2 C1 2 DMF (4 mL, 3:1 v/v) at 0 oC was added 1 mL oftrifluoroacetic acid. The homogeneous solution was stirred an additional 40 min. at this temperature, warmed 30 to ambient temperature over a 30 min. period and concentrated in vacuo to give 4 mg (100% yield) of 4-(2- {[4-(carbamoylmethoxy)phenyl]amino}(1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine trifluoroacetate. 'H NMR (DMSO-d 6 , 300 WO 00/47578 -222- PCT/US99/18065 MHz) 6 2.75 (s, 3H), 4.21(d, 2H, J=5.7 Hz), 6.64 (dd, 1H, J=2.4, 8.2 Hz), 6.97 (dd, 1H, J=1.1, 8.2 Hz), 7.16 (s, 1H), 7.22 (m, 1H), 7.60-7.63 (m, 1H), 7.69-7.72 (m, 1H), 7.88 (t, 1H, J=2.1 Hz), 8.42 (s, 1H); Mass Spectrum (ESI) m/z calcd. for
C,,
7
H,,
7 NsO 5 2
S
3 , 419.6 (M+H), found 420.1. 5 Example 192 a) Isopropyl 5-methyl-4-{2-J(3,4,5-trimethoxyphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrobromide: Isopropyl-4-(2-bromoacetyl)-5 methylthiophene-2-carboxylate (84 mg, 0.27 mmol) was allowed to react with 3,4,5 10 trimethoxyphenylthiourea (66.5 mg) as described in Example 154, step (a) to give 68 mg (48% yield) of isopropyl 5-methyl-4-{2-[(3,4,5-trimethoxyphenyl)amino](1,3 thiazol-4-yl)}thiophene-2-carboxylate hydrobromide. Mass Spectrum (ESI) m/z calcd. For C 21
H
2 4
N
2 0 5
S
2 , 448.56 (M+H), found 449.0. b) S-Methyl-4-{2-[(3,4,5-trimethoxyphenyl)amino](1,3-thiazol-4 15 yl)}thiophene-2-carboxamidine hydrochloride: Isopropyl 5-methyl-4-{2- [(3,4,5 trimethoxyphenyl)amino](1,3-thiazol-4-yl) } thiophene-2-carboxylate hydrobromide (59 mg, 0.11 mmol) was treated as described in Example 154, step (b) to give 24.4 mg (50% yield) of 5-methyl-4- {2-[(3,4,5-trimethoxyphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.81 20 (s, 3H), 3.61 (s, 3H), 3.77 (s, 6H), 7.04 (s, 2H), 7.09 (s, 1H), 8.40 (s, 1H); Mass Spectrum (ESI) m/z calcd. for CIH 20
N
4 0 3
S
2 , 404.5 (M+H), found 405.2. Example 193 a) Isopropyl 5-methyl-4-{2-[(4-phenoxyphenyl)amino](1,3-thiazol-4 25 yl)}thiophene-2-carboxylate hydrobromide: Isopropyl- 4-(2-bromoacetyl)-5 methylthiophene-2-carboxylate (91 mg, 0.29 mmol) was allowed to react with 4 phenoxyphenylthiourea (72.6 mg) as described in Example 154, step (a) to give 115 mg (75% yield) of isopropyl 5-methyl-4- {2-[(4-phenoxyphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxylate hydrobromide. 'H NMR (DMSO-d 6 , 300 MHz) 8 1.28 30 (d, 6H, J=6.2 Hz), 2.70 (s, 3H), 6.06 (quintet, 1H, J=6.2 Hz), 6.92-7.09 (m, 5H), 7.15 (s, 1H), 7.30-7.37 (m, 2H), 7.56-7.70 (m, 2H), 7.98 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 24
H
22
N
2 0 3
S
2 , 450.6 (M+H), found 451.2, 409.2 [-CH(CH 3
)
2
].
WO 00/47578 -223- PCT/US99/18065 b) 5-Methyl-4-{2-[(4-phenoxyphenyl)amino](1,3-thiazol-4-yl)}thiophene-2 carboxamidine hydrochloride: Isopropyl 5-methyl-4-{2-[(4 phenoxyphenyl)amino](1,3-thiazol-4-yl)}thiophene-2-carboxylate hydrobromide (95.5 mg, 0.17 mmol) was treated as described in Example 154, step (b) to give 23.8 5 mg (32% yield) of 5-methyl-4-{2-[(4-phenoxyphenyl)amino](1,3-thiazol-4 yl)}thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.76 (s, 3H), 6.95-7.12 (m, 6H), 7.34-7.39 (m, 2H), 7.72-7.78 (m, 2H), 8.33 (s, 1H), 8.98 (bs, 3H), 10.29 (bs, 1H); Mass Spectrum (ESI) m/z calcd. for C 21
H,
8
N
4 0 2
S
3 , 406.5 (M+H), found 407.2. 10 Example 194 a) Isopropyl 5-methyl-4-[2-(phenylamino) (1,3-thiazol-4-yl)]-thiophene-2 carboxylate hydrobromide: Isopropyl 4-(2-bromoacetyl)-5-methylthiophene-2 carboxylate (64 mg, 0.21 mmol) was allowed to react with phenylthiourea (32.1 mg) 15 as described in Example 154, step (a) to give 80 mg (87% yield) of isopropyl 5 methyl-4-[2-(phenylamino)(1,3-thiazol-4-yl)]-thiophene-2-carboxylate hydrobromide. Mass Spectrum (ESI) m/z calcd. for C,H, 8 sN 2 0 2
S
2 , 358.5 (M+H), found 359.2. b) 5-Methyl-4-[2-(phenylamino)(1,3-thiazol-4-yl)]thiophene-2 carboxamidine hydrochloride: Isopropyl 5-methyl-4-[2-(phenylamino)(1,3-thiazol 20 4-yl)]-thiophene-2-carboxylate hydrobromide (74 mg, 0.16 immol) was treated with phenylthiourea (24.3 mg) as described in Example 154, step (b) to give 15 mg (28% yield) (of 5-methyl-4-[2-(phenylamino)(1,3-thiazol-4-yl)]thiophene-2-carboxamidine hydrochloride, which was further purified by recrystallization from methanol-water. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.79 (s, 3H), 6.96 (t, 1H, J=7.2 Hz), 7.09 (s, 1H), 25 7.33 (t, 2H, J=7.5 Hz), 7.71 (d, 2H, J=7.7 Hz), 8.39 (s, 1H), 8.95 (bs, 2H), 9.33 (bs, 2H), 10.37 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C 15
HI
4
N
4
S
3 , 314.4 (M+H), found 315.2. Example 195 a) Methyl 4-(4-isoxazol-5-yl(1,3-thiazol-2-yl))-5-methylthiothiophene-2 30 carboxylate: Methyl 4-(aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (872 mg, 2.51 immol) was allowed to react with 2-bromo-1-isoxazol-5-ylethan-1-one (737 mg, prepared from from isoxazole-5-carbonyl chloride [Maybridge Chemicals, WO 00/47578 -224- PCT/US99/18065 Cornwall, UK] as described in Example 177, step (a) as described in Example 154, step (a) to give 704 mg (83% yield) of methyl 4-(4-isoxazol-5-yl(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.75 (s, 3H), 3.85 (s, 3H), 6.93 (d, 1H, J=1.8 Hz), 8.22 (s, 1H), 8.38 (s, 1H), 8.70 (d, 1H, J=1.8 5 Hz). b) 4-(4-Isoxazol-S-yl(1,3-thiazol-2-yl))-S-methylthiothiophene-2 carboxamidine hydrochloride: Methyl 4-(4-isoxazol-5-yl(1,3-thiazol-2-yl))-5 methylthiothiophene-2-carboxylate (350 mg, 1.03 mmol) was treated as described in Example 154, step (b) to give 290 mg (78% yield) of 4-(4-isoxazol-5-yl(1,3-thiazol-2 10 yl))-5-methylthiothiophene-2-carboxamidine hydrochloride, of which an aliquot was further purified by recrystallization from methanol-isopropanol-water (3:1:0.2, v/v/v). 'H NMR (DMSO-d 6 , 300 MHz) 5 2.79 (s, 3H), 6.93 (d, 1H, J=1.9 Hz), 8.45 (s, 1H), 8.74 (m, 2H), 9.23 (bs, 2H), 9.53 (bs, 2H); Mass Spectrum (MALDI-TOF, CHCA matrix) m/z calcd. for C1 2
HI
0
N
4 0S 3 , 322.4 (M+H), found 323.3. 15 Example 196 a) Methyl 4-[4-(2-hydroxyphenyl) (1,3-thiazol-2-yl)]-5-methylthiothiophene 2-carboxylate: Methyl 4-(aminothioxomethyl)-5-methylthiothiophene-2-carboxylate (808 mg, 3.26 mmol) was allowed to react with 2-(2-bromoacetyl)hydroxybenzene (925 mg, prepared from 2-(chlorocarbonyl)phenyl acetate [Aldrich Chemicals, 20 Milwaukee, WI] as described in Example 177, step (a)) as described in Example 154, step (a) to give 433 mg (37% yield) of methyl 4-[4-(2-hydroxyphenyl)(1,3-thiazol-2 yl)]-5-methylthiothiophene-2-carboxylate. 'H NMR(DMSO-d 6 , 300 MHz) 8 2.77 (s, 3H), 3.86 (s, 3H), 6.91-7.00 (m, 2H), 7.23 (m, 1H), 8.14-8.19 (m, 2H), 8.24 (s, 1H); Mass Spectrum (ESI) m/z calcd. for C, 6
HI
3 NO3S 3 , 363.48 (M+H), found 364.2. 25 b) 4-[4-(2-Hydroxyphenyl)(1,3-thiazol-2-yl)]-5-methylthiothiophene-2 carboxamidine hydrochloride: Methyl 4-[4-(2-hydroxyphenyl)(1,3-thiazol-2-yl)]-5 methylthiothiophene-2-carboxylate (400 mg, 1.1 mmol) was treated as described in Example 154, step (b) to give 173 mg (41% yield) of 4-[4-(2-hydroxyphenyl)(1,3 thiazol-2-yl)]-5-methylthiothiophene-2-carboxamidine hydrochloride. 1 H NMR 30 (DMSO-d 6 , 300 MHz) 8 2.81 (s, 3H), 6.92-7.02 (m, 2H), 7.22 (m, 1H), 8.20 (dd, 1H, J=1.7, 7.8 Hz), 8.27 (s, 1H), 8.65 (s, 1H), 9.00 (bs, 2H), 9.41 (bs, 2H), 10.58 (s, 1H); Mass Spectrum (ESI) m/z calcd. for CjsH 3
N
3 0S 3 , 347.48 (M+H), found 348.2.
WO 00/47578 -225- PCT/US99/18065 Example 197 5-Methylthio-4-(6-quinolylamino)thiophene-2-carboxamidine hydrochloride a) Methyl 5-methylthio-4-(6-quinolylamino)thiophene-2 5 carboxylate: To an oven-dried glass vial with stir bar was added a mixture of 65.2 mg (0.244 mmol) of methyl 4-bromo-5-methylthiothiophene-2 carboxylate (as prepared in Example 241, step (a)), 5.2 mg (9.5 mol %) of palladium (II) acetate, 22.2 mg (14.6 mol %) of racemic-2,2' bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), 125 mg (0.384 mmol) of 10 cesium carbonate and 50.3 mg (0.349 mmol) of 6-aminoquinoline. The vial was transferred to a glove bag, flushed with dry argon and anhydrous toluene (488 pL) was added. The vial was capped with a Teflon-lined screw cap and heated at 100 0 C for 48 h. To the cooled suspension was added ethyl acetate (4 mL), the mixture filtered (Celite), washing with ethyl acetate (2 x 2 mL), and 15 the solvents removed in vacuo. The resulting residue was purified by chromatography on a 10-g silica SPE column with a gradient of 5-12 % ethyl acetate-CH 2 C1 2 to afford 53.3 mg (66%) of the title compound as a pale yellow resin. 'H-NMR (CDCl 3 , 400 MHz) 8 8.77 (dd, 1H, J = 4.2, 1.6 Hz), 8.04 (d, 1H, J = 9.4 Hz), 8.02 (d, 1H, J = 8.4 Hz), 7.90 (s, 1H), 7.41 (dd, 1H, J = 9.0, 20 2.6 Hz), 7.36 (dd, 1H, J = 8.3, 4.2 Hz), 7.27 (d, 1H, J = 2.6 Hz), 3.92 (s, 3H) and 2.45 (s, 3H). Mass spectrum (ESI, m/z): Calcd. For C 16 Hs 15
N
2 0 2
S
2 , 331.1 (M+H), found 331.2. b) 5-Methylthio-4-(6-quinolylamino)thiophene-2-carboxamidine hydrochloride: Trimethylaluminum (2.0 M in toluene, 0.76 mL, 1.52 mmol) 25 was added dropwise to a suspension of ammonium chloride (85.6 mg, 1.60 mmol) in anhydrous toluene (0.76 mL) under Ar at 0OC. The mixture was stirred at 25 0 C for 30 min and then 50.2 mg (0.152 mmol) of methyl 5 mehtylthio-4-(6-quinolylamino)thiophene-2-carboxylate (as prepared in previous step) was added. The reaction mixture was heated slowly to 100 0 C 30 and stirred for 4 h. The cooled mixture was added to a vigorously stirred slurry of silica gel (3 g) in chloroform (15 mL). The suspension was filtered (Celite) washing with 25% MeOH-CH 2
C
2 (2 x 5 mL), 50% MeOH-CH 2
C
2 (2 x 5 mL) and 75% MeOH-CH 2 C1 2 (2 x 5 mL). The combined washings were concentrated and the resulting residue was purified on a 5-g silica SPE column WO 00/47578 -226- PCT/US99/18065 with a gradient of 10-15% MeOH-CH 2 C1 2 to afford 42.2 mg (79%) of the title compound as a yellow solid. 'H-NMR (DMSO-d 6 , 400 MHz) 5 9.39 (br s, 2H), 9.12 (br s, 2H), 8.63 (dd, 1H, J = 4.2, 1.6 Hz), 8.44 (s, 1H), 8.16 (m, 2H), 7.89 (d, 1H, J = 8.5 Hz), 7.54 (dd, 1H, J = 9.1, 2.6 Hz), 7.39 (dd, 1H, J = 8.3, 5 4.2 Hz), 7.20 (d, 1H, J = 2.5 Hz) and 2.55 (s, 3H). Mass spectrum (ESI, m/z): Called. For C 15
HI
4
N
4
S
2 , 315.1 (M+H), found 315.2. Example 198 5-Methylthio-4-[(3-phenylphenyl)aminofthiphene-2-carboxamidine 10 hydrochloride a) Methyl 5-methylthio-4-J(3-phenylphenyl)amino]thiophene-2 carboxylate: The same procedure as in Example 197, step (a), was followed using 62.2 mg (0.233 mmol) of methyl 4-bromo-5-methylthiothiophene-2 15 carboxylate (as prepared in Example 241, step (a)), 4.7 mg (9.0 mol %) of palladium (II) acetate, 20.0 mg (13.8 mol %) of racemic-BINAP, 140 mg (0.430 mmol) of cesium carbonate, 48.2 mg (0.285 mmol) of 3-aminobiphenyl and 466 .L of toluene, and chromotographed as before using 20-40%
CH
2 C1 2 -hexane to afford 52.3 mg (63%) of the title compound as a yellow 20 resin. 'H-NMR (CDCl 3 , 400 MHz) 8 7.81 (s, 1H), 7.61 (m, 2H), 7.46 (m, 2H), 7.38 (m, 2H), 7.21 (m, 2H), 7.03 (m, 1H), 6.22 (s, 1H), 3.90 (s, 3H), 2.43 (s, 3H). Mass spectrum (ESI, m/z): Calcd. For Cl 9
H
17
NO
2
S
2 , 356.1 (M+H), found 356.2. b) 5-Methylthio-4-[(3-phenylphenyl)aminofthiophene-2 25 carboxamidine hydrochloride: The same procedure as in Example 197, step (b) was followed using 46.4 mg (0.131 mmol) of methyl 5-methylthio-4 [(3-phenylphenyl)amino]thiophene-2-carboxylate (as prepared in previous step), 0.76 mL of trimethylaluminum (2.0 M in toluene, 1.57 mmol), 87.7 mg of ammonium chloride (1.64 mmol) and 0.79 mL of toluene, and purified on a 30 5-g silica SPE column with 5-10% MeOH-CH 2 C1 2 to afford 46.8 mg (95%) of the title compound as a yellow foam. 'H-NMR (DMSO-d 6 , 400 MHz) 8 9.04 (br s, 4H), 8.10 (s, 1H), 8.06 (s, 1H), 7.62 (m, 2H), 7.46 (m, 2H), 7.35 (m, 2H), 7.19 (t, 1H, J = 1.9 Hz), 7.12 (d, 1H, J = 8.2 Hz), 6.95 (dd, 1H, J = 7.8, 1.9 Hz), 2.53 (s, 3H). Mass spectrum (ESI, m/z): Calcd. For C 18 sH 17
N
3
S
2 , 35 340.1 (M+H), found 340.2.
WO 00/47578 -227- PCT/US99/18065 Example 199 5-Methylthio-4-(3-quinolylamino)thiophene-2-carboxamidine hydrochloride a) Methyl 5-methylthio-4-(3-quinolylamino)thiophene-2 5 carboxylate: The same procedure as in Example 197, step (a) was followed using 104 mg (0.389 mmol) of methyl 4-bromo-5-methylthiothiophene-2 carboxylate (as prepared in Example 241, step (a)), 7.1 mg (8.1 mol %) of palladium (II) acetate, 29.3 mg (12.1 mol%) of racemic-BINAP, 192 mg (0.589 mmol) of cesium carbonate, 70.5 mg (0.489 mmol) of 3 10 aminoquinoline and 778 pL of toluene, and chromatographed as before using 3-8% ethyl acetate-CH 2 C1 2 to afford 34.4 mg (27%) of the title compound as a yellow resin. 'H-NMR (CDCl 3 , 400 MHz) 8 8.73 (d, 1H, J = 2.5 Hz), 8.04 (d, 1H, J = 8.2 Hz), 7.85 (d, 1H, J = 4.0 Hz), 7.71 (d, 1H, J = 7.9 Hz), 7.62 (m, 1H), 7.56 (m, 2H), 6.34 (s, 1H), 3.93 (s, 3H) and 2.46 (s, 3H). Mass spectrum 15 (ESI, m/z): Calcd. For C 16
H
1 4
N
2 0 2
S
2 , 331.1 (M+H), found 331.3. b) 5-Methylthio-4-(3-quinolylamino)thiophene-2-carboxamidine hydrochloride: The same procedures as in Example 197, step (b) was followed using 32.3 mg (0.0977 mmol) of methyl 5-methylthio-4-(3 quinolylamino)thiophene-2-carboxylate (as prepared in previous step), 0.586 20 mL of trimethylaluminum (2.0 M in toluene, 1.17 mmol) and 65.8 mg of ammonium chloride (1.26 mmol) and 0.59 mL of toluene and purified on a 5-g silica SPE column with 5-12% MeOH-CH 2 C1 2 to afford, after concentration once from MeOH-MeCN (1:1), 17.3 mg (51%) of the title compound as a light tan crystalline solid. 'H-NMR (DMSO-d 6 , 400 MHz) 8 9.09 (br s, 4H), 8.79 25 (s, 1H), 8.56 (s, 1H), 8.12 (s, 1H), 7.89 (m, 1H), 7.79 (m, 1H), 7.56 (s, 1H), 7.50 (m, 2H) and 2.55 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for
C
15
HI
4
N
4
S
2 , 315.1 (M+H), found 315.4. Example 200 30 5-Methylthio-4-(pyrimidin-2-ylamino)thiophene-2-carboxamidine hydrochloride a) Methyl 5-methylthio-4-(pyrimidin-2-ylamino)thiophene-2 carboxylate: The same procedure as in Example 197, step (a) was followed 35 using 50.9 mg (0.191 mmol) of methyl 4-bromo-5-methylthiothiophene-2- WO 00/47578 -228- PCT/US99/180 6 5 carboxylate (as prepared in Example 241, step (a)), 2.7 mg (6.3 mol %) of palladium (II) acetate, 11.3 mg (9.5 mol %) of racemic-BINAP, 101 mg (0.310 mmol) of cesium carbonate, 25.9 mg (0.270 mmol) of 2 aminopyrimidine and 381 pL of dioxane, and chromatographed as before 5 using 5-10% ethyl acetate-hexane to afford 16.7 mg (31%) of the title compound as a yellow crystalline solid: 1H-NMR (CDCl3, 400 MHz) 5 8.72 (s, 1H), 8.49 (d, 1H, J = 4.8 Hz), 6.80 (t, 1H, J = 4.8 Hz), 3.92 (s, 3H), 2.42 (s, 3H) and 1.28 (br s, 2H). Mass spectrum (ESI, m/z): Calcd. for CIIHIIN 3 0 2 S2, 282.0 (M+H), found 282.3. 10 b) 5-Methylthio-4-(pyrimidin-2-ylamino)thiophene-2 carboxamidine hydrochloride: The same procedure as in Example 197, step (b) was followed using 15.2 mg (0.0540 mmol) of methyl 5-methylthio-4 (pyrimidin-2-ylamino)thiophene-2-carboxylate (as prepared in previous step), 0.324 mL or trimethylaluminum (2.0 M in toluene, 0.648 mmol) and 36.4 mg 15 of ammonium chloride (0.680 mmol) and 0.32 mL of toluene, and purified on a 2-g silica SPE column with 5-15% MeOH-CH 2 C12 to afford, after concentration once from MeOH-MeCN (1:10), 11.4 mg (70%) of the title compound as a light yellow crystalline solid. 'H-NMR (DMSO-d 6 , 300 MHz) 8 9.24 (br s, 2H), 8.85 (br s, 2H), 8.45 (d, 1H, J = 4.8 Hz), 8.25 (s, 1H), 6.87 (t, 20 1H, J = 4.8 Hz) and 2.53 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for CloHIINsS 2 , 266.1 (M+H), found 266.2. Example 201 4-[( 4 -Cyclohexylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 25 hydrochloride a) Methyl 4-[(4-cyclohexylphenyl)aminof-5-methllylthiothlliophene -2-carboxylate: The same procedure as in Example 197, step (a) was followed using 122 mg (0.457 mmol) of methyl 4-bromo-5-methylthiothiophene-2 30 carboxylate (as prepared in Example 241, step (a)), 9.9 mg (9.7 mol %) of palladium (II) acetate, 42.3 mg (14.9 mol %) of racemic-BINAP, 206 mg (0.632 mmol) of cesium carbonate, 102 mg (0.582 mmol) of 4 cyclohexylaniline and 913 pL of toluene, and chromatographed as before using 20-40% CH 2 C1 2 -hexane to afford 73.8 mg (45%) of the title compound 35 as a light green resin: 1H-NMR (CDCl 3 , 400 MHz) 8 7.74 (s, 1H), 7.15 (d, WO 00/47578 -229- PCT/US99/18065 2H, J = 8.4 Hz), 6.98 (d, 2H, J = 8.4 Hz), 6.12 (s, 1H), 3.88 (s, 3H), 2.48 (inm, 1H), 2.39 (s, 3H), 1.87 (min, 4H), 1.76 (br d, 1H, J = 12.5 Hz), 1.41 (min, 4H) and 1.28 (min, 1H). Mass spectrum (ESI, m/z): Calcd. for C 19
H
23
NO
2
S
2 , 362.1 (M+H), found 362.4. 5 b) 4-[(4-Cyclohexylphenyl)amino]-5-methylthiothiophene-2 carboxamidine hydrochloride: The same procedure as in Example 197, step (b) was followed using 70.2 mg (0.194 mmol) of methyl 4-[(4 cyclohexylphenyl)amino]-5-methylthiothiophene-2-carboxylate (as prepared in previous step), 0.970 mL or trimethylaluminum (2.0 M in toluene, 1.94 10 mmol), 109 mg of ammonium chloride (2.04 mmol) and 0.97 mL of toluene, and purified on a 10-g silica SPE column with 4-8% MeOH-CH 2
CI
2 to afford 57.7 mg (78%) of the title compound as a yellow foam. 'H-NMR (DMSO-d 6 , 400 MHz) 8 8.45 (br s, 4H), 7.97 (s, 1H), 7.86 (s, 1H), 7.08 (d, 2H, J = 8.5 Hz), 6.92 (d, 2H, J = 8.5 Hz), 2.48 (s, 3H), 1.65-1.85 (min, 5H) and 1.35 (inm, 15 5H). Mass spectrum (ESI, m/z): Calcd. for C 18
H
23
N
3
S
2 , 346.1 (M+H), found 346.4. Example 202 Methyl 4-amino-5-methylthiothiophene-2-carboxylate 20 To a pressure tube (Ace Glass, Vineland, NJ) containing 1.0 g (4.30 mmol) of 5-(methoxycarbonyl)-2-methylthiothiophene-3-carboxylic acid (as prepared in Example 95), 1.01 mL (1.1 equiv, 4.73 mmol) of diphenylphosphoryl azide, and 1.57 mL (2.1 equiv, 9.03 mmol) of NN 25 diisopropylethylamine was charged 7 mL of t-butanol. The resultant mixture was sealed and heated to 80oC in an oil bath for 6 h. The dark reaction mixture was cooled to ambient temperature and concentrated in vacuo. The crude oil was dissolved in 3 mL of CH 2 C1 2 and then treated with 2 mL of 1:1
CH
2 C1 2 -trifluoroacetic acid followed by 0.5 mL H 2 0. After 6 h, the mixture 30 was concentrated in vacuo, dissolved in 50 mL of CH 2 Cl 2 , washed with sat'd. NaHCO 3 , dried (Na 2
SO
4 ), and eluted through a pad of silica gel with 50% ethyl acetate-hexanes. The solvent was concentrated in vacuo and the crude amine was purified by preparative thin layer chromatography (20% ethyl acetate-hexanes, 2000 tm SiO 2 gel) to yield 210 mg (24%) of methyl 4- WO 00/47578 -230- PCT/US99/18065 amino-5-methylthiothiophene-2-carboxylate as a honey-colored oil. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.28 (s, 3H), 3.77 (s, 3H), 5.36 (bs, 2H), 7.24 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C 7
H
9
NO
2
S
2 , 204.02 (M+H), found 204.0. 5 Example 203 Methyl 4-[(aminothzioxomethyl)amino]-5-methylthiothiophene-2-carboxylate To a stirring 5 mL biphasic CH 2 Cl 2 -NaHCO 3 (1:1, v/v) mixture of 98 10 mg (0.48 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate was added 43 pL (1.2 equiv, 0.57 mmol) of thiophosgene (Aldrich Chemical, Milwaukee, WI). The reaction was stirred vigorously overnight, diluted with
CH
2 C1 2 (50 mL), and the layers separated. The organic layer was washed with NaHCO 3 (lxl5 mL), brine (lxl5mL), and dried (Na 2
SO
4 ). Concentration of 15 the solvent in vacuo yielded the crude isothiocyanate, which was dissolved in 5 mL of 2M NH 3 in MeOH and stirred overnight. Thereaction was concentrated to V2 volume and filtered. The filtered solids were washed with acetone and dried, yielding 79.8 mg (63.4%) of methyl 4 [(aminothioxomethyl)amino]-5-methylthiothiophene-2-carboxlyate as a light 20 tan solid. IH NMR (DMSO-d 6 , 300 MHz) 8 2.51 (s, 3H), 3.81 (s, 3H), 7.41 (bs, 2H), 8.03 (s, 1H) and 9.27 (bs, 1H). Mass spectrum (ESI, m/z): Calcd. for CsHI 0
N
2 0 2
S
3 , 263.00 (M+H), found 263.2. Example 204 25 5-Methylthio-4-[(4-phenyl(1,3-thiazol-2-yl))amino]thiophene- 2 carboxamidine a) Methyl 5-methylthio-4-J(4-phenyl(1,3-thiazol-2 yl)aminofthiophene-2-carboxylate: To a 25-mL round bottom flask 30 containing 40 mg (0.15 mmol) of methyl 4-[(aminothioxomethyl)amino]-5 methylthiothiophene-2-carboxylate and 30.3 mg (1 equiv, 0.15 mmol) of bromoacetophenone was added 2 mL of acetone, and the resultant mixture was heated to reflux for 18 h. The reaction was cooled to room temperature and filtered to give 50 mg (92%) of methyl 5-methylthio-4-[(4-phenyl(1,3-thiazol 35 2-yl))amino]thiophene-2-carboxylate, which was used without further WO 00/47578 -231- PCT/US99/18065 purification. 'H NMR (DMSO-d 6 , 300 MHz) 5 2.49 (s, 3H), 3.84 (s, 3H), 7.09 (s, 1H), 7.26-7.48 (min, 3H), 7.85 (min, 2H), 8.63 (s, 1H), 10.06 (bs, 1H). Mass spectrum (ESI, m/z): Calcd. for C1 6 H1 4
N
2 0 2
S
3 , 363.03 (M+H), found 363.4. b) 5-Methylthio-4-J(4-phenyl(1,3-thiiazol-2-yl)aminofthiophene 5 2-carboxamidine: Using a procedure similar to that of Example 154, step (b), 47 mg (0.13 mmol) of methyl 5-methylthio-4-[(4-phenyl(1,3-thiazol-2 yl))amino]thiophene-2-carboxylate was allowed to react with 0.5 mL (8 equiv, 1.04 mmol) of the AlMe 3
/NH
4 Cl reagent and purified by preparative thin layer chromatography (20% MeOH-CHCl 3 -sat'd. NH 3 , 500 [m SiO 2 gel plate) to 10 give 19 mg (42%) of 5-mehtylthio-4-[(4-phenyl(1,3-thiazol-2 yl))amino]thiophene-2-carboxamidine as a yellow solid. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.43 (s, 3H), 7.27-7.42 (m, 4H), 7.90 (d, 2H, J = 7.1 Hz), 8.41 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for CI 5
HI
4
N
4
S
3 , 347.05 (M+H), found 347.1. 15 Example 205 5-Methylthio-4-{[4-(4-phenylphenyl)(1,3-thiazol-2-yl)]amino}thiophene-2 carboxamidine 20 a) Methyl 5-methylthio-4-{[4-(4-phenylphenyl)(1,3-thiazol-2 yl)]amino}thiophene-2-carboxylate: Using a procedure similar to Example 204, step (a) 53 mg (0.2 mmol) of methyl 4-[(aminothioxomethyl)amino]-5 methylthiothiophene-2-carboxylate was allowed to react with 55.6 mg (0.2 mmol) of 4-phenyl-bromoacetophenone for 3 h to afford 57 mg (65%) of 25 methyl 5-methylthio-4- { [4-(4-phenylphenyl)(1,3-thiazol-2 yl)]amino}thiophene-2-carboxylate. 1H NMR (DMSO-d 6 , 300 MHz) 8 2.51 (s, 3H), 3.86 (s, 3H), 6.93 (s, 1H rotomer), 7.10 (s, 1H rotomer), 7.27 (s, 1H rotomer), 7.37-7.50 (min, 3H rotomer), 7.72-7.76 (min, 4H rotomer), 8.4 (d, 2H, 8.4 Hz), 8.66 (s, 1H rotomer), 10.10 (bs, 1H). Mass spectrum (ESI, m/z): 30 Calcd. for C 22
H,
8
N
2 0 2
S
3 , 439.06 (M+H), found 439.2. b) 5-Methylthio-4-{[4-(4-phenylphenyl(1,3-thiazol-2 yl)]amino}thiophene-2-carboxamidine: Using a procedure similar to that of Example 154, step (b), 57 ming (0.12 mmol) of methyl 5-methylthio-4- { [4-(4 phenylphenyl)(1,3-thiazol-2-yl)]amino}thiophene-2-carboxylate was allowed 35 to react with 6.7 equiv (0.87 mmol) of the AlMe 3
/NH
4 Cl reagent and purified WO 00/47578 -232- PCT/US99/18065 by preparative thin layer chromatography (20% MeOH-CHCl 3 -sat'd. NH 3 , 500 jIm SiO 2 plate) to give 20.7 mg (40.7%) of 5-methylthio-4-{ [4-(4 phenylphenyl)(1,3-thiazol-2-yl)]amino } thiophene-2-carboxamidine. 1H NMR (DMSO-d 6 , 400 MHz) 8 2.51 (s, 3H), 6.93 (s, 1H), 7.10 (s, 1H), 7.27 (s, 1H), 5 7.35-7.50 (m, 4H), 7.72-7.76 (m, 4H), 7.94-7.96 (m, 2H), 8.66 (s, 1H), 10.11 (bs, 1H). Mass spectrum (ESI, m/z): Calcd. for C 2 1 HIsN 4
S
3 , 423.08 (M+H), found 423.2. Example 206 10 4-[(5-Methyl-4-phenyl(1,3-thiazol-2-yl))amino]-5-methylthiothiophene-2 carboxamidine a) Methyl-4-J(5-methlyl-4-phenyl(1,3-thiazol-2-yl)aminof-5 methylthiothiophene-2-carboxylate: Using a procedure similar to Example 15 204, step (a), 51 mg (0.19 mmol) of methyl 4-[(aminothioxomethyl)amino]-5 methylthiothiophene-2-carboxylate was allowed to react with 41.4 mg (0.38 mmol) of 2-bromopropiophenone (Aldrich Chemical Co., Milwaukee, WI) in 2 mL of DMF for 4 h. Concentration in vacuo of the reaction mixture afforded 73 mg (100%) of methyl-4-[(5-methyl-4-phenyl(1,3-thiazpl-2 20 yl))amino]-5-methylthiothiophene-2-carboxylate. Mass spectrum (ESI, m/z): Calcd. for CI 7
H
16
N
2 0 2
S
3 , 377.05 (M+H), found 377.2. b) 4-[(5-Methyl-4-phenyl(1,3-thiazol-2-yl))amino]-5 methylthiothiophene-2-carboxamidine: Using a procedure similar to Example 154, step (b), 73 mg (0.19 mmol) of methyl-4-[(5-methyl-4 25 phenyl(1,3-thiazol-2-yl))amino]-5-methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (1.5 mmol) of the AIMe 3
/NH
4 Cl reagent and purified by preparative thin layer chromatography (20 %-MeOH-CHCl 3 -sat'd.
NH
3 , 500 pm SiO 2 plate) to afford 17.9 mg (26 %) of 4-[(5-methyl-4 phenyl(1,3-thiazol-2-yl))amino]-5-methylthiothiophene-2-carboxamidine. 'H 30 NMR (DMSO-d 6 , 300 MHz): 8 2.40 (s, 3H), 2.51 (s, 3H rotomer), 2.73 (s, 3H rotomer), 7.29-7.44 (m, 2H rotomer), 7.64-7.73 (m, 3H rotomer), 7.95 (s, 1H rotomer), 8.06 (s, 1H rotomer). Mass spectrum (ESI, m/z): Calcd. for C1 6
HI
6
N
4
S
3 , 361.06 (M+H), found 361.2.
WO 00/47578 -233- PCT/US99/18065 Example 207 4-{[4-Hydroxy-4-(trifluoromethyl) (1,3-thiazolin-2-yl)]amino}-5 methlylthiothiophene-2-carboxamidine 5 a) Methyl 4-{[4-hydroxy-4-(trifluoromethyl)(1,3-thiazolin-2 yl)]amino}-5-methylthiothiophene-2-carboxylate: Using a procedure similar to Example 204, step (a), 56 mg (0.21 mmol) of methyl 4 [(aminothioxomethyl)amino]-5-methylthiothiophene-2-carboxylate was allowed to react with 40 mg (0.21 mmol) of bromotrifluoroacetone (Aldrich 10 Chemical Co., Milwaukee, WI) to afford 40.3 mg (54 %) of methyl 4-{[4 hydroxy-4-(trifluoromethyl)(1,3-thiazolin-2-yl)]amino}-5 methylthiothiophene-2-carboxylate. Mass spectrum (ESI, m/z): Calcd. for C IH IF 3
N
2 0 3
S
3 , 373.00 (M+H), found 373.0. b) 4-{[4-Hydroxy-4-(trifluoromethyl) (1,3-thiazolin-2-yl)]amino} 15 5-methliylthiothiophene-2-carboxamidine: Using a procedure similar to Example 154, step (b), 40 mg (0.11 mmol) of methyl 4-{ [4-hydroxy-4 (trifluoromethyl)(1,3-thiazolin-2-yl)]amino }-5-methylthiothiophene-2 carboxylate was allowed to react with 8 equiv (0.89 mmol) of the AIMe 3
/NH
4 Cl reagent and purified by preparative thin layer chromatography 20 (20 %-MeOH-CHCl 3 -sat'd. NH 3 , 500 pm SiO 2 plate) to afford 11 mg (28 %) of 4- { [4-hydroxy-4-(trifluoromethyl)(1,3 -thiazolin-2-yl)] amino } -5 methylthiothiophene-2-carboxamidine as a ca. 1:1 mixture of cyclized aminal and open imine tautomers. 'H NMR (DMSO-d 6 , 300 MHz) 8 2.73 (s, 3H tautomer), 2.89 (s, 3H tautomer), 3.36 (d, 2H, J=6.5 Hz), 3.62 (d, 2H, J=7.2 25 Hz), 7.95 (s, 1H), 8.36 (bs, 2H), 9.79 (bs, 1H). Mass spectrum (ESI, m/z): Calcd. for CIoHIIF 3
N
4 0S 3 , 357.01 (M+H), found 357.2. Example 208 5-Methylthio-4-(2-naphthylamino) thiophene-2-carboxamidine 30 a) Methyl 5-methylthio-4-(2-naphthylamino)thiophene-2 carboxylate: To an oven-dried round bottom flask equipped with Teflon coated stir bar and rubber septum was added 190 mg (0.93 mmol) of methyl 4 amino-5-methylthiothiophene-2-carboxylate, 320 mg (2 equiv, 1.86 mmol) of 35 2-napthalene boronic acid (Lancaster Synthesis, Windham, NH), and 168 mg WO 00/47578 -234- PCT/US99/18065 (1 equiv, 0.93 mmol) of Cu(OAc) 2 (Aldrich Chemical Co., Milwaukee, WI). The flask was flushed with Ar, then charged with 4 mL CH 2 C1 2 followed by 259 pL (2 equiv, 1.86 mmol) of NEt 3 . The mixture was stirred vigorously for 48 h and then filtered through a small pad of SiO 2 , eluting with 50 % ethyl 5 acetate-hexanes. Concentration of the solvent in vacuo, and purification of the residue by preparative thin layer chromatography (25 % ethyl acetate-hexanes, 1000 pM SiO 2 plate) afforded 170 mg (55 %) of methyl 5-methylthio-4-(2 naphthylamino)thiophene-2-carboxylate and 54 mg (28 %) of recovered methyl 4-amino-5-methylthiothiophene-2-carboxylate. 'H NMR (CDCl 3 , 400 10 MHz) 5 2.43 (s, 3H), 3.92 (s, 3H), 6.29 (s, 1H), 7.21 (dd, 1H, J= 2.35, 8.7 Hz), 7.33-7.37 (m, 2H), 7.45 (min, 1H), 7.71 (d, 1H, J=8.2 Hz), 7.78 (min, 2H), 7.88 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C 7 Hi 5 N0 2
S
2 , 330.06 (M+H), found 330.1. b) 5-Methylthio-4-(2-naphthylamino)thiophene-2 15 carboxamidine hydrochloride: Using a procedure similar to Example 154, step(b),730 mg (2.21 mmol) of methyl 5-methylthio-4-(2 naphthylamino)thiophene-2-carboxylate was allowed to react with 8 equiv (17.7 mmol) of the AlMe 3
/NH
4 Cl reagent and purified by preparative thin layer chromatography (20 %-MeOH-CHCl 3 -sat'd. NH 3 , 1000 Pm SiO 2 plate) 20 to afford 5-methylthio-4-(2-naphthylamino)thiophene-2-carboxamidine, which was dissolved in 4 mL of dry MeOH, cooled to 0 0 C and carefully treated with 1.6 mL(1.5 equiv, 3.31 mmol) of 2M HCI in ether. The reaction was stored at 5 0 C overnight, then concentrated in vacuo with toluene (3x1 0mL) and then hexanes (2x 10 mL). The yellow solid was dried under vacuum to afford 415 25 mg (53.6 %) of 5-methylthio-4-(2-naphthylamino)thiophene-2-carboxamidine hydrochloride. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.53 (s, 3H), 7.20 (d, 1H, J=2.2 Hz), 7.24-7.31 (min, 2H), 7.38 (min, 1H), 7.69 (d, 1H, 8.1 Hz), 7.75-7.79 (min, 2H), 8.13 (s, 1H), 8.24 (s, 1H), 9.06 (bs, 2H), 9.33 (bs, 2H). Mass spectrum (ESI, m/z): Calcd. for C1 6
HI
5
N
3
S
2 , 314.08 (M+H), found 314.5. 30 WO 00/47578 -235- PCT/US99/18065 Example 209 4-J(4-Chlorophenyl)aminof-5-methylthiothiophene-2-carboxamidine a) Methyl 4-[(4-chlorophenyl)aminof-5-methylthiothiophene-2 5 carboxylate: Using a procedure similar to Example 208, step (a), 66.6 mg (0.32 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate was allowed to react with 100 mg (2 equiv, 0.64 mmol) of 4-chlorophenyl boronic acid to give 11.8 mg (11.7 %) of methyl 4-[(4-chlorophenyl)amino]-5 methylthiothiophene-2-carboxylate and 21 mg (31.5 %) of unreacted starting 10 material. 'H NMR (CDCl 3 , 400 MHz) 8 2.41 (s, 3H), 3.89 (s, 3H), 6.09 (bs, 1H), 6.94 (d, 2H, J=8.6 Hz), 7.25 (d, 2H, J=8.6 Hz), 7.70 (s, 1H). b) 4-[(4-chlorophenyl)aminoJ-5-methylthiothiophene-2 carboxamidine hydrochloride: Using a procedure similar to Example 154, step (b), 11.8 mg (0.037 mmol) of methyl 4-[(4-chlorophenyl)amino]-5 15 methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (2.96 mmol) of the AlMe 3
/NH
4 Cl reagent to afford 13 mg (100 %) of 4-[(4 chlorophenyl)amino]-5-methylthiothiophene-2-carboxamidine. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.41 (s, 3H), 6.91-6.95 (m, 2H), 7.10-7.13 (m, 2H), 7.64 (s, 1H), 7.93 (s, 1H), 8.67 (bs, 2H), 9.11 (bs, 2H). Mass spectrum (ESI, 20 m/z): Calcd. for C1 2 H1 2 C1N 3
S
2 , 298.02 (M+H), found 298.1. Example 210 4-[(3-Methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 25 a) Methyl 4-[(3-methylphenyl)amino]-5-methylthiothiophene-2 carboxylate: Using a procedure similar to Example 208, step (a), 55.7 mg (0.27 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate was allowed to react with 73.4 mg (2 equiv, 0.54 mmol) of 3-methylphenyl boronic acid to give 29.2 mg (36.8 %) of methyl 4-[(3-methyl)amino]-5 30 methylthiothiophene-2-carboxylate. 1H NMR (CDCl 3 , 400 MHz) 8 2.35 (s, 3H), 2.40 (s, 3H), 3.89 (s, 3H), 6.11 (bs, 1H), 6.80-6.86 (m, 3H), 7.20 (m, 1H), 7.77 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C1 4 HIsNO 2
S
2 , 294.06 (M+H), found 294.1.
WO 00/47578 -236- PCT/US99/18065 b) 4-J(3-Methlzylphenyl)aminof-5-methylthiothiophene-2 carboxamidine: Using a procedure similar to Example 154, step (b), 29.2 mg (0.098 mmol) of methyl 4-[(3-methyl)amino]-5-methylthiothiophene-2 carboxylate was allowed to react with 8 equiv (0.78 mmol) of the 5 AlMe 3
/NH
4 Cl reagent and purified by preparative thin layer chromatography (20 %-MeOH-CHCl 3 -sat'd. NH 3 , 500 Im SiO 2 plate) to afford 27 mg (100 %) of 4-[(3-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine. IH NMR (CDCl 3 , 400 MHz) 8 2.24 (s, 3H), 2.50 (s, 3H), 6.65 (d, 1H, J=7.3 Hz), 6.74-6.76 (min, 2H), 7.10 (min, 1H), 7.88 (s, 1H), 7.97 (s, 1H), 9.07 (bs, 3H). 10 Mass spectrum (ESI, m/z): Calcd. for Cl 3 Hi 5
N
3
S
2 , 278.08 (M+H), found 278.2. Example 211 4-[(3-Methoxyphenyl)aminof-5-methylthliothiophene-2-carboxamidine 15 a) Methyl-4-[(3-methoxyphenyl)amino]-S-methylthiothiophene 2-carboxylate: Using a procedure similar to Example 208, step (a), 73.2 mg (0.35 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate was allowed to react with 109 mg (2 equiv, 0.70 mmol) of 3-methoxyphenyl 20 boronic acid to give 25.2 mg (23 %) of methyl 4-[(3-methoxyphenyl)amino] 5-methylthiothiophene-2-carboxylate. 'H NMR (CDCl 3 , 400 MHz) 8 2.40 (s, 3H), 3.81 (s, 3H), 3.89 (s, 3H), 6.12 (s, 1H), 6.43-6.63 (min, 2H), 7.20 (min, 1H), 7.78 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for Cl 4
H]
5
NO
3
S
2 , 310.06 (M+H), found 310.1. 25 b) 4-[(3-Methylphenyl)amino]-5-methylthiothiophene-2 carboxamidine hydrochloride: Using a procedure similar to Example 154, step (b), 25.2 mg (0.081 mmol) of methyl 4-[(3-methyl)amino]-5 methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (0.64 mmol) of the AIMe 3
/NH
4 Cl reagent to afford 27 mg (100 %) of 4-[(3 30 methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine. 'H NMR (DMSO, 400 MHz) 8 2.49 (s, 3H), 3.71 (s, 3H), 6.41 (dd, 1H, J=2.1, 8.0 Hz), 6.49 (m, 1H), 6.50-6.54 (min, 1H), 7.12 (min, 1H), 7.97 (s, 1H), 8.01 (s, 1H), 8.88 (bs, 2H), 9.23 (bs, 2H). Mass spectrum (ESI, m/z): Calcd. for Ci 3 HIsN 3 0S 2 , 294.07 (M+H), found 294.1.
WO 00/47578 -237- PCT/US99/18065 Example 212 4-{[3-(Methylethyl)phenyl]amino}-5-methylthiothiophene-2-carboxamidine a) Methyl 4-{[3-(methylethyl)phenyl]amino}-5 5 methylthiothiophene-2-carboxylate: Using a procedure similar to Example 208, step (a), 74.4 mg (0.36 mmol) of methyl 4-amino-5-methylthiothiophene 2-carboxylate was allowed to react with 118 mg (2 equiv, 0.72 mmol) of 3 isopropylphenyl boronic acid to give 22.6 mg (19.5 %) of methyl 4-[(3 methylethylphenyl)amino]-5-methylthiothiophene-2-carboxylate. IH NMR 10 (CDCl 3 , 400 MHz) 8 1.27 (d, 6H, J=6.9 Hz), 2.40 (s, 3H), 2.89 (m, 1H), 3.88 (s, 3H), 6.15 (s, 1H), 6.86-6.89 (m, 3H), 7.24 (m, 1H), 7.77 (s, 1H). b) 4-{J3-(Methylethyl)phenyl]amino}-5-methylthiothiophene-2 carboxamidine: Using a procedure similar to Example 154, step (b), 22.6 mg (0.07 mmol) of methyl 4- { [3-(methylethyl)phenyl]amino}-5 15 methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (0.56 mmol) of the AlMe 3
/NH
4 Cl reagent to afford 18.9 mg (78.8 %) of ) 4-{ [3 (methylethyl)phenyl]amino } -5-methylthiothiophene-2-carboxamidine. IH NMR (DMSO-d 6 , 400 MHz) 8 1.18 (d, 6H, J=9.2 Hz), 2.51 (s, 3H), 2.81 (m, 1H), 6.71-6.77 (m, 2H), 6.85 (s, 1H), 7.14 (m, 1H), 7.98 (s, 1H), 8.32 (s, 1H), 20 8.88 (bs, 2H), 9.23 (bs, 2H). Mass spectrum (ESI, m/z): Calcd. for
CI
5
HI
9
N
3
S
2 , 306.11 (M+H), found 306.2. Example 213 5-Methylthio-4-[(3-nitrophenyl)amino]thiophene-2-carboxamidine 25 a) Methyl 5-methylthio-4-J(3-nitrophenyl)amino]thiophene-2 carboxylate: Using a procedure similar to Example 208, step (a), 74.4 mg (0.36 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate was allowed to react with 120 mg (2 equiv, 0.72 mmol) of 3-nitrophenyl boronic 30 acid to give 14.5 mg (12.4 %) of methyl 5-methylthio-4-[(3-nitrophenyl) amino]thiophene-2-carboxylate. 'H NMR (CDCl 3 , 400 MHz) 8 2.45 (s, 3H), 3.93 (s, 3H), 6.21 (s, 1H), 7.41-7.47 (m, 2H), 7.73-7.78 (m, 3H). b) S-Methylthio-4-[(3-nitrophenyl)amino]thiophene-2 carboxamidine: Using a procedure similar to Example 154, step (b), 14.5 mg WO 00/47578 -238- PCT/US99/18065 (0.04 mmol) of methyl 5-methylthio-4-[(3-nitrophenyl)amino]thiophene-2 carboxylate was allowed to react with 8 equiv (0.35 mmol) of the AlMe 3
/NH
4 C1 reagent to afford 4.3 mg (34.8 %) of 5-methylthio-4-[(3 nitrophenyl)amino]thiophene-2-carboxamidine. Mass spectrum (ESI, m/z): 5 Calcd. for C1 2
H
12
N
4 0 2
S
2 , 309.05 (M+H), found 309.2. Example 214 4-{[4-(Methylethyl)phenyl]amino}-5-methylthiothiophene-2-carboxamidine 10 a) Methyl 4-{[4-(methylethyl)phenyl]amino}-5 methylthiothiophene-2-carboxylate: Using a procedure similar to Example 208, step (a), 74.4 mg (0.36 mmol) of methyl 4-amino-5-methylthiothiophene 2-carboxylate was allowed to react with 118 mg (2 equiv, 0.72 mmol) of 4 isopropylphenyl boronic acid to give 14.5 mg (12.5 %) of methyl 4-[(4 15 methylethylphenyl)amino]-5-methylthiothiophene-2-carboxylate. 'H NMR (CDCl 3 , 400 MHz) 8 1.26 (d, 6H, J=6.2 Hz), 2.39 (s, 3H), 2.89 (m, 1H), 3.89 (s, 3H), 6.98-7.01 (m, 2H), 7.17-7.19 (m, 2H), 7.73 (s, 1H). b) 4-{[4-(Methylethyl)phenyl]amino}-5-methylthiothiophene-2 carboxamidine: Using a procedure similar to Example 154, step (b), 14.5 mg 20 (0.045 mmol) of methyl 4- { [4-(methylethyl)phenyl]amino}-5 methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (0.36 mmol) of the AlMe 3
/NH
4 Cl reagent to afford 11.4 mg (74 %) of 4- {[4 (methylethyl)phenyl]amino } -5-methylthiothiophene-2-carboxamidine. 'H NMR (DMSO-d 6 , 400 MHz) 8 1.17 (d, 6H, J=9.2 Hz), 2.51 (s, 3H), 2.81 (m, 25 1H), 6.92 (d, 2H, J=11.4 Hz), 7.10 (d, 2H,J=11.2 Hz), 7.88 (s, 1H), 7.96 (s, 1H), 8.89 (bs, 2H), 9.22 (bs, 2H). Mass spectrum (ESI, m/z): Calcd. for
C
5 sHI 9
N
3
S
2 , 306.11 (M+H), found 306.2. Example 215 30 4-J(3,4-Dimethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine a) Methyl 4-[(3,4-dimethylphenyl)amino]-S methylthiothiophene-2-carboxylate: Using a procedure similar to Example 208, step (a), 74.4 mg (0.36 mmol) of methyl 4-amino-5-methylthiothiophene- WO 00/47578 -239- PCT/US99/18065 2-carboxylate was allowed to react with 108 mg (2 equiv, 0.72 mmol) of 3, 4 dimethylphenyl boronic acid to give 135.9 mg (32.4 %) of methyl 4-[(3,4 dimethylphenyl)amino]-5-methylthiothiophene-2-carboxylate. 'H NMR (CDCl 3 , 400 MHz) 8 2.24 (s, 3H), 2.26 (s, 3H), 2.38 (s, 3H), 3.88 (s, 3H), 6.11 5 (bs, 1H), 6.80 -6.84 (m, 2H), 7.07 (d, 1H, J=7.9 Hz), 7.71 (s, 1H). b) 4-[(3,4-Dimethliylphenyl)amino]-5-methylthiothiophene-2 carboxamidine: Using a procedure similar to Example 154, step (b), 35.6 mg (0.116 mmol) of methyl 4-[(3,4-dimethylphenyl)amino]-5 methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (0.93 10 mmol) of the AlMe 3
/NH
4 C1 reagent to afford 26.1 mg (68.5 %) of 4-[(3,4 dimethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.13 (s, 3H), 2.16 (s, 3H), 2.51 (s, 3H), 6.69-6.78 (m, 2H), 6.99 (d, 1H, J=10.8 Hz), 7.76 (s, 1H), 7.91 (s, 1H), 8.82 (bs, 2H), 9.17 (bs, 2H). Mass spectrum (ESI, m/z): Calcd. for Cl 4
H,
7
N
3
S
2 , 292.09 (M+H), 15 found 292.2. Example 216 5-Methylthio-4-[(4-phenylphenyl)amino]thliiophene-2-carboxamidine 20 a) Methyl 5-methylthio-4-J(4-phenylphenyl)amino]thiophene-2 carboxylate: Using a procedure similar to Example 208, step (a), 74.4 mg (0.36 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate was allowed to react with 142.5 mg (2 equiv, 0.72 immol) of 4-phenylphenyl boronic acid to give 24.5 mg (19.1 %) of methyl 4-[(4-phenylphenyl)amino] 25 5-methylthiothiophene-2-carboxylate. 'H NMR (CDCl 3 , 400 MHz) 8 2.45 (s, 3H), 3.92 (s, 3H), 6.38 (bs, 1H), 7.08-7.14 (m, 2H), 7.33 (m, 1H), 7.43-7.46 (m, 2H), 7.54-7.60 (m, 4H), 7.82 (s, 1H). b) 5-Methylthio-4-[(4-phenylphenyl)amino]thiophene-2 carboxamidine: Using a procedure similar to Example 154, step (b), 24.5 mg 30 (0.07 immol) of methyl 4-[(4-phenylphenyl)amino]-5-methylthiothiophene-2 carboxylate was allowed to react with 8 equiv (0.56 mmol) of the AlMe 3
/NH
4 Cl reagent to afford 16.9 mg (64.1 %) of 5-methylthio-4-[(4 phenylphenyl)amino]thiophene-2-carboxamidine. ' H NMR (DMSO-d 6 , 400 MHz) 8 2.51 (s, 3H), 7.03 (d, 2H, J=8.6 Hz), 7.26-7.61 (m, 7H), 8.04 (s, 1H), WO 00/47578 240- PCT/US99/18065 -240 8.15 (s, 1H), 8.88 (bs, 2H), 9.25 (bs, 2H). Mass spectrum (ESI, m/z): Calcd. for CI 8 sHI 7
N
3
S
2 , 340.09 (M+H), found 340.2. Example 217 5 4-J(3-Fluoro-4-phenylphenyl)amino]-5-methylthiothiophene-2 carboxamidine a) Methyl 4-[(3-fluoro-4-phenylphenyl)aminof-5 methylthiothiophene-2-carboxylate: Using a procedure similar to Example 10 208, step (a), 74.4 mg (0.36 mmol) of methyl 4-amino-5-methylthiothiophene 2-carboxylate was allowed to react with 155.5 mg (2 equiv, 0.72 mmol) of 3 fluoro-4-phenylphenyl boronic acid to give 50.6 mg (41.6 %) of methyl 4-[(3 fluoro-4-phenylphenyl)amino]-5-methylthiothiophene-2-carboxylate. 'H NMR (CDCl 3 , 400 MHz) 8 2.44 (s, 3H), 3.91 (s, 3H), 6.19 (s, 1H), 6.78-6.86 (m, 15 2H), 7.32-7.39 (m, 2H), 7.73-7.47 (m, 2H), 7.55 (d, 1H, J=6.9 Hz), 7.82 (s, 1H). b) 4-[(3-Fluoro-4-phenylphenyl)amino]-5-methylthiothiophene 2-carboxamidine: Using a procedure similar to Example 154, step (b), 50.6 mg (0.13 mmol) of methyl 4-[(3-fluoro-4-phenylphenyl)amino]-5 20 methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (1.08 mmol) of the AlMe 3
/NH
4 CI reagent to afford 39 mg (76.1 %) of 4-[(3-fluoro 4-phenylphenyl)amino]-5-methylthiothiophene-2-carboxamidine. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.51 (s, 3H), 6.75-6.87 (m, 2H), 7.30-7.50 (m, 6H), 8.06 (s, 1H), 8.37 (s, 1H), 8.90 (bs, 2H), 9.27 (bs, 2H). Mass spectrum (ESI, 25 m/z): Calcd. for CI 8
HI
6
FN
3
S
2 , 358.08 (M+H), found 358.2. Example 218 4-(2H-Benzo[d]l,3-dioxolen-5-ylamino)-5-methylthiothiophene-2 carboxamidine 30 a) Methyl 4-(2H-benzo[d]Jl,3-dioxolen-5-ylamino)-5 methylthiothiophene-2-carboxylate: Using a procedure similar to Example 208, step (a), 74.4 mg (0.36 mmol) of methyl 4-amino-5-methylthiothiophene 2-carboxylate was allowed to react with 119.4 mg (2 equiv, 0.72 mmol) of 35 3,4-methylenedioxyphenyl boronic acid to give 24.4 mg (20.9 %) of methyl 4 (2H-benzo[d] 1,3-dioxolen-5-ylamino)-5-methylthiothiophene-2-carboxylate.
WO 00/47578 -241- PCT/US99/18065 'H NMR (CDC1 3 , 400 MHz) 8 2.39 (s, 3H), 3.87 (s, 3H), 5.96 (s, 2H), 6.00 (bs, 1H), 6.52 (dd, 1H, J=2.3, 8.3 Hz), 6.63 (d, 1H, J=2.2 Hz), 6.76 (d, 1H, J=8.3 Hz), 7.59 (s, 1H). b) 4-(2H-Benzo[d]Jl,3-dioxolen-5-ylamino)-5 5 methylthiothiophene-2-carboxamidine: Using a procedure similar to Example 154, step (b), 24.4 mg (0.075 mmol) of methyl 4-(2H-benzo[d]1,3-dioxolen-5 ylamino)-5-methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (0.6 mmol) of the AlMe 3
/NH
4 Cl reagent to afford 7.7 mg (29.7 %) 4 (2H-benzo[d] 1,3-dioxolen-5-ylamino)-5-methylthiothiophene-2 10 carboxamidine. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.51 (s, 3H), 5.95 (s, 2H), 6.46 (dd, 1H, J=3.0, 11.2 Hz), 6.65 (d, 1H, J=2.8 Hz), 6.79 (d, 1H, J=l 1.0 Hz), 7.80 (s, 1H), 7.87 (s, 1H), 8.91 (bs, 2H), 9.24 (bs, 2H). Mass spectrum (ESI, m/z): Calcd. for C1 3
HI
3
N
3 0 2
S
2 , 308.05 (M+H), found 308.2. 15 Example 219 4-[(4-Butylphenyl)amino]-5-methliylthiothiophene-2-carboxamidine a) Methyl 4-[(4-butylphenyl)amino]-5-methylthliiothiophene-2 carboxylate: Using a procedure similar to Example 208, step (a), 74.4 mg 20 (0.36 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate was allowed to react with 128 mg (2 equiv, 0.72 mmol) of 4-butylphenyl boronic acid to give 22.2 mg (18.3 %) of methyl 4-[(4-butylphenyl)amino]-5 methylthiothiophene-2-carboxylate. 'H NMR (CDCl 3 , 400 MHz) 8 0.97 (t, 2H, J=7.4 Hz), 1.38 (m, 2H), 1.59 (m, 2H obscured by water), 2.39 (s, 3H), 25 2.58 (t, 2H, J=7.6 Hz), 3.90 (s, 3H), 6.12 (bs, 1H), 6.97 (d, 2H, J=8.2 Hz), 7.12 (d, 2H, J=8.4 Hz), 7.73 (s, 1H). b) 4-[(4-Butylphenyl)amino]-5-methylthiothiophene-2 carboxamidine: Using a procedure similar to Example 154, step (b), 22.2 mg (0.06 mmol) of methyl 4-[(4-butylphenyl)amino]-5-methylthiothiophene-2 30 carboxylate was allowed to react with 8 equiv (0.52 mmol) of the AlMe 3
/NH
4 CI reagent to afford 18.9 mg (88 %) of 4-[(4-butylphenyl)amino] 5-methylthiothiophene-2-carboxamidine. 'H NMR (DMSO-d 6 , 400 MHz) 6 0.89 (t, 2H, J=9.7 Hz), 1.23-1.33 (m, 2H), 1.51 (m, 2H), 2.47-2.50 (m, 2H obscured by DMSO-d 6 ), 2.51 (s, 3H), 6.90 (d, 2H, J=11.3 Hz), 7.05 (d, 2H, WO 00/47578 -242- PCT/US99/18065 J=11.2 Hz), 7.86 (s, 1H), 7.94 (s, 1H), 8.78 (bs, 2H), 9.21 (bs, 2H). Mass spectrum (ESI, m/z): Calcd. for C 16
H
2 1
N
3
S
2 , 320.13 (M+H), found 320.2. Example 220 5 5-Methylthio-4-Jbenzylamino]thiophene-2-carboxamidine a) Methyl-5-methylthio-4-[benzylamino]thiophene-2 carboxylate: To a 2-dram vial equipped with a stir bar and septum cap was weighed 60 mg (0.29 mmol) of methyl 4-amino-5-methylthiothiophene-2 10 carboxylate and 30.7 mg (0.29 mmol) of benzaldehyde. The vial was charged with 1 mL CH 2 C1 2 -DMF (2:1, v/v) and 135 mg (2.2 equiv, 0.63 mmol) of NaHB(OAc) 3 was added. The reaction was flushed with Ar and allowed to stir for 48 h. At this time 2 mL of CH 3 OH was added, the reaction stirred an additional 15 min then diluted with 20 ml of CH 2 C1 2 . The organic layer was 15 washed with water (2x20 mL), dried (Na 2
SO
4 ), and concentrated in vacuo into an oven-dried 2 dram vial to give the crude methyl-5-methylthio-4 [benzylamino]thiophene-2-carboxylate together with unreduced imine. The crude reaction mixture was converted to the amidine without further purification. Mass spectrum (ESI, m/z): Calcd. for C 14 Hs 15
NO
2
S
2 , 294.06 20 (M+H), found 292.2 (imine), 294.2. b) 5-Methylthio-4-[benzylaminoJthiophene-2-carboxamidine: To a 2-dram vial containing a stir bar and methyl-5-methylthio-4 [benzylamino]thiophene-2-carboxylate (assume 0.29 mmol) was added 2 mL of toluene, followed by 8 equiv (2.32 mmol) of the AlMe 3
/NH
4 CI reagent. The 25 resultant yellow mixture was heated to 110 0 C for 3 h, cooled to ambient temperature, and then added to a slurry of 1 g of SiO 2 gel in 10 mL of CHCl 3 . After stirring for 15 min, the slurry was eluted through a 15-mL sintered glass funnel containing a pad of silica gel with 50 % CHCl 3 - CH 3 OH. The solvent was removed in vacuo and the residue was triturated with 10 % CH 3 OH 30 CHCl 3 and filtered. Removal of the solvent in vacuo gave the crude product which was purified by preparative thin layer chromatography (500 [m SiO 2 , 20 % CH 3 OH-CHCl 3 -sat'd. NH 3 ) 14.8 mg (18.3 % from methyl 4-amino-5 methylthiothiophene-2-carboxylate) of 5-methylthio-4-[benzylamino] thiophene-2-carboxamidine. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.49 (s, 3H), WO 00/47578 -243- PCT/US99/18065 4.35 (d, 2H, J=6.7 Hz), 5.91 (t, 1H, J=6.8 Hz), 7.20-7.38 (m, 6H). Mass spectrum (ESI, m/z): Calcd. for C1 3 Hi 5
N
3
S
2 , 278.08 (M+H), found 278.3. Example 221 5 4-(Indan-5-ylamino)-5-methylthiothiophene-2-carboxamidine a) Methyl 4-(indan-5-ylamino)-5-methylthiothiophene-2 carboxylate: Using the procedure described in Example 220, step (a), 60 mg (0.29 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate, 42.3 10 mg (0.29 mmol) of 5-indancarboxaldehyde, and 135 mg (2.2 equiv, 0.63 mmol) of NaHB(OAc) 3 were allowed to react to give methyl 4-(indan-5 ylamino)-5-methylthiothiophene-2-carboxylate. Mass spectrum (ESI, m/z): Calcd. for C 17 H1 9
NO
2
S
2 , 334.09 (M+H), found 332.3 (imine), 333.4. b) 4-(Indan-5-ylamino)-5-methylthiothiophene-2 15 carboxamidine: Using the procedure described in Example 220, step (b), 22.0 mg (27.3 % from methyl 4-amino-5-methylthiothiophene-2-carboxylate) of 4 (indan-5-ylamino)-5-methylthiothiophene-2-carboxamidine was obtained. 'H NMR (DMSO-d 6 , 400 MHz) 8 1.94-2.01 (m, 2H), 2.49 (s, 3H), 2.77-2.82 (m, 4H), 4.29 (d, 2H, J=5.6 Hz), 5.78 (t, 1H, J=8.1 Hz), 7.08 (d, 1H, J=7.8 Hz), 20 7.14 (d, 1H, J=7.5 Hz), 7.20 (s, 1H), 7.23 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for Cl 6
HI
9
N
3
S
2 , 318.11 (M+H), found 318.3. Example 222 4-(2,3-Dihydrobenzo[b]furan-5-ylamino)-5-methylthiothiophene-2 25 carboxamidine a) Methyl 4-(2,3-dihliydrobenzo[bffuran-5-ylamino)-5 methylthiothiophene-2-carboxylate: Using the procedure described in Example 220, step (a), 60 mg (0.29 mmol) of methyl 4-amino-5 30 methylthiothiophene-2-carboxylate, 42.9 mg (0.29 mmol) of 2,3 dihydrobenzo[b]furan-5-carboxaldehyde, and 135 mg (2.2 equiv, 0.63 mmol) of NaHB(OAc) 3 were allowed to react to give methyl 4-(2,3 dihydrobenzo[b]furan-5-ylamino)-5-methylthiothiophene-2-carboxylate. Mass spectrum (ESI, m/z): Calcd. for C1 6
H,
7
NO
3
S
2 , 336.07 (M+H), found 334.3 35 (imine), 335.3.
WO 00/47578 -244- PCT/US99/18065 b) 4
-(
2 ,3-Dihydrobenzo[bfuran-5-ylamino)-S methylthiothiophene-2-carboxamidine: Using the procedure described in Example 220, step (b), 21.8 mg (23.5 % from methyl 4-amino-5 methylthiothiophene-2-carboxylate) of 4-(2,3-dihydrobenzo[b]furan-5 5 ylamino)-5-methylthiothiophene-2-carboxamidine was obtained. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.49 (s, 3H), 3.13 (t, 2H, J=8.7 Hz), 4.24 (d, 2H, J=6.6 Hz), 4.48 (t, 2H, J=8.7Hz), 5.69 (t, 1H, J=6.7 Hz), 6.68 (d, 1H, J=12.4 Hz), 7.06 (d, 1H, J=7.4 Hz), 7.21 (s, 1H), 7.26 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C1 5
H,
7
N
3 0S 2 , 320.09 (M+H), found 320.3. 10 Example 223 5-Methylthio-4-J(2-phenylimidazol-4-yl)amino]thliiophene-2-carboxamidine a) Methyl 5-methylthio-4-[(2-phenylimidazol-4-yl) 15 amino]thiophene-2-carboxylate: Using the procedure described in Example 220, step (a), 60 mg (0.29 mmol) of methyl 4-amino-5-methylthiothiophene-2 carboxylate, 49.9 mg (0.29 mmol) of 4-formyl-2-phenylimidazole, and 135 mg (2.2 equiv, 0.63 mmol) of NaHB(OAc) 3 were allowed to react to give) methyl 5-methylthio-4-[(2-phenylimidazol-4-yl)amino]thiophene-2 20 carboxylate. Mass spectrum (ESI, m/z): Calcd. for CI 7 H1 7
N
3 0 2
S
2 , 360.08 (M+H), found 360.0. b) 5-Methylthio-4-J(2-phenylimnidazol-4-yl)amino]thiophene-2 carboxamidine: Using the procedure described in Example 220, step (b), 30.9 mg (30 % from methyl 4-amino-5-methylthiothiophene-2-carboxylate) of 5 25 methylthio-4-[(2-phenylimidazol-4-yl)amino]thiophene-2-carboxamidine was obtained. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.49 (s, 3H), 4.30-4.38 (m, 3H), 7.09 (bs, 1H), 7.32 (min, 1H), 7.40-7.44 (mn, 3H), 7.90-7.95 (mn, 3H), 8.43 (bs, 3H). Mass spectrum (ESI, m/z): Calcd. for Cl 6
HI
7
N
5
S
2 , 344.10 (M+H), found 344.2. 30 WO 00/47578 -245- PCT/US99/18065 Example 224 5-Methylthio-4-[(2-quinolylmethyl)aminofthiophene-2-carboxamidine a) Methyl 5-methylthio-4-[(2-quinolylmethyl)aminofthiophene 5 2-carboxylate: Using the procedure described in Example 220, step (a), 60 mg (0.29 mmol) of methyl 4-amino-5-methylthiothiophene-2-carboxylate, 45.5 mg (0.29 mmol) of 2-quinolinecarboxaldehyde, and 135 mg (2.2 equiv, 0.63 mmol) of NaHB(OAc) 3 were allowed to react to give methyl 5 methylthio-4-[(2-quinolylmethyl)amino]thiophene-2-carboxylate. Mass 10 spectrum (ESI, m/z): Calcd. for C 1 7 HI6N 2 0 2
S
2 , 345.07 (M+H), found 343.3 (imine), 345.2. b) 5-Methylthio-4-[(2-quinolylmethyl)aminoJthiophene-2 carboxamidine: Using the procedure described in Example 220, step (b), 2.5 mg (2.6 % from methyl 4-amino-5-methylthiothiophene-2-carboxylate) of 5 15 methylthio-4-[(2-quinolylmethyl)amino]thiophene-2-carboxamidine was obtained. Mass spectrum (ESI, m/z): Calcd. for C 1 6
HI
6
N
4
S
2 , 329.09 (M+H), found 329.3. Example 225 20 4-{[(3-Hydroxyphenyl)methyl]amino}-5-methylthiothiophene-2 carboxamidine a) Methyl 4-{[(3-hydroxyphenyl)methylfamino}-5 methylthiothiophene-2-carboxylate: Using the procedure described in 25 Example 220, step (a), 61.6 mg (0.30 mmol) of methyl 4-amino-5 methylthiothiophene-2-carboxylate, 49.5 mg (0.30 mmol) of 3 acetoxybenzaldehyde, and 135 mg (2.2 equiv, 0.63 mmol) of NaHB(OAc) 3 were allowed to react to give methyl 4-{ [(3-hydroxyphenyl)methyl]amino}-5 methylthiothiophene-2-carboxylate. Mass spectrum (ESI, m/z): Calcd. for 30 C 14
HI
6
NO
3
S
2 , 352.07 (M+H), found, 350.2 (imine), 352.1. b) Methyl 4-{[(3-hydroxyphenyl)methyl]amino}-5 methylthiothiophene-2-carboxylate: Using the procedure described in Example 220, step (b), 7.9 mg (8.9 % from methyl 4-amino-5 methylthiothiophene-2-carboxylate) of methyl 4-{ [(3-hydroxyphenyl) WO 00/47578 -246- PCT/US99/18065 methyl]amino}-5-methylthiothiophene-2-carboxylate; Mass spectrum (ESI, m/z): Calcd. for C1 3
HI
5
N
3 0S 2 , 294.07 (M+H), found 294.3. Example 226 5 5-Methylthio-4-(phenylcarbonylamino)thiophene-2-carboxamidine a) Methyl 5-methylthio-4-(phenylcarbonylamino)thiophene-2 carboxylate: To a stirring solution of 114 mg (0.55 mmol) of methyl 4-amino 5-methylthiothiophene-2-carboxylate in 4 mL of CH 2 C1 2 at 0 oC was added 10 142 pL (1.5 equiv, 0.82 mmol) of N,N-diisopropylethylamine via syringe, followed by 71.3 pL (1.1 equiv, 0.61 mmol) of benzoyl chloride. The reaction was allowed to warm to room temperature, and stirred an additional 4h. At this time the reaction was partitioned in 40 mL of 1:1 CH 2 Cl 2 -sat'd. NaHCO 3 (v/v) and the organic layer was separated, washed with 20 mL of brine, dried 15 (Na 2
SO
4 ), and concentrated in vacuo to afford 113 mg (66.8 %) of methyl 5 methylthio-4-(phenylcarbonylamino)thiophene-2-carboxylate which was used without further purification. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.55 (s, 3H), 3.83 (s, 3H), 7.47-7.56 (min, 2H), 7.64 (min, 1H), 7.88 (s, 1H), 7.93-7.99 (min, 2H), 10.12 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C1 4
HI
3 NO3S 2 , 308.04 20 (M+H), found 308.2. b) 5-Methylthio-4-(phenylcarbonylamino)thiophene-2 carboxamidine: Using a procedure similar to Example 154, step (b), 100 mg (0.32 mmol) of methyl 4- { [4-(methylethyl)phenyl] amino }-5 methylthiothiophene-2-carboxylate was allowed to react with 8 equiv (2.58 25 mmol) of the AlMe 3
/NH
4 Cl reagent to afford 95.4 mg (100 %) of 5 methylthio-4-(phenylcarbonylamino)thiophene-2-carboxamidine. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.59 (s, 3H), 7.30-7.64 (min, 3H), 7.98-8.00 (min, 2H), 8.23 (s, 1H), 9.19 (bs, 2H), 9.41 (bs, 2H), 10.35 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for Ci 3 HI4N 3 0S 2 , 292.06 (M+H), found 292.2. 30 Example 227-240: To each of a series of 2-dram vials equipped a with a stir bar and Teflon septum was added between 0.3 and 0.6 mmol an acid chloride (1 equiv), followed by 1 equiv of methyl 4-amino-5 methylthiothiophene-2-carboxylate as a 1IM CH 2 C1 2 solution. An additional 2 mL of CH 2 C1 2 was charged into each vial, followed by 1.5 equiv of NN- WO 00/47578 -247- PCT/US99/18065 diisopropylethylamine. Each vial was swept with Ar and allowed to stir for 3 h. At this time 4 mL of sat'd. NaHCO 3 was added to each vial and stirring was continued for 5 min. The aqueous layers were removed by pipette and Na 2
SO
4 was added to each vial. The vials were allowed to stand overnight, and the 5 contents then eluted through 5-g silica gel (SPE column) cartridges with 0.5 % MeOH-CH 2 C1 2 . The amides were concentrated in vacuo into pre-weighed 2 dram vials equipped with a stir bar and Teflon septum for the subsequent amidination reactions. The vials were purged with Ar and charged with 2 mL of toluene, followed by 8 equiv of the AlMe 3
/NH
4 CI reagent as a 1M solution 10 in toluene. The reactions were heated to 110 0 C in a heating block for 3 h. They were then cooled to ambient temperature and each was added by pipette to a slurry of 1.5 g silica gel in 15 mL of CH 2 C1 2 . Each slurry was vigorously stirred for 15 min, at which time they were filtered through a 15 mL sintered glass funnel containing 20 cm of silica gel with 50 % CHCl 3 -MeOH. The 15 yellow fractions were collected and concentrated in vacuo. The solids were triturated with 10 % MeOH-CHCl 3 and filtered. Concentration in vacuo yielded the crude amidines, which were purified by preparative thin layer chromatography (20 % MeOH-CHCl 3 -sat'd. NH 3 , 500 [m SiO 2 ) to afford the amidines as their respective free bases.
WO 00/47578 -248- PCT/US99/18065 Example Acid Chloride Amidine Product % Yield a 227 cinnamoyl chloride 4-((2E)-3-phenylprop-2-enoylamino)- 15.3 % 5-methylthiothiophene-2 carboxamidine 228 4-chlorobenzoyl chloride 4-[(4-chlorophenyl)carbonylamino]-5- 44.6 % methylthiothiophene-2-carboxamidine 229 cyclohexoyl chloride 4-(cyclohexylcarbonylamino)-5- 17.8 % methylthiothiophene-2-carboxamidine 230 3-nitro-4-methylbenzoyl methyl 4-[(4-methyl-3- 8.8 % chloride nitrophenyl)carbonylamino]-5 methylthiothiophene-2-carboxylate 231 2-furoyl chloride 4-(2-furylcarbonylamino)-5- 13.3 % methylthiothiophene-2-carboxamidine 232 2,2-dimethyl-propanoyl 4-(2,2-dimethylpropanoylamino)-5- 8.5 % chloride methylthiothiophene-2-carboxamidine 233 5-(3,5-dichloro- 4- { [5-(3,5-dichlorophenoxy)(2- 22.9 % phenoxy)furan-2- furyl)]carbonylamino}-5 carbonyl chloride methylthiothiophene-2-carboxamidine 234 1-napthoyl chloride 5-methylthio-4- 3.1 % (naphthylcarbonylamino)-thiophene-2 carboxamidine 235 2-quinolinyl chloride 5-methylthio-4-(2-quinolylcarbonyl- 6.8 % amino)thiophene-2-carboxamidine 236 3-methoxybenzoyl 4-[(3-methoxyphenyl)carbonylamino]- 6.8 % chloride 5-methylthiothiophene-2 carboxamidine 237 2-(2,5- 4-[2-(2-hydroxy-5- 18.3 % dimethoxyphenyl)acetyl methoxyphenyl)acetylamino]-5 chloride methylthiothiophene-2-carboxamidine 238 4-ethoxybenzoyl 4-[(4-ethoxyphenyl)carbonylamino]-5- 34 % chloride methylthiothiophene-2-carboxamidine 239 2-phenoxyacetyl 5-methylthio-4-(2- 10 % chloride phenoxyacetylamino)-thiophene-2 carboxamidine 240 3-methylbenzoyl 4-[(3-methylphenyl)carbonylamino]-5- 21.1 % chloride methylthiothiophene-2-carboxamidine " Yield calculated from starting methyl 4-amino-5-methylthiothiophene-2-carboxylate.
WO 00/47578 -249- PCT/US99/18065 Example 227 4-((2E)-3-Phenylprop-2-enoylamino)-5-methylthiothiophene-2 carboxamidine 5 a) Methyl 4-((2E)-3-phenylprop-2-enoylamino)-5 methylthiothiophene-2-carboxylate: yield: 100%. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.49 (s, 3H), 3.83 (s, 3H), 7.12 (d, 1H, J=15.7 Hz), 7.41-7.66 (m, 6H), 8.24 (s, 1H), 9.92 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C1 6
H
5 sNO 3
S
2 , 334.06 (M+H), 334.1. 10 b) 4-((2E)-3-Phenylprop-2-enoylamino)-5-methlylthiothiophene 2-carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 8 2.54 (s, 3H), 7.13 (d, 1H, J=15.7 Hz), 7.41-7.51 (m, 3H), 7.59-7.66 (m, 2H), 8.40 (s, 1H), 8.81 (bs, 3H), 10.02 (bs, 1H). Mass spectrum (ESI, m/z): Calcd. for C1 5 HIsN 3 0S 2 , 318.07 (M+H), 318.2. 15 Example 228 4-[(4-Chlorophenyl)carbonylaminof-5-methylthiothiophene-2-carboxamidine a) Methyl 4-J(4-chlorophenyl)carbonylamino]-5 20 methylthiothiophene-2-carboxylate: yield: 53%. 1H NMR (DMSO-d 6 , 400 MHz) 8 2.55 (s, 3H), 3.83 (s, 3H), 7.62 (d, 2H, J=8.5 Hz), 7.87 (s, 1H), 7.97 (d, 2H, J=8.5 Hz), 10.21 (s, 1H). b) 4-J(4-Chlorophenyl)carbonylamino]-5-methylthiothiophene 2-carboxamidine: IH NMR (DMSO-d 6 , 400 MHz) 8 2.59 (s, 3H). 7.63-7.66 25 (m, 2H), 7.98-8.01 (m, 2H), 8.99 (bs, 2H), 9.33 (bs, 2H), 10.39 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for Cl 3
HI
2
CIN
3 0S 2 , 326.02 (M+H), found 326.2. Example 229 4-(Cyclohexylcarbonylamino)-5-methylthiothiophene-2-carboxamidine 30 a) Methyl 4-(cyclohexylcarbonylamino)-5-methylthliothiophene 2-carboxylate: yield: 69.9%. IH NMR (DMSO-d 6 , 400 MHz) 8 1.22-1.81 (m, 11H), 2.51 (s, 3H), 3.82 (s, 3H), 7.97 (s, 1H), 9.55 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for Cl 4
HI
9
NO
3
S
2 , 314.09 (M+H), found 314.2.
WO 00/47578 -250- PCT/US99/18065 b) 4-(Cyclohexylcarbonylamino)-5-methylthiothiophene- 2 carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 8 2.59 (s, 3H), 7.63-7.66 (m, 2H), 7.98-8.01 (m, 2H), 8.99 (bs, 2H), 9.33 (bs, 2H), 10.39 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for CI 3
H
20
N
3 0S 2 , 298.10 (M+H), found 298.2. 5 Example 230 Methyl 4-[(4-methyl-3-nitrophenyl)carbonylamino]-5-methylthiothiophene-2 carboxylate 10 a) Methyl 4-[(4-methyl-3-nitrophenyl)carbonylamino]-5 methylthiothiophene-2-carboxylate: yield: 80%. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.56 (s, 3H), 2.61 (s, 3H), 3.82 (s, 3H), 7.70 (d, 1H, J=8.1 Hz), 7.86 (s, 1H), 8.19 (dd, 1H, J=1.7, 8.0 Hz), 8.56 (d, 1H, J=1.7 Hz), 10.41 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C 5 sHI 4
N
2 0 5
S
2 , 367.42 (M+H), found 15 367.2. b) Methyl 4-[(4-methyl-3-nitrophenyl)carbonylamino]-5 methylthiothiophene-2-carboxylate: 'H NMR (DMSO-d 6 , 400 MHz) 8 2.47 (s, 3H), 2.61 (s, 3H), 7.12 (bs, 3H), 7.69-7.73 (m, 2H), 8.20 (dd, 1H, J=1.6, 7.9 Hz), 8.57 (d, 1H, J=1.6 Hz). Mass spectrum (ESI, m/z): Calcd. for 20 Cl 4
HI
4
N
4 0 3
S
2 , 351.06 (M+H), found 351.2. Example 231 4-(2-Furylcarbonylamino)-5-methylthiothiophene-2-carboxamidine 25 a) Methyl 4-(2-furylcarbonylamino)-5-methylthiothiophene-2 carboxylate: yield: 100%; 1H NMR (DMSO-d 6 , 400 MHz) 8 2.54 (s, 3H), 3.83 (s, 3H), 6.71 (dd, 1H, J=1.8, 3.4 Hz), 7.33 (d, 1H, J=3.5 Hz), 7.87 (s, 1H), 7.95 (m, 1H), 9.93 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C1 2
HIINO
4
S
2 , 298.02 (M+H), found 298.3. 30 b) 4-(2-Furylcarbonylamino)-5-methylthiothiophene-2 carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 6 2.51 (s, 3H), 6.71 (dd, 1H, J=1.8, 3.5 Hz), 7.18 (bs, 3H), 7.32 (d, 1H, J=3.4 Hz), 7.79 (s, 1H), 7.96 (m, 1H). Mass spectrum (ESI, m/z): Calcd. for CIIHIIN 3 0 2
S
2 , 282.04 (M+H), found 282.2. 35 WO 00/47578 -251- PCT/US99/18065 Example 232 4-(2,2-Dimethylpropanoylamino)-5-methylthiothiophene-2-carboxamidine a) Methyl 4-(2,2-dimethylpropanoylamino)-5 5 methylthiothiophene-2-carboxylate: yield: 93.4%. 'H NMR (DMSO-d 6 , 400 MHz) 8 1.23 (s, 9H), 2.51 (s, 3H), 3.81 (s, 3H), 7.74 (s, 1H), 9.04 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for Cl 2 H1 7
NO
3
S
2 , 288.07 (M+H), found 288.1. b) 4-(2,2-Dimethylpropanoylamino)-5-methylthiothiophene-2 10 carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 8 1.24 (s, 9H), 2.55 (s, 3H), 8.05 (s, 1H), 9.0 (bs, 3H), 9.1 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for
CI
1
HI
7
N
3 0S 2 , 272.09 (M+H), found 272.2. Example 233 15 4-{[5-(3,5-Dichlorophenoxy)(2-furyl)]carbonylamino}-5 methylthiothIiophene-2-carboxamidine a) Methyl 4-{[5-(3,5-dichlorophenoxy)(2-furyl)]carbonylamino} 5-methylthiothiophene-2-carboxylate: yield: 96.9%. Mass spectrum (ESI, 20 m/z): Calcd. for C, 8 sH1 3
CI
2 NOsS 2 , 457.97 (M+H), found 457.9. b) 4-{[5-(3,5-Dichlorophenoxy)(2-furyl)]carbonylamino}-5 methylthiothiophene-2-carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 8 2.53 (s, 3H), 6.12-6.17 (m, 1H), 6.79 (d, 1H, J=1.8 Hz), 7.40-7.43 (m, 2H), 7.70 (m, 1H), 8.13 (s, 1H), 8.92 (bs, 2H), 9.21 (bs, 1H), 10.06 (s, 1H). Mass 25 spectrum (ESI, m/z): Calcd. for C1 7 HI4C1 2
N
3 0 3
S
2 , 441.99 (M+H), found 442.2. Example 234 5-Methylthio-4-(naphthylcarbonylamino) thiophene-2-carboxamidine 30 a) Methyl 5-methylthio-4-(naphthylcarbonylamino)thiophene-2 carboxylate: yield: 80.8 %. 'H NMR (DMSO-d 6 , 400 MHz) 8 7.59-7.67 (m, 3H), 7.80 (d, 1H, J=6.8 Hz), 8.02-8.34 (m, 4H), 10.38 (s, 1H). b) 5-Methylthio-4-(naphthylcarbonylamino)thiophene-2 35 carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 8 2.50 (s, 3H), 7.60 (m, 3H), WO 00/47578 -252- PCT/US99/18065 7.76 (d, 1H, J=6.7 Hz), 7.94 (s, 1H), 8.03 (d, 1H, J=6.8 Hz), 8.09 (d, 1H, 8.3 Hz), 8.30 (d, 1H, J=8.8 Hz). Mass spectrum (ESI, m/z): Calcd. for
CI
7
HI
5
N
3 0S 2 , 342.07 (M+H), found 342.2. 5 Example 235 5-Methylthio-4-(2-quinolylcarbonylamino) thiophene-2-carboxamidine a) Methyl 5-methylthio-4-(2-quinolylcarbonylamino)thiophene 2-carboxylate: yield: 80.9 %. 1H NMR (DMSO-d 6 , 400 MHz) 8 2.59 (s, 3H), 10 3.86 (s, 3H), 8.03-8.06 (m, 3H), 8.24-8.29 (m, 3H), 9.58 (s, 1H), 10.63 (s, 1H). b) 5-Methylthio-4-(2-quinolylcarbonylamino)thiophene-2 carboxamidine: 'H NMR (DMSO-d6, 400 MHz) 8 2.53 (s, 3H), 7.21 (bs, 3H), 7.74 (s, 1H), 7.96-7.98 (m, 2H), 8.19-8.22 (m, 4H), 9.77 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for C16HI 4
N
4 0S 2 , 343.45 (M+H), found 343.1. 15 Example 236 4-[(3-Methoxyphenyl)carbonylaminof-5-methylthiothiophene-2 carboxamidine 20 a) Methyl 4-[(3-methoxyphenyl)carbonylamino]-5 methylthiothiophene-2-carboxylate: yield: 90.3 %. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.55 (s, 3H), 3.83 (s, 3H), 3.85 (s, 3H), 7.19 (m, 1H), 7.39-7.59 (m, 3H), 7.85 (s, 1H), 10.09 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for Ci 5 sH1 5
NO
4
S
2 , 338.05 (M+H), found 338.3. 25 b) 4-[(3-Methoxyphenyl)carbonylamino]-5-methylthiothiophene 2-carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 8 2.58 (s, 3H), 3.84 (s, 3H), 7.19 (dd, 1H, J=2.1, 8.1 Hz), 7.45-7.57 (m, 3H), 8.15 (s, 1H), 9.11 (bs, 4H), 10.32 (bs, 1H). Mass spectrum (ESI, m/z): Calcd. for C1 4 Hi 5
N
3 0 2
S
2 , 322.07 (M+H), found 322.2. 30 Example 237 4-[2-(2, 5-Dimethoxyphenyl)acetylaminof-5-methylthiothiophene-2 carboxamidine 35 a) Methyl 4-J2-(2,5-dimethoxyphenyl)acetylamino]-5 methylthiothiophene-2-carboxylate: 'H NMR (DMSO-d 6 , 400 MHz) 8 2.47 WO 00/47578 -253- PCT/US99/18065 (s, 3H), 3.67 (s, 2H), 3.70 (s, 3H), 3.75 (s, 3H), 3.80 (s, 3H), 6.81 (dd, 1H, J=3.0, 8.8 Hz), 6.87 (d, 1H, J=3.0 Hz), 6.93 (d, 1H, J=8.9 Hz), 8.04 (s, 1H), 9.62 (s, 1H); b) 4-[2-(2,5-Dimethoxyphenyl)acetylamino]-5 5 methylthiothiophene-2-carboxamidine: 'IH NMR (DMSO-d 6 , 400 MHz) 8 2.38 (s, 3H), 3.66 (s, 2H), 3.70 (s, 3H), 3.76 (s, 3H), 6.81 (dd, 1H, J=3.3, 8.0 Hz), 6.88-6.94 (m, 2H), 7.91 (s, 1H), 9.42 (bs, 1H). Example 238 10 4-J(4-Ethoxyphenyl)carbonylamino]-5-methylthiothiophene-2-carboxamlidine a) Methyl 4-[(4-ethoxyphenyl)carbonylamino]-5 methylthiothiophene-2-carboxylate: 'H NMR (DMSO-d 6 , 400 MHz) 8 1.36 (t, 3H, J=7.0 Hz), 2.54 (s, 3H), 3.83 (s, 3H), 4.13 (q, 2H, J=7.0 Hz), 7.05 (d, 15 2H, J=8.8 Hz), 7.87 (s, 1H), 7.93 (d, 2H, J=8.8 Hz), 9.93 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for CI 6
HI
7
NO
4
S
2 , 352.07 (M+H), found 352.2. b) 4-[(4-Ethoxyphenyl)carbonylamino]-5-methylthiothiophene 2-carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 6 1.36 (t, 3H, J=7.0 Hz), 2.55 (s, 3H), 4.13 (q, 2H, J=7.0 Hz), 7.04-7.08 (m, 2H), 7.94-7.97 (m, 2H), 20 8.09 (s, 1H);8.73 (bs, 3H), 10.01 (bs, 1H). Mass spectrum (ESI, m/z): Calcd. for CI 5 sHI7N 3 0 2
S
2 , 336.08 (M+H), found 336.2. Example 239 5-Methylthio-4-(2-phenoxyacetylamino) thiophene-2-carboxamidine 25 a) Methyl 5-methylthio-4-(2-phenoxyacetylamino)thiophene-2 carboxylate: yield: 79 %. 'H NMR (DMSO-d 6 , 400 MHz) 6 2.48 (s, 3H), 3.82 (s, 3H), 4.78 (s, 2H), 6.97-7.02 (m, 2H), 7.31-7.35 (m, 2H), 8.05 (s, 1H), 9.80 (s, 1H). 30 b) 5-Methlylthliio-4-(2-phenoxyacetylamino)thiophene-2 carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz): 8 2.52 (s, 3H), 4.81 (s, 2H), 6.97-7.04 (m, 3H), 7.31-7.35 (m, 2H), 8.26 (s, 1H), 8.84 (bs, 4H). Mass spectrum (ESI, m/z): Calcd. for C1 4 H15N 3 0 2
S
2 , 322.43 (M+H), found 322.2.
WO 00/47578 -254- PCT/US99/18065 -254 Example 240 4-[J(3-Methylphenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine a) Methyl 4-[(3-methylphenyl)carbonylamino]-5 5 methylthiothiophene-2-carboxylate: yield: 79 %. 'H NMR (DMSO-d 6 , 400 MHz) 8 2.40 (s, 3H), 2.55 (s, 3H), 3.83 (s, 3H), 4.78 (s, 2H), 7.42-7.43 (m, 2H), 7.47-7.77 (m, 2H), 7.86 (s, 1H), 10.06 (s, 1H). Mass spectrum (ESI, m/z): Calcd. for CI 5 HIsNO 3
S
2 , 322.06 (M+H), found 322.2. b) 4-[(3-Methylphenyl)carbonylamino]-5-methylthiothiophene 10 2-carboxamidine: 'H NMR (DMSO-d 6 , 400 MHz) 2 2.40 (s, 3H), 2.55 (s, 3H), 7.43-7.44 (m, 2H), 7.75-7.78 (m, 2H), 8.05 (s, 1H), 8.52 (bs, 3H), 10.12 (bs, 1H). Mass spectrum (ESI, m/z): Calcd. for Cl 4
HI
5
N
3 0S 2 , 306.07 (M+H), found 306.2. 15 Example 241 a) Methyl 4-bromo-5-methylthiothiophene-2-carboxylate: To a stirred solution of 4-bromo-5-methylthiothiophene-2-carboxylic acid (87 mmol), prepared according to the procedure of Kleemann, et al., EP 0676395A2, in dry methanol (750 mL) was added thionyl chloride (7 mL, 96 20 mmol) dropwise. After stirring for 10 min at room temperature, the solution was heated to reflux and stirred 7.5 h. The solution was cooled and the solvents were removed in vacuo. The resulting solid was dissolved in dichloromethane (1500 mL) and washed with saturated sodium bicarbonate (2 x 300 mL), water (300 mL), saturated brine (300 mL), and dried over 25 anhydrous sodium sulfate. The solvents were removed in vacuo. The resulting solid was recrystallized twice from hexane/ethyl acetate to give methyl 4-bromo-5-methylthiothiophene-2-carboxylate (4.4 g, 19 %). 'H NMR (CDCl 3 , 400 MHz) 8 7.66 (s, 1H), 3.90 (s, 3H), 2.60 (s, 3H). b) Methyl 5-methylthio-4-{[3-(phenylmethoxy)phenyl]amino} 30 thiophene-2-carboxylate: A dry mixture of 60 mg (0.225 mmol) of methyl 4 bromo-5-methylthiothiophene-2-carboxylic acid, as prepared in the previous step, 3.0 mg (6 mole %) of palladium (II) acetate (Aldrich Chemical Co., Milwaukee, WI), 12.6 mg (9 mole %) of racemic-2,2' bis(diphenylphosphino)-1,1 '-binaphthyl (Strem, Newburyport, MA), 110 mg WO 00/47578 -255- PCTIUS99/18065 (0.34 mmol, 1.5 eq) of cesium carbonate (Aldrich Chemical Co., Milwaukee, WI), and 54 mg (0.29 mmol, 1.3 eq) of 3-benzyloxyaniline (Aldrich Chemical Co., Milwaukee, WI) was added to an oven-dried 1-dram glass vial. This vial was flushed with dry argon in a glove bag, dry toluene (450 ptL, 0.5 M) was 5 added, and the assembly was heated at 100C for 36 h. To the cooled suspension ethyl acetate (4 mL) was added, the mixture passed through 1 inch of Celite, washed with ethyl acetate (2 x 4 mL) and the solvents removed in vacuo. Purification by preparative thin-layer chromatography (1:1 dichloromethane/hexanes) gave 13 mg of the title compound (15 %) as a pale 10 yellow solid. 'H-NMR (CDCl 3 , 400 MHz) 8 7.77 (s, 1H), 7.47-6.59 (min, 9H), 6.11 (s, 1H), 5.07 (s, 2H), 3.89 (s, 3H), 2.47 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 20
H
1 9 NO3S 2 , 386.1 (M+H), found 386.3. c) 5-Methylthio-4-{[3-(phenylmethoxy)phenyl]amino}thiophene 2-carboxamidine: Trimethylaluminum (2.0 M in toluene, 2 mL) was added 15 dropwise over 10 min to a suspension of ammonium chloride (216 mg) in toluene (2 mL), stirred under dry nitrogen at 0 0 C. After the mixture was stirred at 25oC for 30 min, when most of the solid had dissolved, this mixture was taken up in a syringe and added to 13 mg (0.03 mmol) of methyl 5 methylthio-4- { [3-(phenylmethoxy)phenyl]amino } thiophene-2-carboxylate. 20 The reaction mixture was heated to reflux in stages and stirred for 2h 10 min. The cooled mixture was poured in to a vigorously stirred slurry of silica gel (2 g) in chloroform (20 mL). To this suspension methanol (50 mL) was added, the mixture was passed through 1 inch of silica gel in a sintered glass Biichner funnel, washed with methanol (50 mL), and the solvents removed in vacuo. 25 The crude product was purified on a 5 g silica gel SPE column washing first with dichloromethane and then eluting the product off with 10 % methanol in dichloromethane. The product was further purified by preparative High Pressure Liquid Chromatography (HPLC) on a Dynamax C 18 column, 60 A pore size, 10 p 1 M particle size, 40 to 100 % methanol over 30 min in 0.1 % 30 trifluoroacetic acid to give 5.4 mg of the title compound (45 %) as a yellow solid. 'H-NMR (CD 3 OD, 400 MHz) 8 7.84 (s, 1H), 7.44-6.60 (min, 9H), 5.08 (s, 2H), 2.48 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for Cl 9
HI
9
N
3 0S 2 , 370.1 (M+H), found 370.2.
WO 00/47578 -256- PCT/US99/18065 Example 242 a) Methyl 5-methylthio-4-[(3-phenoxyphenyl)aminofthiophene 2-carboxylate: A stirred suspension of 80 mg (0.299 mmol) of methyl 4 bromo-5-methylthiothiophene-2-carboxylate and 72 mg (0.389 mmol, 1.3 eq) 5 of 3-phenoxyaniline (Aldrich, Milwaukee, WI) was treated as in Example 241, step (b). Further purification of the product by preparative thin layer chromatography eluting with 10 % ethyl acetate in hexane gave 36 mg of the title compound (32 %) as a yellow oil. 'H-NMR (CDCl 3 , 400 MHz) 6 7.76 (s, 1H), 7.40-6.65 (min, 9H), 6.26 (s, 1H), 3.89 (s, 3H), 2.40 (s, 3H). Mass 10 spectrum (ESI, m/z): Calcd. for Cl 9
H
1 7 NO3S 2 , 372.1 (M+H), found 372.2. b) 5-Methylthio-4-[(3-phenoxyphenyl)amino]thiophene-2 carboxamidine: Methyl 5-methylthio-4-[(3-phenoxyphenyl)amino]thiophene 2-carboxylate (36 mg, 0.097 mmol) was treated as in Example 241, step (c), but without HPLC purification to give 30 mg of the title compound (86 %) as 15 an orange glass. 'H-NMR (CDCl 3 , 400 MHz) 6 9.28 (s, 2H), 8.11 (s, 2H), 7.99 (s, 1H), 7.34-6.50 (min, 9H), 6.29 (s, 1H), 2.35 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for CI 8
H
1 7
N
3 0S 2 , 356.1 (M+H), found 356.2. Example 243 20 a) S-Methylthio-4-[(4-phenoxyphenyl)amino]thiophene-2 carboxamidine: A stirred suspension of 80 mg (0.299 mmol) of methyl 4 bromo-5-methylthiothiophene-2-carboxylate and 72 mg (0.389 mmol, 1.3 eq) of 4-phenoxyaniline (Aldrich, Milwaukee, WI) was treated as in Example 241, step (b). Further purification of the product by preparative thin layer 25 chromatography eluting with 10 % ethyl acetate in hexane gave 53 mg of the title compound (48 %) as a yellow oil. 'H-NMR (CDCl 3 , 400 MHz) 8 7.70 (s, 1H), 7.34-7.00 (min, 9H), 6.11 (s, 1H), 3.89 (s, 3H), 2.42 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for Cl 9
H
17
NO
3
S
2 , 372.1 (M+H), found 372.1. b) 5-Methylthio-4-[(4-phenoxyphenyl)amino]thiophene-2 30 carboxamidine: Methyl 5-methylthio-4-[(4-phenoxyphenyl)amino]thiophene 2-carboxylate (53 mg, 0.14 mmol) was treated as in Example 241, step (c), but without HPLC purification to give 58 mg of the title compound (quantitative yield) as an orange glass. 'H-NMR (CDCl 3 , 400 MHz) 8 8.89 (s, 2H), 8.59 (s, WO 00/47578 -257- PCT/US99/18065 2H), 8.00 (s, 1H), 7.25-6.87 (m, 9H), 6.20 (s, 1H), 2.27 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for CisH17N 3 OS2, 356.1 (M+H), found 356.2. Example 244 5 a) Methyl 4-[(2-methoxyphenyl)amino]-5-metihylthiothiophene 2-carboxylate: A stirred suspension of 103 mg (0.386 mmol) of methyl 4 bromo-5-methylthiothiophene-2-carboxylate and 57 mg (0.46 mmol, 1.2 eq) of 2-methoxyaniline (Aldrich, Milwaukee, WI) was treated in a manner similar to Example 241, step (b) to give 78 mg the title compound (65 %) as a 10 yellow oil. 'H-NMR (CDCl 3 , 400 MHz) 8 7.82 (s, 1H), 7.12-6.52 (min, 4H), 6.52 (s, 1H), 3.92 (s, 3H), 3.87 (s, 3H), 2.40 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 1 4 HIsNO 3
S
2 , 310.1 (M+H), found 310.2. b) 4-[(2-Methoxyphenyl)aminof-5-methylthiothiophene- 2 carboxamidine: Methyl 4-[(2-methoxyphenyl)amino]-5 15 methylthiothiophene-2-carboxylate (78 mg, 0.25 mmol) was treated as in Example 241, step (c), but without HPLC purification to give 75 mg of the title compound (quantitative yield) as an orange glass. 'H-NMR (CD 3 OD, 400 MHz) 8 7.91 (s, 1H), 7.15-6.93 (min, 4H), 3.93 (s, 3H), 2.48 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 13 HIsN 3 0S 2 , 294.1 (M+H), found 294.2. 20 Example 245 a) Methyl 4-[(2-methylphenyl)aminof-5-methylthiothiophene- 2 carboxylate: A dry mixture of 100 mg (0.374 mmol) of methyl 4-bromo-5 methylthiothiophene-2-carboxylate, 51 mg (14.9 mole %) of 25 tris(dibenzylideneacetone)dipalladium (Lancaster, Pelham, NH), 52 mg (22.3 mole %) of racemic-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (Strem, Newburyport, MA), 183 mg of (0.56 mmol, 1.5 eq) cesium carbonate (Aldrich Chemical Co., Milwaukee, WI), and 71 pL (0.49 mmol, 1.3 eq) of 2 methylaniline (Aldrich Chemical Co., Milwaukee, WI) was added to an oven 30 dried 1-dram glass vial. This vial was flushed with dry argon in a glove bag, dry toluene (750 pL, 0.5 M) was added, and the assembly was heated at 100 0 C for 40 h. To the cooled suspension ethyl acetate (4 mL) was added, the mixture passed through 1 inch of Celite, washed with ethyl acetate (2 x 4 mL) and the solvents removed in vacuo. Purification by preparative thin-layer WO 00/47578 -258- PCT/US99/18065 chromatography (1:1 dichloromethane/hexanes) gave 67 mg of the title compound (61 %) as a yellow oil. 1 H-NMR (CDCl 3 , 400 MHz) 8 7.64 (s, 1H), 7.23-6.94 (min, 4H), 5.91 (br s, 1H), 3.88 (s, 3H), 2.41 (s, 3H), 2.31 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 14
H
15
NO
2
S
2 , 294.1 (M+H), found 5 294.2. b) 4-[(2-Methylphenyl)aminof-S-methylthiothiophene-2 carboxamidine: Methyl 4-[(2-methylphenyl)amino]-5-methylthiothiophene 2-carboxylate (67 mg, 0.23 mmol) was treated as in Example 241, step (c), but without HPLC purification to give 20 mg of the title compound (30 %) as a 10 yellow glass. 'H-NMR (CD 3 OD; 400 MHz) 8 7.56 (s, 1H), 7.24-6.99 (min, 4H), 2.49 (s, 3H), 2.29 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 1 3
H
1 5
N
3
S
2 , 278.1 (M+H), found 278.2. Example 246 15 a) Methyl 4-[(3-chlorophenyl)aminof-5-methylthiothiophene-2 carboxylate: A stirred suspension of 80 mg (0.299 mmol) of methyl 4-bromo 5-methylthiothiophene-2-carboxylate and 41 iL (0.389 mol, 1.3 eq) of 3 chloroaniline (Aldrich, Milwaukee, WI) was treated in a manner similar to Example 241, step (b) to give 47 mg of the title compound (50 %) as a yellow 20 oil. 'H-NMR (CDCl 3 , 400 MHz) 8 7.75 (s, 1H), 7.23-6.89 (min, 4H), 6.10 (s, 1H), 3.89 (s, 3H), 2.42 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for
C
13
H
12
NO
2
S
2 C1, 314.0 (M+H), found 314.1. b) 4-[(3-Chlorophenyl)aminof-5-methylthiothiophene-2 carboxamidine: Methyl 4-[(3-chlorophenyl)amino]-5-methylthiothiophene-2 25 carboxylate (47 mg, 0.15 mmol) was treated as in Example 241, step (c) to give 33 mg of the title compound (75 %) as a light yellow solid. 1 H-NMR (DMSO-d 6 , 400 MHz) 8 9.22 (s, 2H), 8.81 (s, 2H), 8.22 (s, 1H), 7.99 (s, 1H), 7.24-6.82 (min, 4H), 2.53 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for
C
12
H
12
N
3
S
2 C1, 298.0 (M+H), found 298.3. 30 Example 247 a) Methyl 4-(methylphenylamino)-5-methylthiothiophene-2 carboxylate: A stirred suspension of 100 mg (0.374 mmol) of methyl 4 bromo-5-methylthiothiophene-2-carboxylate and 72 pL (0.487 mmol, 1.3 eq) WO 00/47578 -259- PCT/US99/18065 of N-methylaniline (Aldrich Chemical Co., Milwaukee, WI) was treated in a manner similar to Example 245, step (a) to give 23 mg of the title compound (21 %) as a yellow oil. 1 H-NMR (CDCl 3 , 400 MHz) 8 7.61 (s, 1H), 7.26-6.68 (mn, 5H), 3.89 (s, 3H), 3.25 (s, 3H), 2.50 (s, 3H). Mass spectrum (ESI, m/z): 5 Calcd. for C 14
H
15
NO
2
S
2 , 294.1 (M+H), found 294.3. b) 4-(Methylphenylamino)-5-methylthiothiophene-2 carboxamidine: Methyl 4-[(2-methylphenyl)amino]-5-methylthiothiophene 2-carboxylate (23 mg, 0.078 mmol) was treated as in Example 241, step (c), but without HPLC purification to give 5.6 mg of the title compound (26 %) as 10 a yellow glass. 'H-NMR (CD 3 OD, 400 MHz) 8 7.83 (s, 1H), 7.24-6.71 (inm, 4H), 3.27 (s, 3H), 2.57 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for
C
13 HIsN 3
S
2 , 278.1 (M+H), found 278.3. Example 248 15 a) Methyl 5-methyl-4-(phenylamino)thiophene-2-carboxylate: A stirred suspension of 400 mg (1.7 mmol) methyl 5-methyl-4-bromo-thiophene 2-carboxylate and 192 ptL (2.1 mmol, 1.25 eq) of aniline (Aldrich, Milwaukee, WI) was treated in a manner similar to Example 241, step (b) to give 66 mg of the title compound (16 %) as a brown glass. 'H-NMR (DMSO-d 6 , 400 MHz) 20 5 7.70 (s, 1H), 7.56 (s, 1H), 7.17 (m, 2H), 6.72 (min, 3H), 3.79 (s, 3H), 2.31 (s, 3H). Mass spectrum (MALDI, gentisic acid matrix, m/z): Calcd. for
C
1 3
H
1 3
NO
2 S, 248.1 (M+H), found 247.5. b) 5-Methyl-4-(phenylamino)thiophene-2-carboxamidine: Methyl 4-(methylphenylamino)-5-methylthiothiophene-2-carboxylate (66 mg, 25 0.27 mmol) was treated as in Example 241, step (c), but without HPLC purification to give 57 mg of the title compound (91 %) as a brown glass. 'H NMR (DMSO-d 6 , 400 MHz) 8 9.17 (s, 2H), 8.85 (s, 2H), 7.98 (s, 1H), 7.85 (s, 1H), 7.21-6.73 (min, 5H), 2.39 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for Cl 2
HI
3
N
3 S, 232.1 (M+H), found 232.2. 30 Example 249 a) Methyl 4-{[4-(dimethylamino)phenyl]amino}-5 methylthiothiophene-2-carboxylate: A stirred suspension of 100 mg (0.267 mmol) methyl 5-methyl-4-bromo-thiophene-2-carboxylate and 66 mg (0.35 WO 00/47578 -260- PCT/US99/18065 mmol, 1.3 eq) of 4-amino-N,N-dimethylaniline (Fluka, Milwaukee, WI) was treated in a manner similar to Example 241, step (b), but eluting with 1:1 ethyl acetate/hexane for preparative thin-layer chromatrography purification, to give 86 mg of the title compound (quantitative yield) as an orange glass. 'H-NMR 5 (CDCl 3 , 400 MHz) 8 7.53 (s, 1H), 7.16 and 6.62 (AB quartet, 4H, J=8.9 Hz), 5.99 (s, 1H), 3.86 (s, 3H), 2.94 (s, 6H), 2.39 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for Cs 15
H
1 8
N
2 0 2
S
2 , 323.1 (M+H), found 323.3. b) 4-{[4-(Dimethylamino)phenyl]amino}-S-methylthiothiophene 2-carboxamidine: Methyl 4-(methylphenylamino)-5-methylthiothiophene-2 10 carboxylate (86 mg, 0.267 mmol) was treated as in Example 241, step (c), but without HPLC purification. This material was further purified by passing through 1 inch of basic alumina and eluting with 10 % methanol in dichloromethane (15 mL) to give 62 mg of the title compound (76 %) as a brown glass. 'H-NMR (DMSO-d 6 , 400 MHz) 8 8.95 (s, 4H), 7.75 (s, 1H), 15 7.56 (s, 1H), 6.97 and 6.72 (AB quartet, 4H, J=8.9 Hz), 2.83 (s, 6H), 2.44 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for C 1 4 HlsN 4
S
2 , 307.1 (M+H), found 307.3. Example 250 20 4-[(4-Ethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine hydrochloride a) Methyl 4-[(4-ethylphenyl)aminol-5-methylthiothiophene-2 carboxylate: To an oven-dried glass vial with stir bar was added a mixture of 25 100 mg (0.374 mmol) of methyl 4-bromo-5-methylthiothiophene-2 carboxylate (as prepared in Example 241, step (a)), 5.8 mg (6.9 mol %) of palladium (II) acetate, 21.7 mg (9.3 mol %) of racemic-2,2' bis(diphenylphosphino)-1,1 '-binaphthyl, 171.5 mg (0.526 mmol) of cesium carbonate and 59 mg (0.487 mmol) of 4-ethylaniline. The vial was transferred 30 to a glove bag, flushed with dry argon and anhydrous toluene (749 PL) was added. The vial was capped with a Teflon-lined screw cap and heated at 100 oC for 48 h. The cooled suspension was filtered (Celite) washing with ethyl acetate (2 x 2 mL), and the solvents removed in vacuo. The resulting residue was purified on 1 mm silica prep plates eluting with 40% methylene chloride 35 hexanes to afford 14 mg (12 %) of methyl 4-[(4-ethylphenyl)amino]-5- WO 00/47578 PCT/US99/18065 -261 methylthiothiophene-2-carboxylate as a pale yellow resin which was used directly in the following step. b) 4-[(4-Ethylphenyl)aminoj-5-methylthiothiophene-2 carboxamidine hydrochloride: Trimethylaluminum (2.0 M in toluene, 0.182 5 mL, 0.363 mmol) was added dropwise to a suspension of ammonium chloride (19 mg, 0.363 mmol) in anhydrous toluene (1 mL) under Ar at 0 0 C. The mixture was stirred at 25 oC for 30 min and then 14 mg (0.036 mmol) of methyl 4-[(4-ethylphenyl)amino]-5-methylthiothiophene-2-carboxylate (as prepared in previous step) was added. The reaction mixture was heated slowly 10 to 100 C and stirred for 4 h. The cooled mixture was added to a vigorously stirred slurry of silica gel (1.3 g) in chloroform (20 mL). The suspension was filtered (silica) washing with 50 % MeOH-CH 2 C1 2 (2 x 50 mL). The washings were concentrated and the resulting residue was purified on a 0.5 mm silica prep plate eluting with a 10 % MeOH-CH 2 C1 2 to afford 8 mg (67 %) of 4-[(4 15 ethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine hydrochloride as a yellow oil. 'H-NMR (CD 3 OD, 400 MHz) 8 7.84 (s, 1H), 7.14 (d, 2H, 8 Hz), 7.01 (d, 2H, 8 Hz), 2.55 (q, 2H, 65.5 Hz), 2.48 (s, 3H), 1.23 (t, 3H, 15.2 Hz). Mass spectrum (ESI, m/z): Calcd. for C 14
H
17
N
3
S
2 , 292.1 (M+H), found 292.5. 20 Example 251 5-Methylthio-4-{[4-(phenylmethoxy)phenyl]amino}thiophene-2 carboxamidine hydrochloride a) Methyl 5-methylthio-4-{[4-(phenylmethoxy)phenyl] 25 amino}thiophene-2-carboxylate: The same procedure as in Example 250, step a was followed using 100 mg (0.374 mmol) of methyl 4-bromo-5 methylthiothiophene-2-carboxylate (as prepared in Example 241, step (a)), 5.5 mg (6.5 mol %) of palladium (II) acetate, 23.6 mg (10.1 mol %) of racemic 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl, 194 mg (0.595 mmol) of cesium 30 carbonate, 97.3 mg (0.488 mmol) of 4-benzyloxyaniline and 749 ptL of toluene, and chromatographed as before using 40 % CH 2 Cl2-hexane to afford 7 mg (5 %) of methyl 5-methylthio-4-{[4-(phenylmethoxy)phenyl] amino}thiophene-2-carboxylate as a yellow resin which was used directly in the following step. 35 b) 5-Methylthio-4-{[4-(phenylmethoxy)phenylfamino}thiophene 2-carboxamidine hydrochloride: The same procedure as in Example 250, WO 00/47578 -262- PCT/US99/18065 step (b) was followed using 7 mg (0.018 mmol) of methyl 5-methylthio-4-{[4 (phenylmethoxy)phenyl]amino}thiophene-2-carboxylate (as prepared in previous step), 0.091 mL of trimethylaluminum (2.0 M in toluene, 0.182 mmol), 10 mg of ammonium chloride (0.182 mmol) and 1 mL of toluene, and 5 purified on a 0.5 mm silica prep plate eluting with 10 % MeOH-CH 2
C
2 to afford 3 mg (41 %) of 5-methylthio-4- { [4 (phenylmethoxy)phenyl]amino}thiophene-2-carboxamidine hydrochloride as a yellow oil. 'H-NMR (CD 3 OD, 400 MHz) 5 7.72 (s, 1H), 7.45 (d, 2H, 7 Hz), 7.39 (t, 2H, 9 Hz), 7.37 (d, 1H, 12 Hz), 7.06 (d, 2H, 12Hz), 6.97 (d, 2H, 10 12Hz), 5.08 (s, 2H), 2.46 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for
C
19
H
19
N
3 0S 2 , 370.1 (M+H), found 370.3. Example 252 5-Methylthio-4-[4-(phenylamino)phenyl]amino}thiophene-2-carboxamidine 15 hydrochloride a) Methyl 5-methylthio-4-{[4-(phenylamnino)phenyl]amino} thiophene-2-carboxylate: The same procedure as in Example 250, step (a) was followed using 100 mg (0.374 mmol) of methyl 4-bromo-5 20 methylthiothiophene-2-carboxylate (as prepared in Example 241, step a), 5.5 mg (6.5 mol %) of palladium (II) acetate, 21.6 mg (9.3 mol %) of racemic 2,2'-bis(diphenylphosphino)-1,1 '-binaphthyl, 173.7 mg (0.533 mmol) of cesium carbonate, 92.3 mg (0.500 mmol) of N-phenyl-1,4-phenylenediamine, and 749 ptL of toluene, and chromatographed as before using 40 % CH 2
C
2 25 hexane to afford 58 mg (42 %) of methyl 5-methylthio-4-{[4 (phenylamino)phenyl]amino}thiophene-2-carboxylate as a brown solid. 'H NMR (DMSO-d 6 , 400 MHz) 8 7.85 (s, 1H), 7.61 (s, 1H), 7.48 (s, 1H), 7.14 (t, 2H, 16 Hz), 6.99 (d, 2H, 16 Hz), 6.90 (q, 4H, 44 Hz), 6.70 (t, 2H, 4 Hz), 3.77 (s, 3H), 2.43 (s, 3H). 30 b) 5-Methylthio-4-[4-(phenylamino)phenyl]amino}thiophene-2 carboxamidine hydrochloride: The same procedure as in Example 250, step (b) was followed using 58 mg (0.156 mmol) of methyl 5-methylthio-4-{[4 (phenylamino)phenyl]amino}thiophene-2-carboxylate (as prepared in previous step), 0.783 mL of trimethylaluminum (2.0 M in toluene, 1.56 mmol), 84 mg 35 of ammonium chloride (1.56 mmol) and 10 mL of toluene, and purified by WO 00/47578 -263- PCT/US99/18065 passing through a pad of silica eluting with 50 % MeOH-CH 2
CI
2 to afford 50 mg (75 %) of the 5-methylthio-4- { [4-(phenylamino)phenyl]amino } thiophene 2-carboxamidine hydrochloride as a brown solid. IH-NMR (DMSO-d 6 , 400 MHz) 6 7.91 (d, 2H, 12 Hz), 7.78 (s, 1H), 7.20 (t, 3H, 12 Hz), 7.04-6.94 (inm, 5 5H), 6.71 (min, 1H), 2.47 (s, 3H). Mass spectrum (ESI, m/z): Calcd. for
CI
8
H
18
N
4
S
2 , 355.1 (M+H), found 355.4. Example 253 10 4-J(4-Methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine hydrochloride a) Methyl 4-[(4-methoxyphenyl)aminof-5-methylthiothiophene 2-carboxylate: To an oven-dried glass vial with stir bar was added a mixture 15 of 120 ming (0.449 mmol) of methyl 4-bromo-5-methylthiothiophene-2 carboxylate (as prepared in Example 241, step (a)), 7.1 mg (7 mol %) of palladium (II) acetate, 29.4 mg (10.5 mol %) of racemic-2,2' bis(diphenylphosphino)-1,1 '-binaphthyl, 205 mg (0.629 mmol) of cesium carbonate and 69.1 mg (0.561 mmol) of p-anisidine. The vial was transferred 20 to a glove bag, flushed with dry argon and anhydrous toluene (0.9 mL) was added. The vial was capped with a Teflon-lined screw cap and heated at 100 oC for 48 h. To the cooled suspension was added ethyl acetate (4 mL), the mixture filtered (Celite) washing with ethyl acetate (2 x 2 mL), and the solvents removed in vacuo. The resulting residue was purified by silica gel 25 preparative thin layer chromatography (40% CH 2 C1 2 in hexane) to afford 83 mg (60 %) of the title compound as a yellow oil. 1 H-NMR (CDCl 3 , 400 MHz) 8 2.39 (s, 3H), 3.82 (s, 3H), 3.87 (s, 3H), 6.03 (s, 1H), 6.89 (mn, 2H), 7.03 (inm, 2H), 7.58 (s, 1H). b) 4-[(4-Methoxyphenyl)amino]-5-methylthiothiophene-2 30 carboxamidine hydrochloride: Trimethylaluminum (2.0 M in toluene, 2 mL, 4 mmol) was added dropwise to a suspension of ammonium chloride (216 mg, 4 mmol) in anhydrous toluene (1 mL) under Ar at room temperature. The mixture was stirred at 25 oC for 30 min and then 80 mg (0.259 mmol) of methyl 4-[(4-methoxyphenyl)amino]-5-methylthiothiophene-2-carboxylate (as 35 prepared in previous step) in anhydrous toluene (1 mL) was added. The WO 00/47578 PCT/US99/18065 -264 reaction mixture was heated slowly to 100 oC and stirred for 2.5 h. The cooled mixture was added to a vigorously stirred slurry of silica gel (3 g) in chloroform (20 mL). The suspension was filtered washing with MeOH (4 x 5 mL) and 50 % MeOH-CH 2 C1 2 (4 x 5 mL). The combined washings were 5 concentrated and the resulting residue was purified on a 2-g silica SPE column with 5 % MeOH-CH 2
CI
2 to afford 50 mg (59 %) of the title compound as an orange solid. 1 H-NMR (DMSO-d 6 ; 400 MHz) 8 2.44 (s, 3H), 3.69 (s, 3H), 6.84 (min, 2H), 6.98 (min, 2H), 7.73 (s, 1H), 7.84 (s, 1H), 9.01 (br s, 2H), 9.24 (br s, 2H). Mass spectrum (ESI, m/z): Calcd. for C 13 Hs 15
N
3 0S 2 , 294.1 (M+H), 10 found 294.2. Example 254 4-[(3-Fluoro-4-methylphenyl)amino]-5-methylthiothiophene-2 15 carboxamidine a) Methyl 4-[(3-fluoro-4-methylphenyl)amino]-5 methylthiothiophene-2-carboxylate: To an oven-dried glass vial with stir bar was added a mixture of 120 mg (0.449 mmol) of methyl 4-bromo-5 20 methylthiothiophene-2-carboxylate (as prepared in Example 241, step (a), 41 mg (10 mol %) of tris-(dibenzylidineacetone)dipalladium, 42 mg (15 mol %) of racemic-2,2'-bis(diphenylphosphino)-1 ,1 '-binaphthyl , 205 mg (0.629 mmol) of cesium carbonate and 70 mg (0.56 mmol) of 3-fluoro-4 methylaniline. The vial was transferred to a glove bag, flushed with dry argon 25 and anhydrous toluene (0.9 mL) was added. The vial was capped with a Teflon-lined screw cap and heated at 100 oC for 48 h. To the cooled suspension was added ethyl acetate (4 mL), the mixture filtered (Celite) washing with ethyl acetate (2 x 2 mL), and the solvents removed in vacuo. The resulting residue was purified by silica gel preparative thin layer 30 chromatography (10% Et 2 0 in hexane) to afford 103 mg (78 %) of the title compound as a yellow oil 'H-NMR (CDCl 3 , 400 MHz) 8 2.22 (d, 3H, J = 1.6 Hz), 2.40 (s, 3H), 3.89 (s, 3H), 6.09 (s, 1H), 6.68 (min, 1H), 6.71 (s, 1H), 7.08 (min, 1H), 7.72 (s, 1H). b) 4-[(3-Fluoro-4-methylphenyl)amino]-5-methylthiotziophene 35 2-carboxamidine: The same procedure as in Example 253, step (b) was WO 00/47578 -265- PCT/US99/18065 followed using 103 mg (0.349 mmol) of methyl 4-[(3-fluoro-4 methylphenyl)amino]-5-methylthiothiophene-2-carboxylate (as prepared in previous step), 2 mL of trimethylaluminum (2.0 M in toluene, 4 mmol), 216 mg of ammonium chloride (4 mmol) and 2 mL of toluene, and purified on a 5 2-g silica SPE column with 5 % MeOH-CH 2
CI
2 to afford 45 mg (44 %) of the title compound as a yellow foam 'H-NMR (DMSO-d 6 ; 400 MHz) 8 2.13 (s, 3H), 2.50 (s, 3H), 6.70 (m, 2H), 7.10 (m, 1H), 7.98 (s, 1H), 8.09 (s, 1H), 9.16 (br s, 4H). Mass spectrum (ESI, m/z): Calcd. for Cl 3
H
14
FN
3
S
2 , 296.1 (M+H), found 296.2. 10 Example 255 4-(Indan-5-ylamino)-5-methylthiothiophene-2-carboxamidine 15 a) Methyl 4-(indan-5-ylamino)-5-methliylthiothiophene-2 carboxylate: The same procedure as in Example 254, step (a) was followed using 120 mg (0.449 mmol) of methyl 4-bromo-5-methylthiothiophene-2 carboxylate (as prepared in Example 241, step (a), 41 mg (10 mol %) of tris (dibenzylidineacetone)dipalladium, 42 mg (15 mol %) of racemic-2,2' 20 bis(diphenylphosphino)-1,1'-binaphthyl, 205 mg (0.629 rmmol) of cesium carbonate and 74.8 mg (0.56 mmol) of 5-aminoindan in 900 pL of toluene, and chromatographed as before using 40 % CH 2
CI
2 -hexane to afford 100 mg (73 %) of the title compound as a yellow resin. 'H-NMR (CDCl 3 , 400 MHz) 8 2.05-2.12 (m, 2H), 2.85-2.90 (m, 4H), 3.86 (s, 3H), 6,09 (s, 1H), 6.82 (d, 1H, J 25 = 8.0 Hz), 6.93 (s, 1H), 7.14 (d, 1H, J = 8.0 Hz), 7.70 (s, 1H). b) 4-(Indan-5-ylamino)-5-methylthiothiophene-2 carboxamidine: The same procedure as in Example 253, step (b) was followed using 100 mg (0.33 mmol) of methyl 4-(indan-5-ylamino)-5 methylthiothiophene-2-carboxylate (as prepared in previous step), 2 mL of 30 trimethylaluminum (2.0 M in toluene, 4 mmol), 216 mg of ammonium chloride (4 mmol) and 2 mL of toluene, and purified on a 2-g silica SPE column with 5 % MeOH-CH 2 C1 2 to afford 65 mg (65 %) of the title compound as a yellow foam. 'H-NMR (DMSO-d 6 , 400 MHz) 8 1.99 (m, 2H), 2.48 (s, 3H), 2.78 (m, 4H), 6.77 (dd, 1H, J = 8.0, 1.78 Hz), 6.86 (s, 1H), 7.08 (d, 1H, J WO 00/47578 -266- PCT/US99/18065 = 8.1 Hz), 7.80 (s, 1H), 7.94 (s, 1H), 9.13 (br s, 4H). Mass spectrum (ESI, m/z): Calcd. for C 15
H
17
N
3
S
2 , 304.1 (M+H), found 304.3. Example 256 5 4
-[(
9 -Ethylcarbazol-3-yl)amino]-5-methylthiothliophene-2-carboxamidine a) Methyl 4-[(9-ethlylcarbazol-3-yl)aminof-5-. methylthiothiophene-2-carboxylate: The same procedure as in Example 254, 10 step (a) was followed using 120 mg (0.449 mmol) of methyl 4-bromo-5 methylthiothiophene-2-carboxylate (as prepared in Example 241, step (a), 41 mg (10 mol %) of tris-(dibenzylidineacetone)dipalladium, 42 mg (15 mol %) of racemic-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl, 205 mg (0.629 mmol) of cesium carbonate and 118 mg (0.56 mmol) of 3-amino-9 15 ethylcarbazole in 900 pL of toluene, and chromatographed as before using 40 % CH 2 Cl 2 -hexane to afford 80 mg (47 %) of the title compound as a yellow resin. 'H-NMR (CDCl 3 , 400 MHz) 8 1.46 (t, 3H, J = 7.2 Hz), 2.44 (s, 3H), 3.85 (s, 3H), 4.39 (q, 2H, J = 7.2 Hz), 6.25 (s, 1H), 7.24 (min, 1H), 7.28 (s, 1H), 7.40 (min, 2H), 7.49 (min, 1H), 7.61 (s, 1H), 7.83 (d, 1H, J = 2.1 Hz), 8.06 (d, 1H, 20 J= 7.8 Hz). b) 4-[(9-Ethylcarbazol-3-yl)amino]-5-methylthiothiophene-2 carboxamidine: The same procedure as in Example 253, step (b) was followed using 80 mg (0.21 mmol) of methyl 4-[(9-ethylcarbazol-3-yl)amino] 5-methylthiothiophene-2-carboxylate (as prepared in previous step), 2 mL of 25 trimethylaluminum (2.0 M in toluene, 4 mmol), 216 mg of ammonium chloride (4 mmol) and 2 mL of toluene, and purified on a 2-g silica SPE column with 5 % MeOH-CH 2 C1 2 to afford 56 mg (70 %) of the title compound as a yellow foam. 'H-NMR (DMSO-d 6 , 400 MHz) 8 1.31 (t, 3H, J = 7.0 Hz), 2.50 (s, 3H), 4.42 (q, 2H, J = 7.0 Hz), 7.14 (min, 1H), 7.27 (dd, 1H, J = 8.7, 2.1 30 Hz), 7.43 (min, 1H), 7.56 (min, 2H), 7.82 (d, 1H, J = 2.0 Hz), 7.87 (s, 1H), 7.92 (s, 1H), 8.10 (d, 1H, J = 7.7 Hz), 9.11 (br s, 4H). Mass spectrum (ESI, m/z): Calcd. for C 20
H
20
N
4
S
2 , 381.1 (M+H), found 381.3.
WO 00/47578 -267- PCT/US99/18065 Examples 257 and 258 5-Methylthio-4-{[(4-phenylphenyl)sulfonyl]amino}thiophene-2 carboxamidine trifluoroacetate 5 4-{Bis[(4-phenylphenyl)sulfonyl]amino}-5-methylthiothiophene-2 carboxamidine trfluoroacetate a) Methyl 5-methylthio-4-J(phenylsulfonyl)amino]thiophene-2 carboxylate and methyl 4-{bis[(4-phenylphenyl)sulfonylfamino}-5 10 methylthio-thiophene-2-carboxylate: To an oven-dried round bottom flask with stir bar was added a mixture of 50 mg (0.24 mmol) of methyl 4-amino-5 methylthiothiophene-2-carboxylate (as prepared in Example 202), 68 mg (0.27 mmol) of 4-biphenylsulfonyl chloride and 50 mg (0.49 mmol) of 4 dimethylaminopyridine. The flask was flushed with dry argon and anhydrous 15 acetonitrile (3 mL) was added. The reaction was refluxed for 3 hours and then the solvent was removed in vacuo. The crude of the reaction was extracted with ethyl acetate (2 x 25 mL) and 1N HCI (50 mL), The organic layer was collected, dried (Na 2
SO
4 ), filtered and concentrated under vacuum to yield a foam that was chromatographed on silica with 30 % ethyl ether-hexane to 20 obtain 143 mg of a mixture of methyl 5-methylthio-4 [(phenylsulfonyl)amino]thiophene-2-carboxylate and methyl 4- {bis[(4 phenylphenyl)-sulfonyl]amino } -5-methylthiothiophene-2-carboxylate. This mixture was used in the next reaction without further purification. Mass spectrum (ESI, m/z): Calcd. for C 19
H
17
NO
4
S
3 , 420.0 (M+H), found 419.7. 25 b) 5-Methylthio-4-{[(4-phenylphenyl)sulfonyl]amino}thiophene 2-carboxamidine trifluoroacetate and 4-{bisf(4-phenylphenyl)sulfonyl] amino}-5-methylthiothiophene-2-carboxamidine trifluoroacetate: Trimethylaluminum (2.0 M in toluene, 1.36 mL, 2.72 mmol) was added dropwise to a suspension of ammonium chloride (155 mg, 2.89 mmol) in 30 anhydrous toluene (2.0 mL) under Ar at 0 oC. The mixture was stirred at 25 oC for 30 min and then 143 mg of a mixture of methyl 5-methylthio-4 [(phenylsulfonyl)amino]thiophene-2-carboxylate and methyl 4- {bis[(4 phenylphenyl)sulfonyl] amino }-5-methylthiothiophene-2-carboxylate (as prepared in previous step) in anhydrous toluene (2.0 mL) was added. The 35 reaction mixture was heated slowly to 100 oC and stirred for 4 h. The cooled mixture was added to a vigorously stirred slurry of silica gel (3 g) in WO 00/47578 -268- PCT/US99/18065 chloroform (15 mL). The suspension was filtered (Celite) washing with 25 % MeOH-CH 2
CI
2 (2 x 5 mL), 50 % MeOH-CH 2
C
2 (2 x 5 mL) and 75 % MeOH-CH 2
CI
2 (2 x 5 mL). The combined washings were concentrated and the resulting residue was purified on a 10-g silica SPE column with a gradient 5 of 10-15 % MeOH-CH 2 C1 2 saturated with ammonia to afford 66 mg of a mixture of the title compounds as a yellow solid. This mixture was chromatographed by preparative reverse phase HPLC performed with a Rainin SD-1 Dynamax system and a 2-in. C18 reverse phase Dynamax 60A column using a gradient of 30% MeOH /0.1% TFA in water to 100% MeOH and a 10 flow rate of 50 mL/ min. to yield 15 mg 5-methylthio-4-{[(4 phenylphenyl)sulfonyl]amino } thiophene-2-carboxamidine trifluoroacetate; mass spectrum (ESI, m/z): Calcd. for C 18
H
17
N
3 0 2
S
3 , 404.0 (M+H), found 404.1; and 11 mg of 4- {bis[(4-phenylphenyl)sulfonyl]amino}-5 methylthiothiophene-2-carboxamidine trifluoroacetate. Mass spectrum (ESI, 15 m/z): Calcd. for C 3 0
H
2 5
N
3 0 4
S
4 , 619.8 (M+H), found 620.2. Examples 259 to 282 The same methods as for Examples 257 and 258 were used to synthesize the 20 following compounds: WO 00/47578 -269- PCT/US99/18065 Mass spec, ESI. m/z Examp Reagent Compound Formula Calc, Found M+H 259 1- 5-Methylthio-4-[(2-naphthylsulfonyl)- Cl16H15N302S3 378.0 378.1 Naphthalenesulfonyl amino]thiophene-2-carboxamidine chloride 260 1- 4-[Bis(2-naphthylsulfonyl)amino]-5- C26H21N304S4 568.0 568.1 Naphthalenesulfonyl methylthiothiophene-2-carboxamidine chloride 261 7-Bromonaphthalene 4-{[(6-Bromo(2- CI6HI4BrN302S3 455.9 * sulfonyl chloride naphthyl))sulfonyl]amino }-5 methylthiothiophene-2-carboxamidine 262 7-Bromonaphthalene 4-{Bis[(6-bromo(2- C26HI9Br2N304S 723.9 * sulfonyl chloride naphthyl))sulfonyl]amino }-5- 4 methylthiothiophene-2-carboxamidine 263 2- 5-Methylthio-4-[(naphthylsulfonyl)- C16H15N302S3 378.0 378.1 Naphthalenesulfonyl amino]thiophene-2-carboxamidine chloride 264 2- 4-[Bis(naphthylsulfonyl)amino]-5- C26H21N304S4 568.7 568.3 Naphthalenesulfonyl methylthiothiophene-2-carboxamidine chloride 265 o-Toluenesulfonyl 4-{[(2-Methylphenyl)sulfonyl]amino}- C13H15N302S3 342.4 342.1 chloride 5-methylthiothiophene-2 carboxamidine 266 o-Toluenesulfonyl 4-{Bis[(2- C20H21N304S4 496.6 496.1 chloride methylphenyl)sulfonyl]amino }-5 methylthiothiophene-2-carboxamidine 267 m-Toluenesulfonyl 4-{ [(3-Methylphenyl)sulfonyl]amino }- C13HI5N302S3 342.0 342.1 chloride 5-methylthiothiophene-2 carboxamidine 268 m-Toluenesulfonyl 4-{Bis[(3- C20H21N304S4 496.6 496.0 chloride methylphenyl)sulfonyl]amino }-5 methylthiothiophene-2-carboxamidine 269 p-Toluenesulfonyl 4-{ [(4-Methylphenyl)sulfonyl]amino }- Cl3H115N302S3 342.0 342.1 chloride 5-methylthiothiophene-2 carboxamidine 270 p-Toluenesulfonyl 4-{Bis[(4- C20H21N304S4 496.6 496.1 chloride methylphenyl)sulfonyl]amino }-5 methylthiothiophene-2-carboxamidine 271 cx-Toluenesulfonyl 5-Methylthio-4- Cl 3H I5N302S3 342.0 342.1 chloride { [benzylsulfonyl]amino }-thiophene-2 carboxamidine SUBSITUTE SHEET (RULE 26) WO 00/47578 -270- PCT/US99/18065 Mass spec, ESI, m/z Examp Reagent Compound Formula Calc, Found M+HI 272 4-Methoxybenzene- 4-{ [(4-Methoxyphenyl)sulfonyl]amino}- Cl3HI5N303S3 358.0 358.1 sulfonyl chloride 5-methylthiothiophene-2-carboxamidine 273 4-Methoxybenzene- 4-{Bis[(4- C20H21N306S4 528.0 528.0 sulfonyl chloride methoxyphenyl)sulfonyl]amino }-5 methylthiothiophene-2-carboxamidine 274 4- 4-{[(4-lodophenyl)sulfonyl]amino}-5- C12H12IN302S3 453.9 454.0 Iodobenzenesulfony methylthiothiophene-2-carboxamidine I chloride 275 3,4- 4-{[(3,4- Cl4HI7N304S3 388.0 388.1 Dimethoxybenzene Dimethoxyphenyl)sulfonyl]amino}-5 sulfonyl chloride methylthiothiophene-2-carboxamidine 276 3,4- 4-{bis[(3,4- C22H25N308S4 588.0 588.1 Dimethoxybenzene Dimethoxyphenyl)sulfonyl]amino}-5 sulfonyl chloride methylthiothiophene-2-carboxamidine 277 2- 4-{ [(2-Chlorophenyl)sulfonyl]amino}-5- C12HI2CIN302S3 361.9 362.1 Chlorobenzenesulfo methylthiothiophene-2-carboxamidine nvl chloride 278 3- Chlorobenzene- 4-{ [(3-Chlorophenyl)sulfonyl]amino}-5- C12H12CIN302S3 361.9 362.1 sulfonyl chloride methylthiothiophene-2-carboxamidine 279 3- Chlorobenzene- 4-{Bis[(3-chlorophenyl)sulfonyl]amino}- C18HI5C12N304S 535.9 537.9 sulfonyl chloride 5-methylthiothiophene-2-carboxamidine 4 280 4- Chlorobenzene- 4-{ [(4-Chlorophenyl)sulfonyl]amino }-5- Cl2HI2CIN302S3 361.9 362.1 sulfonyl chloride methylthiothiophene-2-carboxamidine 281 4- Chlorobenzene- 4-{Bis[(4-chlorophenyl)sulfonyl]amino}- C18HI5C12N304S 535.9 * sulfonyl chloride 5-methylthiothiophene-2-carboxamidine 4 282 Benzenesulfonyl 5-Methylthio-4-[(phenylsulfonyl)amino]- Cl 2H I3N302S3 328.0 328.1 chloride thiophene-2-carboxamidine Mass spec, ESI, m/z Examp Reagent Compound Formula Calc, Found M+H 283 Benzenesulfonyl 4-[Bis(phenylsulfonyl)amino]-5- Cl8H I7N304S4 468.0 467.9 chloride methylthiophene-2-carboxamidine 284 4-tert- 4-({[4-(Tert- C16H21N302S3 384.0 384.2 Butylbenzene- butyl)phenyl]sulfonyl}amino)-5 sulfonyl chloride methylthiothiophene-2-carboxamidine 285 4-tert- 4-(Bis{ [4-(tert- C26H33N304S4 580.1 580.2 Butylbenzene- butyl)phenyl]sulfonyl } amino)-5 sulfonyl chloride methylthiothiophene-2-carboxamidine 286 Trans-3-styrene 4-{[((lE)-2- C14H15N302S3 354.0 * sulfonyl chloride Phenylvinyl)sulfonyl]amino}-5 methylthiothiophene-2-carboxamidine 287 4-benzensulfonyl- 5-Methylthio-4-({ [4-(phenylsulfonyl)(2- C 6H I5N304S5 473.9 474.1 thiophene-2- thienyl)]sulfonyl } amino)thiophene-2 sulfonyl chloride carboxamidine * Mass spectral data inconclusive. 5 SUBSTITUTE SHEET (RULE 26) WO 00/47578 -271- PCT/US99/18065 Example 288 5-Methylthio-4-phenoxythiophene-2-carboxamidine trifluoroacetate. 5 a) Methyl 5-methylthio-4-phenoxythiophene-2-carboxylate: To an oven-dried round bottom flask with stir bar was added a mixture of 100 mg (0.37 mmol) of methyl 4-bromo-5-methylthiothiophene-2-carboxylate (as prepared in Example 241), 20 mg of Cu (0) (Brewster, R.Q. and Groening T., Organic Syntheses, Vol. II, Note 1, pp 445-446) and 42 mg (0.46 mmol) of 10 phenol. The flask was flushed with dry argon and anhydrous tetrahydrofuran (5 mL) was added. The reaction was refluxed for 48 hours and then the solvent was removed in vacuo. The resulting residue was purified on a 10-g silica SPE column with a gradient of 50-100 % CH 2
CI
2 -hexane to yield 48 mg of methyl 5-methylthio-4-phenoxythiophene-2-carboxylate (37%). 'H-NMR (CDC1 3 , 15 400 MHz) 8 7.39 (s, 1H), 7.32 (min, 2H), 7.09 (mn, 2H), 6.97 (d, 1H, J = 8.4 Hz), 3.86 (s, 3H) and 2.49 (s, 3H). b) 5-Methylthlzio-4-phenoxythiophene-2-carboxamidine trifluoroacetate: The same procedure as in Example 257, step (b) was followed using 48.0 mg (0.17 mmol) of methyl 5-methylthio-4 20 phenoxythiophene-2-carboxylate (as prepared in the step before), 78 mg of ammonium chloride (1.5 mmol), 0.68 ml of trimethylaluminum (2.0 M in toluene, 1.3 mmol) and 3 ml of anhydrous toluene and chromatographed as before using preparative reverse phase HPLC performed with a Rainin SD-1 Dynamax system and a 2-in. C 18 reverse phase Dynamax 60A column using a 25 gradient of 30% MeOH /0.1% TFA in water to 100% MeOH and a flow rate of 50 mL/ min. ' 1 H-NMR (CD 3 OD, 400 MHz) 8 7.66 (s, 1H), 7.39 (t, 2H, J = 7.5 Hz), 7.17 (t, 2H, J= 7.4 Hz), 7.02 (d, 1H, J= 7.7 Hz) and 2.58 (s, 3H). Mass spectrum (ESI, m/z): Calcd. C 1 2
H
12
N
2 0S 2 , 265.0 (M+H), found 262.2. 30 Example 289 5-Methylthio-4-(phenylsulfonyl)thiophene-2-carboxamidine trifluoroacetate. a) 4-Bromo-5-methylthiothiophene-2-carboxylic acid: To 1.0 g 35 (3.7 mmol) of methyl 4-bromo-5-methylthiothiophene-2-carboxylate (as prepared in Example 241, step (a) dissolved in 25 ml of MeOH was added 450 WO 00/47578 -272- PCT/US99/18065 mg of NaOH dissolved in 10 ml of H 2 0. The reaction was stirred for 5 hours at room temperature, and then the solvents were removed under vacuum. The residue of the reaction was extracted with ethyl acetate (2 x 50 mL) and IN HC1. The organic layer was collected, dried (Na 2
SO
4 ), filtered and 5 concentrated under vacuum to yield 833 mg (89%) of 4-bromo-5 methylthiothiophene-2-carboxylic acid as a white solid. b) 5-Methylthio-4-(phenylsulfonyl)thiophene-2-carboxylic acid: To an oven-dried round bottom flask with stir bar was added 100 mg (0.39 mmol) 4-bromo-5-methylthiothiophene-2-carboxylic acid (as prepared in 10 Example before). The flask was flushed with dry argon and anhydrous tetrahydrofuran (3 mL) was added. Then the solution was cooled at -78 oC before adding 511 tL of tert-butyl lithium (0.87 mmol, 1.7 M in tetrahydrofuran). The mixture was stirred for a period of 45 minutes and 77 mg of benzenesulfonyl flouride (0.39 mmol) was added and the reaction was 15 allowed to rise to room temperature. The reaction was stirred for 12 hours and then quenched carefully with H20. The solvents were removed under vacuum and the residue of the reaction was extracted with ethyl acetate (2 x 50 ml) and 1N HC1. The organic layer was collected, dried (Na 2
SO
4 ), filtered and concentrated under vacuum to yield 130 mg of a solid. This solid was used in 20 the next step without further purification. c) Methyl 5-methylthio-4-(phenylsulfonyl)thiophene-2 carboxylate: To a solution of 25 mg of the mixture from the previous step dissolved in 3 mL of MeOH wa added dropwise 397 pL of trimethylsilyldiazomethane (0.79 mmol, 2 M solution in hexanes) and the 25 reaction was stirred for a period of 1 hour. The solvents were removed under vacuum. The resulting residue was purified on a 10-g silica SPE column with a gradient of 50-100 % ethyl acetate-hexane to yield 13.8 mg of methyl 5 methylthio-4-(phenylsulfonyl)thiophene-2-carboxylate. Mass spectrum (ESI, m/z): Calcd. C 1 3
H
12 0 4
S
3 , 329.0 (M+H), found 329.0. 30 d) 5-Methylthio-4-(phenylsulfonyl)thiophene-2-carboxamidine trifluoroacetate: The same procedure as in Example 257, step (b) was followed using 13.8 mg (0.044 mmol) of methyl 5-methylthio-4 (phenylsulfonyl)thiophene-2-carboxylate (as prepared in the step before), 20 mg of ammonium chloride (0.376 mmol), 0.176 ml of trimethylaluminum (2.0 WO 00/47578 -273- PCT/US99/18065 M in toluene, 0.353 mmol) and 3 ml of anhydrous toluene and chromatographed as before by preparative reverse phase HPLC performed with a Rainin SD-1 Dynamax system and a 2-in. C18 reverse phase Dynamax 60A column using a gradient of 30% MeOH /0.1% TFA in water to 100% 5 MeOH and a flow rate of 50 mL/ min to yield 2.3 mg of 5-methylthio-4 (phenylsulfonyl)thiophene-2-carboxamidine. 'H-NMR (CD30D, 400 MHz) 6 8.42 (s, 1H), 8.04 (m, 2H), 7.70 (m, 2H), 7.62 (m, 1H) and 2.70 (s, 3H). Mass spectrum (ESI, m/z): Calcd. C 12
H
12
N
2 0 2
S
3 , 313.0 (M+H), found 313.2.
WO 00/47578 PCT/US99/18065 -274 5 Example 290 Tablet Preparation Tablets containing 25.0, 50.0, and 100.0 mg, respectively, of the following active compounds are prepared as illustrated below: a. 4-(4-methylthiazol-2-yl)-5-methylthiothiophene-2 10 carboxamidine; b. 4-[4-(4-phenylphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine. TABLET FOR DOSES CONTAINING FROM 25-100 MG OF THE ACTIVE COMPOUND Amount-mg 15 Active Compound 25.0 50.0 100.00 Microcrystalline cellulose 37.25 100.0 200.0 Modified food corn starch 37.25 4.25 8.5 Magnesium stearate 0.50 0.75 1.5 All of the active compound, cellulose, and a portion of the cornstarch -0 are mixed and granulated to 10% corn starch paste. The resulting granulation is sieved, dried and blended with the remainder of the corn starch 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. 15 Example 291 Intravenous Solution Preparation An intravenous dosage form of the above-indicated active compounds is prepared as follows: WO 00/47578 PCT/US99/18065 -275 5 Active Compound 0.5-10.0 mg Sodium Citrate 5-50 mg Citric Acid 1-15 mg Sodium Chloride 1-8 mg Water for Injection (USP) q.s. to 1 ml 0 Utilizing the above quantities, the active compound is dissolved at room temperature in a previously prepared solution of sodium chloride, citric acid, and sodium citrate in Water for Injection (USP, see page 1636 of United States Pharmacopeia/National Formulary for 1995, published by United States Pharmacopeial Convention, Inc., Rockville, Maryland (1994). L5 Example 292 In vitro Inhibition of Purified Enzymes Reagents: All buffer salts were obtained from Sigma Chemical Company (St. Louis, MO), and were of the highest purity available. The enzyme substrates, N-benzoyl-Phe-Val-Arg-p-nitroanilide (Sigma B7632), 0 N-benzoyl-Ile-Glu-Gly- Arg-p-nitroanilide hydrochloride (Sigma B2291), N-p-tosyl-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) were obtained from Sigma. N-Succinyl-Ala-Ala-Pro-Arg p-nitroanilide (BACHEM L-1720) and N-succinyl-Ala-Ala- Pro-Val '5 p-nitroanilide (BACHEM L-1770) were obtained from BACHEM (King of Prussia, PA). Human a-thrombin, human factor Xa and human plasmin were obtained from Enzyme Research Laboratories (South Bend, Indiana). Bovine a-chymotrypsin (Sigma C4129), bovine trypsin (Sigma T8642) and human 0 kidney cell urokinase (Sigma U5004) were obtained from Sigma. Human leukocyte elastase was obtained from Elastin Products (Pacific, MO). Ki Determinations: All assays are based on the ability of the test compound to inhibit the enzyme catalyzed hydrolysis of a peptide p-nitroanilide substrate.
WO 00/47578 PCT/US99/18065 -276 5 In a typical Ki determination, substrate is prepared in DMSO, and diluted into an assay buffer consisting of 50 mM HEPES, 200 mM NaC1, pH 7.5. The final concentrations for each of the substrates is listed below. In general, substrate concentrations are lower than the experimentally determined value for Km. Test compounds are prepared as a 1.0 mg/mL solution in DMSO. 10 Dilutions are prepared in DMSO yielding 8 final concentrations encompassing a 200 fold concentration range. Enzyme solutions are prepared at the concentrations listed below in assay buffer. In a typical Ki determination, into each well of a 96 well plate is pipetted 280 pL of substrate solution, 10 pL of test compound solution, and 15 the plate allowed to thermally equilibrate at 37 0 C in a Molecular Devices plate reader for > 15 minutes. Reactions were initiated by the addition of a 10 pL aliquot of enzyme and the absorbance increase at 405 nm is recorded for 15 minutes. Data corresponding to less than 10% of the total substrate hydrolysis were used in the calculations. The ratio of the velocity (rate of change in 20 absorbance as a function of time) for a sample containing no test compound is divided by the velocity of a sample containing test compound, and is plotted as a function of test compound concentration. The data are fit to a linear regression, and the value of the slope of the line calculated. The inverse of the slope is the experimentally determined Ki value. 25 Thrombin: Thrombin activity was assessed as the ability to hydrolyze the substrate N-succinyl-Ala-Ala-Pro-Arg-p-nitroanilide. Substrate solutions were prepared at a concentration of 32 pM (32 pM<<Km = 180 pM) in assay buffer. Final DMSO concentration was 4.3%. Purified human a-thrombin was diluted into assay buffer to a concentration of 15 nM. Final reagent 30 concentrations were: [thrombin] = 0.5 nM, [substrate N-succinyl-Ala-Ala-Pro-Arg-p-nitroanilide] = 32 pM. Factor X [FXa]: FXa activity was assessed as the ability to hydrolyze the substrate N-benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide hydrochloride. Substrate solutions were prepared at a concentration of 51 pM (51 << Km = 1.3 mM) in 35 assay buffer. Final DMSO concentration was 4.3%. Purified activated human WO 00/47578 PCT/US99/18065 -277 5 Factor X was diluted into assay buffer to a concentration of 300 nM. Final reagent concentrations were: [FXa] = 10 nM, [N-benzoyl-Ile-Glu-Gly-Arg p-nitroanilide hydrochloride] = 51 iM. Plasmin: Plasmin activity was assessed as the ability to hydrolyze the N-p-Tosyl-Gly-Pro-Lys-p-nitroanilide. Substrate solutions were prepared at a 10 concentration of 37 gM (37 tM<< Km = 243 [M) in assay buffer. Final DMSO concentration was 4.3%. Purified human plasmin was diluted into assay buffer to a concentration of 240 nM. Final reagent concentrations were: [Plasmin] = 8 nM, [N-p-Tosyl-Gly-Pro-Lys-p-nitroanilide] = 37 pIM. Chymotrypsin: Chymotrypsin activity was assessed as the ability to 15 hydrolyze N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide. Substrate solutions were prepared at a concentration of 14 iM (14 pM<< Km = 62 piM) in assay buffer. Final DMSO concentration was 4.3%. Purified bovine chymotrypsin was diluted into assay buffer to a concentration of 81 nM. Final reagent concentrations were: [Chymotrypsin] = 2.7 nM, 20 [N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide] = 14 pM. Trypsin: Trypsin activity was assessed as the ability to hydrolyze N-benzoyl-Phe-Val-Arg-p-nitroanilide. Substrate solutions were prepared at a concentration of 13 pM (13 pM<< Km = 291 IM) in assay buffer. Final DMSO concentration was 4.3%. Purified bovine trypsin was diluted into 25 assay buffer to a concentration of 120 nM. Final reagent concentrations were: [Trypsin] = 4 nM, [N-benzoyl-Phe-Val-Arg-p-nitroanilide] = 13 pjM. Elastase: Elastase activity was assessed as the ability to hydrolyze N-succinyl-Ala-Ala-Pro-Val-p-nitroanilide. Substrate solutions were prepared at a concentration of 19 [M (19 pM<< Km = 89 pM) in assay buffer. Final 30 DMSO concentration was 4.3%. Purified human leukocyte elastase was diluted into assay buffer to a concentration of 750 nM. Final reagent concentrations were: [Elastase] = 25 nM, [N-succinyl-Ala-Ala-Pro-Val-p-nitroanilide] = 19 pM. Urokinase: Urokinase activity was assessed as the ability to hydrolyze 35 N-CBZ-Val-Gly-Arg-p-nitroanilide. Substrate solutions were prepared at a WO 00/47578 PCT/US99/18065 -278 5 concentration of 100 pM (100 pM <Km = 1.2mM) in assay buffer. Final DMSO concentration was 4.3%. Purified human kidney urokinase was diluted into assay buffer to a concentration of 1.2 tM. Final reagent concentrations were: [Urokinase] = 40 nM, and N-CBZ-Val-Gly-Arg-p-nitroanilide] = 100 mM. 0 The results of exemplary assays are shown in the following table.
WO 00/47578 PCT/US99/18065 -279 5 Protease Inhibition Data Protease Ki Example# micromolar Trypsin 0.858 8 Trypsin 0.474 52 10 Factor Xa 2.73 94 Factor Xa 3.00 119 Chymo- 4.90 11 trypsin tPA 9.49 1 15 Plasmin 7.31 12 C1S 0.940 283 Additionally, the following compounds have Ki values in the range of 0.016 to 3.5 micromolar for uPA: Ex. # 28, 40, 53, 79, 84, 89, 131, 138, 139, 140, 143, 145, 172, 187, 20 200, 204, 206, 208, 213, 220, 222, 223, 227, 233, 235, 239, 260, 281 and 288. The results indicate that the compounds of the present invention are inhibitors of proteases, including urokinase. Having now fully described this invention, it will be understood to those of ordinary skill in the art that the same can be performed within a wide 25 and equivalent range of conditions, formulations, and other parameters without affecting the scope of the invention or any embodiment thereof. All patents and publications cited herein are fully incorporated by reference herein in their entirety.

Claims (76)

1. A compound of Formula I: 2 RI I\ R3 x NR 4 or a solvate, hydrate or pharmaceutically-acceptable salt thereof, wherein: X is O, S or NR 7 , where R 7 is hydrogen, alkyl, aralkyl, 10 hydroxy(C 2 . 4 )alkyl, or alkoxy(C 2 - 4 )alkyl; Y is a direct covalent bond, CH 2 or NH; Z is NR'R 6 , hydrogen or alkyl, provided that Y is NH whenever Z is hydrogen or alkyl; R' is hydrogen, amino, hydroxy, halogen, cyano, Cl- 4 alkyl or -CH 2 R, 15 where R is hydroxy, amino or C 1 . 3 alkoxy; R 2 and R 3 are independently: i. hydrogen; ii. halogen; iii. hydroxy; 20 iv. nitro; v. cyano; vi. amino, monoalkylamino, dialkylamino, monoarylamino, diarylamino, monoalkylmonoarylamino, monoaralkylamino, diaralkylamino, monoalkylmonoaralkylamino, monoheterocycleamino, diheterocycleamino, 25 monoalkylmonoheterocycleamino, alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, alkylsulfonylamino, aralkylsulfonylamino, aralkenylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, di(aralkylsulfonyl)amino, di(aralkenylsulfonyl)amino, di(arylsulfonyl)amino, or di 30 (heteroarylsulfonyl)amino, formylamino, alkanoylamino, alkenoylamino, WO 00/47578 PCT/US99/1806 5 -281 5 alkynoylamino, aroylamino, aralkanoylamino, aralkenoylamino, heteroaroylamino, heteroaralkanoylamino, H(S)CNH-, or thioacylamino, wherein any of the aryl or heteroaryl containing groups can be optionally substituted on the aromatic ring and wherein any of the heterocycle containing groups can be optionally ring substituted; 10 vii. aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl, acyl, aminoacyl, monoarylaminocarbonyl, diarylaminocarbonyl or monoalkylmonoarylaminocarbonyl; viii. aminothiocarbonyl, monoalkylaminothiocarbonyl, dialkylaminothiocarbonyl, thioacyl or aminothioacyl; 15 ix. aminocarbonylamino, mono- and dialkylaminocarbonylamino, mono- and diarylaminocarbonylamino, or mono- and diaralkylaminocarbonylamino; x. aminocarbonyloxy, mono- and dialkylaminocarbonyloxy, mono- and diarylaminocarbonyloxy, mono- and diaralkylaminocarbonyloxy; 20 xi. aminosulfonyl, mono- and dialkylaminosulfonyl, mono- and diarylaminosulfonyl, or mono- and diaralkylaminosulfonyl; xii. alkoxy, or alkylthio, wherein the alkyl portion of each group may be optionally substituted, xiii. aralkoxy, aryloxy, heteroaryloxy, aralkylthio, arylthio, or 25 heteroarylthio, wherein the aryl portion of each group can be optionally substituted; xiv. alkylsulfonyl, wherein the alkyl portion can be optionally substituted; xv. aralkylsulfonyl, aralkenylsulfonyl, arylsulfonyl or 30 heteroarylsulfonyl, wherein the aryl portion of each group can be optionally substituted; xvi. alkenyl, or alkynyl; xvii. optionally substituted aryl; xviii. optionally substituted alkyl; 35 xix. optionally substituted aralkyl; WO 00/47578 PCT/US99/18065 -282 5 xx. optionally substituted heterocycle; or xxi. optionally substituted cycloalkyl; and R 4 , R 5 and R 6 are independently hydrogen, CI- 4 alkyl, aryl, hydroxyalkyl, aminoalkyl, monoalkylamino(C 2 - 1 0 )alkyl, dialkylamino(C 2 10 )alkyl, carboxyalkyl, cyano, amino, alkoxy, or hydroxy, or -CO 2 Rw, where 10 Rw is alkyl, cycloalkyl, phenyl, benzyl, Rf or 0 Rh 0or Rg 0 Rd R e where Rd and Re are independently hydrogen, C 1 . 6 alkyl, C 2 - 6 alkenyl or phenyl, R' is hydrogen, C- 6 alkyl, C 2 . 6 alkenyl or phenyl, R9 is hydrogen, C 1 . 6 alkyl, C 2 - 6 alkenyl 15 or phenyl, and Rh is aralkyl or Cl- 6 alkyl; provided that at least one of R 2 or R 3 is selected from the group consisting of: (a) an optionally substituted alkyl group, preferably C-C 6 alkyl, more preferably C 1 -C 3 ; (b) alkoxy, aryloxy, alkylthio or arylthio, any of which is optionally 20 substituted; (c) optionally substituted C 6 -Cl 4 aryl, or optionally substituted aralkyl, except that R 3 is not nitrophenyl or aminophenyl, when R 1 and R 2 are both hydrogen or methyl; (d) optionally substituted heterocycle; and 25 (e) optionally substituted cycloalkyl.
2. A compound of claim 1, wherein R 2 or R 3 is alkyl, cycloalkyl, alkoxy, alkylthio or alkylsulfonyl, and the alkyl portion of said alkyl, cycloalkyl, alkoxy, alkylthio or alkylsulfonyl is optionally substituted with 1 to 4 substituents selected from the group consisting of halogen, hydroxy, thiol, 30 amino, monoalkylamino, dialkylamino, formylamino, acylamino, aminoacyl, WO 00/47578 PCT/US99/18065 -283 5 monoalkylaminocarbonyl, dialkylaminocarbonyl, thiocarbonylamino, thioacylamino, aminothiocarbonyl, alkoxy, aryloxy, aminocarbonyloxy, monoalkylaminocarbonyloxy, dialkylaminocarbonyloxy, monoarylaminocarbonyloxy, diarylaminocarbonyloxy, monoaralkylaminocarbonyloxy, diaralkylaminocarbonyloxy, alkylsulfonyl, 10 arylsulfonyl, aralkylsulfonyl, alkylsulfonylamino, arylsulfonylamino, aralkylsulfonylamino, alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, monoalkylaminothiocarbonyl, dialkylaminothiocarbonyl, aralkoxy, carboxy, carboxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, nitro, cyano, trifluoromethyl, alkylthio and arylthio. 15
3. A compound of claim 1, wherein R 3 is optionally substituted alkyl or alkylthio.
4. A compound of claim 2, wherein said 1 to 4 substituents are selected from the group consisting of chloro, hydroxy, amino, mono(Cl- 4 )alkylamino, di(C _ 4 )alkylamino, formylamino, C 2 . 6 acylamino, 20 aminocarbonyl, C 2 - 8 aminoacyl, C 2 - 6 thioacylamino, aminothiocarbonyl, C 2 - 8 aminothioacyl, C1- 6 alkoxy, C 6 - 1 4 aryloxy, carboxy, carboxy(C. 6 )alkyl, C 2 -8 alkoxycarbonyl, nitro, cyano, trifluoromethyl, C- 6 alkylthio, C 6 - 14 arylthio, Cl-6 aralkylsulfonylamino, Cl.6 arylsulfonylamino, monoalkylaminocarbonyloxy, dialkylaminocarbonyloxy, mono(C 6 . 1 0 )arylaminocarbonyloxy, di(C 6 25 1 )arylaminocarbonyloxy, monoaralkylcarbonyloxy, diaralkylcarbonyloxy, C 1 _ 6 alkoxycarbonylamino, C 7 -C 5 aralkoxycarbonylamino, and C 6 -C 0 aryloxycarbonylamino.
5. A compound of claim 1, wherein at least one of R 2 and R 3 is aryl, aralkoxy, arylthio, aralkyl, aryloxy, aralkylthio, aralkylsulfonyl, 30 arylsulfonyl, heterocycle or heterocycloalkyl optionally substituted with 1 to 4 substituents selected from the group consisting of halogen, hydroxy, thiol, amino, monoalkylamino, dialkylamino, formylamino, acylamino, aminoacyl, WO 00/47578 PCT/US99/18065 -284 5 mono alkylaminocarbonyl, dialkylaminocarbonyl, thiocarbonylamino, thioacylamino, aminothiocarbonyl, alkoxy, aryloxy, aminocarbonyloxy, mono alkylaminocarbonyloxy, dialkylaminocarbonyloxy, monoarylaminocarbonyloxy, diarylaminocarbonyloxy, monoaralkylaminocarbonyloxy, diaralkylaminocarbonyloxy, alkylsulfonyl, 10 arylsulfonyl, aralkylsulfonyl, alkylsulfonylamino, arylsulfonylamino, aralkylsulfonylamino, alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, mono alkylaminothiocarbonyl, dialkylaminothiocarbonyl, aralkoxy, carboxy, carboxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, nitro, cyano, trifluoromethyl, alkylthio and arylthio. 15 6. A compound of claim 5, wherein said 1 to 4 substitutents are selected from the group consisting of chloro, hydroxy, amino, mono(Cl- 4 )alkylamino, di(Cl 4 )alkylamino, formylamino, C 2 - 6 acylamino, aminocarbonyl, C 2 - 8 aminoacyl, C 3 . 7 cycloalkyl, C 1 - 6 alkyl, Cl- 6 alkoxy, C 6 .1 4 aryloxy, carboxy, carboxy(C 1 _ 6 )alkyl, C 2 - 8 alkoxycarbonyl, nitro, cyano, 20 trifluoromethyl, Cl.
6 alkylthio, C 6 - 1 4 arylthio, C 6 - 14 aryl, tetrazolyl, thienyl, 3,4 methylenedioxy, 3,4-ethylenedioxy, 3,4-propylenedioxy, CI- 6 alkylsulfonylamino, C,_-6 aralkylsulfonylamino, Cl- 6 arylsulfonylamino, mono or dialkylaminocarbonyloxy, mono- or di- C 6 -1 0 arylaminocarbonyloxy, mono or diaralkylcarbonyloxy, Cl- 6 alkoxycarbonylamino, C 7 -C 1 5 25 aralkoxycarbonylamino, C6-C 0 aryloxycarbonylamino, C 2 . 6 thioacylamino, aminothiocarbonyl, and C 2 . 8 aminothioacyl.
7. A compound of claim 1, wherein X is sulfur or oxygen; Y is a covalent bond or -NH-; 30 R' is hydrogen, amino, hydroxy or halogen; one of R 2 or R 3 is hydrogen, Cl. 6 alkylthio, CI. 6 alkyl, or C 1 -6 alkoxy, and the other of R 2 or R 3 is aminoacyl, acylamino, aminosulfonyl, sulfonylamino, aminocarbonylamino, alkoxycarbonylamino, optionally WO 00/47578 PCT/US99/18065 -285 5 substituted oxazolyl, optionally substituted isoxazolyl, optionally substituted benzothienyl, optionally substituted furanyl, optionally substituted pyrazolyl or optionally substituted pyridyl.
8. A compound of claim 7, wherein R 4 , R 5 and R 6 are hydrogen.
9. A compound of claim 1, wherein 0 X is sulfur or oxygen; Y is a covalent bond or -NH-; Z is NRR 6 ; R' is hydrogen, amino, hydroxy or halogen; one of R 2 and R 3 is hydrogen, CI- 6 alkylthio, C 1 . 6 alkyl or Ci- 6 15 alkoxy, and the other of R 2 and R 3 is =Ar R8 R where Ar is phenyl, thiazolyl, thiazolinyl, oxazolyl, isothiazolyl, isoxazolyl, furanyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, thienyl, 20 tetrazolyl, pyrrolyl, pyrazolyl, oxadiazolyl, oxazolinyl, isoxazolinyl, imidazolinyl, triazolyl, pyrrolinyl, benzothiazolyl, benzothienyl, benzimidazolyl, 1,3-oxazolidin-2-onyl, and imidazolin-2-onyl; R 8 and R 9 are independently selected from the group consisting of hydrogen, halogen, amino, mono(C 1 - 4 )alkylamino, arylamino, mono C6-14 25 arylamino, di(C 6 - 1 4 )arylamino, mono(C 6 .- 1 4 )ar(C 16 )alkylamino, di(C 6 - 4 )ar(C, 6 )alkylamino, di(Cl- 4 )alkylamino, formylamino, C 2 . 6 acylamino, aminocarbonyl, C 2 - 8 aminoacyl, C 2 - 6 thioacylamino, aminothiocarbonyl, C 2 - 8 aminothioacyl, C 1 - 6 alkyl, C 3 - 6 cycloalkyl, Cl- 6 alkoxy, carboxy, carboxy(C 1 . 6 )alkyl, C 2 - 8 alkoxycarbonyl, nitro, cyano, trifluoromethyl, 30 tetrazolyl, thienyl, C 6 - 1 4 aryloxy, C 1 _ 6 alkylthio, C 6 - 1 4 arylthio, C 6 1 4 aryl, and C 6 -, 4 ar(C, 1 - 6 )alkyl, wherein the aryl portions of any of said groups may be optionally substituted with 1 to 3 substituents independently selected from the WO 00/47578 PCT/US99/18065 -286 5 group consisting of halogen, hydroxy, amino, mono(Cl 4 )alkylamino, di(C 1 . 4 )alkylamino, formylamino, Cl- 4 acylamino., Cl- 4 aminoacyl, mono(C 1 4 )alkylaminocarbonyl, di(Cl- 4 )alkylaminocarbonyl, thiocarbonylamino, C1, 4 thioacylamino, aminothiocarbonyl, Cl- 4 alkoxy, C 6 - 0 loaryloxy, aminocarbonyloxy, mono(Cl- 4 )alkylaminocarbonyloxy, di(C1-4) 10 alkylaminocarbonyloxy, mono(C 6 -o 10 )arylaminocarbonyloxy, di(C 6 1 )arylaminocarbonyloxy, mono(C 4 - 1 2 )aralkylaminocarbonyloxy, di(C 4 1 2 )aralkylaminocarbonyloxy, Cl- 4 alkylsulfonyl, C 6 .-o 10 arylsulfonyl, (C 7 -12) aralkylsulfonyl, CI. 4 alkylsulfonylamino, C 6 .- 1 0 arylsulfonylamino, (C 7 .- 12) aralkylsulfonylamino, Cl- 4 alkoxycarbonylamino, C 7 . 1 2 aralkoxycarbonylamino, 15 C 6 . 1 0 aryloxycarbonylamino, mono(C - 4 )alkylaminothiocarbonyl, di(C 1 4 )alkylaminothiocarbonyl, C 7 - 1 2 aralkoxy, carboxy, carboxy(C- 4 )alkyl, C 1 4 alkoxycarbonyl, Cl. 4 alkoxycarbonylalkyl, nitro, cyano, trifluoromethyl, CI 4 alkylthio, C 6 . 1 0 arylthio, 3,4-methylenedioxy, 3,4-ethylenedioxy, and 3,4 propylenedioxy; and 20 R 4 , R 5 , R 6 are independently hydrogen, Cl. 4 alkyl, amino, C 1 4 alkoxy or hydroxy.
10. A compound of Claim 9, wherein X is sulfur; Y is a covalent bond; 25 Z is NR'R 6 ; R' is hydrogen; R 2 is R' =Arl R9 where 30 Ar is phenyl, thiazolyl, oxazolyl, pyridyl or imidazolyl; R 8 and R 9 are independently selected from the group consisting of hydrogen and C 6 - 1 0 aryl optionally substituted with 1 to 3 substituents independently selected from the group consisting of chloro, hydroxy, C 1 . 4 WO 00/47578 PCT/US99/18065 -287 5 alkyl, C 3 . 6 cycloalkyl, Cl. 4 alkoxy, amino, carboxy, phenyl, napthyl, biphenyl, hydroxyphenyl, methoxyphenyl, chlorophenyl, dichlorophenyl, aminophenyl, carboxyphenyl, nitrophenyl, 3,4-ethylenedioxy, 3,4-methylenedioxy, and 3,4 propylenedioxy; R is methylthio or methyl; and 10 R 4 , R 5 , R 6 are hydrogen.
11. A compound of claim 1, wherein X is sulfur; Y is a direct covalent bond; Z is NR'R 6 ; 15 R 1 is hydrogen; R 2 is alkyl, ar(alkyl), alkylsulfonyl, -SO 2 -alkyl, amido, amidino, or R8 where 20 Ar is an aromatic or heteroaromatic group selected from the group consisting of phenyl, thiazolyl, oxazolyl, imidazolyl and pyridyl; R' and R 9 are independently selected from the group consisting of hydrogen, carboxy, phenyl, naphthyl, alkyl, pyridyl, oxazolyl, furanyl, cycloalkyl and amino, any of which may be optionally substituted with 1 to 3 25 substituents independently selected from the group consisting of halogen, alkyl, haloalkyl, alkaryl, heteroaryl, phenyl, naphthyl, alkoxy, aryloxy, hydroxy, amino nitro, thiophenyl, benzothiophenyl, fluorenyl, 3,4 ethylenedioxy, 3,4-methylenedioxy, 3,4-propylenedioxy, arylsulfonamido, alkylsulfonamido and aryloxy, each of said 1 to 3 substituents may be further 30 optionally substituted with one or more groups selected from alkoxy, haloalkyl, halogen, alkyl, amino, acetyl, hydroxy, dialkylamino, dialkylamino acyl, monoalkylaminoacyl, -SO 2 -heteroaryl, -SO 2 -aryl, or aryl; WO 00/47578 PCT/US99/18065 -288 5 R 3 is -SO 2 -alkyl, trifluoromethyl, S(O)-alkyl, hydrogen, alkoxy, alkylthio, alkyl, aralkyllthio; and R 4 , R 5 , R 6 are hydrogen.
12. A compound of claim 11, wherein Ar is thiazolyl and at least one of R" 7 and R 18 is phenyl. 10
13. A compound of claim 11 or 12, wherein said thiazolyl is thiazol-2-yl.
14. A compound of claim 13, wherein R 2 is a 4-phenylthiazol-2-yl group, wherein said phenyl is further optionally substituted.
15. A compound of claim 11 or 12, wherein said thiazolyl is 15 thiazol-4-yl.
16. A compound of claim 15, wherein R 2 is a 2-aminothiazol-4-yl group.
17. A compound of claim 11, wherein said oxazolyl is oxazol-2-yl.
18. A compound of claim 11, wherein said oxazolyl is oxazol-4-yl. 20
19. A compound of claim 11, wherein R 3 is methylthio.
20. A compound of Formula I 2 R1 R 4 x NR 4 WO 00/47578 PCT/US99/18065 -289 5 or a solvate, hydrate or pharmaceutically acceptable salt thereof, wherein X is sulfur; Y is a covalent bond; Z is NR'R 6 ; R' is hydrogen; 10 R 2 is R 8 R 9 where Ar is phenyl, thiazolyl, or oxazolyl; R 8 and R 9 are independently selected from the group consisting 15 of hydrogen and C 6 -, 0 aryl optionally substituted with 1 to 3 substituents independently selected from the group consisting of chloro, hydroxy, Cl-4 alkyl, C 1 - 4 alkoxy, phenyl, 3,4-ethylenedioxy, 3,4-methylenedioxy, and 3,4 propylenedioxy; R 3 is methylthio; and 20 R 4 , R 5 , R 6 are hydrogen.
21. A compound of Formula I 2 R R R I x NR 4 or a solvate, hydrate or pharmaceutically acceptable salt thereof, wherein X is sulfur; 25 Y is a covalent bond; R' is hydrogen; R 2 iS WO 00/47578 PCT/US99/18065 -290 Ar R 9 where Ar is thiazolyl; R 8 and R 9 are independently selected from the group consisting of hydrogen and C 6 . 1 0 aryl substituted with a sulfonamide group; 10 R 3 is methylthio; and R 4 , R 5 , R 6 are hydrogen.
22. A compound of claim 21, wherein said sulfonamide group is a C 6 -1 0 arylsulfonamide, alkylsulfonamide, alkoxysulfonamide or heteroarylsulfonamide. 15
23. A compound of claim 22, wherein said sulfonamide group is selected from the group consisting of 4-methylphenylsulfonamide, methylsulfonamide, phenylsulfonamide, trifluoromethylsulfonamide, 4-fluorophenylsulfonamide, 4-chlorophenylsulfonamide, 3-chlorophenylsulfonamide, 4-methoxysulfonamide, 2,4 20 difluorophenylsulfonamide, 2-(thiophene)sulfonamide, 2-(5-chlorothiophene) sulfonamide, butylsulfonamide, and isopropylsulfonamide.
24. A compound of Formula I 2 x NR 4 or a solvate, hydrate or pharmaceutically acceptable salt thereof, wherein 25 X is sulfur; Y is a covalent bond; Z is NR'R 6 ; WO 00/47578 PCT/US99/18065 -291 5 R 1 is hydrogen; R 2 is R' Rs .R9 where Ar is thiazolyl; 10 R 8 and R 9 are independently selected from the group consisting of hydrogen and C 6 - 10 aryl substituted with a group selected from -OCH 2 C(O)-alkoxy, -OCH 2 C(O)-amino, -OCH 2 C(O)-NH-alkyl or -OCH 2 C(O)-N(alkyl) 2 ; R 3 is methylthio; and 15 R 4 , R 5 , R 6 are hydrogen.
25. A compound of Formula III, O 11 / G,, R,, / III G!-H2)7( N\ NH A S NH 2 or a salt thereof, wherein A is methylthio or methyl; 20 G' is -0-, -S-, -NH-, or a covalent bond; n is an integer from 1-10; m is an integer from 0-1; and R' and R" are independently selected from hydrogen, alkyl, aryl or aralkyl, or R' and R" taken together with the N atom to which they are WO 00/47578 PCT/US99/18065 -292 5 attached form a 3-8 membered heterocyclic ring optionally containing an additional O, N, or S atom.
26. A compound according to claim 25, wherein said 3-8 membered heterocyclic ring contains an additional N atom, said additional N atom optionally substituted by hydrogen, CI 4alkyl, C 6 -1 0 aryl, C6- 0 ar(CI- 4 )alkyl, 10 Cl- 6 alkoxy, alkoxycarbonyl or benzyloxycarbonyl.
27. A compound according to claim 25, wherein said 3-8 membered heterocyclic ring is piperazinyl, pyrrolidinyl, piperidinyl or morpholinyl, which is further optionally substituted by 1-4 substituents selected from halogen, hydroxy, amino, monoalkylamino, dialkylamino, 15 formylamino, acylamino, aminoacyl, monoalkylaminocarbonyl, dialkylaminocarbonyl, thiocarbonylamino, thioacylamino, aminothiocarbonyl, alkoxy, aryloxy, aminocarbonyloxy, monoalkylaminocarbonyloxy, dialkylaminocarbonyloxy, monoarylaminocarbonyloxy, diarylaminocarbonyloxy, monoarakylaminocarbonyloxy, 20 diaralkylaminocarbonyloxy, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, alkylsulfonylamino, arylsulfonylamino, arakylsulfonylamino, alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, mono alkylaminothiocarbonyl, dialkylaminothiocarbonyl, aralkoxy, carboxy, carboxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, nitro, cyano, 25 trifluoromethyl, alkylthio and arylthio.
28. A compound of claim 11, wherein Ar is thiazolyl; and one of R 8 and R 9 is hydrogen and the other of R 8 and R 9 is amino, said amino group optionally substituted with halogen, hydroxy, amino, 30 monoalkylamino, dialkylamino, formylamino, acylamino, aminoacyl, monoalkylaminocarbonyl, dialkylaminocarbonyl, thiocarbonylamino, thioacylamino, aminothiocarbonyl, alkoxy, aryloxy, aminocarbonyloxy, WO 00/47578 PCT/US99/18065 -293 5 monoalkylaminocarbonyloxy, dialkylaminocarbonyloxy, monoarylaminocarbonyloxy, diarylaminocarbonyloxy, monoaralkylaminocarbonyloxy, diaralkylaminocarbonyloxy, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, alkylsulfonylamino, arylsulfonylamino, aralkylsulfonylamino, alkoxycarbonylamino, aralkoxycarbonylamino, 10 aryloxycarbonylamino, monoalkylaminothiocarbonyl, dialkylaminothiocarbonyl, aralkoxy, carboxy, carboxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, nitro, cyano, trifluoromethyl, alkylthio and arylthio.
29. A compound of Formula IV R"' I HN N S_ IV NH A S NH 2 15 wherein A is methylthio or methyl; and R'" is hydrogen, C 6 - 1 4 aryl, C, 6 _alkyl, alkoxy (C 6 . 1 4 )aryl, amino(C 6 - 1 4 )aryl, monoalkylamino(C 6 - 1 4 )aryl, dialkylamino(C 6 - 14 )aryl, C 61 - 0 ar(C _ 6 )alkyl, C 1 . 6 alk(C 6 -1 4 )aryl, amino(Cl- 6 )alkyl, monoalkylamino (Ci- 6 )alkyl, dialkylamino 20 (Cl- 6 )alkyl, hydroxy(C 6 -1 4 )aryl, or hydroxy(C- 6 )alkyl, any of which is further optionally substituted by 1-4 non-hydrogen substituents selected from halogen, hydroxy, amino, monoalkylamino, dialkylamino, formylamino, acylamino, aminoacyl, mono- or di- alkylaminocarbonyl, thiocarbonylamino, thioacylamino, aminothiocarbonyl, alkoxy, aryloxy, aminocarbonyloxy, mono 25 or di-alkylaminocarbonyloxy, mono- or diarylaminocarbonyloxy, mono- or diarakylaminocarbonyloxy, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, WO 00/47578 PCT/US99/18065 -294 5 alkylsulfonylamino, arylsulfonylamino, arakylsulfonylamino, alkoxycarbonylamino, aralkoxycarbonylamino, aryloxycarbonylamino, mono or di- alkylaminothiocarbonyl, aralkoxy, carboxy, carboxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, nitro, cyano, trifluoromethyl, alkylthio and arylthio. 10
30. A compound of claim 29 which is one of: 4-{2-[(3 methoxyphenyl)amino](1,3-thiazol-4-yl)}-5-methylthiothiophene-2 carboxamidine, 4- {2-[(4-methoxyphenyl)amino](1,3-thiazol-4-yl)} -5 methylthiothiophene-2-carboxamidine, 4-(2-{ [4 (dimethylamino)phenyl]amino } (1,3-thiazol-4-yl))-5-methylthiothiophene-2 15 carboxamidine, 4- { 2-[(4-chloro-2-methylphenyl)amino](1,3-thiazol-4-yl) }-5 methylthiothiophene-2-carboxamidine, 4- { 2-[(diphenylmethyl)amino](1,3 thiazol-4-yl) }-5-methylthiothiophene-2-carboxamidine, 5-methylthio-4- {2-[(3 phenylpropyl)amino](1,3-thiazol-4-yl) } thiophene-2-carboxamidine, 5 methylthio-4- {2-[(2,4,5-trimethylphenyl)amino](1,3-thiazol-4-yl) } thiophene 20 2-carboxamidine, 4- {2-[(2-fluorophenyl)amino](1,3-thiazol-4-yl) }-5 methylthiothiophene-2-carboxamidine, 4- {2-[(3-chloro-2 methylphenyl)amino](1,3-thiazol-4-yl) } -5-methylthiothiophene-2 carboxamidine, 4-(2- { [2-(methylethyl)phenyl] amino } (1,3-thiazol-4-yl))- 5 methylthiothiophene-2-carboxamidine, 5-methylthio-4-(2- { [4 25 (phenylmethoxy)phenyl]amino } (1,3-thiazol-4-yl))thiophene-2-carboxamidine, 4- {2-[(2-bromophenyl)amino](1,3-thiazol-4-yl) } -5-methylthiothiophene-2 carboxamidine, 4- {2-[(2,6-dichlorophenyl)amino](1,3-thiazol-4-yl) } -5 methylthiothiophene-2-carboxamidine, 4-{2-[(2-bromo-4 methylphenyl)amino](1,3-thiazol-4-yl)} -5-methylthiothiophene-2 30 carboxamidine, 5-methylthio-4- {2-[(2-morpholin-4-ylethyl)amino](1,3 thiazol-4-yl)}thiophene-2-carboxamidine, 4- {2-[(2,3 dichlorophenyl)amino](1,3-thiazol-4-yl) } -5-methylthiothiophene-2 carboxamidine, 5-methylthio-4- { 2- [(3,4,5-trimethoxyphenyl)amino](1,3 thiazol-4-yl) } thiophene-2-carboxamidine, WO 00/47578 PCT/US99/18065 -295 5 5-methylthio-4- { 2-[(2-piperidylethyl)amino](1,3-thiazol-4-yl) } thiophene-2 carboxamidine, 4-(2- { [(4-methylphenyl)methyl]amino } (1,3-thiazol-4-yl))-5 methylthiothiophene-2-carboxamidine, 4-(2- { [4-(4 chlorophenoxy)phenyl]amino}(1,3-thiazol-4-yl))-5-methylthiothiophene-2 carboxamidine 4-(2-{ [4-phenoxyphenyl]amino} (1,3-thiazol-4-yl))-5 10 methylthiothiophene-2-carboxamidine, 5-methylthio-4-(2- { [4 (phenylamino)phenyl]amino } (1,3-thiazol-4-yl))thiophene-2-carboxamidine, 5 methylthio-4-(2-{ [4-benzylphenyl]amino}(1,3-thiazol-4-yl))thiophene-2 carboxamidine, 5-methylthio-4-(2- { [4-(piperidylsulfonyl)phenyl]amino}(1,3 thiazol-4-yl))thiophene-2-carboxamidine 5-methylthio-4-[2-(3 15 quinolylamino)(1,3-thiazol-4-yl)]thiophene-2-carboxamidine, 5-methylthio-4 [2-(2-naphthylamino)(1,3-thiazol-4-yl)]thiophene-2-carboxamidine, 4-[2-(2H benzo[3,4-d] 1,3-dioxolan-5-ylamino)(1,3-thiazol-4-yl)]-5 methylthiothiophene-2-carboxamidine, 4-{2-[(7-bromofluoren-2 yl)amino](1,3-thiazol-4-yl) } -5-methylthiothiophene-2-carboxamidine, 4- { 2 20 [(4-cyclohexylphenyl)amino](1,3-thiazol-4-yl) } -5-methylthiothiophene-2 carboxamidine, 5-methylthio-4-(2- { [4-(phenyldiazenyl)phenyl] amino } (1,3 thiazol-4-yl))thiophene-2-carboxamidine, 5-methylthio 4-(2- { [3 (hydroxymethyl)phenyl]amino}(1,3-thiazol-4-yl))-thiophene-2-carboxamidine, 4- [2-( { 3- [(3-methylpiperidyl)methyl]phenyl } amino)(1,3-thiazol-4-yl)]-5 25 methylthiothiophene-2-carboxamidine, 4-{2-[(3-hydroxyphenyl)amino](1,3 thiazol-4-yl)}-5-methylthiothiophene-2-carboxamidine, 4-(2- { [4 (carbamoylmethoxy)phenyl]amino } (1,3-thiazol-4-yl))-5-methylthiothiophene 2-carboxamidine, 5-methyl-4- {2-[(3,4,5-trimethoxyphenyl)amino](1,3-thiazol 4-yl) } thiophene-2-carboxamidine, 5-methyl-4- {2-[(4 30 phenoxyphenyl)amino](1,3-thiazol-4-yl) } thiophene-2-carboxamidine, 5 methyl-4-[2-(phenylamino)(1,3-thiazol-4-yl)]thiophene-2-carboxamidine, 4 (4-isoxazol-5-yl(1,3-thiazol-2-yl))-5-methylthiothiophene-2-carboxamidine. WO 00/47578 PCT/US99/18065 -296 5
31. A compound of claim 1 which is one of: 4-[4-(4-chlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-phenyl-5-methylthiothiophene-2-carboxamidine; 4-[4-(2,4-dichlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 0 4-(4-methylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-[4-(3-methoxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(3-hydroxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-(4-phenylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-[4-(4-nitrophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 5 4-[4-(3,4-ethylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-[4-(3,4-propylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-[4-(4-(naphth-2-yl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 0 and 4-isopropylsulfonyl-5-methylthiothiophene-2-carboxamidine; or a hydrate, solvate or pharmaceutically acceptable salt thereof.
32. A compound of claim 11, which is one of: 4-phenyl-5-methylthiothiophene-2-carboxamidine; 5 4-[4-(4-chlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(4-phenylphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(3-methoxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(3-hydroxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-(4-phenylthiazol--2-yl)-5-methylthiothiophene-2-carboxamidine; 0 4-[4-(4-nitrophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(3,4-ethylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-[4-(4-methoxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; WO 00/47578 PCT/US99/18065 -297 5 4-[4-(3,4-propylenedioxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-isopropylsulfonyl-5-methylthiothiophene-2-carboxamidine; 4-(4-methylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4- [4-(2,4-dichlorophenyl)thiazol-2-yl]-5 -methylthiothiophene-2 0 carboxamidine; 4-(2-naphthylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-[4-(4-chloro-3 -methylphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-(5 -methyl-4-phenylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 5 4- [4-(4-chloro-3-nitrophenyl)thiazol-2-yl] -5 -methylthiothiophene-2 carboxamidine; 4-(5-phenyloxazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4- [4-(3 -fluoro-5 -trifluoromethylphenyl)-5-methylthiazol-2-yl]-5 methylthiothiophene-2-carboxamidine; o 4-[4-(3 ,5-bis(trifluoromethyl)phenyl)-5-methyl-thiazol-2-yI]-5 methylthiothiophene-2-carboxamidine; 4- [4-(3 -fluoro-5-trifluoromethylphenyl)thiazol-2-yl] -5-methylthiothiophene-2 carboxamidine; 4- [4-(3 -bromophenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 5 4- [4-(3 ,4-methylenedioxyphenyl)thiazol-2-yI] -5-methylthiothiophene-2 carboxamidine; 4 -[4-(4-methylphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(3 ,5-bis(trifluoromethyl)phenyl)thiazol-2-yl]-5 -methylthiothiophene-2 carboxamidine; o 4- [4-(2-methoxyphenyl)thiazol-2-yl]-5 -methylthiothiophene-2-carboxamidine; 4 -(4-phenylimidazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4- [4-(2,4-dimethoxyphenyl)thiazol-2-yI] -5 -methylthiothiophene-2 carboxamidine; 4 -(4-benzylthiazol-2-yI)-5-methylthiothiophene-2-carboxamidine; WO 00/47578 PCT/US99/18065 -298 5 4-[4-(3 ,4-dichlorophenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-[4-(3 -methylphenyl)thiazol-2-yl]-5-methylthiothiophene-2-carboxamidine; 4-[4-(3 ,5 -dimethoxyphenyl)thiazol-2-yl] -5-methylthiothiophene-2 carboxamidine; 10 4- [4-(2-methylphenyl)thiazol-2-y-5-methylthiothiophene-2-carboxamidine; 4-[4-(2,5-dimethoxyphenyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-(4,5-diphenyl)thiazol-2-yl-5 -methylthiothiophene-2-carboxamidine; 4-(2-phenyl)thiazol-4-yl-5 -methylthiothiophene-2-carboxamidine; 15 4- [4-(2-chloro-3-pyridyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 4-[4-(phenoxymethyl)thiazol-2-yl] -5-methylthiothiophene-2-carboxamidine; 4-(4-cyclohexylthiazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4- [4-(4-chlorophenyl)thiazol-2-yI] -5-methylthiothiophene-2-carboxamidine; 20 4- [4-(2-hydroxyphenyl)thiazol-2-yl] -5-methylthiothiophene-2-carboxamidine; 4- [4-(3 -trifluoromethoxyphenyl)thiazol-2-yl]-5 -methylthiothiophene-2 carboxamidine; 4-[4-(2-chloro-4-pyridyl)thiazol-2-yl]-5-methylthiothiophene-2 carboxamidine; 25 4-(5-phenyl-2-pyridyl)-5-methylthiothiophene-2-carboxamidine; 4- [2-(2-chlorophenylamino)thiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; 4-[2-(3-methoxyphenylamino)thiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; 30 4-[2-(phenylamino)thiazol-4-yl]-5 -methylthiothiophene-2-carboxamidine; 4-[2-(2,5 -dimethoxyphenylamino)thiazol-4-yI]-5 -methylthiothiophene-2 carboxamidine; 4-(2-aminothiazol-4-yl)-5-methylthiothiophene-2-carboxamidine; 4-[2-(4-chloro-2-methylphenylamino)thiazol-4-yl]-5-methylthiothiophene-2 35 carboxamidine; WO 00/47578 PCT/US99/18065 -299 5 4- [2-(4-dimethylaminophenylamino)thiazol-4-yl] -5-methylthiothiophene-2 carboxamidine; 4-[2-(4-methoxyphenylamino)thiazol-4-yl]-5 -methylthiothiophene-2 carboxamidine; 4- [4-(4-hydroxy-3-methoxyphenyl)thiazol-2-yl] -5-methylthiothiophene-2 10 carboxamidine; 4- [4-(3 -hydroxy-4-methoxyphenyl)thiazol-2-yl] -5-methylthiothiophene-2 carboxamidine; 4- [2-(2-fluorophenylamino)thiazol-4-yl] -5-methylthiothiophene-2 carboxamidine; 15 4- [2-(2,4,5-trimethylphenyl)aminothiazol-4-yl]-5 -methylthiothiophene-2 carboxamidine; 4- [2-(3 -chloro-2-methylphenyl)aminothiazol-4-yl] -5 -methylthiothiophene-2 carboxamidine; 4- [2-(2-isopropylphenyl)aminothiazol-4-yl]-5-methylthiothiophene-2 20 carboxamidine; 4- [2-(4-benzyloxyphenyl)aminothiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; 4-[2-(2-bromophenyl)aminothiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; 25 4- [2-(2, 5-dichlorophenyl)aminothiazol-4-yl]-5 -methylthiothiophene-2 carboxamidine; 4-[2-(2-bromo-4-methylphenyl)aminothiazol-4-yl] -5-methylth-iothiophene-2 carboxamidine; 4- [2-(2,3 -dichlorophenyl)aminothiazol-4-yl]-5-methylthiothiophene-2 30 carboxamuidine; 4- [2-(3 ,4,5-trimethoxyphenyl)aminothiazol-4-yl] -5-methylthiothiophene-2 carboxamidine; 4-[2-(2-piperidinylethyl)aminothiazol-4yl]-5-methylthiothiophene-2 carboxamidine; WO 00/47578 PCT/US99/18065 -300 4-[2-(4-methylphenyl)aminothiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; 4-(4-phenyloxazol-2-yl)-5-methylthiothiophene-2-carboxamidine; 4-[2-(diphenylmethyl)aminothiazol-4-yl]-5-methylthiothiophene-2 5 carboxamidine; 4-[2-(3-phenylpropyl)aminothiazol-4-yl]-5-methylthiothiophene-2 carboxamidine; or a solvate, hydrate or pharmaceutically acceptable salt hereof.
33. A method of treating a disease selected from benign prostatic 10 hypertrophy, prostatic carcinoma, tumor metastisis, restenosis and psoriasis, comprising administering to a patient in need of such treatment an effective amount of a compound of Formula I: 2 R1I X NR 4 or a solvate, hydrate or pharmaceutically-acceptable salt thereof, wherein: 15 X is oxygen, sulfur or NRT; R 7 is hydrogen, alkyl, aralkyl, hydroxy(C 3 - 4 )alkyl, alkoxy(C 3 - 4 )alkyl; Y is a covalent bond, CH 2 or NH; Z is NRR 6 ; 20 R 1 is a hydrogen, amino, hydroxy, halogen, cyano, Cl- 4 alkyl, -CH 2 R where R is hydroxyamino, or Ci- 3 alkoxy; R 2 and R 3 are independently: i. hydrogen; ii. halogen; 25 iii. hydroxy; iv. nitro; WO 00/47578 PCT/US99/18065 -301 v. cyano; vi. amino, monoalkylamino, dialkylamino, monoarylamino, diarylamino, monoalkylmonoarylamino, monoaralkylamino, diaralkylamino, alkarylamino, alkoxycarbonylamino, aralkoxycarbonylamino, 5 aryloxycarbonylamino, alkylsulfonylamino, aralkylsulfonylamino, arylsulfonylamino, formylamino, acylamino, H(S)CNH-, or thioacylamino; vii. aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl, acyl, arylaminocarbonyl, or aminoacyl; viii. aminothiocarbonyl, monoalkylaminothiocarbonyl, 0 dialkylaminothiocarbonyl, thioacyl, or aminothioacyl; ix. aminocarbonylamino, monoalkylaminocarbonylamino, dialkylaminocarbonylamino, monoarylaminocarbonylamino, diarylaminocarbonylamino, monoaralkylaminocarbonylamino, or diaralkylaminocarbonylamino, 5 x. aminocarbonyloxy, monoalkylaminocarbonyloxy, dialkylaminocarbonyloxy, monoarylaminocarbonyloxy, diarylaminocarbonyloxy, monoaralkylaminocarbonyloxy, or diaralkylaminocarbonyloxy, xi. aminosulfonyl, monoalkylaminosulfonyl, 0 dialkylaminosulfonyl, monoarylaminosulfonyl, diarylaminosulfonyl, or monoaralkylaminosulfonyl, or diaralkylaminosulfonyl, xii. alkoxy or alkylthio, wherein said alkyl portion of said alkoxy or alkylthio group may be optionally substituted; xiii. aralkoxy, aryloxy, aralkylthio, or arylthio, wherein the 5 aryl portion of said aralkoxy, aryloxy, aralkylthio or arylthio group may be optionally substituted; xiv. alkylsulfonyl, wherein the alkyl portion may be optionally substituted; xv. aralkylsulfonyl, or arylsulfonyl, wherein the aryl portion 0 of each group can be optionally substituted, xvi. alkenyl or alkynyl; WO 00/47578 PCT/US99/18065 -302 xvii. optionally substituted aryl; xviii. optionally substituted alkyl; xix. optionally substituted aralkyl; xx. optionally substituted heterocycle; or 5 xxi. optionally substituted cycloalkyl; and R 4 , R 5 and R 6 are independently hydrogen, C 1 . 4 alkyl, aryl, hydroxyalkyl, aminoalkyl, monoalkylamino(C 2 - 1 0 )alkyl, dialkylamino(C 2 - 1 0 )alkyl, carboxyalkyl, cyano, amino, alkoxy, or hydroxy.
34. A method according to claim 33, wherein said effective amount 10 is between about 0.01 and about 50 milligrams per kilogram per day.
35. A method according to claim 34, wherein said effective amount is between about 0.1 and about 20 milligrams per kilogram per day.
36. A pharmaceutical composition comprising a compound according to claim 1 or claim 11, or a pharmaceutically acceptable ester, salt or 15 ether thereof, and a pharmaceutically acceptable carrier.
37. A pharmaceutical composition according to claim 36, wherein said compound is present in an amount between 0.01 and 100 milligrams.
38. A method of inhibiting a protease selected from the group consisting of leukocyte neutrophil elastase, chymotrypsin, trypsin, pancreatic 20 elastase, cathepsin G, thrombin, urokinase, factor Xa, plasmin, thermolysin, C-1 esterase, C-3 convertase, acrosin, thrombin, kallikreins, and pepsin, comprising contacting said protease with a compound according to claim 1 or claim 11.
39. A method according to claim 38 wherein said protease is 25 trypsin, chymotrypsin, plasmin or urokinase. WO 00/47578 PCT/US99/18065 -303
40. A method of treating adult respiratory distress syndrome, wound healing, gout, rheumatoid arthritis, reperfusion damage, atherosclerosis, restenosis, neoplasia, metastasis, emphysema, Alzheimer's disease, pancreatitis, benign prostatic hypertrophy, prostatic carcinoma, 5 psoriasis or Parkinson's disease, comprising administering to a patient in need of such treatment an effective amount of a compound according to claim 1 or claim 11.
41. A pharmaceutical composition according to claim 36, suitable for parenteral, oral, subcutaneous, intravenous, intramuscular, intraperitoneal, 10 transdermal, buccal or ocular administration.
42. A process for forming a compound of Formula I 2 R 1 X NR 4 or a solvate, hydrate or pharmaceutically-acceptable salt thereof, wherein: X is oxygen, sulfur or NR'; 5 R 7 is hydrogen, alkyl, aralkyl, hydroxy(C 2 .- 4 )alkyl, or alkoxy(C 2 - 4 )alkyl; Y is a covalent bond, CH 2 or NH; R 1 is a hydrogen, amino, hydroxy, halogen, cyano, C- 1 . 4 alkyl or -CH 2 R where R is hydroxyamino or C,- 3 alkoxy; !0 R 2 and R 3 are independently: i. hydrogen; ii. halogen; iii. hydroxy; iv. nitro; WO 00/47578 PCT/US99/18065 -304 v. cyano; vi. amino, monoalkylamino, dialkylamino, monoarylamino, diarylamino, monoalkylmonoarylamino, monoaralkylamino, diaralkylamino, alkarylamino, alkoxycarbonylamino, aralkoxycarbonylamino, 5 aryloxycarbonylamino, alkylsulfonylamino, aralkylsulfonylamino, arylsulfonylamino, formylamino, acylamino, H(S)CNH-, or thioacylamino; vii. aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl, acyl, arylaminocarbonyl, or aminoacyl; viii. aminothiocarbonyl, monoalkylaminothiocarbonyl, 10 dialkylaminothiocarbonyl, thioacyl, or aminothioacyl; ix. aminocarbonylamino, monoalkylaminocarbonylamino, dialkylaminocarbonylamino, monoarylaminocarbonylamino, diarylaminocarbonylamino, monoaralkylaminocarbonylamino, or diaralkylaminocarbonylamino, 15 x. aminocarbonyloxy, monoalkylaminocarbonyloxy, dialkylaminocarbonyloxy, monoarylaminocarbonyloxy, diarylaminocarbonyloxy, monoaralkylaminocarbonyloxy, or diaralkylaminocarbonyloxy, xi. aminosulfonyl, monoalkylaminosulfonyl, 20 dialkylaminosulfonyl, monoarylaminosulfonyl, diarylaminosulfonyl, or monoaralkylaminosulfonyl, or diaralkylaminosulfonyl, xii. alkoxy or alkylthio, wherein said alkyl portion of said alkoxy or alkylthio group may be optionally substituted; xiii. aralkoxy, aryloxy, aralkylthio, or arylthio, wherein the 25 aryl portion of said aralkoxy, aryloxy, aralkylthio or arylthio group may be optionally substituted; xiv. alkylsulfonyl, wherein the alkyl portion may be optionally substituted; xv. aralkylsulfonyl, or arylsulfonyl, wherein the aryl portion 30 of each group can be optionally substituted, xvi. alkenyl or alkynyl; WO 00/47578 PCT/US99/18065 -305 xvii. optionally substituted aryl; xviii. optionally substituted alkyl; xix. optionally substituted aralkyl; xx. optionally substituted heterocycle; or 5 xxi. optionally substituted cycloalkyl; and R 4 , R 5 and R 6 are independently hydrogen, C1 4 alkyl, aryl, hydroxyalkyl, aminoalkyl, monoalkylamino(C 2 -o 10 )alkyl, dialkylamino(C 2 o 0 )alkyl, carboxyalkyl, cyano, amino, alkoxy, hydroxy or hydrazino; said process comprising: 10 (a) adding a Lewis acid to a suspension of anhydrous ammonium chloride in an aprotic solvent stirred under an inert atmosphere at a temperature near OoC to form a mixture; (b) allowing said mixture to warm to room temperature with stirring and thereafter stirring said mixture until substantially all of the solid 15 has dissolved; (c) adding to said mixture a compound of formula V 2 R' R I R3 V R 8 0 'X r X 0 wherein R'-R 3 , R', X and Y are as defined above; and R' is selected from alkyl and aryl; 20 (d) heating said mixture at reflux for a predetermined period of time and thereafter allowing said mixture to cool to room temperature.
43. A process according to claim 42, wherein said Lewis acid is trimethylaluminum or triethylaluminum.
44. A process according to claim 42, wherein said aprotic solvent is 25 selected from toluene, benzene, xylene, or mesitylene. WO 00/47578 PCT/US99/18065 -306
45. A pharmaceutical composition suitable for oral administration according to claim 41, wherein said compound is present in an amount between 25 milligrams and 100 milligrams.
46. A pharmaceutical composition suitable for parenteral 5 administration according to claim 41, wherein said compound is present in an amount between 0.5 milligrams and 10 milligrams.
47. A compound of claim 1, wherein X is sulfur.
48. A compound of claim 1, wherein Y is a covalent bond.
49. A compound of claim 1, wherein R 1 is hydrogen. 10
50. A compound of claim 1, wherein R 4 , R 5 and R 6 are all hydrogen.
51. A compound of claim 1, wherein R 3 is methylthio or methyl.
52. A compound of claim 1, wherein the aryl portion of R' is optionally substituted with 1 to 4 substituents selected from the group 15 consisting of halogen, C 1 _ 6 alkyl, Ci- 6 alkoxy, hydroxy, nitro, trifluoromethyl, C 6 -1 0 aryl, C 6 -1 0 aryloxy, C6- 1 0 arylmethoxy, C 1 - 6 aminoalkyl, carboxy, 3,4 methylenedioxy, 3,4-ethylenedioxy, 3,4-propylenedioxy, amino, mono(Ci 6 )alkylamino, di(Ci- 6 )alkylamino, mono(C 6 .- 10 o)arylamino, di(C 6 -o 10 )arylamino, C 1 6 alkylsulfonylamino, C 6 - 1 0 arylsulfonylamino, C,_ 8 acylamino, C _ 8 20 alkoxycarbonyl, C,- 6 alkanoylamino, C 6 - 1 4 aroylamino, C. 6 hydroxyalkyl, methylsulfonyl, phenylsulfonyl, thienyl and tetrazole. WO 00/47578 PCT/US99/18065 -307
53. A compound of claim 52, wherein any of said C 6 -1 0 aryl, C 6 - 10 aryloxy, C 6 10 arylmethoxy is further substituted by one or two of chloro, halogen, C, 6 _ alkyl, C,. 6 alkoxy, phenyl, hydroxy, nitro, trifluoromethyl, carboxy, 3,4-methylenedioxy, 3,4-ethylenedioxy, 3,4-propylenedioxy or 5 amino.
54. A compound of claim 52, wherein said optional substituent is thienyl further substituted by one or two of chloro, amino, methyl, methoxy or hydroxy.
55. A compound of claim 1, wherein R 2 is C, 1 6 alkylsulfonylamino, 10 C 6 - 1 0 ar(C- 1 . 6 )alkylsulfonylamino, C 6 -o 10 ar(C 2 - 6 )alkenylsulfonylamino, C 6 - 1 0 arylsullfonylamino, heteroarylsulfonylamino, di(C 6 - 1 0 ar(C 1 . 6 )alkylsulfonyl)amino, di(C 6 - 1 0 (ar(C 2 - 6 )alkenylsulfonyl)amino , di(C 6 , 0 arylsulfonyl)amino or di-(heteroarylsulfonyl)amino, wherein any of the aryl or heteroaryl containing groups is optionally substituted on the aromatic ring. 5
56. A compound of claim 55, wherein R 2 is C 6 - 10 arylsullfonylamino, di(C 6 - 1 0 arylsulfonyl)amino, C 6 - 10 ar(C,_3) alkylsulfonylamino, di(C 6 . 1 0 ar(C 1 - 3 )alkylsulfonyl)amino or thienylsulfonylamino, wherein any of the aryl or thienyl containing groups is further optionally substituted. 0
57. A compound of claim 1, wherein R2 is biphenylsulfonylamino, bis(biphenylsulfonyl)amino, naphth-2-ylsulfonylamino, di(naphth-2 ylsulfonyl)amino, 6-bromonaphth-2-ylsulfonylamino, di(6-bromonaphth-2 ylsulfonyl)amino, naphth-1-ylsulfonylamino, di(naphth-1-ylsulfonyl)amino, 2 methylphenylsulfonylamino, di-(2-methylphenylsulfonyl)amino, 3 5 methylphenylsulfonylamino, di-(3-methylphenylsulfonyl)amino, 4 methylphenylsulfonylamino, di-(4-methylphenylsulfonyl)amino, benzylsulfonylamino, 4-methoxyphenylsulfonylamino, di-(4- WO 00/47578 PCT/US99/18065 -308 methoxyphenylsulfonyl)amino, 4-iodophenylsulfonylamino, di-(4 iodophenylsulfonyl)amino, 3,4-dimethoxyphenylsulfonylamino, bis-(3,4 dimethoxyphenylsulfonyl)amino, 2-chlorophenylsulfonylamino, di-(2 chlorophenylsulfonyl)amino, 3-chlorophenylsulfonylamino, di-(3 5 chlorophenylsulfonyl)amino, 4-chlorophenylsulfonylamino, di-(4 chlorophenylsulfonyl)amino, phenylsulfonylamino, di-(phenylsulfonyl)amino, 4-tert-butylphenylsulfonylamino, di-(4-tert-butylphenylsulfonyl)amino, 2 phenylethenylsulfonylamino or 4-(phenylsulfonyl)thien-2-ylsulfonylamino.
58. A compound of claim 1, wherein R 2 is amino, 0 mono(C 1 . 6 )alkylamino, di(Cl- 6 )alkylamino, mono(C 6 . 1 - 0 io)arylamino, di(C 6 - 1 0 )arylamino, mono(C 6 )alkylmono(C 6 - o)arylamino, monoar(Cl- 6 )alkylamino, di(C 6 - 1 0 )ar(Cl- 6 )alkylamino, mono(Cl- 6 )alkylmono(C 6 . o)ar(CI- 6 )alkylamino, monoheteroarylamino, diheteroarylamino, mono(C, 6 )alkylmonoheteroarylamino, wherein any of the aryl or heteroaryl 5 containing groups is optionally substituted on the aromatic ring.
59. A compound of claim 58, wherein R 2 is mono(C 6 - 1 0 )arylamino, mono(Cl- 6 )alkylmono(C 6 -10)arylamino, mono(C 6 - 1 0 )ar(C - 3 )alkylamino, mono(C 1 - 6 )alkylmono(C 6 -o 10 )ar(Cl- 3 )alkylamino, monoheteroarylamino or mono(C - 6 )alkylmonoheteroarylamino. 0
60. A compound of claim 59, wherein R 2 is 1,3-thiazol-2-ylamino, imidazol-4-ylamino, quinolin-2-ylamino or quinolin-6-ylamino.
61. A compound of claim 58, wherein R 2 is anilino, naphtha-2 ylamino, naphth-1-ylamino, 4-(biphenyl)thiazol-2-ylamino, 4-(phenyl)thiazol 2-ylamino, 4-phenyl-5-methylthiazol-2-ylamino, 4-hydroxy-4 5 trifluoromethylthiazol-2-ylamino, 3-phenylphenylamino, pyrimidin-2-ylamino, 4-isopropylphenylamino, 3-isopropylphenylamino, 4-phenylphenylamino, 3 fluoro-4-phenylphenylamino, 3,4-methylenedioxyphenylamino, n- WO 00/47578 PCT/US99/18065 -309 butylphenylamino, N-methyl-N-(2-methylphenyl)amino, 3-nitrophenylamino, 4-methoxyphenylamino, 3-methoxyphenylamino, 2-methoxyphenylamino, 2 methylphenylamino, 3-methylphenylamino, 3,4-dimethylphenylamino, 3 chlorophenylamino, 4-chlorophenylamino, 4-(3-fluoro-4-methylphenyl)amino, 5 4-(indan-5-yl)amino, benzylamino, indanylmethylamino, 2,3 dihydrobenzofuranylmethylamino, 2-phenylimidazol-5-ylamino, 3 hydroxybenzylamino, 3-phenoxyphenylamino, 4-phenoxyphenylamino, 3-benzyloxyphenylamino, 4-benzyloxyphenylamino, quinolin-6-ylamino, quinolin-3-ylamino, 4-(phenylamino)phenylamino, 4-(4 10 ethylphenyl)phenylamino, 4-(dimethylamino)phenylamino, 4-cyclohexylphenylamino, 4-(9-ethylcarbazol-3-yl)amino, 4-(t butyl)phenylamino or 4-methylthiophenylamino.
62. A compound of claim 1, wherein R 2 is alkanoylamino, alkenoylamino, aroylamino, aralkanoylamino, aralkenoylamino, 15 heteroaroylamino, or heteroarylalkanoylamino, any of which is optionally substituted on the aromatic ring.
63. A compound of claim 62, wherein R 2 is (C 6 - 0 o)arylcarbonylamino, C 6 - 0 oar(CI- 3 )alkylcarbonylamino, C 6 l 0 ar(C 2 - 3 )alkenylcarbonylamino, C 6 -1 0 aryloxy(C, 3 )alkylcarbonylamino, C 3 . 20 8 cycloalkylcarbonylamino, Cl 6 alkylcarbonylamino or heteroarylcarbonylamino.
64. A compound of claim 63, wherein R 2 is furanylcarbonylamino, and quinolinylcarbonylamino.
65. A compound of claim 63, wherein R 2 is 25 3-hydroxyphenylcarbonylamino, 2-phenylethenylcarbonylamino, phenylcarbonylamino, cyclohexylcarbonylamino, 4-methyl-3 nitrophenylcarbonylamino, furan-2-ylcarbonylamino, tert-butylcarbonylamino, WO 00/47578 PCT/US99/18065 -310 5-(3,5-dichlorophenoxy)furan-2-ylcarbonylamino, naphth-1-ylcarbonylamino, quinolin-2-ylcarbonylamino. 4-ethoxyphenylcarbonylamino, phenoxymethylcarbonylamino or 3-methylphenylcarbonylamino.
66. A compound of claim 1, wherein R 2 is C 6 - 1 0 aryloxy, 5 C 6 - 10 ar(Cl- 6 )alkoxy, C 6 - 1 0 arylsulfonyl, C 6 - 10 ar(Cl- 6 )alkylsulfonyl or C 6 - 10 ar(C 2 - 6 )alkenylsulfonyl, any of which is optionally substituted on the aromatic ring.
67. A compound of claim 66, wherein R 2 is C 6 - 1 0 aryloxy or C 6 - 1 0 arylsulfonyl. 10
68. A compound of claim 67, wherein R 2 is phenoxy, naphthyloxy, phenylsulfonyl or naphthylsulfonyl.
69. A compound of claim 1, which is one of: 5-methylthio-4-(6-quinolylamino)thiophene-2-carboxamidine; 15 5-methylthio-4-[(3-phenylphenyl)amino]thiophene-2-carboxamidine; 5-methylthio-4-(3-quinolylamino)thiophene-2-carboxamidine; 5-methylthio-4-(pyrimidin-2-ylamino)thiophene-2-carboxamidine; 4-[(4-cyclohexylphenyl)amino]-5-methylthiothiophene-2-carboxamidine; methyl 4-amino-5-methylthiothiophene-2-carboxylate; 20 methyl 4-[(aminothioxomethyl)amino]-5-methylthiothiophene-2-carboxylate; 5-methylthio-4-[(4-phenyl(1,3-thiazol-2-yl))amino]thiophene-2 carboxamidine; 5-methylthio-4- { [4-(4-phenylphenyl)( 1,3-thiazol-2-yl)]amino } thiophene-2 carboxamidine; 25 4-[(5-methyl-4-phenyl(1,3-thiazol-2-yl))amino]-5-methylthiothiophene-2 carboxamidine; 4- { [4-hydroxy-4-(trifluoromethyl)(1,3-thiazolin-2-yl)]amino}-5 methylthiothiophene-2-carboxamidine; WO 00/47578 PCT/US99/1 8065 -311 5 -methylthio-4-(2-naphthylamino)thiophene-2-carboxamidine; 4- [(4-chlorophenyl)amino]l-5 -methylthiothiophene-2-carboxamidine; 4-[(3-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 4- [(3 -methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 5 4- { [3 -(methylethyl)phenyl] amino)} -5 -methylthiothiophene-2-carboxamidine; 5-methylthio-4- [(3 -nitrophenyl)amino]thiophene-2-carboxamidine; 4- {[4-(methylethyl)phenyl] amino)}-5 -methylthiothiophene-2-carboxamidine; 4-[(3 ,4-dimethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 5-methylthio-4- [(4-phenylphenyl)amino]thiophene-2-carboxamidine; 0 4-[(3-fluoro-4-phenylphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 4-(2H-benzo [d] 1,3 -dioxolen-5-ylamino)-5-methylthiothiophene-2 carboxamidine; 4- [(4-butylphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 5 -methylthio-4- [benzylamino]thiophene-2-carboxamidine; 5 4-(indan-5-ylamino)-5-methylthiothiophene-2-carboxamidine; 4-(2,3 -dihydrobenzo [b]furan-5-ylamino)-5-methylthiothiophene-2 carboxamidine; 5-methylthio-4- [(2-phenylimidazol-4-yl)amino]thiophene-2-carboxamidine; 5-methylthio-4- [(2-quinolylmethyl)amino]thiophene-2-carboxamidine; 0 4-f{ [(3 -hydroxyphenyl)methyl] amino } -5-methylthiothiophene-2 carboxamidine; 5-methylthio-4-(phenylcarbonylamino)thiophene-2-carboxamiline; 4 -(( 2 E)-3-phenylprop-2-enoylamino)-5-methylthiothiophene-2.carboxamidine; 4 -[( 4 -chlorophenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine; 5 4 -(cyclohexylcarbonylamino)-5-methylthiothiophene-2-carboxamidine; methyl 4- [(4-methyl-3 -nitrophenyl)carbonylamino]-5 -methylthiothiophene-2 carboxylate; 4-(2-furylcarbonylamino)-5-methylthiothiophene-2-carboxamidine; 4 -(2,2-dimethylpropanoylamino)-5-methylthiothiophene-2-carboxamidine; 0 4-({[5-(3 ,5-dichlorophenoxy)(2-furyl)]carbonylamino }-5 -methyithiothiophene 2-carboxamidine; WO 00/47578 PCT/US99/18065 -312 5-methylthio-4-(naphthylcarbonylamino)-thiophene-2-carboxamidine; 5-methylthio-4-(2-quinolylcarbonyl-amino)thiophene-2-carboxamidine; 4-[(3-methoxyphenyl)carbonylamino]-5-methylthiothiophene-2 carboxamidine; 5 4-[2-(2-hydroxy-5-methoxyphenyl)acetylamino]-5-methylthiothiophene-2 carboxamidine; 4-[(4-ethoxyphenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine; 5-methylthio-4-(2-phenoxyacetylamino)-thiophene-2-carboxamidine; 4-[(3-methylphenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine; 10 5-methylthio-4- { [3-(phenylmethoxy)phenyl]amino}thiophene-2 carboxamidine; 5-methylthio-4-[(3-phenoxyphenyl)amino]thiophene-2-carboxamidine; 5-methylthio-4-[(4-phenoxyphenyl)amino]thiophene-2-carboxamidine; 4- [(2-methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 15 4-[(2-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 4-[(3-chlorophenyl)amino]-5-methylthiothiophene-2-carboxamidine; 4-(methylphenylamino)-5-methylthiothiophene-2-carboxamidine; 5-methyl-4-(phenylamino)thiophene-2-carboxamidine; 4- { [4-(dimethylamino)phenyl]amino} -5-methylthiothiophene-2 z0 carboxamidine; 4- [(4-ethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 5-methylthio-4- { [4-(phenylmethoxy)phenyl]amino}thiophene-2 carboxamidine; 5 -methylthio-4- { [4-(phenylamino)phenyl]amino } thiophene-2-carboxamidine; 15 4- [(4-methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 4- [(3-fluoro-4-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine; 4-(indan-5-ylamino)-5-methylthiothiophene-2-carboxamidine; 4- [(9-ethylcarbazol-3-yl)amino]-5-methylthiothiophene-2-carboxamidine; 5-methylthio-4- { [(4-phenylphenyl)sulfonyl]amino}thiophene-2 carboxamidine; WO 00/47578 PCT/US99/18065 -313 4- {bis[(4-phenylphenyl)sulfonyl]amino } -5-methylthiothiophene-2 carboxamidine; 5-methylthio-4-[(2-naphthylsulfonyl)-amino]thiophene-2-carboxamidine; 4-[bis(2-naphthylsulfonyl)amino]-5-methylthiothiophene-2-carboxamidine; 5 4- { [(6-bromo(2-naphthyl))sulfonyl]amino}-5-methylthiothiophene-2 carboxamidine; 4- {bis [(6-bromo(2-naphthyl))sulfonyl]amino }-5-methylthiothiophene-2 carboxamidine; 5-methylthio-4-[(naphthylsulfonyl)-amino]thiophene-2-carboxamidine; 10 4-[bis(naphthylsulfonyl)amino]-5-methylthiothiophene-2-carboxamidine; 4- { [(2-methylphenyl)sulfonyl]amino} -5-methylthiothiophene-2 carboxamidine; 4- {bis[(2-methylphenyl)sulfonyl]amino } -5-methylthiothiophene-2 carboxamidine; 15 4- { [(3 -methylphenyl)sulfonyl]amino} -5 -methylthiothiophene-2 carboxamidine; 4- {bis [(3 -methylphenyl)sulfonyl] amino } -5-methylthiothiophene-2 carboxamidine; 4- { [(4-methylphenyl)sulfonyl]amino} -5-methylthiothiophene-2 20 carboxamidine; 4- {bis [(4-methylphenyl)sulfonyl]amino } -5-methylthiothiophene-2 carboxamidine; 5-methylthio-4- { [benzylsulfonyl]amino } -thiophene-2-carboxamidine; 5-methylthio-4-phenoxythiophene-2-carboxamidine; or 25 5-methylthio-4-(phenylsulfonyl)thiophene-2-carboxamidine; or a salt, solvate, hydrate or prodrug thereof.
70. A compound of Formula I: WO 00/47578 PCT/US99/18065 -314 2 R ZY X NR 4 or a solvate, hydrate or pharmaceutically-acceptable salt thereof, wherein: X is oxygen or sulfur; Y is a covalent bond or NH; 5 Z is NR'R 6 ; R 1 is a hydrogen, amino, hydroxy or halogen; R 2 is alkylsulfonylamino, aralkylsulfonylamino, aralkenylsulfonylamino, arylsulfonylamino, LO heteroarylsulfonylamino, di(aralkylsulfonyl)amino, di(aralkenylsulfonyl)amino, di(arylsulfonyl)amino, or di(heteroarylsulfonyl)amino, wherein any of the aryl or heteroaryl containing groups are optionally substituted on the aromatic ring; or 5 amino, monoalkylamino, dialkylamino, monoarylamino, diarylamino, monoalkylmonoarylamino, monoaralkylamino, diaralkylamino, monoalkylmonoaralkylamino, monoheterocycleamino, diheterocycleamino, monoalkylmonoheterocycleamino, .0 wherein any of the aryl or heterocycle containing groups is optionally substituted; or alkanoylamino, alkenoylamino, alkynoylamino, aroylamino, aralkanoylamino, aralkenoylamino, heteroaroylamino, 5 heteroarylalkanoylamino, any of which is optionally substituted on the aromatic ring; or alkoxy, alkylthio, aryloxy, aralkoxy, arylthio, WO 00/47578 PCT/US99/18065 -315 aralkylthio, arylsulfonyl, aralkylsulfonyl, aralkenylsulfonyl, any of which is optionally substituted; or alkoxycarbonylamino, aralkoxycarbonylamino, 5 aryloxycarbonylamino, wherein any of the aryl containing groups is optionally substituted on the aromatic ring; or formylamino, H(S)CNH-, or thioacylamino; R 3 is hydrogen, C 1 _ 6 alkyl, C- 6 alkyl optionally substituted with 0 OH, NH 2 , COOH, aminocarbonyl, or C 1 - 6 alkoxy.
71. A pharmaceutical composition comprising a compound according to claim 70, or a pharmaceutically acceptable ester, salt or ether thereof, and a pharmaceutically acceptable carrier.
72. A pharmaceutical composition according to claim 71, wherein .5 said compound is present in an amount between 0.01 and 100 milligrams.
73. A method of inhibiting a protease selected from the group consisting of leukocyte neutrophil elastase, chymotrypsin, trypsin, pancreatic elastase, cathepsin G, thrombin, urokinase, factor Xa, plasmin, thermolysin, C-1 esterase, C-3 convertase, acrosin, thrombin, kallikreins, and pepsin, 0 comprising contacting said protease with a compound according to 70.
74. A method according to claim 73 wherein said protease is trypsin, chymotrypsin, plasmin or urokinase.
75. A method of treating adult respiratory distress syndrome, wound healing, gout, rheumatoid arthritis, reperfusion damage, Z5 atherosclerosis, restenosis, neoplasia, metastasis, emphysema, Alzheimer's disease, pancreatitis, benign prostatic hypertrophy, prostatic carcinoma, WO 00/47578 PCT/US99/18065 -316 psoriasis or Parkinson's disease, comprising administering to a patient in need of such treatment an effective amount of a compound according to claim 70.
76. A compound of claim 70, which is one of: 5-methylthio-4-(6-quinolylamino)thiophene-2-carboxamidine 5 5-methylthio-4-[(3-phenylphenyl)amino]thiophene-2-carboxamidine 5-methylthio-4-(3-quinolylamino)thiophene-2-carboxamidine 5-methylthio-4-(pyrimidin-2-ylamino)thiophene-2-carboxamidine 4-[(4-cyclohexylphenyl)amino]-5-methylthiothiophene-2-carboxamidine methyl 4-amino-5-methylthiothiophene-2-carboxylate 10 methyl 4-[(aminothioxomethyl)amino]-5-methylthiothiophene-2-carboxylate 5-methylthio-4-[(4-phenyl(1,3-thiazol-2-yl))amino]thiophene-2-carboxamidine 5-methylthio-4- { [4-(4-phenylphenyl)(1,3 -thiazol-2-yl)]amino } thiophene-2 carboxamidine 4- [(5-methyl-4-phenyl(1,3-thiazol-2-yl))amino]-5-methylthiothiophene-2 15 carboxamidine 4- { [4-hydroxy-4-(trifluoromethyl)(1,3-thiazolin-2-yl)]amino}-5 methylthiothiophene-2-carboxamidine 5-methylthio-4-(2-naphthylamino)thiophene-2-carboxamidine 4-[(4-chlorophenyl)amino]-5-methylthiothiophene-2-carboxamidine 20 4-[(3-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4-[(3-methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4- { [3-(methylethyl)phenyl]amino }-5-methylthiothiophene-2-carboxamidine 5-methylthio-4-[(3-nitrophenyl)amino]thiophene-2-carboxamidine 4- { [4-(methylethyl)phenyl]amino } -5-methylthiothiophene-2-carboxamidine 25 4-[(3,4-dimethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 5-methylthio-4-[(4-phenylphenyl)amino]thiophene-2-carboxamidine 4-[(3-fluoro-4-phenylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4-(2H-benzo[d] 1,3-dioxolen-5-ylamino)-5-methylthiothiophene-2 carboxamidine 30 4 -[( 4 -butylphenyl)amino]-5-methylthiothiophene-2-carboxamidine WO 00/47578 PCT/US99/18065 -317 5-methylthio-4- [benzylamino]thiophene-2-carboxamidine 4-(indan-5-ylamino)-5 -methylthiothiophene-2-carboxamidine 4-(2,3 -dihydrobenzo [b]furan-5 -ylamino)-5-methylthiothiophene-2 carboxamidine 5 5-methylthio- 4 -[(2-phenylimidazol-4-yl)amino]thiophene2carboxamidine 5-methylthio-4- [( 2 -quinolylmethyl)amino]thiophene-2-carboxamidine 4-f{ [(3 -hydroxyphenyl)methyl] amino 1 -5 -methylthiothiophene-2 carboxamidine 5-methylthio-4-(phenylcarbonylamino)thiophene-2-carboxamidine 10 4 -(( 2 E)- 3 -phenylprop- 2 -enoylamino)-5-methylthiothiophene2carboxamidine 4 -[( 4 -chlorophenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine 4-(cyclohexylcarbonylamino)-5 -methylthiothiophene-2-carboxamidine methyl 4-[(4-methyl-3 -nitrophenyl)carbonylamino]-5-methylthiothiophene-2. carboxylate 15 4 -( 2 -furylcarbonylamino)-5-methylthiothiophene-2carboxamidine 4 -( 2 , 2 -dimethylpropanoylamino)-5-methylthiothiophene-2.carboxamidine 4- {[5 -(3 ,5-dichlorophenoxy)(2-fiiryl)]carbonylamino }-5 -methylthiothiophene 2-carboxamidine 5 -methylthio-4-(naphthylcarbonylamnino)-thiophene-2-carboxamidine W 5 -methylthio- 4 -(2-quinolylcarbonyl-amino)thiophene-2-carboxarnidine 4- [( 3 -methoxyphenyl)carbonylamin o]-5 -methylthiothiophene-2 carboxamidine 4 -[ 2 -( 2 -hydroxy-5-methoxyphenyl)acetylamino]-5-methylthiothiophene-2 carboxamidine :5 4 -[( 4 -ethoxyphenyl)carbonylamino]-5-methylthiothiophene-2-carboxamidine 5-methylthio-4-(2-phenoxyacetylamino)-thiophene-.2carboxamidine 4-[(3 -methylphenyl)carbonylamino]-5 -methylthiothiophene-2-carboxamidine 5 -methylthio-4-1{[3 -(phenylmethoxy)phenyl] amino)} thiophene-2 carboxamidine 0 5-methylthio-4-[(3-phenoxyphenyl)amino]thiophene-2-carboxamidine 5-methylthio-4-[(4-phenoxyphenyl)amino]thiophene2carboxamidine WO 00/47578 PCT/US99/1 8065 -318 4-[(2-methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4- [(2-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4- [(3 -chlorophenyl)amino]-5-methylthiothiophene-2-carboxamidine 4-(methylphenylamino)-5-methylthiothiophene-2-carboxamidine 5 5 -methyl-4-(phenylamino)thiophene-2-carboxamidine 4- { [4-(dimethylamino)phenyl] amino)} -5 -methylthiothiophene-2 carboxamidine 4-[(4-ethylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 5 -methylthio-4- { [4-(phenylmethoxy)phenyl]amino }thiophene-2 10 carboxamidine 5-methylthio-4- {[4-(phenylamino)phenyl] amino }thiophene-2-carboxamidine 4-[(4-methoxyphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4- [(3 -fluoro-4-methylphenyl)amino]-5-methylthiothiophene-2-carboxamidine 4-(indan-5-ylamino)-5-methylthiothiophene-2-carboxamidine 15 4-[(9-ethylcarbazol-3-yl)amino]-5-methylthiothiophene-2-carboxamidine 5-methylthio-4- {[(4-phenylphenyl)sulfonyl]aminol}thiophene-2-carboxamidine 4- f{bis [(4-phenylphenyl)sulfonyl] amino } -5 -methylthiothiophene-2 carboxamidine 5 -methylthio-4- [(2-naphthylsulfonyl)-amino]thiophene-2-carboxamidine !0 4- [bis(2-naphthylsulfonyl)amino]-5-methylthiothiophene-2-carboxamidine 4- { [(6-bromo(2-naphthyl))sulfonyl]amino } -5-methylthiothiophene-2 carboxamidine 4-f{ bis [(6-bromo(2-naphthyl))sulfonyl]amino }-5-methylthiothiophene-2 carboxamidine 5 5-methylthio-4-[(naphthylsulfonyl)-amino]thiophene-2-carboxamidine 4- [bis(naphthylsulfonyl)amino] -5-methylthiothiophene-2-carboxamidine 4-f{ [(2-methylphenyl)sulfonyl] amino)} -5 -methylthiothiophene-2 carboxamidine 4- f bis [(2-methylphenyl)sulfonyl]amino I -5-methylthiothiophene-2 carboxamidine WO 00/47578 PCT/US99/18065 -319 4- { [(3 -methylphenyl)sulfonyl]amino} -5-methylthiothiophene-2 carboxamidine 4- { bis[(3-methylphenyl)sulfonyl]amino } -5-methylthiothiophene-2 carboxamidine 5 4- { [(4-methylphenyl)sulfonyl]amino } -5-methylthiothiophene-2 carboxamidine 4- {bis[(4-methylphenyl)sulfonyl]amino } -5-methylthiothiophene-2 carboxamidine 5-methylthio-4- { [benzylsulfonyl]amino } -thiophene-2-carboxamidine L0 5-methylthio-4-phenoxythiophene-2-carboxamidine 5-methy1thio-4-(phenylsulfonyl)thiophene-2-carboxamidine or salts or prodrugs thereof
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Families Citing this family (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003529536A (en) * 1999-02-09 2003-10-07 ３−ディメンショナル ファーマシューティカルズ， インコーポレイテッド Methods of treating C1s-mediated diseases and conditions, and compounds and compositions therefor
CA2484822A1 (en) * 2002-05-28 2003-12-04 3-Dimensional Pharmaceuticals, Inc. Novel thiophene amidines, compositions thereof, and methods of treating complement-mediated diseases and conditions
DK1569912T3 (en) 2002-12-03 2015-06-29 Pharmacyclics Inc 2- (2-hydroxybiphenyl-3-yl) -1h-benzoimidazole-5-carboxamidine derivatives as factor VIIa inhibitors.
US7482376B2 (en) * 2003-07-03 2009-01-27 3-Dimensional Pharmaceuticals, Inc. Conjugated complement cascade inhibitors
JP2007523868A (en) * 2003-07-10 2007-08-23 アキリオン ファーマシューティカルズ，インコーポレーテッド Substituted arylthiourea derivatives useful as inhibitors of viral replication
US8685369B2 (en) 2004-02-12 2014-04-01 Molecular Insight Pharmaceuticals, Inc. Technetium- and rhenium-bis (heteroaryl) complexes, and methods of use thereof
EP2344453B1 (en) * 2008-10-09 2016-12-28 The U.S.A. As Represented By The Secretary, Department Of Health And Human Services Activators of human pyruvate kinase
EP2451796B1 (en) 2009-07-08 2013-04-17 Dermira (Canada), Inc. Tofa analogs useful in treating dermatological disorders or conditions
US20130109672A1 (en) 2010-04-29 2013-05-02 The United States Of America,As Represented By The Secretary, Department Of Health And Human Service Activators of human pyruvate kinase
CN103562202B (en) * 2011-01-25 2016-09-14 密执安大学评议会 BCL-2/BCL-XL inhibitor and their Therapeutic Method of use
AR086744A1 (en) * 2011-06-28 2014-01-22 Nippon Soda Co HETEROCICLIC COMPOUND CONTAINING NITROGEN AND FUNGICIDE FOR USE IN AGRICULTURE AND GARDENING
EP2606726A1 (en) * 2011-12-21 2013-06-26 Bayer CropScience AG N-Arylamidine-substituted trifluoroethylsulfide derivatives as acaricides and insecticides
TWI644899B (en) 2013-02-04 2018-12-21 健生藥品公司 Flap modulators
JP6424173B2 (en) * 2013-02-04 2018-11-14 ヤンセン ファーマシューティカ エヌ．ベー． FLAP regulator
CN111918865B (en) * 2018-03-30 2023-10-27 住友化学株式会社 Heterocyclic compound and harmful arthropod control composition containing same
CN109020837A (en) * 2018-07-27 2018-12-18 广东省石油与精细化工研究院 A kind of preparation method of 2- substituted-phenyl-B amidine hydrochloric acid salt

Family Cites Families (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4220654A (en) * 1979-06-04 1980-09-02 Merck & Co., Inc. Cyclic imidazole cyanoguanidines
US4424367A (en) * 1982-08-19 1984-01-03 Norwich Eaton Pharmaceuticals, Inc. 5-(4-Aminophenyl)-2-thiophenecarboximidamide hydrochloride hemihydrate
JPS59139357A (en) * 1983-01-28 1984-08-10 Torii Yakuhin Kk Amidine derivative
DE3427865A1 (en) * 1984-07-27 1986-02-06 Torii & Co., Tokio/Tokyo Amidino compounds, process for their preparation and pharmaceutical compositions containing these compounds
US5084466A (en) * 1989-01-31 1992-01-28 Hoffmann-La Roche Inc. Novel carboxamide pyridine compounds which have useful pharmaceutical utility
US5340833A (en) * 1992-05-01 1994-08-23 Eisai Co., Ltd. Urokinase inhibitors
GB9312761D0 (en) * 1993-06-21 1993-08-04 Wellcome Found Amino acid derivatives
PT724435E (en) * 1993-10-21 2002-11-29 Searle & Co UTEIS AMIDINE DERIVATIVES AS NITRIC OXIDE INHIBITORS
ZA951617B (en) * 1994-03-04 1997-02-27 Lilly Co Eli Antithrombotic agents.
US5612353A (en) * 1995-06-07 1997-03-18 Rhone-Poulenc Rorer Pharmaceuticals Inc. Substituted (sulfinic acid, sulfonic acid, sulfonylamino or sulfinylamino) N-[(aminoiminomethyl)phenylalkyl]-azaheterocyclylamide compounds
TW414795B (en) * 1996-07-01 2000-12-11 Yamanouchi Pharma Co Ltd A thiophene derivative and the pharmaceutical composition
DE19632773A1 (en) * 1996-08-14 1998-02-19 Basf Ag New thrombin inhibitors
PE121699A1 (en) * 1997-02-18 1999-12-08 Boehringer Ingelheim Pharma BICYCLE HETERO CYCLES DISSTITUTED AS INHIBITORS OF THROMBIN
WO1999037611A1 (en) * 1998-01-26 1999-07-29 Basf Aktiengesellschaft Heterocyclic amidines as callicrein protease inhibitors
CA2317761A1 (en) * 1998-01-26 1999-07-29 Basf Aktiengesellschaft Thrombin inhibitors
NZ506507A (en) * 1998-02-09 2003-08-29 Dimensional Pharm Inc Heteroaryl amidine, methylamidine or guanidine derivatives useful as urokinase inhibitors
AU1917999A (en) * 1998-03-31 1999-10-18 Warner-Lambert Company Quinoxalinones as serine protease inhibitors such as factor xa and thrombin
KR20010042296A (en) * 1998-03-31 2001-05-25 로즈 암스트롱, 크리스틴 에이. 트러트웨인 Benzoxazinones/Benzothiazinones as Serine Protease Inhibitors
JP2002509928A (en) * 1998-03-31 2002-04-02 ワーナー−ランバート・カンパニー Quinolones as serine protease inhibitors

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