AU726595B2 - Biphenylsulfonamides and derivatives thereof that modulate the activity of endothelin - Google Patents

Description

S F Ref: 392218D1

AUSTRALIA

PATENTS ACT 1990 COMPLETE SPECIFCATION FOR A STANDARD PATENT

ORIGINAL

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Name and Address of Applicant: Texas Biotechnology Corporation Suite 1920 7000 Fannin Street Houston Texas 77030 UNITED STATES OF AMERICA 9* 9 See.

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Actual Inventor(s): Address for Service: Invention Title: Ming Fai Chan, Bore Gowda Raju, Adam Kois, Erik Joel Verner, Chengde Wu, Rosario Silverstre Castillo, Venkatachalapathi Yalamoori, Vltukudi Narayanaiyengar Balaji and Timothy Kogan Spruson Ferguson, Patent Attorneys Level 33 St Martins Tower, 31 Market Street Sydney, New South Wales, 2000, Australia Biphenylsulfonamides and Derivatives Thereof that Modulate the Activity of Endothelin The following statement is a full description of this invention, including the best method of performing it known to me/us:- 5845 1 Biphenylsulfonamides and Derivatives Thereof That Modulate the Activity of Endothelin Related Applications The subject matter of each US patent numbers 5,594,021, 5,571,821, 5,591,761, 5,464,853 and 5,514,691 is incorporated herein in its entirety.

These documents are the issued US patents resulting from US application Serial No. 08/447,223, 08/247,072, 08/222,287 08/142,159 and 08/142,552, respectively.

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(The next page is page 4) [R:\LIBA]02587.doc:TLT FIELD OF THE INVENTION The present invention relates to the compounds that modulate the activity of the endothelin family of peptides. In particular, the invention relates to the use of sulfonamides and sulfonamide pro-drugs as endothelin agonisis S and antagonists.

BACKGROUND OF THE INVENTION The vascular endothelium releases a variety of vasoactive substances, .including the endothelium-derived vasoconstrictor peptide, endothelin (ET) (see, e.g, Vanhoutte et al. (1986) Annual Rev. Physiol. 48: 307-320; Furchgott and Zawadski (1980) Nature 288: 373-376). Endothelin, which was originally 15 identified in the culture supernatant of porcine aortic endothelial cells (see, Yanagisawa et al. (1988) Nature 332: 411-415), is a potent twenty-one amino acid peptide vasoconstrictor. It is the most potent vasopressor known and is produced by numerous cell types, including the cells of the endothelium, trachea, kidney and brain. Endothelin is synthesized as a two hundred and-three 20 amino acid precursor preproendothelin that contains a signal sequence which is cleaved by an endogenous protease to produce a thirty-eight (human) or thirtynine (porcine) amino acid peptide. This intermediate, referred to as big endothelin, is processed in vivo to the mature biologically active form by a putative endothelin-converting enzyme (ECE) that appears to be a metaldependent neutral protease (see, Kashiwabara et al. (1989) FEBS Lttrs.

247: 337-340). Cleavage is required for induction of physiological responses (see, von Geldern et a. (1991) Peptide Res. 4: 32-35). In porcine aortic endothelial cells, the thirty-nine amino acid intermediate, big endothelin, is hydrolyzed at the Trp2'-Val 2 2 bond to generate endothelin-1 and a C-terminal fragment. A similar cleavage occurs in human cells from a thirty-eight amino acid intermediate. Three distinct endothelin isopeptides, endothelin-1, endothelin-2 and endothelin-3, that exhibit potent vasoconstrictor activity have been identified.

The family of three isopeptides endothelin-1, endothelin-2 and endothelin- 3 are encoded by a family of three genes (see, Inoue et al. (1989) Proc. Natl.

Acad. Sci. USA 86: 2863-2867; see, also Saida et al (1989)J. Biol. Chem.

264: 14613-14616). The nucleotide sequences of the three human genes are highly conserved within the region encoding the mature 21 amino acid peptides and the C-terminal portions of the peptides are identical. Endothelin-2 is (Trp 6 ,Leu 7 endothelin-1 and endothelin-3 is (Thr 2 ,Phe 4 ,Thrs,Tyr6,Lys',Tyr4) 10 endothelin-1. These peptides are, thus, highly conserved at the C-terminal ends.

Release of endothelins from cultured endothelial cells is modulated by a variety of chemical and physical stimuli and appears to be regulated at the level of transcription and/or translation. Expression of the gene encoding endothelin-1 is increased by chemical stimuli, including adrenaline, thrombin and Ca 2 15 ionophore. The production and release of endothelin from the endothelium is stimulated by angiotensin II, vasopressin, endotoxin, cyclosporine and other factors (see, Brooks et al. (1991) Eur. J. Pharm. 194:115-117), and is inhibited by nitric oxide. Endothelial cells appear to secrete short-lived endotheliumderived relaxing factors (EDRF), including nitric oxide or a related substance 20 (Palmer t al. (1987) Nature 327: 524-526), when stimulated by vasoactive agents, such as acetylcholine and bradykinin. Endothelin-induced vasoconstriction is also attenuated by atrial natriuretic peptide (ANP).

The endothelin peptides exhibit numerous biological activities in vitro and in vivo. Endothelin provokes a strong and sustained vasoconstriction in vivo in rats and in isolated vascular smooth muscle preparations; it also provokes the release of eicosanoids and endothelium-derived relaxing factor (EDRF) from perfused vascular beds. Intravenous administration of endothelin-1 and in vitro addition to vascular and other smooth muscle tissues produce long-lasting pressor effects and contraction, respectively (see, Bolger et a. (1991) Can.

J. Physiol. Pharmacol. 69: 406-413). In isolated vascular strips, for example, endothelin-1 is a potent (ECso 4 x 1010 slow acting, but persistent, contractile agent. In vivo, a single dose elevates blood pressure in about twenty to thirty minutes. Endothelin-induced vasoconstriction is not affected by antagonists to known neurotransmitters or hormonal factors, but is abolished by calcium channel antagonists. The effect of calcium channel antagonists however, is most likely the result of inhibition of calcium influx, since calcium influx appears to be required for the long-lasting contractile response to endothelin.

Endothelin also mediates renin release, stimulates ANP release and induces a positive inotropic action in guinea pig atria. In the lung, endothelin-1 acts as a potent bronchoconstrictor (Maggi al. (1989) Eur. J. Pharmacol 160: 179-182). Endothelin increases renal vascular resistance, decreases renal blood flow, and decreases glomerular filtrate rate. It is a potent mitogen for glomerular mesangial cells and invokes the phosphoinoside cascade in such cells (Simonson al. (1990) J. lin. Invest. 85: 790-797).

rn There are specific high affinity binding sites (dissociation constants in the 15 range of 2-6 x 10-0 M) for the endothelins in the vascular system and in other tissues, including the intestine, heart, lungs, kidneys, spleen, adrenal glands and brain. Binding is not inhibited by catecholamines, vasoactive peptides, n neurotoxins or calcium channel antagonists. Endothelin binds and interacts with receptor sites that are distinct from other autonomic receptors and voltage 20 dependent calcium channels. Competitive binding studies indicate that there are multiple classes of receptors with different affinities for the endothelin isopeptides. The sarafotoxins, a group of peptide toxins from the venom of the snake Ajctais en si that cause severe coronary vasospasm in snake bite victims, have tructural and functional homology to endothelin-1 and bind competitively to the same cardiac membrane receptors (Kloog t al. (1989) Trends Pharmacol Sci. L: 212-214).

Two distinct endothelin receptors, designated ETA and ET, have been identified and DNA clones encoding each receptor have been isolated (Arai et al (1990) Nature 348: 730-732; Sakurai et al. (1990) Nature 3le8: 732-735).

Based on the amino acid sequences of the proteins encoded by the cloned DNA, it appears that each receptor contains seven membrane spanning domains and exhibits structural similarity to G-protein-coupled membrane proteins.

Messenger RNA encoding both receptors has been detected in a variety of tissues, including heart, lung, kidney and brain. The distribution of receptor subtypes is tissue specific (Martin et l. (1989) Bichem

B

Corminun_ 162: 130-137).

ET-

Commun. 162: 130-137). ETA receptors appear to be selective for endothelin-l and are predominant in cardiovascular tissues.

ET

8 receptors are predominant in noncardiovascular tissues, including the central nervous system and kidney, and interact with the three endothelin isopeptides (Sakurai et al. (1990) Nature 348: 732-734). In addition, ET receptors occur on vascular smooth muscle, are linked to vasoconstriction and have been associated with cardiovascular, renal and central nervous system diseases; whereas ET, receptors are located on the vascular endothelium, linked to vasodilation Takayanag 1991) FE Lttrs. 2: 103-106) and have been associated with bronchoconstrictive disorders.

By virtue of the distribution of receptor types and the differential affinity *5 of each isopeptide for each receptor type, the activity of the endothelin isopeptides varies in different tissues. For example, endothelin-1 inhibits 251s labelled endothelin-1 binding in cardiovascular tissues forty to seven hundred times more potently than endothelin-3. 1 25 1-labelled endothelin-1 binding in noncardiovascular tissues, such as kidney, adrenal gland, and cerebellum, is i20 inhibited to the same extent by endothelinl and endothelin-3, which indicates that ETA receptors predominate in cardiovascular tissues and ETB receptors predominate in non-cardiovascular tissues.

Endothelin plasma levels are elevated in certain disease states (see, e., International PCT Application WO 94/27979, and U.S. Patent No. 5,382,569, which disclosures are herein incorporated in their entirety by reference). Endothelin-1 plasma levels in healthy individuals, as measured by radioimmunoassay (RIA), are about 0.26-5 pg/ml. Blood levels of endothelin- and its precursor, big endothelin, are elevated in shock, myocardial infarction, vasospastic angina, kidney failure and a variety of connective tissue disorders. In patients undergoing hemodialysis or kidney transplantation or suffering from cardiogenic shock, myocardial infarction or pulmonary hypertension levels as high as pg/ml have been observed (see, Stewart et al. (1991) Annals Internal Med 114: 464-469). Because endothelin is likely to be a local, rather than a systemic, regulating factor, it is probable that the levels of endothelin at the endothelium/smooth muscle interface are much higher than circulating levels.

Elevated levels of endothelin have also been measured in patients suffering from ischemic heart disease (Yasuda et al. (1990) Amer. Heart J.

119:801-806, Ray et al. (1992) Br. Heart J. 67:383-386). Circulating and tissue endothelin immunoreactivity is increased more than twofold in patients with advanced atherosclerosis (Lerman et al. (1991) New Enql. J. Med.

325:997-1001). Increased endothelin immunoreactivity has also been asscciated a" 10 with Buerger's disease (Kanno et al. (1990) J. Amer. Med. Assoc. 264:2868) and Raynaud's phenomenon (Zamora et al. (1990) Lancet 336 1144-1147).

Increased circulating endothelin levels were observed in patients who underwent percutaneous transluminal coronary angioplasty (PTCA) (Tahara et al. (1991) Metab. Clin. Exp 40:1235-1237; Sanjay et al. (1991) Circulation 84(Suol.

41:726), and in individuals (Miyauchi et al. (1992) Jpn. J. Pharmacol.58:279P; Stewart et al. (1991) Ann.lnternal Medicine 114:464-469) with pulmonary hypertension. Thus, there is clinical human data supporting the correlation between increased endothelin levels and numerous disease states.

Endothelin agonists and antagonists Because endothelin is associated with certain disease states and is implicated in numerous physiological effects, compounds that can interfere with Sor potentiate endothelin-associated activities, such as endothelin-receptor interaction and vasoconstrictor activity, are of interest. Compounds that exhibit endothelin antagonistic activity have been identified. For example, a fermentation product of Streptomyces misakiensis, designated BE-18257B, has been identified as an ETA receptor antagonist. BE-18257B is a cyclic pentapeptide, cyclo(D-Glu-L-Ala-allo-D-lle-L-Leu-D-Trp), which inhibits 1251labelled endothelin-1 binding in cardiovascular tissues in a concentrationdependent manner (IC 5 s 1.4 pM in aortic smooth muscle, 0.8 pM in ventricle membranes and 0.5 pM in cultured aortic smooth muscle cells), but fails to inhibit binding to receptors in tissues in which ET, receptors predominate at concentrations up to 100/ M. Cyclic pentapeptides related to BE-18257B, such as cyclo(D-Asp-Pro-D-Val-Leu-D-Trp) (BQ-123), have been synthesized and shown to exhibit activity as ETA receptor antagonists (see, U.S. Patent No.

5,114,918 to Ishikawa t al.; see, also, EP Al 0436 189 to BANYU PHARMACEUTICAL CO., LTD (October 7, 1991)). Studies that measure the inhibition by these cyclic peptides of endothelin-1 binding to endothelin-specific receptors indicate that these cyclic peptides bind preferentially to ETA receptors.

Other peptide and non-peptidic ETA antagonists have been identified (see, 5,352,800, 5,334,598, 5,352,659, 5,248,807, 5,240,910, 5,198,548, S. 5,187,195, 5,082,838). These include other cyclic pentapeptides, 10 acyltripeptides, hexapeptide analogs, certain antraquinone derivatives, indanecarboxylic acids, certain N-pyriminylbenzenesulfonamides, certain benzenesulfonamides, and certain naphthalenesulfonamides (Nakajima et al.

(1991) J. Antibiot. 44:1348-1356; Miyata et al. (1992) J. Antibiot. 45:74-8; S: Ishikawa et al. (1992) J.Med. Chem. 35:2139-2142; U.S. Patent No.

15 5,114,918 to Ishikawa et al.; EP Al 0 569 193; EP Al 0 558 258; EP Al 0 436 189 to BANYU PHARMACEUTICAL CO., LTD (October 7, 1991); Canadian Patent Application 2,067,288; Canadian Patent Application 2,071,193;

U.S.

Patent No. 5,208,243; U.S. Patent No. 5,270,313; U.S. Patent No. 5,464,853 to Chan et al; Cody et al. (1993) Med. Chem. Res. 3:154-162; Miyata era.

20 (1992) Antibiot 45:1041-1046; Miyata et al. (1992) J. Antibiot 45:1029- 1040, Fujimoto et al. (1992) FEBS Lett. 305:41-44; Oshashi et al. (1002) J.

Antibiot 45:1684-1685; EP Al 0 496 452; Clozel et al. (1993) Nature 365:759- 761; International Patent Application W093/08799; Nishikibe et l. (1993) Life Sci. 52:717-724; and Benigni et l. (1993) Kidney Int. 44:440-444). In general, the identified compounds have activities in in vitro assays as ETA antagonists at concentrations on the order of about 50-100 pM or less. A number of such compounds have also been shown to possess activity in in vivo animal models.

Very few selective ET, antagonists have been identified.

Endothelin antagonists and agonists as therapeutic agents It has been recognized that compounds that exhibit activity at ICso or EC 5 s concentrations on the order of 10- or lower in standard in vitro assays that assess endothelin antagonist or agonist activity have pharmacological utility (see, U.S. Patent Nos. 5,352,800, 5,334,598, 5,352,659, 5,248,807, 5,240,910, 5,198,548, 5,187,195, 5,082,838). By virtue of this activity, such compounds are considered to be useful for the treatment of hypertension such as peripheral circulatory failure, heart disease such as angina pectoris, cardiomyopathy, arteriosclerosis, myocardial infarction, pulmonary hypertension, vasospasm, vascular restenosis, Raynaud's disease, cerebral stroke such as cerebral arterial spasm, cerebral ischemia, late phase cerebral spasm after subarachnoid hemorrhage, asthma, bronchoconstriction, renal failure, particularly post-ischemic renal failure, cyclosporine nephrotoxicity such as acute renal S 10 failure, colitis, as well as other inflammatory diseases, endotoxic shock caused by or associated with endothelin, and other diseases in which endothelin has been implicated.

In view of the numerous physiological effects of endothelin and its association with certain diseases, endothelin is believed to play a critical role in these 15 pathophysiological conditions (see, Saito et al. (1990) Hyertension 734-738; Tomita et al. (1989) N. Enql. J. Med. 321: 1127; Kurihara et al.

(1989) J. Cardiovasc. Pharmacol. 13(Supl. S13-S17; Doherty (1992) J.

Med. Chem. 35: 1493-1508; Morel et al. (1989) Eur. J. Pharmacol. 167: 427- 428). More detailed knowledge of the function and structure of the endothelin S. 20 peptide family should provide insight in the progression and treatment of such conditions.

To aid in gaining further understanding of and to develop treatments for endothelin-mediated or related disorders, there is a need to identify compounds that modulate or alter endothelin activity. Identification of compounds that modulate endothelin activity, such as those that act as specific antagonists or agonists, may not only aid in elucidating the function of endothelin, but may yield in therapeutically useful compounds. In particular, compounds that specifically interfere with the interaction of endothelin peptides with the ETA or ET, receptors should be useful in identifying essential characteristics of endothelin peptides, should aid in the design of therapeutic agents, and may be useful as disease specific therapeutic agents.

11 Therefore, it is an object herein to provide compounds that have the ability to modulate the biological activity of one or more of the endothelin isopeptides. It is another object to provide compounds that have use as specific endothelin antagonists. It is also an object to use compounds that specifically interact with or inhibit the interaction of endothelin peptides with ETA or ETB receptors. Such compounds should be useful as therapeutic agents for the treatment of endothelin-mediated diseases and disorders and also for the identification of endothelin receptor subtypes.

Summary of the Invention Sulfbnamides and methods for modulating the interaction of an endothelin peptide I0 with ETA and/or ETB receptors are provided. In particular, sulfonamides and methods for inhibiting the binding of an endothelin peptide to ETA or ETo receptors are provided. The methods are effected by contacting the receptors with one or more sulfonamides prior to, simultaneously with, or subsequent to contacting the receptors with an endothelin peptide.

A first aspect of the present invention provides a compound that has formula I: Ar 2 SO- N- Ar 1 2

H

or a pharmaceutically acceptable salt, acid or ester thereof, wherein: Ar l is a five or six membered aromatic or heteroaromatic ring, preferably isoxazolyl, pyridazinyl, thiazolyl, pyrimidinyl or phenyl or is a bicyclic or tricyclic carbon or heterocyclic ring; and 2( Ar has formula: S****13 R 2 6 wherein: Ar 2 is substituted with one or more than one substituent, each of which is selected independently from the selections set forth for R 26 and R' 3 in which R 26 and R 1 3 are each -i independently selected from H, OH, OHNH, NH 2

NO

2 halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, alkoxy, alkylamino, dialkylamino, alkylthio, haloalkoxy, haloalkyl, alkylsufinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsufinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, carbonyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, [R:\LIBW]00623.docaak Ila in which the alkyl, alkenyl and alkynyl portions contain from 1 up to about 14 carbon atoms, preferably from 1 to 6 atoms, and are either straight or branched chains or cyclic, and the aryl portions contain from about 4 to about 16 carbons, preferably 4 to 10 carbons, with the proviso that if there is only one R 1 3 it is not hydrogen, and with the proviso that the compound of formula is a 4 -biphenylsulfonamide that is unsubstituted at the 2 or 6 position on the sulfonamide-linked phenyl group.

The sulfonamides are substituted or unsubstituted monocyclic or polycyclic aromatic or heteroaromatic sulfonamides, such as benezene sulfonamides, naphthalene sulfonamides and thiophene sulfonamides. Particularly preferred sulfonamides are N-isoxazolyl I0 sulfonamides. More particularly preferred among such sulfonamides are those in which Ar 2 is a heterocycle that contains one ring, multiple rings or fused rings, typically two or three rings and one or two heteroatoms in the ring or rings.

S.The sulfonamides have formula 1:

(I)

H

in which Ar' is a substituted or unsubstituted aryl group with one or more substituents, including an alkyl group, an aryl group, a substituted aryl group, a nitro group, an amino o group or a halide or is an alkyl group. In particular, Ar' is alkyl or is a five or six membered substituted or unsubstituted aromatic or heteroaromatic ring, particularly 3- or 5- isoxazolyl and pyridazinyl, and also including thiazolyl, including 2-thiazolyl, pyrimidinyl, including 2 2 -pyrimidinyl, or a [R:\LIBW]00623.doc:aak -12substituted benzene groups, including aryloxy substituted benzene groups or is a bicyclic or tricyclic carbon or heterocyclic ring.

Ar' is, in certain embodiments, selected from groups such as:

R

R NN N o R R R N and R is selected from H, NH 2 halide, pseudohalide, alkyl, alkylcarbonyl, formyl, an aromatic or heteroaromatic group, alkoxyalkyl, alkylamino, alkylthio, arylcarbonyl, aryloxy, arylamino, arylthio, haloalkyl, haloaryl, carbonyl, in which the aryl and alkyl portions, are unsubstituted or substituted with any of the preceeding groups, and straight or branched chains of from about 1 up to about 10-12 carbons, preferably, 1 to about 5 or 6 carbons. R is preferably H, NH 2 halide, CH 3 CH30O or another aromatic group. Ar 2 is any group such that the resulting sulfonamide inhibits binding by 50%, compared to binding in the absence of the sulfonamide, of an endothelin peptide to an endothelin receptor 25 at a concentration of less than about 100 pM, except that Ar 2 is not phenyl or naphthyl when Ar' is N-(5-isoxazolyl) or N-(3-isoxazolyl) unless the isoxazole is a 4 -halo-isoxazole, a 4-higher alkyl (Ce to C, 5 )-isoxazole, or the compound is a 4biphenyl that is unsubstituted at the 2 or 6 position on the sulfonamide-linked phenyl group.

In the embodiments described in detail herein, Ar' is an isoxazole and compounds are represented by the formulae II: -13- R' R 2 R' R 2 5I1 Ar- so- N oN Ar SON 0 H

H

in which R 1 and R 2 are either (ii) or (iii) as follows: R' and R 2 are each independently selected from H, NH 2

NO

2 halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, alkoxy, alkylamino, alkylthio, alkyloxy, haloalkyl, alkylsufinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsufinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, 15 alkylcarbonyl, aminocarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, in which the alkyl, alkenyl and alkynyl portions contain from 1 up to about 14 carbon atoms and are either straight or branched chains or cyclic, and the aryl portions contain from about 4 to about 16 carbons, except that R 2 is not halide or pseudohalide; or, 20 (ii) R' and R 2 together form where n is 3 to 6; or, (iii) R' and R 2 together form 1,3-butadienyl, and with the above proviso that Ar 2 is not phenyl or naphthyl when Ar 1 is isoxazolyl) or N-(3-isoxazolyl) unless the isoxazole is a 4 -halo-isoxazole, a 4higher alkyl to C 5 s)-isoxazole, or the compound is a 4 -biphenylsulfonamide that is unsubstituted at the 2 or 6 position on the sulfonamide-linked phenyl group.

In preferred embodiments herein, R' and R 2 are each selected independently from among alkyl, lower alkenyl, lower alkynyl, lower haloalkyl, halide, pseudohalide or H, except that R 2 is not halide; Ar 2 is any group such that the resulting sulfonamide inhibits binding by compared to binding in the absence of the sulfonamide, of an endothelin peptide to an endothelin receptor at a concentration of less than about 100 pM, with the above proviso. In particular, Ar 2 is a substituted or unsubstituted group selected from among groups including, but not limited to, the following: naphthyl, phenyl, biphenyl, quinolyl, styryl, thienyl, furyl, isoquinolyl, pyrrolyl, benzofuranyl, pyridinyl, thion 'aphthalyl, indolyl, alkyl, and alkenyl. It is understood that the positions indicated for substituents, including the sulfonamide groups, may be varied. Thus, for example, compounds herein encompass groups that include thiophene-3-sulfonam ides and thiophene-2-sulfonamides.

In certain embodiments described in detail herein, Ar 2 is a 4-biphenyl or is a single ring heterocycle, particularly a 5-membered ring, or is a fused bicyclic or tricyclic heterocycle that contains one or or more, particularly one, heteroatom selected from S, 0 and NR 42 in the ring, where

R

4 contains up to about 9 10 carbon atoms, preferably 1 to 10, more preferably 1 to 6 and is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, 9 9 cycloalkyl, cycloalkenyl, cycloalkynyl, C(0)R 15 and S(0)nR' in which n is 0-2;

R"

5 is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, S..aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl; R 42 and R' 5 are unsubstituted or 15 are substituted with one or more substituents each selected independently from Z, which is hydrogen, halide, pseudoahlide, alkyl, alkoxy, alkenyl, alkynyl, aryl, .heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, OH, CN,

*.C(O)R

1 6 CO R 16 SH, S(O),,R 1 in which n is 0-2, NHOH, NR 12 NON,

OR

16 Rl 2 NC.OR 16 and CONR1 2 R1 6 R 1 6 is hydrogen, alkyl, alkenyl, alkynyl, aryl, 20 alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl;

R'

2 ,which is selected independently from R 2 and Z, is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl,

C(O)R

1 and S(O),R 17 in which n is 0-2; and is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; each of R 42

R

12 R's and R' may be further substituted with the any of the groups set forth for Z.

In preferred embodiments herein, R 42 is aryl, such as phenyl or alkyl phenyl, hydrogen or loweralkyl.

Thus, in the compounds provided herein Ar 2 includes thienyl, furyl and pyrrolyl, benzofuryl, benzopyrolyl, benzothienyl, benzo[blfuryl, benzofblthienyl, and indolyl (benzofblpyrrolyl) and 4-biphenyl, and Ar 1 is preferably isoxazolyl) o .r N-(3-isoxazolyl). The sulfonamides are N-isoxazolyl sulfonamides and the compounds have either have formula Ill: R2R 53 Ir' RrR: S i SO- N. 2 x H H in which X is S, 0 or NR"1 in which R"1 contains up to about 30 carbon atoms, preferably 1 to 1 0, more preferably 1 to 6 and is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, A. cycloalkenyl, cycloalkynyl, C(0)R' 5 and S(O)rR' 5 in which n is 0-2; R1 5 is 0 9.

hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, 15 cycloalkyl, cycloalkenyl, cycloalkynyl; R" and R" 5 are unsubstituted or are substituted with one or more substituents each selected independently from Z, 0 which is hydrogen, halide, pseudoahlide, alkyl, alkoxy, alkenyl, alkynyl, aryl, 0 a, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, OH, CN, G(O)R 1 6 C0 2 R 1 6 SH, S(O),R 1 6 in which n is 0-2, NHOH, NR' 2

NO

2

N

3

OR'",

R1 2 NCOR16 and CONR1 2 R1 6 R 6 is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; R',which is selected independently from R" and Z, is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl,

C(O)R'

7 and 7 in which n is 0-2; and R, 7 is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; each of R1 2 R 15 and R 16 may be further substituted with the any of the groups set forth for Z, and R" is preferably hydrogen, aryl, such as phenyl or alkyl phenyl, loweralkyl; or the compounds are are 4 -biphenylsulfonides in which Ar' is preferably isoxazolyl) or N-(3-isoxazolyl.

Among the embodiments described in detail herein, Ar 2 is thienyl, furyl, pyrrolyl or a group that is a derivative or analog, as described below, of a thienyl, furyl or pyrrolyl group, including benzofbj derivatives such as a benzo[bIthienyl, Ar' is N-(5-isoxazolyl) or N-(3-isoxazolyl). Ar 2 has the formula

IV:

-16x x

AB

in which X is 0, S or NR"1, where R 1 is as defined above; that can be substituted at any or all positions or is an analog or derivative of the V"096groups of formula (IV) in which the substituents form fused aromatic, aliphatic "00060or heterocyclic rings; and RI and R 10 are each independently selected as 0 04follows from Mi or (ii): (i)M R' n R1 0 which each contain hydrogen orup to about 15 carbon atoms, generally up to about 30, more generally 20 or fewer, are each independently selected from hydrogen, halide, pseudohalide, alkyl, alkoxy, alkenyl, alkynyl, aryl, aryloxy, heterocycle, aralkyl, aralkoxy, cycloalkyl, '00:46cycloalkenyl, cycloalkynyl, OH, CN, C(O)R' 8

(OAC)CH=CHR

1 8 C0 2 R S H,

(CHA)C(O)(CH

2 1

(CH

2 )r(CH=CH).(CH 2 )nR' 8

(CH

2 =CH).(CH 2 )nR 18 20 (CH 2 )r(CH =CH),C(O)(CH 2 (CHA)NH(CH =CH) (CH 2 4,R' 8 C =N(OH)(CH 2 )rR 1 0664(CH2,C=

CH).NH(CH

2 )nR 1

C(O)(CH

2 )rNH(CH 2 )nRjI,

(CH

2 )rNH(CH 2 )nR1B, (CH 2 8 S(O)mR 1 8 in which m is 0-2, s, n and r are e-ah independently 0 to 6, preferably 0-3, HNOH, NR1 8

R

1 9

NO

2

N

3

OR"

8

R

19

NCOR'

8 and CONRI'R' 8 in which R" 9 is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, alkoxy, aryloxy, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, C(O)R 20 S(O)nR 2 1 in which n is 0-2; and RIB and R 20 are independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, heterocycle, alkoxy, aryloxy, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; and any of the groups set forth for R1, RI and RIO are unsubstituted or substituted with any substituents set forth for Z, which is hydrogen, halide, pseudoahlide, alkyl, alkoxy, alkenyl, alkynyl, aryl, aryloxy, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, OH, CN,

C(O)R

21 C0 2

R

21 SH, S(O)nR 2 in which n is 0-2, NHOH, NR 22

R

21

NO

2

N

3

OR

21 R 2 2 NCOR 21 and C0NR 22

R

21 R 22 is selected from hydrogen, alkyl, alkenyl, alkynyl, -17aryl, alkylaryl, heterocycle, aralkyl, alkoxy, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, C(0)R 23 and S(O).R 2 3 in which n is 0-2; and R21 and R 23 are independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl, with the proviso that if R 8 is NR'BR" 9

OR

18 R1SNCOR 8 and CONR1 9

R

1 8 C0 2

R

18

(CH

2 ),NH(CH CH).(CH,)nR'8,

(CH

2 CH),NH(CH 2

),R

1 8

(CH

2 ),C(O)NH(CH,)nR18'

C(O)(CH

2 )rNH(CH2),R',

(CH

2

),NH(CH

2 )R or (CH 2 8 and R 1 8 is an aryl group containing 5 or 6 members, then the aryl group has at 0 least two substituents, and preferably one substituent at the 2 -position relative to the linkage to the thienyl, furyl or pyrrolyl; any two of R 8

R

9 and R'o with the carbon to which each is S attached form an aryl, aromatic ring, heteroaromatic ring, carbocyclic or heterocyclic ring, which is saturated or unsaturated, containing from about 3 to about 16 members, preferably 3 to about 10 members, more preferably 5 to 7 members that is substituted with one or more substituents, each substituent is independently selected from Z; the other of R 8

R

9 and R'o is selected as in and the heteroatoms are NR", O, or S, with the proviso that Ar 2 is not 5-halo-3loweralkylbenzo[bthienyl, 5-halo-3-loweralkylbenzofb]furyl, 5-halo-3loweralkylbenzo[b]pyrrolyl.

In the embodiments provided herein, the alkyl, alkynyl and alkenyl portions of each listed substituent are straight or branched chains, acyclic or cyclic, and preferably have from about 1 up to about 10 carbons; in more preferred embodiments they have from 1-6 carbons. The aryl, alicyclic aromatic rings and heterocyclic groups can have from 3 to 16, generally, 3-7, more often 5-7 members in the rings, and may be single or fused rings. The ring size and carbon chain length are selected up to an amount that the resulting molecule binds and retains activity as an endothelin antagonist or agonist, such that the resulting compound inhibits binding by 50%, compared to binding in the absence of the sulfonamide, of an endothelin peptide to an endothelin receptor at a concentration of less than about 100 pM.

In preferred embodiments of interest herein,

R

9 and R' i are hydrogen, halide or methyl, more preferably hydrogen or halide, and R 8 is selected from -18-

CO

2

R'

8

(CH,),C(O)(CH

2 CHS(CH 2 8 C= N(OH)(CH 2 ),R'1 8

CH).(CH

2

)R'

8

(CH

2

CH)C(O)(CH

2 8

(CH

2 ),NH(CH= CH)(CH)nR' 8

(CH

2

CH),NH(CH

2 8

(CH

2 )rC(O)NH(CH 2

C(O)(CH,),NH(CH

2 R 8 with the proviso that if R 8 is C0 2

R

1 8

(CH

2 )rC(O)NH(CH 2 )nR',

C(O)(CH

2

)NH(CH

2

R

1 8 (CH),C(O)NH(CH)

R'

8 or (CH 2 rR'" 8 and R'" 8 is phenyl, the phenyl group is substituted at at least two positions, and preferably, at least one of those positions is ortho.

In the preferred compounds,

R'

18 is aryl orheteroaryl, preferably having or 6 members in the ring, more preferably phenyl or pyrimidinyl, most preferably phenyl.

In the most preferred compounds herein, is phenyl, which is substituted at more than one position, and most preferably at least one .substituent is at the ortho position, R 9 and R' 0 are each hydrogen, halide or S* 15 loweralkyl, preferably hydrogen, and R 8 is C(O)NHR'",

C(O)CH

2

R'

8

(CH,)R'

8 whith the proviso that if R 8 is C(O)NHR 18 then the phenyl group must have at east two substituents, preferably one of the substituents is in the ortho positio..

In other preferred embodiments, Ar 2 is a benzolblthienyl, benzolb]furyl, or indolyl (benzofb]pyrrolyl), with the proviso that the benzene ring is substituted 20 and the substituents are other than 5 halo, 3-loweralkyl. Preferred substituents on the benzene ring, include, but are not limited to, one or more selected from alkylenedioxy, particularly methylenedioxy, preferably 3 4 -methylenedioxy, ethylenedioxy, aryl, particularly phenyl, dimethylamino, diethylamino, benzyl, alkoxy, particularly lower alkoxy, such as methoxy and ethoxy, halide, and alkyl, preferably loweralkyl.

In the preferred compounds herein,

R

2 is preferably, selected from among alkyl, lower alkenyl, lower alkynyl, lower haloalkyl or H; and R' is halide or loweralkyl, and more preferablly, R' is bromide or chloride, methyl or ethyl. In the most active compounds provided herein, as evidenced by Ln vitro binding assays, R' is bromide or chloride. For use in vivo R' is preferably chloride.

Of the compounds described herein, those that inhibit or increase an endothelin-mediated activity by about 50% at concentrations of less than about -19pM are preferred. More preferred are those that inhibit or increase an endothelin-mediated activity by about 50% at concentrations of less than about 1 pM, more preferably less than about 0.1 pM, even more preferably less than about 0.01 pM, and most preferably less than about 0.001 pM. It is noted that, as described below, the ICso concentration determined in the in vitro assays is a non-linear function of incubation temperature. The preferred values recited herein refer to the assays that are performed at 40 C. When the assays.

are performed at 240 C, somewhat higher (see, Table 1) ICs, concentrations are observed. Accordingly, the preferred ICso concentrations are about S 10 higher.

Also among the most preferred compounds for use in methods provided herein, are those that are ETA selective, they interact with ETA receptors at substantially lower concentratons (at an ICso at least about 10-fold lower, preferably 100-fold lower) than they interact with ET, receptors. In particular, compounds that interact with ETA with an IC 50 of less than about 10 pM, preferably less than 1 pM, more preferably less than 0.1 pM, but with ET, with an ICo 0 of greater than about about 10 pM or compounds that interact with ET, with an ICso of less than about 10 pM, preferably less than 1 pM, more preferably less than 0.1 pM, but with ETA with an IC 5 s of greater than about 20 pM are preferred.

Preferred compounds also include compounds that are ET, receptor selective or that bind to ET B receptors with an ICso of less than about 1 pM. ET, selective compounds interact with ET, receptors at ICso concentrations that are at least about 10-fold lower than the concentrations at which they interact with ETA receptors. In these compounds,

R

2 is selected from among alkyl, lower alkenyl, lower alkynyl, lower haloalkyl, halide or H; and R' is halide or loweralkyl, and in preferred embodiments,

R

1 is bromide or chloride, preferabaly chloride; R 9 and R 1 0 are selected independently from hydrogen, loweralkyl, preferably methyl or ethyl, or halide, and R 8 which is the substituent at the 5-position (see, formulae III and IV), is aryl or a heterocycle, particularly phenyl and isoxazolyl, which are unsubstituted or substituted with Z, which is preferably loweralkyl or halide.

Pharmaceutical compositions formulated for administration by an appropriate route and means containing effective concentrations of one or more of the compounds provided herein or pharmaceutically acceptable salts or acids thereof that deliver amounts effective for the treatment of hypertension, stroke, asthma, shock, ocular hypertension, glaucoma, renal failure, inadequate retinal perfusion and other conditions that are in some manner mediated by an endothelin peptide or that involve vasoconstriction or whose symptoms can be.

ameliorated by administration of an endothelin antagonist or agonist, are also provided. Particularly preferred compositions are those that deliver amounts effective for the treatment of hypertension or renal failure. The effective S.amounts and concentrations are effective for ameliorating any of the symptoms of any of the disorders.

Methods for inhibiting binding of an endothelin peptide to an endothelin receptor are provided. These methods are practiced by contacting the receptor with one or more of the compounds provided herein simultaneously, prior to, or subsequent to contacting the receptor with an endothelin peptide.

Methods for treatment of endothelin-mediated disorders, including but not limited to, hypertension, asthma, shock, ocular hypertension, glaucoma, inadequate retinal perfusion and other conditions that are in some manner 20 mediated by an endothelin peptide, or for treatment of disorder that involve vasoconstriction or that are ameliorated by administration of an endothelin antagonist or agonist are provided.

In particular, methods of treating endothelin-mediated disorders by administering effective amounts of the sulfonamides, prodrugs or other suitable derivatives of the sulfonamides are provided. In particular, methods for treating endothelin-mediated disorders, including hypertension, cardiovascular diseases, cardiac diseases including myocardial infarction, pulmonary hypertension, erythropoietin-mediated hypertension, respiratory diseases and inflammatory diseases, including asthma, bronchoconstriction, ophthalmologic diseases, gastroenteric diseases, renal failure, endotoxin shock, menstrual disorders, obstetric conditions, wounds, anaphylactic shock, hemorrhagic shock, and other diseases in which endothelin mediated physiological responses are implicated, by -21administering effective amounts of one or more of the compounds provided herein in pharmaceutically acceptable carriers are provided. Preferred methods of treatment are methods for treatment of hypertension and renal failure.

More preferred methods of treatment are those in which the compositions contain at least one compound that inhibits the interaction of endothelin-1 with ETA receptors at an ICo 5 of less than about 10 pM, and preferably less than about 5 more preferably less than about 1 even more preferably less than 0.1 pM, and most preferably less than 0.05 pM Other preferred methods are those in which the compositions contain one or more compounds that is (are) ETA selective or one or more compounds that is ((are) ET, selective. Methods in which the compounds are ETA selective are for treatment of disorders, such as hypertension; and methods in which the compounds are ET, selective are for treatment of disorders, such as asthma, e'l* that require bronchodilation.

In practicing the methods, effective amounts of compositions containing therapeutically effective concentrations of the compounds formulated for oral, intravenous, local and topical application for the treatment of hypertension, cardiovascular diseases, cardiac diseases, including myocardial infarction, respiratory diseases, including asthma, inflammatory diseases, ophthalmologic 20 diseases, gastroenteric diseases, renal failure, immunosuppressant-mediated renal vasoconstriction, erythropoietin-mediated vasoconstriction, endotoxin shock, anaphylactic shock, hemorrhagic shock, pulmonary hypertension, and other diseases in which endothelin mediated physiological responses are implicated are administered to an individual exhibiting the symptoms of one or more of these disorders. The amounts are effective to ameliorate or eliminate one or more symptoms of the disorders.

Methods for the identification and isolation of endothelin receptor subtypes are also provided. In particular, methods for detecting, distinguishing and isolating endothelin receptors using the disclosed compounds are provided.

In particular, methods are provided for detecting, distinguishing and isolating endothelin receptors using the compounds provided herein.

-22- In addition, methods for identifying compounds that are suitable for use in treating particular diseases based on their preferential affinity for a particular endothelin receptor subtype are also provided.

Articles of manufacture containing packagirimaterial, a compound provided herein, which is effective for ameliorating the symptoms of an endothelin-mediated disorder, antagonizing the effects of endothelin or inhibiting binding of an endothelin peptide to an ET receptor with an ICso of less than about 10 pM, within the packaging material, and a label that indicates that the compound or salt thereof is used for antagonizing the effects of endothelin, treating an endothelin-mediated disorder, or inhibiting the binding of an endothelin peptide to an ET receptor are provided.

Also provided herein are substituted biphenyl sulfonamides that have ET, activity.

DETAILED DESCRIPTION OF PREFERRED

EMBODIMENTS

Definitions Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs. All patents and publications referred to herein are incorporated by reference.

20 As used herein, endothelin (ET) peptides include peptides that have substantially the amino acid sequence of endothelin-1, endothelin-2 or endothelin-3 and that act as potent endogenous vasoconstrictor peptides.

As used herein, an endothelin-mediated condition is a condition that is caused by abnormal endothelin activity or one in which compounds that inhibit endothelin activity have therapeutic use. Such diseases include, but are not limited to hypertension, cardiovascular disease, asthma, inflammatory diseases, ophthalmologic disease, menstrual disorders, obstetric conditions, gastroenteric disease, renal failure, pulmonary hypertension, endotoxin shock, anaphylactic shock, or hemorrhagic shock. Endothelin-mediated conditions also include conditions that result from therapy with agents, such as erythropoietin and immunosuppressants, that elevate endothelin levels.

-23- As used herein an effective amount of a compound for treating a particular disease is an amount that is sufficient to ameliorate, or in some manner reduce the symptoms associated with the disease. Such amount may be administered as a single dosage or may be administered according to a regimen, whereby it is effective. The amount may cure the disease but, typically, is administered in order to ameliorate the symptoms of the disease.

Typically, repeated administration is required to achieve the desired amelioration of symptoms.

As used herein, an endothelin agonist is a compound that potentiates or 10 exhibits a biological activity associated with or possessed by an endothelin peptide.

As used herein, an endothelin antagonist is a compound, such as a drug or an antibody, that inhibits endothelin-stimulated vasoconstriction and contraction and other endothelin-mediated physiological responses. The antagonist may act by interfering with the interaction of the endothelin with an endothelin-specific receptor or by interfering with the physiological response to or bioactivity of an endothelin isopeptide, such as vasoconstriction. Thus, as used herein, an endothelin antagonist interferes with endothelin-stimulated vasoconstriction or other response or interferes with the interaction of an 20 endothelin with an endothelin-specific receptor, such as ETA receptors, as assessed by assays known to those of skill in the art.

The effectiveness of potential agonists and antagonists can be assessed using methods known to those of skill in the art. For example, endothelin agonist activity can be identified by its ability to stimulate vasoconstriction of isolated rat thoracic aorta or portal vein ring segments (Borges et al (1989) "Tissue selectivity of endothelin" Eur. J. Pharmacol. 165: 223-230). Endothelin antagonist activity can be assessed by the ability to interfere with endothelininduced vasoconstriction. Exemplary assays are set forth in the EXAMPLES. As noted above, the preferred ICso concentration ranges are set forth with reference to assays in which the test compound is incubated with the ET receptor-bearing cells at 40 C. Data presented for assays in which the incubation step is performed at the less preferred 240 C are identified. It is understood that for -24purposes of comparison, these concentrations are somewhat higher than the concentrations determined at 40 C.

As used herein, the biological activity or bioactivity of endothelin includes any activity induced, potentiated or influenced by endothelin in vivo. It also includes the ability to bind to particular receptors and to induce a functional response, such as vasoconstriction. It may be assessed by in vivo assays or by in vitro assays, such as those exemplified herein. The relevant activities include, but are not limited to, vasoconstriction, vasorelaxation and bronchodilation. For example, ET, receptors appear to be expressed in vascular 10 endothelial cells and may mediate vasodilation and other such responses; whereas ETA receptors, which are endothelin-1-specific, occur on smooth muscle and are linked to vasoconstriction Any assay known to those of skill in the art to measure or detect such activity may be used to assess such activity (see, Spokes et al. (1989) J. Cardiovasc. Pharmacol. 13(Suppl. 5):5191- S192; Spinella et al. (1991) Proc. Natl. Acad. Sci. USA 88: 7443-7446; Cardell et al. (1991) Neurochem. Int. 18:571-574); and the Examples herein).

As used herein, the ICso refers to an amount, concentration or dosage of a particular test compound that achieves a 50% inhibition of a maximal response, such as binding of endothelin to tissue receptors, in an assay that e 20 measures such response.

As used herein, EC 5 0 refers to a dosage, concentration or amount of a particular test compound that elicits a dose-dependent response at 50% of maximal expression of a particular response that is induced, provoked or potentiated by the particular test compound.

As used herein a sulfonamide that is ETA selective refers to sulfonamides that exhibit an ICo 5 that is at least about 10-fold lower with respect to ET, receptors than ET, receptors.

As used herein, a sulfonamide that is ET, selective refers to sulfonamides that exhibit an IC5o that is at least about 10-fold lower with respect to ET, receptors than ETA receptors.

As used herein, pharmaceutically acceptable salts, esters or other derivatives of the compounds include any salts, esters or derivatives that may be readily prepared by those of skill in this art using known methods for such derivatization and that produce compounds that may be administered to animals or humans without substantial toxic effects and that either are pharmaceutically active or are prodrugs. For example, hydroxy groups can be esterified or etherified.

As used herein, treatment means any manner in which the symptoms of a conditions, disorder or disease are ameliorated or otherwise beneficially altered. Treatment also encompasses any pharmaceutical use of the compositions herein, such as use as contraceptive agents.

10 As used herein, amelioration of the symptoms of a particular disorder by administration of a particular pharmaceutical composition refers to any S" lessening, whether permanent or temporary, lasting or transient that can be attributed to or associated with administration of the composition.

As used herein, substantially pure means sufficiently homogeneous to appear free of readily detectable impurities as determined by standard methods of analysis, such as thin layer chromatography (TLC), gel electrophoresis and *high performance liquid chromatography (HPLC), used by those of skill in the art to assess such purity, or sufficiently pure such that further purification would not detectably alter the physical and chemical properties, such as enzymatic and 20 biological activities, of the substance. Methods for purification of the compounds to produce substantially chemically pure compounds are known to those of skill in the art. A substantially chemically pure compound may, however, be a mixture of stereoisomers. In such instances, further purification might increase the specific activity of the compound.

As used herein, biological activity refers to the in vivo activities of a compound or physiological responses that result upon in vivo administration of a compound, composition or other mixture. Biological activity, thus, encompasses therapeutic effects and pharmaceutical activity of such compounds, compositions and mixtures.

As used herein, a prodrug is a compound that, upon in vivo administration, is metabolized or otherwise converted to the biologically, pharmaceutically or therapeutically active form of the compound. To produce a -26prodrug, the pharmaceutically active compound is modified such that the active compound will be regenerated by metabolic processes. The prodrug may be designed to alter the metabolic stability or the transport characteristics of a drug, to mask side effects or toxicity, to improve the flavor of a drug or to alter other characteristics or properties of a drug. By virtue of knowledge of pharmacodynamic processes and drug metabolism in vivo, those of skill in this art, once a pharmaceutically active compound is known, can design prodrugs of the compound (see, Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392). For example, 10 succinyl-sulfathiazole is a prodrug of 4-amino-N-(2-thiazoyl)benzenesulfonamide (sulfathiazole) that exhibits altered transport characteristics.

As used herein, acid isostere means a group that is significantly ionized at physiological pH. Examples of suitable acid isosteres include sulfo, phosphono, alkylsulfonylcarbamoyl, tetrazolyl, arylsulfonylcarbamoyl or heteroarylsulfonylcarbamoyl.

As used herein, halo or halide refers to the halogen atoms; F, Cl, Br and I.

As used herein, pseudohalides are compounds that behave substantially similar to halides. Such compounds can be used in the same manner and treated in the same manner as halides in which X is a halogen, such as Cl or 20 Br). Pseudohalides include, but are not limited to cyanide, cyanate, thiocyanate, selenocyanate and azide.

As used herein, haloalkyl refers to a loweralkyl radical in which one or more of the hydrogen atoms are replaced by halogen including, but not limited to, chloromethyl, trifluoromethyl, 1-chloro-2-fluoroethyl and the like.

As used herein, alkyl means an aliphatic hydrocarbon group that is a straight or branched chain preferably having about 1 to 12 carbon atoms in the chain. Preferred alkyl groups are loweralkyl groups which are alkyls containing 1 to about 6 carbon atoms in the chain. Branched means that one or more loweralkyl groups such as methyl, ethyl or propyl are attached to a linear alkyl chain. The alkyl group may be unsubstituted or independently substituted by one or more groups, such as, but not limited to: halo, carboxy, formyl, sulfo, sulfino, carbamoyl, amino and imino. Exemplary alkyl groups include methyl, -27ethyl, propyl, methanoic acid, ethanoic acid, propanoic acid, ethanesulfinic acid and ethane sulfonic acid.

As used herein the term lower describes alkyl, alkenyl and alkynyl groups containing about 6 carbon atoms or fewer. It is also used to describe aryl groups or heteroaryl groups that contain 6 or fewer atoms in the ring.

Loweralkyl, lower alkenyl, and lower alkynyl refer to carbon chains having less than about 6 carbons. In preferred embodiments of the compounds provided herein that include alkyl, alkenyl, or alkynyl portions include loweralkyl, lower alkenyl, and lower alkynyl portions.

10 As used herein, alkenyl means an aliphatic hydrocarbon group containing a carbon-carbon double bond and which may be straight or branched chained Shaving from about 2 to about 10 carbon atoms in the chain. Preferred alkenyl groups have 2 to about 4 carbon atoms in the chain. Branched means that one or more loweralkyl or lower alkenyl groups are attached to a linear alkenyl chain.

The alkenyl group may be unsubstituted or independently substituted by one or more groups, such as halo, carboxy, formyl, sulfo, sulfino, carbamoyl, amino and imino. Exemplary alkenyl groups include ethenyl, propenyl, carboxyethenyl, carboxypropenyl, sulfinoethenyl and sulfonoethenyl.

As used herein, alkynyl means an aliphatic hydrocarbon group containing 20 a carbon-carbon triple bond and which may be straight or branched having-about 2 to 10 carbon atoms in the chain. Branched means that one or more loweralkyl, alkenyl or alkynyl groups are attached to a linear alkynyl chain. An exemplary alkynyl group is ethynyl.

As used herein, aryl means an aromatic monocyclic or multicyclic hydrocarbon ring system containing from 3 to 15 or 16 carbon atoms, preferably from 5 to 10. Aryl groups include, but are not limited to groups, such as phenyl, substituted phenyl, napthyl, substituted naphthyl, in which the substitunent is loweralkyl, halogen, or lower alkoxy. Preferred aryl groups are lower aryl groups that contain less than 7 carbons in the ring structure.

As used herein, the nomenclature alkyl, alkoxy, carbonyl, etc. are used as is generally understood by those of skill in this art. For example, as used herein alkyl refers to saturated carbon chains that contain one or more carbons; the -28chains may be straight or branched or include cyclic portions or be cyclic. As used herein, alicyclic refers to aryl groups that are cyclic.

As used herein, cycloalkyl refers to saturated cyclic carbon chains; cycloalkyenyl and cycloalkynyl refer to cyclic carbon chains that include at least one unsaturated double or triple bond, respectively. The cyclic portions of the carbon chains may include one ring or two or more fused rings.

As used herein, cycloalkenyl means a non-aromatic monocyclic or multicyclic ring system containing a carbon-carbon double bond and having about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings 10 include cyclopentenyl or cyclohexenyl; preferred is cyclohexenyl. An exemplary S multicyclic cycloalkenyl ring is norbornylenyl. The cycloalkenyl group may be independently substituted by one or more halo or alkyl.

As used herein, "haloalkyl" refers to a loweralkyl radical in which one or more of the hydrogen atoms are replaced by halogen including, but not limited to, chloromethyl, trifluoromethyl, 1-chloro-2-fluoroethyl and the like.

As used herein, "haloalkoxy" refers to RO- in which R is a haloalkyl group.

As used herein, "carboxamide" refers to groups of formula RpCONH 2 in which R is selected from alkyl or aryl, preferably loweralkyl or lower aryl and p is 0 or 1.

As used herein, "alkylaminocarbonyl" refers to -C(O)NHR in which R is hydrogen, alkyl, preferably loweralkyl or aryl, preferably lower aryl.

As used herein "dialkylaminocarbonyl" as used herein refers to -C(O)NR'R in which R' and R are independently selected from alkyl or aryl, preferably loweralkyl or loweraryl; "carboxamide" refers to groups of formula NR'COR.

As used herein, "alkoxycarbonyl" as used herein refers to -C(O)OR in which R is alkyl, preferably loweralkyl or aryl, preferably lower aryl.

As used herein, "alkoxy" and "thioalkoxy" refer to RO- and RS-, in which R is alkyl, preferably loweralkyl or aryl, preferably lower aryl.

As used herein, "haloalkoxy" refers to RO- in which R is a haloalkyl group.

As used herein, "aminocarbonyl" refers to -C(O)NH,.

-29- As used herein, "alkylaminocarbonyl" refers to -C(O)NHR in which R is alkyl, preferably loweralkyl or aryl, preferably lower aryl.

As used herein, "alkoxycarbonyl" refers to -C(0)OR in which R is alkyl, preferably loweralkyl.

As used herein, cycloalkyl refers to satured cyclic carbon chains; cycloalkyenyl and cycloalkynyl refer to cyclic carbon chains that include at least one unsaturated triple bond. The cyclic portions of the carbon chains may include one ring or two or more fused rings.

As used herein, alkylenedioxy means an -O-alkyl-O- group in which the 10 alkyl group is as previously described. A replacement analog of alkylenedioxy means an alkylenedioxy in which one or both of the oxygen atoms is replaced by a similar behaving atom or group of atoms such as, S, N, NH, Se. An exemplary replacement alkylenedioxy group is ethylenebis(sulfandiyl). Alkylenethioxyoxy is -S-alkyl-O- -O-alkyl-S- and alkylenedithioxy is -S-alkyl-S-.

As used herein, heteroaryl means an aromatic monocyclic or fused ring Ssystem in which one or more of the carbon atoms in the ring system is(are) replaced by an element(s) other than carbon, for example nitrogen, oxygen or sulfur. Preferred cyclic groups contain one or two fused rings and include from about 3 to about 7 members in each ring. Similar to "aryl groups", the 20 heteroaryl groups may be unsubstituted or substituted by one or more substituents. Exemplary heteroaryl groups include pyrazinyl, pyrazolyl, tetrazolyl, furanyl, or 3-)thienyl, or 4-)pyridyl, imidazoyl, pyrimidinyl, isoxazolyl, thiazolyl, isothiazolyl, quinolinyl, indolyl, isoquinolinyl, oxazolyl and 1, 2 ,4-oxadiazolyl. Preferred heteroaryl groups include 5 to 6 -membered nitrogen-containing rings, such as pyrmidinyl.

As used herein, alkoxycarbonyl means an alkyl-O-CO- group. Exemplary alkoxycarbonyl groups include methoxy- and ethoxycarbonyl.

As used herein, carbamoyl means -CONH,. As with all groups described herein, these groups may be unsubstituted or substituted. Substituted carbamoyl includes groups such as -CONY 2

Y

3 in which Y 2 and Y 3 are independently hydrogen, alkyl, cyano(loweralkyl), aryalkyl, heteroaralkyl, carboxy(loweralkyl), carboxy(aryl substituted loweralkyl), carboxy(carboxy substituted loweralkyl), carboxyihydroxy substituted loweralkyl), carboxy(heteroaryl substituted loweralkyl), carbamoyi(loweralkyl,) alkoxycarbonyl(loweralkyl) or alkoxycarbonyl(aryl substituted loweralkyl), provided that only one of Y' and Y' may be hydrogen and when one of Y' and y 3 is carboxy(loweralkyl), carboxy(aryl substituted Joweralkyl), carbamoyl(loweralkyl), alkoxycarbonyl(loweralkyl) or alkoxycarbonyl(aryl substituted loweralkyl) then the other of Y' and Y' is hydrogen or alkyl.

Preferred for Y' and y 3 are independently hydrogen, alkyl, cyano(Ioweralkyl), aryalkyl, heteroaralkyl, carboxy(loweralkyl), carboxy(aryl substituted loweralkyl) and carbamoyl(loweralkyl).

used herein, any corresponding

N-(

4 -halo3methy-5-isoxazoly, N-(4hafo-5-methyl-3isoxazolyl),

N-(

3 ,4-dimethyl-5-.isoxazolyl), N-(4-halo-5-methyl.3 isoxazolyl),

N-(

4 -halo-3methyl-5isoxazolyl), S-dimethyl-3-isoxazolyl) derivative thereof refers to compounds in which Ar' is the same as the compound specifcally set forth, but Ar' is NW( 4 -halo3m ethyl-5-isoxazolyl), N-(4halo-5-methyI3isoxazlyl),

N-(

3 ,4-dimethy.5.isoxazolyl),

N-(

4 -halo-5-methyl.3 isoxzoll),N-( 4 -halo.3-methyl.5.isoxazolyl), rN(,-iehl3ioaoy)i which halo is any halide, peferably Cl or Br.

As used herein, the abbreviations for any protective groups, amino acids and other compounds, are, unless indicated otherwise, in accord with their common usage, recognized abbreviations, or the IUPAC-JUB Commission on Biochemical Nomenclature (see, (1972) Biochemn. 1192-4) A C m p o u d s f r u s i n t r e a i n g n d o t e i m e d i 9 t4 d d i s e a s e A. Compoundanehds for uei treating endothelin-mediated diseases usn the compounds of formulae I and 11 are provided. In particular, in the compounds provided herein, Ar 2 is thienyl, furyl, pyrrolyl or a group, such as benzofuryl, thionaphthyl or indolyl, that is a derivative or analog, as described below, of a thienyl, furyl or pyrrolyl group or a 4-biphenyl group, Ar' is preferably N-(5-isoxazolyl) or N-(3-isoxazolyl).

-31- 1. Ar 2 is a thiophene, pyrrole, furan, benzotbjthiophene, indolyl (benzo[blpyrrole), or benzo~blfuran Among the compounds provided herein are those represented by the formula V: R 1 0 RI R2 Ri SO-N 0 NS-N 0 14 31 2 1

H

2 H R0 X R R R*

R

R R

R

2 R

R

2 14 3j 14 315 2 CS0-N 0 1SO- N N r 1 X R X I N' *R

HH

in which R' and R 2 are either GOi or (iii) as follows: Wi R' and R 2 are each independently selected fromr H, NH 2

NO

2 halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heterolaryl, alkoxy, alkylamino, alkylthio, haloalkoxy, haloalkyl, alkylsufinyl, alkylsulfonyl, aryloxy, S arylamino, arylthio, arylsufinyl, arylsulfonyl, aminocarbonyl, arylaminocarbonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, in which the alkyl, alkenyl and alkynyl portions are either straight or branched chains that contain from 1 up to about 10 carbon atoms, and the aryl portions contain from about 4 to about 14 carbons, except the R 2 is not halide, pseudohalide or higher alkyl; or, 00i R' and R 2 together form -(CH 2 where n is 3 to 6; or, (iii) R 1 and R 2 together form 1 ,3-butadienyl; and X is S, 0 or NR" in which R" contains up to about 30 carbon atoms, preferably 1 to 10, more preferably 1 to 6 and is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl,

C(O)R'

5 and S(0)nR's in which n is 0-2; R' 5 is -32hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl; R" and RI 5 are unsubstituted or are substituted with one or more substituents each selected independently from Z, which is hydrogen, halide, pseudoahlide, alkyl, alkoxy, alkenyl, alkynyl, aryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, OH, CN,

C(O)R'

6 C0 2

R

16 SH, S(O)nR' in which n is 0-2, NHOH, NR' 2

R

1

NO

2

N

3

OR'

6 R1 2 NCOR 1 6 and CONR1 2 R 16 R 16 is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalky"nyl; R1 2 which is selected independently from and Z, is selected from hydrogen, alkyl, alkenyi, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, C(O)R" and S(O)nR' 7 in which n is 0-2; and R' 7 is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; each of R 11 R 1 2

R"

5 and R 16 may be further substituted with the any of the groups set forth for Z, and R 1 is preferably *o 15 hydrogen, aryl, such as phenyl or alkyl phenyl, loweralkyl; and

R

9 and R' 0 which each contain hydrogen or up to about 50 carbon atoms, 00% generally up to about 30, more generally 20 or fewer, are each independently 00 selected as described above, and more preferably from or (ii) as follows: Mi RI and R 10 are selected from hydrogen, halide, pseudohalide, alkjI, 20 alkoxy, alkenyl, alkynyl, aryl, aryloxy, heterocycle, aralkyl, aralkoxy, cycloalkyl, o. cycloalkenyl, cycloalkynyl, OH, CN, C(O)R 18 (OAC)CH =CHR' 8 C0 2

R

18

SH,

(CH

2 )rC(O)(CH 2 )nR' 8

(CH

2 )r(CH=CH),(CH 2 )rR' 8

(CH

2 )rC(O)(CH =CH),(CH 2 0

,RI

8

(CH

2 )r(CH =CH) 8

C(O)(CH

2

(CH

2 ),NH(CH =CH),(CH 2 8

C=N(OH)(CHA)R',

(CH

2

=CH),NH(CH

2

(CH

2 )rC(O)NH(CH 2

C(O)(CH

2

),NH(CH

2 8

(CH

2

),NH(CH

2

),R

18

(CH

2 )rR'B, S(O)mR 18 in which mn is 0-2, s, n and r are each independently 0 to 6, preferably 0-3, HNOH, NR' 8

NO

2

N

3

OR'

8

R'

9

NCOR'

8 and CONR' 9 in which R' 9 is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, alkoxy, aryloxy, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, C(O)R 20 S(O),R 20 in which n is 0-2; and R' 8 and R 20 are independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, heterocycle, alkoxy, aryloxy, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; Re is selected from C(O)R' 8

(OAC)CH=CHR

18

CO

2 R's,

(CH

2 (CH 2 8

(CH

2 =CH)S

(CH

2 18

(CH

2 )rC(0) (CH =CH),(CH 2 8

(CH

2 =CH),C(0)(CH 2 (CH 2 )rNH(CH

=CH),(CH

2 0 C =N(OH)(CH 2 ),R18

(CH

2

=CH),NH(CH

2

,R

18

(CH

2 )rC(O)NH(CH 2 )nRB, C(0)(CHA)NH(CH 2

I',

(CH

2

),NH(CH

2 )nR18, (CH 2 )rR 8 1 in which mn is 0-2, s, n and r are each independently 0 to 6, preferably 0-3, in which

R"

8 is aryl, preferably phenyl, with the proviso that, if Re is (CH 2

),C(O)NH(CH

2 )nR18 C(O)(CH 2 )rNH(CH 2 )nRlf,

(CH

2 )rNH(CH 2 )nR 18, (GH 2 )rR'a, particularly if r is 0 and/or n is 0, and We 8 is aryl, particularly phenyl, then R" 8 must have two or more substituents, with 10 preferably at feast one ortho substituent; where any of the groups set forth for Re, R' and R' 0 are unsubstituted or substituted with any substituents set forth for Z, which is hydrogen, halide, pseudoahlide, alkyl, alkoxy, alkenyl, alkynyl, aryl, aryloxy, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, OH, CN, C(O)R 21 C0 2

R

21

SH,

15 S(0),R 21 in which n is 0-2, NHOH,

NR

22

R

2

NO

2 OR 21

R

22 NCOR 2 1 and C0NR 22

R

21 R 1 2 is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, alkoxy, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, C(0)R 23 and S(0),,R 23 in which n is 0-2; and R 2 1 and R 2 are independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, 20 aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; or G any two of R 8 R' and R' 0 form an aryl, aromatic ring, heteroaromatic ring, carbocyclic or heterocyclic ring, which is saturated or unsaturated, containing from about 3 to about 1 6 members, preferably 3 to about 10 members, more preferably 5 to 7 members that is substituted with one or more substituents, each substituent being independently selected from Z; the other of Re, R' and 13 10 is selected as from the groups set forth for R 9 and RIO in Mi; and the heteroatoms are NR 11 0, or S, with the proviso that Ar 2 is not halo- 3 -loweralkylbenzo~blthienl 5-halo- 3 -loweralkylbenzofbjfuryl, 5-halo-3loweralkylbenzo[blpyrrolyl.

In these embodiments, Ar 2 is, thus, represented by the formulae

(IVA

and IVB): R' P-'R x x A

B

that can be substituted at any or all positions or is an analog of compounds of.

formula (IV) in which the substituents form fused aromatic, aliphatic or heterocyclic rings; and in which X is NR", O, or S, and which is hydrogen or contains up to about 30 carbon atoms, preferably 1 to 10, more preferably 1 S to 6, and is selected as defined above. R 8

R

9

R'

1 are selected as described above.

In the embodiments provided herein, when R 8

R

9 and R'O are selected as in above, R 8 is preferably selected from among (CH 2

),C(O)(CH

2

)R'

8

(CH

2

),NH(CH

2 )nR 1 8

(CH

2 )rNH(CH 2

)R'

8 (CH)r(CH= CH),(CH2)nR, 8

(CH

2 )rC(0)(CH=

CH),(CH

2

(CH

2 )r(CH= 8

(CH

2

),(CH=CH),NH(CH

2 8 C= N(OH)(CH 2

,R'

8

(CH

2 ),C(0)NH(CH 20 C(0)(CH 2

),NH(CH

2

),R

8

(CH

2 )rNH(CH CH),(CH 2

)R

8

(CH

2 ),C(0)NH(CH 2 nR 8

NH

2 )rNH(CH 2 )nR' (CH 2 8 with the proviso that if R 8 is

(CH

2 rC(0)NH(CH,),R 18

(CH,),C(O)NH(CH

2

)R'

8 or (CH 2 8 and R 1 8 is phenyl, the phenyl group is substituted at least two positions, and preferably, at least one of those positions is ortho.

In preferred of these compounds,

R'

8 is aryl or heteroaryl, preferably having 5 or 6 members in the ring, more preferably phenyl or pyrimidinyl, most preferably phenyl. R 9 and R 1 are preferably hydrogen, halide, loweralkyl, or halo loweralkyl The more preferred compounds provided herein are compounds in which the alkyl, alkynyl and alkenyl portions are straight or branched chains, acyclic or cyclic, and have from about 1 up to about 10 carbons; in certain of the more preferred embodiments they have from 1-6 carbons, and they can have fewer than 6 carbons. The aryl, homocyclic and heterocyclic groups can have from 3 to 16, generally, 3-7, more often 5-7 members in the rings, and may be single or fused rings. The ring size and carbon chain length are selected such that the resulting molecule exhibits activity as an endothelin antagonist or agonist as evidenced by in vitro or in vivo tests, particularly the tests exemplified herein.

In any of the above preferred embodiments: R 1 and R 2 are preferably selected independently from alkyl, lower alkenyl, lower alkynyl, lower haloalkyl, halide, pseudohalide and H, except that R 2 is not halide or pseudohalide, and in preferred embodiments is also not higher alkyl.

In preferred embodiments: X is S, O, NR 1 in which R" is aryl, hydrogen, or loweralkyl, preferably, a substituted or unsubstituted aryl, particularly phenyl, 10 preferably unsubstituted or substituted with loweralkyl or halogen hydrogen or loweralkyl; R 1 is hydrogen, halide, pseudohalide, loweralkyl or lower haloalkyl, most preferably halide; R 2 is hydrogen, loweralkyl or lower haloalkyl.

The aryl groups are unsubstituted or is substituted with groups such as alkyl, alkoxy, alkoxyalkyl, halogen, alkylenedioxy, particularly methylene dioxy, 15 amino, nitro and other such groups. The alkyl substituents are preferably loweralkyl, more preferably containing 1-3 carbons.

In more preferred embodiments, two of R 9 and R'o are hydrogen, halide or loweralkyl and R 8 is C(O)NHR' 8 or C(O)CH 2

R

18 in which R' 8 is a phenyl group that is substituted at least two positions, most preferably at least one S 20 substitutent at the ortho position and also 3,4 or 4,5 alkylenedioxy substituents. In more preferred of these embodiments X is S.

In all embodiments, R 1 is preferably halide, H, CH 3 or C 2 Hs, and R 2 is H,

CH

3

C

2

H

5

C

2

F

5 or CF 3 In yet more preferred embodiments, R 1 preferably Br, Cl or CH 3

R

2 is H, CH 3

CZH

5 or CF 3 In other embodiments two of R 8

R

9 and R'O form a ring so that Ar 2 is benzo[b]thienyl, benzo[b]furyl, or indolyl, with the proviso that there is one or more substituents and they are other than 5-halo and 3-loweralkyl, and the other of R 8

R

9 and R' is selected from aryl, (CH 2 8

C(O)R

18

CO

2

R'

8

NR'

8

R

9

SH,

S(O),R'

8 in which n is 0-2, HNOH, NO 2

N

3

OR'

8

R'

9

NCOR'

1 and CONR' 9

R'.

Ar 2 may be further substituted with any of the groups set forth for R 8

R

9 and

R'

1 and are preferably selected from among alkyl, alkoxy, alkoxyalkyl, aryl, alkylaryl, aminoalkyl, arylamino, aryl-substituted amino, and NR".

-36- In embodiments in which ET 9 antagonists are desired, it is preferred that

R

8 and R 1 0 are H or loweralkyl and R 9 includes heterocyclic or aromatic ring of preferably from 3 to 14, more preferably, 5 to 7, members in the ring. In particular, if X is S, R 8 and Ro 1 are H or loweralkyl, and R 9 includes an aryl group, particularly a substituted phenyl, such as a 2-loweralkyl substituent. The aryl portion is substituted with groups such as alkyl, alkoxy, alkoxyalkyl, halogen, alkylenedioxy, particularly methylenedioxy, amino, nitro and other such groups. The alkyl substituents are preferably loweralkyl, more preferably containing 1-3 carbons.

10 If X is NR", then R" is aryl, particularly unsubstituted phenyl or substituted phenyl, such as isopropylphenyl.

Other preferred compounds, which are ET, active, are those in which Ar 2 has formula IVB in which R 9 is aryl or Z-substituted aryl, particularly phenyl, and SZ is loweralkyl or loweralkoxy.

15 In all embodiments of all of the compounds herein R' is preferably halide or loweralkyl, most preferably Br, and the compounds are, with reference to formulae IV, 2- or 3-sulfonamides, particularly thiophene sulfonamides. In certain embodiments provided herein, Ar 2 is a benzo[b]thienyl, benzo[b]furyl or indolyl (benzo[b]pyrrolyl) group and the compounds provided herein are prefera- 20 bly benzo[b]thienyl-, benzo[b]furyl- or indolylsulfonamides. Benzo[b]thiophene, benzo[b]furyl and indolyl 2- or 3 -sulfonamides are among the compounds preferred herein. The benzo[b]thiophene, benzo[b]furyl and indolyl 2- or 3-sulfonamides provided herein are selected with the proviso that the benzene group has at least one substituent and that substituent is other than 5-halo and 3loweralkyl.

Compounds of particular interest include those of formula III in which Ar 2 is a phenyl-, benzothienyl, benzofuryl or indolyl [benzopyrrolyll group or in which Ar 2 is a substituted phenylaminocarbonylthienyl, substituted phenylaminocarbonylfuryl, substituted aminocarbonylpyrrolyl group in which there are at least two substitutents or Ar 2 is phenylacetylthiophene, phenylacetylfuran, or phenylacetylpyrrole, is an acetoxystyrylthiophene, acetoxystyrylfuran or acetoxystyrylpyrrole.

-37- The most preferred compounds provided herein have an IC 50 for ETA, receptors in the assays exemplified herein less than 0.1 pM, more prefereably less than 0.01 pjM, and more preferably less than 0.001 (see, Table 1 for representative experimental results), when measured at 40 C, as described in the Examples. When measured at 240 C, the 'C 50 concentrations are somewhat higher to 10-fold; see, Table 1 for some comparative values).

Among the preferred compounds of interest herein are those in which Ar 2 has formula VI: x R 3 2 33

R

R 3 :in which M is (CH- 2 )mC(0)(CH- 2 )r (CH 2 )mC(O)NH(CH 2

(CH

2

)(CH=GH)(CH

2 )r, a. (CH 2 )rnC(O)(CH 2

),NH(CH

2

(CH

2 )m(CH=CH)(CH 2 r' C =N(OH)(CH 2

(CH

2 11 C(O) (CH=CH),NH(CH 2

CH(OH)(CH

2

CH(CH

3

)C(O)(CH

2 )r,

CH(CH

3

)C(O)(CH

2 )m,(CH=CH)(CH 2

(CH-

2

(CH

2 )TO, C(O)O, in which m,s and r are each independently 0 to 6, preferably 0 to 3, more preferably M is

:(CH

2 )mnC(O)(CH 2

(CH

2 )mC(0)NH(CH 2

(CH

2 )m(CH CH)(CH 2

(CH

2 )mC(O) (CH 2 5

NH(CH

2

(GH

2 )m,(CH =CH)(CH 2 C =N(OH)(CH 2 )r,

CH(OH)(CH

2

(CH

2

(CH

2 )4O, COO; R 31 R 32

R

33 R 34 and R 35 are each independently selected from Mi or (ii) as follows: Mi R 31 R 32 R 33 R 34 and R 35 are each independently selected from among H, OH, NHR 3 8 C0NR UR 39

NO

2 cyano, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, alkoxy, alkylamino, alkylthic, haloalkyl, alkylsulfinyl, alkylsulfonyl, alkoxycarbonyl, alkylcarbonyl, alkenylthio, alkenylamino, alkenyloxy, alkenyl sulfinyl, alkenylsulfonyl, alkoxycarbonyl, arylaminocarbonyl, alkylaminocarbonyl, aminocarbonyl, (alkylaminocarbony~alkyl, carboxyl, carboxyalkyl, carboxyalkenyl, alkylsulfonylaminoalkyl, cyanoalkyl, acetyl, acetoxyalkyl, hydroxyalkyl, alkyoxyalkoxy, hydroxyalkyl, (acetoxy)alkoxy, (hydroxy)alkoxy and formyf; or (ii) at least two of R 31

R

2

R

3 3

R

3 and R" 3 which substitute adjacent carbons on the ring, together form alkylenedioxy, alkylenethioxyoxy or alkylenedithioxy

-S-(CH

2 where n is 1 to 4, preferably 1 or which is unsubstituted or substituted by replacing one or more hydrogens with halide, loweralkyl, loweralkoxy or halo loweralkyl, and the others of R 31

R

3 2

R

33

R

34 and R 3 are selected as in and

R

3 8 and R 39 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, haloalkyl alkylaryl, heterocycle, arylalkyl, arylalkoxy, alkoxy, aryloxy, cycloalkyl, cycloalkenyl and cycloalkynyl, and is preferably hydrogen, 10 loweralkyl, loweralkoxy and lowerhaloalkyl, with the proviso that when M is

(CH,

2

)C(O)NH(CH

2 then at least two of R 31

R

3 2

R

3 3 R and R 35 are not hydrogen.

M is most preferably

H

1 N "d 0 nd In general, however, in all of these compounds in which Ar 2 has formula V or VI or in which R 8 includes an aryl group, regardless of the selection of M,it is preferred that the aryl substituent have more than one substituent or at least one substituent in the ortho position. Aryl is preferably phenyl that is preferably substituted at the ortho position and, more preferably at at least one additional position, particularly 4 and 6, or adjacent positions, such as 3,4 or 4,5 when the subsitutents are linked to form an alkylenedioxy (or analog thereof in which one or both oxygens is(are) replaced with S.

In all compounds, at least one of R 31 and R 35 is other than hydrogen.

In more preferred compounds, M is C(O)CH 2 C(O)NH, -CH=CH-, CHCHC(O)(CH),,

CH

2 CHC(O)CH,, and most preferably has formula VII: -39- W R 32 in which W is CH 2 or NH.

:40,10 M is even more preferably selected from among: *Y N 0 0 0 0 sob 0 *in which R 40 is preferably hydrogen, alkyl, alkoxy, alkoxyalkyl, haloalkyl, and more preferably loweralkyl, loweralkoxy, or halo loweralkyl, and is more preferably hydrogen or loweralkyl, particularly methyl or ethyl, and is most pree.bl hydrogen.

M is most preferably: N 09 0 0 an In preferred compounds

R"

1 R1 2 R 1 3

RU

4 and R 3 1 are selected from or (ii): R 31

R

32

R

33 R 34 and R" 5 are each independently selected from loweralkyl, haloloweralkyl, phenyl, alkoxy, loweralkylsulfonylaminoloweralky, cyanoloweralkyl, acetyl, loweralkoxycarbonyl, cyano, OH, acetoxyloweralkyl, hydroxy lowerallkyl, acetoxy loweralkoxy or loweralkoxycarbonyl; or (ii) R 32 and R 33 or R 33 and R 34 form alkylene dioxy, preferably methylenedioxy, and the others of R 31 R 32

R

33

RU

4 and R 3 1 are selected as in *4 *e 9 4 4 S *4S*

S.

4 5 0*6 4 4 *5*4

S

.4.4 4@ 4 p.

4 5 50e4 9 .4 S S S #4 *5 94 @9 In preferred embodiments, R 31 R 33 R 3 r are other then hydrogen and are preferably loweralkyl or lower alkoxy, or R 3 1 or R 35 is other than hydrogen, preferably loweralkyl or lower alkoxy, and R 3 2 and R 33 or R 3 3 and R 34 form methyl enedioxy.

In all embodiments, preferred substituents also can be determined by reference to Table 1, which sets forth exemplary compounds.

Preferred compounds are those of Table 1 that have the highest activities, and preferred substituents are those on the compounds with the highest activites.

TABLE 1 10 COMPOUND ETA(IJM)* ET a(WK.

N-(3,4-dimethyl-5-isoxazolyl)-2- 0.167 16.6 methylbenzo~blthiophene-3-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-2- 0.0486 methylbenzofbjthiophene-3-sulfonamide 15 N-(4-bromo-3-methyl-5-isoxazolyl)-2- 0.0067 5.13 ethylbenzofblthiophene-3-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-2.n- 0.01 82 1 benzylbenzo(blthiophene-3-suifonamide N-(4-bromo-3-methyl-5-isoxazolyl)-2- 0.0226 -3 butylbenzo [bjthiophene-3-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-2-i- 0.005 5.7 pro pyl benzo thiophene-3-sulfonamide 0.03' 10.71 N-(4-bromo-3-methyl-5-isoxazolyl)-2-n- 0.024 7.95 propylbenzo[blthiophene-3-sulfonamide 0.074' 16.61 N-(4-bromo-3-methyl-5-isoxazolyl)-2-(4- 0.048' 11 ethylbenzyl)benzo~blthiophene-3-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-2-[3,4. 0.0015t±0.0014 0.324 ±0.78 (methylenedioxy)benzyllbenzofblthiophene-3-sulfon- 0.0074±t0.00 11' 0.939 t 0.262' amide N-(4-bromo-3-methyl-5-isoxazolyl)-2-(3,4,5- 0.013' 1.2' trim ethoxybenzyl)-benzo (b)-thiophene-3.sulfo namide N-(4-bromo-3-methyl-5-isoxazolyl)-2-ethyl-5- 1.89 0.43 1' 54.3:t 2.6' methylbenzo~bjthiophene-3-sulfonamide N-(4-chloro-3-methyl-5-isoxazolyl)-2-[(3,4-methy- 0.011 0.005' 0.936 ±0.095' Ienedioxy)benzylbenzofblthiophene-3-sulfonamideT PCr/US96I04759 -41- 0 0 0000 *0 *0 0 0 0* *0 0 0* 0 0* 0 0* *0 0 COMPOUND ET,

ET

8 (pM)-

N-(

4 -bromo-3-methy5isoxazoyl)2(3i4. 0.021 ±0.017? 2.94 ±1.32' dimethoxybenzyl) benzo[blthiophene-3-sulfonamide

N-(

4 -bromo-3-methyl5-isoxazolyl)-5-(benzo[blthien- 16? 0.80, 2-yl)thiophene-2- sulfonamide

N-(

4 -bromo-3-methy.5.isoxazolyl).2-(4- .01 1. methoxybenzyl) benzof bjthiophene-3sulfonamide 001 N-(4-bromo-3-methyl5isoxozoli).2-(2- .9 2.2t methoxybenzyf)-benzofbjthiophene-3sufonamide0.9 N-(3,4-d im ethyl- 5-isoxazolyl)-2-.(4- 0.21' 4.7? ch lo robenzyl) be nzof [b thio phen e- 3-sulf onam ideI

N-(

4 -chloro-3-methyI5isoxazoy)2(4- 04.041?1 .3' d im ethyl ami no benzyl) benzo (bthiop hene-3-s u fona- 0.014 0O.477 mide

N-(

4 -chloro-3-methyl.5-isoxazolyj) 2 0.15' 22' ethylbenzo[bjfuran-3-sulfonamide

N-(

4 -chloro-3-m ethyl..5.isoxazoly)2phenyl ben- 0.932' 46.8? zofb~thiophene sufonamide

N-(

4 -ch loro-3-m ethyl 5.isoxazoy) 6methoxy-2 20s" 2.39' 3 4 -(methylenedioxy) benzyll benzo [blthiophene-3 sulfonamide

N-(

4 -chloro-5methyl3isoxazolyl)-2[3,4(methyl- 0.0055' 0.364' enedioxy) benzyllbenzo[bjthiophene-3sulfonamide N-(4-chloro-3-methyl-5.isoxazolyl).2- .315.

methoxycarbonylthiophene3sulf onamide 0615.

N-(4-bromo-3-methyl.s.isoxazoly).4-(4- 0.962' 0.435' propylphenyl)thiophene.2.sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-3 0.0801' 3.68' (phenylthio)thiophene-2sulfonamide (phenylaminocarbonyl)thiophene.2-sufonamide016>10

N-(

4 -chloro- 3 -m ethyl-5.isoxazoly)2[(4-toly) amino- 0.00116 2.93 carbonyllthiophene-3-sulfonamide 0.0 105' 14'

N-(

4 -bromo-3-methyl-5.isoxazolyi)5( 4 8.69 0.363 methoxyphenyl)thiophene-2suffonamide 26.3' 2.4' N-(4-bromo-3-methyl-5-isoxazoly).5-( 3 3.26 0.776 methoxyphenyl)thiophene-2sulfonamide 23.4' 4.7' -42- I

~COMPON

N- f4-b romo -3-m ethyyJl.5 -is oxazoly,)..

5 3 thienl~thiphee2 sulfonmd

N-(

4 -bromo-3-methyI..5.isoxazolyI)- 3 methylthiophene.2.sulfonamide

N-(

4 -bromo3methy-5-isoxazoly,)- 3 amnocarbnyljthiophenesulfonamid

N-(

4 -bromo3.methy5.isoxazoy) dhI (pheneylthiophene2sufonamide 10 .brm-dimethy.5.isoxazoy) 2 4mtypey meh!heyaminocarbonyljthiophene3-lfoamd p.:h ienlhohn--ufnmd

N-(

4 -bromo3methyf5isoxazolyl)- 2 .hydroxybenzyl)thiophene3sufonmde

N-(

4 -bromo-5-methyl.3.isoxazoy) 5 4 methylphenyl)thiophene-2sulfonamide *:20 N-(4-.bromo..3-methy,..5..soxazolyl).5-

N-(

4 -bromo.3methyI5isoxazoy) 5 4 (trifiuorom ethy) phenyljthiophene2sulfn de N,N'-bis 3 -f(4-bromo-3-methy..5.

isoxazolyl)aminosulfony)then2-y urea

N-(

4 -bromo.-3-methy5isoxazolyf)- 2 NN'-bis{3-f3,4-dimethy..5 aminophenyl)thiophene2sulf onamide bis (trifourometh) henvflthiphen 2 ffo ide a. a. COMPOUND ETA ET* gB N-(4-bromo-3-methyl-5isoxazoly)5(3,3- 1.12 24.0 dimethylbutyn-1 -yl)thiophene-2-sulfonamjde 7.24' 35 N-(4-bromo-3-methyl.5isoxazolyl).5(2. 0.38 1 1.097 methoxyphenyl)thiophene-2 sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl).5-(2. 0.432 0.31 3 tolyl)thiophene-2-sufonamide

NL-(

4 -bromo-3-methyI-Bisoxazoly)-.2[(3- 0.062' >1loo t carboxyphenyl)aminocarbonyllthiophene3sulfonamide N- 4 -bromo-3-m ethyl- 5-isoxazolyl)-24f2 -0.21' carboyxyiphenyl)aminocarbonyi-thiophene-3sulfonamideI

N-(

4 -bromo-3-methy-5-isoxazolyi)-2 0. 84? 100? (aminocarbonyl)thiophene-3sufonamide N-(4-bromo-3-methy..5isoxazoly).2.(5dimethyl am ino- 1 -naphthyl) sulf onyl- 0.97' 3.9? aminocarbonyllthiophene-3sulfonamide

N-(

4 -bromo-3methyl5isoxazoy)5(5-methy-2- 17' 0.21' thienyl)thiophene-2-sulfonamide N-(4-brom o- 3methyl5 isoxazolyl)2-f(3,4- 0.017' 9.8' m ethylened io xyp henyl) am inocarbonyllthio phene-3 sulfonamide

N-(

4 -bromo-3methy-5isoxazoyi)2-(3,4-.03 m ethylenedio xy) pheno xycarbo nyl Ithiop hene3su 0073 fonamide N-(4-bromo-3-methyl-5isoxazolyi)..3.[(3,4 0.50, 79' methyienedioxy)phenyljthiophene-2sufonamide N-(4-bromo-3-methyl-5-isoxazolyt)..3.((3,4 8.11 3.2' m ethylenedioxy) benzyllthiophene-2-sulf onamide N-(4-bromo-3-methyl-5isoxazolyi)-3 1.6' 39' benzylthiophene-2-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-5(3- 15' 4.2' tolyl~thio phene- 2-sulffonamide N-(4-bromo-3-methyI-5isoxazoy)2[(3,4 0.27' 7.7' m ethylenedio xy) benzyllthiophene-3-suffonamide N-(4-bromo-3-methyl-5-isoxazolyi)-2.[(3,4 2.0' methylenedio xy) benzoyl )thiop hene.3-su lfon amid e -44- I

COMPOUND

N-(4-bromo-3-methyI..5.isoxazoyi) 2 2 hydroxyphenyl)aminocarbonylthiophene3sulfona mide N-(3,4-dimenthyi..5.isoxazoy).2f 3 (methylenedioxy)phenoxycarbonylthiophene 3 sul fonamide

N-(

4 bromo3methy5isoxazoy)5(5ethythn 2 yI)thiophene-2- sulfonamide

N-(

4 -bromo-3-methyi.5.isoxazoy) 2 t( 3 ,4- 10 methylenedioxy)benzoyjaminocarbony ,Jtohnh 3 *sulfonamide

N-(

4 -chloro-3-methyI..5.isoxazolyl)..2.[ 3 4 (methylenedioxy)phenoxycarbonyllthiophene 3 slfonamide .*915

N-(

4 -bromo-3-methy-5-isoxazolyl) 5 1- Ietyylthiophene.2-sulfonamide 4 chloro3methyl5isoxazolyl)5(5ethlthin 2 *yl)thiophene-2- sulfonamide

N-(

4 -bromo-3-.methyl.5.isoxazoyi)- 2 t( 3 4 methylenedioxy)phenyaceyllthiophene 3 slfoa mide

S.N-(

4 -bromo.3-methyl..5.isoxazoli)- 2 134- *9(methylenedioxy)phenoxycarbonylaminolthiophene

N-(

4 -bromo-3methyl.5isoxazoyi) 2 2 -choo3,4 methylenedioxy)cinnaxmetyltphenene-3-sunam n

N-(

4 bromo3-methyl5isoxazoyl)5(1trnsphthI) mthyenedioxsulfonamyde ohn-3slonmd N-(4-bromo-3-methyI..5.isoxazoiy) 5 npty nitrophenyl)thiophene-2-sufonamide N-(4-bromo-3..methyl.5..soxzy) 2 f( 3 4 1methylenedioxy)phenylureidothiophene 3 suffo mide

N-(

4 -chloro-3..methyl..5.isoxazolyl)..

2 3 4 (methylenedioxy)phenylacetylthiophene3sulfona mide I

COMPOUND

N-(4-bromo-3-methyl5isoxazoly) 5 4 methyoxycarbonylphenyl)thiophene-2sulfonamide N-(4-bromo3methyI.5soxazoly)5( 4 carboxyphenyl)thiophene.2-sufonamide N-(4-bromo-3-methyI..5.isoxazoy) 5 4 tolyl)aminocarbonyl)thiophene-2-sulfonamide S 4 -bromo-3-methyi.5.isoxazoly)5( 2 5.55methyfuranyl)thiophene-2sulfonamide

N-(

4 -choro3methyI5.isoxazoyi)2f 3 4 10 (methylenedioxy) benzyloxycarbonylthiophene3sul.

:fonamide

N-(

4 -chloro-3-methy5isoxazoy)2[ 2 3 4 methylenedioxyphenyl)]ethoxycarbony,3-sulfonamide

N-(

4 -choro-3methy5isoxazoy) 2 4 3 4 S. *methyienedioxybenzy!)piperazin-1 ylJcarbonyllthiophene-3-sulfonamide

N-(

4 -bromo-3-methyI..5.isoxazolyl)a.N-( 4 -chloro-3-methyI..5.isoxazolyi)2{l -cyano- 1- 3 4 -methylenedioxy)phenyllacetyIlthiophene-3sul fonamide N-(4-bromo3methy5.isoxazoiy)2l( 3 4 methylenedioxy)phenethyllthiophene-3suffonamide

N-(

4 -chloro-3-methyI..5.isoxazolyl)-2 f3dimethylamno)phenoxycarbonylthiophene3sulfonamide

N-(

4 -bromo-3-methyI5isoxazoy)-l -methylindole-2sulfonamide

N-(

4 -chloro-3-methyI..5-isoxozoly-2- (cyclohexyloxycarbonyl)thiophene-3sulfonamide

N-(

4 choro3methy5isoxazoyi).2B-dx( 3 4 methylenedioxy)phenylethylthiophene 3 slfoa mide N-4boo3mty--soaoy)2croy- methylidole-3-suffonamide -46- COMPOUND ETA(WM).

JES_(YM)'

N-(4-chloro-3-methyl-s..isoxazoly).2.fpj.

oxacyclohexyl)oxycarbonyllthiophene.3suffonamide 1.37' 1.37? I

S

S

S.

S

4 -(romethyl..isoxyazoylcty2f( 4 hne3 methylphieoycroyjhohn~ufnmd

N-(

4 -chloro-3-methyI..5.isoxazoiy)(2[4(methyeny)peyllopeet ulfohjesufamd

N-(

4 -cbroo3-methyl5.isoxazoyl)-23f(4mtylpheinoxyametyl mthihne sulfonamide

N-(

4 -bhroo3-methyl..5.isoxazolyl)-2f( 4 mehhnmethethy~aryllthiophene3 sulfonamide 1.8' 32.5' 31.3' 14.7' 6.023' 5.29? 18.6' 122' 9.7' 0.043' 10. 1' 1.64? 22.8'1

N-(

4 -bromo-3-methy;..5.isoxazoly)3(4-methyltrn-trltiphn--ufnmd

N-(

4 -bromo-3-methyI..5.isoxazoyi)3(4-methylphenethyl)thiophene-2-sulfonamide

N-(

4 -bromo-3-methyl.5.isoxazofyl).2 1(4-methyl- 8 mjd N-(4-bromo-3-methyl5.isoxazoly).2-[( 3 methoxyphenyl)acetyllthiophene-3sufonamide

N-(

4 -bromo-3-methyl..5-isoxazoyl)3(4-methylphenethyl)-5-(4-tolyl)thiophene.2-sulfonamide

N-(

4 -bromo-3-methyl-5.isoxazolyi) 3 4 methylbenzyl)..5(4tolyl)thiophene-2sulfonamide

N-(

4 -bromo-3-methy;..5..soxazoy)3(4-methyltrans-styl)--(4-toly)thiophene-2-sulfonamide 0.94' 0.347' 9.4' 0. 198' 9.13' 0.030' 19.1' 6.1' 4.69' 6.9' 2.09' 1.56' 1.58' -47-

U

U.

U U

U.

U

U.

U

U

U.

U

U.

U

U..

S

a. U U

U.

U. U U U

S.

COMPOUND ETA (IyM)- ET, (pM).

N-4clr--ehl5ioxzli--8g(tyee 0. 128' 2.091 dioxy)- 3 ,4-(methyenedioxy)phenethyflthiophene-3 sulfonmd sufonamide

N-(

4 ych2o[g d ethyl-..ioaoy){hdo 20.? 310 N-{ 4 -chloro-3methyl5isoxazolyl)-2-t(dmethyl 0.56 92'0 ai-34-(methylenedioxy)phennoaoyflthiophene- 3 sulfonamide N 4 -b lrom o 3 m ethyl. 5 isoxazo y).

3 a yd o y -2 5 0 [(metthylenedioxy)phenyllaetyl~thiophene-3l 053?96 fonamide

N-(

4 -bromo-3-methyl5.isoxazoyi)2[( 4 methyl)(cinnamyl)] thiophene-3-suffonamide N-(4-chloro-3-methyl.5.isoxazolyl)..

2 4 ,5 dimethoxy-2-methoxycarbonylphenyl) aminocarbonyllthlophene-.3.suffonamide

N-(

4 -chloro-3-methyl..5.isoxazoyl)2[(2-methyl- 1, 3 4 -thiadiazo-5-yl)aminocarbonyllthiophene3sulfonamide

N-(

4 -chloro-3-methyl.5.isoxazolyl) 5dimethoxy-2,4,5..dimethoxy2methoxycarbonyl)phenyllphenylaminocarbonyllthiophene3sulfonamide

N-(

4 -chloro.3methyl.5isoxazoly) 2-{f2-carboxyl- 4 ,5-(methylenedioxy)phenyllaminocarbonyqthiophene-3-sulfonamide

N-(

4 -bromo-3-methyl-5.isoxazolyl)-3 [3,4 (methylenedioxy)phenethyllthiophene-2sufonamide

N-(

4 -bromo-3-methyl.5.isoxazoly) 3 f 3 4 (methylenedioxy)..ransstyryllthiophene-2sufonamide 0.26' 0. 413' 0.55' 0.13' 3.80' 1.43' 0.236' 1 18' 0.218' -48- (4bo3meth~indoypyljoamolinocarbonylthioph 3 shenamylide ee3sufnmd N-(3 4 -dchylo.3omethyi)-2isoxactyl-24meth (methylpenedyl)pnaminocarbonylpheeesujfon ***mide

N-(

4 choro3methy5.isoxazoy) 2 2 co 4 5 10 dimethoxyphenyl)aminocarbonyjthiophe 3 sffn d N-3 4d meh l5is x zla 2 -olactlp e y) tho.. *-uloamd N-34di.h a.sxzoy)2[34( atyee to. amd

N-(

4 -chloro-3..methyI..5isoxazoli).

2 2 4 doy4 dmethoxheyphemnicarbinylcarbophne-3-shiophe to 0.mide

N(

4 choro.3methy..5-ioxsoyl-2zo2yano-4,5- (mridyfenminocarboenylamnroy)thiophene-3-

N(

4 choro3methyisoxazoyzo-2 )2fehl 4 ~.methyleeiyphenylaminocarbonylthiophene-3 a a mide d 4 choro3.methyi..5.soaoiyI)2- 2 (meth ido-enediy hphenyloamibollh cabnltpee3sulfonamide N-(3 4 -hlorohy.methy..5xaioxai)- 2 2 carbonhelceyl)thiophene -3sulfonammde

N-(

4 chloro3.methy.5.isoxazoyl I2( 2 4 diehyl

N-(

4 -bromo-3-methyl.5.isoxazolyl) 2 2 4 dim ethylphenylacetyl)thiophen3sulfide -49- *9 .9 9 9.99 9* a a 909.

a.

S

9 a.

9 99 *9 10 15 20 COMPOUND ETA_(PM)- ET 5 (.UM) N-(4-chloro-3-methy-5..isoxazoly,)-2-[3,4. 0.0073t 9.2' (methylenedioxy) ]phenylaminocarbonyl-3thiophenesulfona mide

N-(

4 -chloro-3-methyl-5isoxazoy).2.2..methy..4,5. 0.0032' 9t (methylenedioxy) phenytacetyllthiophene-3 sulfonamide N-(4-chloro-3-methyI..5.isoxazoly)2.(3, 4 05 25.7' (methylenedioxy)-6(2acetoxyethyl)phenylamino 0005 carbonyIlthiophene-3-sulfonamide N-(4-chloro-3-methyI5-isoxazolyl)-2[3,4- .056 16.8' (methylenedioxy)-6-(2-hydroxyethyl)pheny .056 aminocarbonyllthiophene-3.sulfonamide

N-(

4 -chloro-3-methy.5isoxazolyi)-2.(3,5-dimethy- 0.045' 17.7? phenylacetyl)thiophene-3sulfonamide

N-(

4 -chloro-3-methyI.5..isoxazoy)2(2,5 0.007' 8T diehypen1 8t.hipen--sfonmd

N-(

4 -chloro-3-methyI-5-isoxazolyl)..2[2- 0.0068' 1 9.8' methanesulfonylaminomethy)4,5 (methylenedioxy)phenylaminocarbonyllthiophene-3 sulfonamide

N-(

4 -chloro-3methy.5-isoxazolyI)-2([2. 0.0038? cyanomethyl.4,5-(methylenedioxy).6-cyanomethylJphenylaminocarbonyl.3-thiophenesulfonamide

N-(

4 -chloro-3-methyI5isoxazolyl)-2.[2-hyroxyproy- 0.0073? 8.3? 4, S-(methylenedioxy)phenylaminocarbonyl..

thiophene-3-sulfonamide

N-(

4 -bromo-3-methyI5isoxazoy)3[2..methyI-4,5 -0.1 -6" (methylenedioxy)cinnamyflthiophene.2-suffonamide

N-(

4 -bromo3-methy-5-isoxazolyl)..3.f2methyl4,5 1 (methylenedioxy)phenethyllthiophene-2sufonamide

N-(

4 -bromo-3-methyl5isoxazoy)3{[2..propyI- 4 ,S (methylenedioxy)phenoxyjmethyllthiophene-2 sulfonamide

N-(

4 -chloro-3-methyi-5.isoxazolyl)-2 [3,4 (methylenedioxy)-6-(2..acetoxyethoxy) lphenylaminocarbonyllthiophene-3-suffonamide

N-(

4 -chloro-3..methyI.5isoxazoly)2[3, 4 (methylenedioxy)-6..(2..hydroxyethoxy) phenylaminocarbonyljthiophene-3sulf onamide -0.02'~ -18,

-I

-0.01 -18, L L 0*

C

0* C

C

a a a I COMPOUND ETA

IET

3

(M

N-(

4 -chloro-3-methyl-5..isoxazolyi)..2.2cyano4,5 (methylenedioxy)phenylacetyglthiophene-3 sulfonamide

N-(

4 -chloro-3-methyl.5.isoxazolyl).2(2- 0.009, 13.8' f(dimethylamino)carbanygmethyl,]4 dioxy)phenylaminocarbonyllthiophene.3 Sulfonamide

N-(

4 -chloro-3-methyl-5.isoxazolyl).2..2methyI 4 ,5 0.794' 6.49' (methylenedioxy)phenylhydroxyiminolthiophene-3 sulfonamide

N-(

4 -bromo-3-methyl5..isoxazoly)2[2methyl- 4 0.0619 t 8.90? (methylenedioxy)phenethyllthiophene-3sulfonamide

N-(

4 -bromo-3-methyl..5.isoxazolyl)3f2- 0.0795' 3.24? (methylenedioxy)cinnamyljthiophene-2sufonamide

N-(

4 -bromo-3-methyl5-isoxazolyl)3{2f[(tetrahydro- 0.0967' 4.14 4 H-pyran-2-ylxoy)methyll-4,5.

(methylenedioxy)cinnamyllthiophene-2sulfonamide

N-(

4 -bromo-3methyl5.isoxazoly)3(2, 4 .0 4.30t dimethylphenethyl)thiophene.2-sulfonamide0.06

N-(

4 -bromo-3-methy..5-isoxazoly).3-(2,4-.10 2.97? dimethylcinnamyl)thiophen-2-sufonamide 010

N-(

4 -bromo3methy5isoxazoly)2(2, 4 -016 2.97? dimethylcinnamyl)thiophene-3sulfonamide 016

N-(

4 -bromo-3-methyg..5.isoxazolyl)3[( 2 4 -036 7.45? dimethylphenoxy)methyllthiophene.2-sulfonamide 036

N-(

4 -bromo3methyl5isoxazoly)22, 4 -038 4.48? dimethylphenoxy)methyljthiophene-3-sufonamide 038 N-(4-chloro-3-methyl-5.,soxazolyl).5- 8..606 (phenylaminocarbonyl)thiophene.2-sulfonamide 2. 06

N-(

4 -chloro-3-methyl..5.isoxazolyl).2-LBacetoxy-2 methyl-4, S-(methylenedioxy)styryljthiophene-3 suffonamide 0.00544 3.74'

N-(

4 -chloro-3methyl5.isoxazolyl)2[( 2 3 4 0.000169? 12.5? trimethoxy-6-cyano) phenylaminocarbonyljthiophene- 3 -sulfonamide

N-(

4 -chloro-3methy5isoxazolyi)2[ 2 .3 .2 (cyano)phenyllbenzo[bjthiophene-3sufonamide 6.3882 -51- .0S COMPOUND ET, ET 8 (UM).

N-(

4 -chloro-3-methyl-.5.isoxazolyi)..2.(34-(methy.. 0.550' 52.6? enedioxy) phenyllbenza[blthiophene-3-sulfonamide N-(4-bromo-3-methyl..5.isoxazolyl)-3-(2. 0.324t 55.1? tolyl)thiophene-2-sulfonamide N-(4-bromo-3-methylg5isoxazolyl).3.(3- 0.832? 21 .2' tolyl)thiophene-2-sulfonamide N-(4-bromo-3-methy..5isoxazoy).3(2. 0.302' 31 00t tolyl)thiophene-2-sulfonamide

N-(

4 -bromo-3-methy-5isoxazoy).3(3. 0.334' methoxypheny)thiophene2sulfonamide N-(4-bromo-3-methyl-5isoxazolyi).3-(3- 1.32' 56.3' methoxyphenyf)thiophene-2-sulfonamide N-(4-bromo-3-methy..5-isoxazolyI)-3(2. 1.71' 59.1' methoxyphenyl)thiophene-2sulfonamide

N-(

4 -bromo-3-methyl-5.isoxazoly!).3-(4. 0.184 43.9' ethyl phenyt) thiophene-2sulf onamide N-(4-bromo-3-methyl.5.isoxazolyl)-3(4- 0.0873 8.48' pro pyl phenyl)thiophene-2sulfonamide N-(4-bromo-3-methy..5isoxazolyl)..3..(4-so. 0.218 28.3' pro py p henyl) thjophene-2suIf onam ide

N-(

4 -bromo-3-methyl-5isoxazolyl).3(4- 0.160 6.11'1_ butyl phenyl)thiophene-2-sulf onamide-

N-(

3 4 -dimethyl-5-isoxazolyI)..2.[2..methyI..4,5 0.00328' 34.3' (methylenedioxy)phenylacetyl]thiophene-3 sulfonamide N-(4-chloro-3-methyl-5.isoxazolyi)..2.(24,6 0.000626t 8.27' trimethylphenylaminocarbonyl)thiophene-3 sulfonamide

N-(

4 -chloro-3methyl5isoxazolyl)-2-(2,4,6tri- 0.000238' 3.82' m ethyl phenylacetyl)thiop hene3sulfonamide

N-(

4 -chloro-5-methyI-3isoxazoly)-.2..(2.methylI4,5- 0.000625' 3.69' (methylenedioxy)phenylacetyllthiophene-3 sulfonamide

N-(

4 -bromo3-methyl5soxazoly).2[2methy;4,5 0.0804' 3.28' (methylenedioxy)cinnamylthiophene3suffonamide N-(4-bromo-3-methyl-5.isox azolyi)2(2,4 0.0555' 3.48' dime hylphenethyl)thiophene-3-sufonamide -62- C. CC

C

4

C

p.

0*

C

CC. C C. C* C C

CC

4. C P.4.

'CCC

a. C S

C.

C. C C C

CC

COMPOUND EA ETg (pM)' N-(4-chloro-3-methyl..s.isoxazlyl)2p[4 0.000266' 9.78' methoxycarbanyl.2, 6-dimethyl)phenylaminocarbonyllthiophene3sufonamide N-(4-chloro-3-methyl.5isoxazoly).2 4.41' 31 %@1001 (phenoxycarbonyl)thiophene3sulfonamide N-(4-bromo-3-methyl.5isoxazoly).2 2.71' 20%@1loot (phenoxycarbonyl)thiophene3sufonamide N-(3,4-dimethyl.5-isoxazolyl)..2{[3,4- 3.61' 30%@ 1 Do" (methylenedioxy)phenoxylcarbonyllthiophene-3.

sulfonamide N-(4-bromo-3-methyl.5..isoxazoly)2[(2- 0.684' 105? methylphenoxy)carbonyllthiophene-3-sufonamide N-(4-bromo-3-methy..isoxazoy)21(3- 1.20' ill, methylphenoxy)carbonyljthiophene-3sulfonamide N-(4-bromo..3.methyl-5-isoxazolyl)-24[(2,4. 0.291' 43.2? dimethylphenoxy)carbonyllthiophene-3sulfonamide N-(4-bromo-3-methyl.5isoxazolyl).2.(2. 0.761' 29%@1l00' methoxylphenoxy)carbonyllthiophene-3sufonamide N-(4-bromo-3..methyl-5-isoxazolyi)-2-[(3- 0.78' methoxylphenoxy)carbonyllthiophene-3.sulfonamide N-(4-bromo-3-methyl..Sisoxazolyl)-2.(4- 1.73' ill, methoxylphenoxy)carbonyllthiophene-3sulfonamide N-(3,4-dimethyl-5-isoxazoly).2.I(4.. tO.324 68.5' methoxylphenoxy)carbonyllthiophene-3sufonamide N-(3,4-dimethyl-5-isoxazolyl)..2-(4- 0.324t 68.5' methytphenoxy)carbonyllthiophene-3sulfonamide N-(4-chloro-3-methyl..5.isoxazolyl)2.j(4- 2.52? 19 100' methylphenoxy)carbonyflthiophene.3-sufonamide N-(4-chloro-3..methyl..5.isoxazolyl)..2-(2,4.. 3.22' 43 100' dimethylphenoxy) carbonyllthiophene-3..sulfonamide N-(3,4-dimethyl5-isoxazoly)2(2,4 0.648? 68.5? dimethyiphenoxy) carbonyllthiophene..3.sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-2.{f2-propyI.4, 5- 0.274' 21 100' (methylenedioxy)phenoxylcarbonylthiophene-3sulfonamide N-(4-chloro-3-methyl-5-isoxazoly).2(3- 0.138' 11.91 methoxycarbonyl-2,4,6trimethylphenylamino.

carbonyl)thiophene-3-sulfanamide -53do 9*06e a *0 0S 0 *000 4 04 4 00 4.

0e 0 00 4* COMPOUND EA ET 8

(UM).

N-(4-bromo-3-methyl.5-isoxazoy)3(24 0. 1002' 60.3' dimethylphenyl)thiophene-2-sulfonamide N -(3,4-dimethyl-5-isoxazolyl).2 2.85' 31 (phenoxycarbonyf)thiophene-3sulfonamide N-(4-bromo-3-methy-5isoxazoyi)3(4-..so. 0.0823' 2.76' butylphenyl)thiophene-2-sulfonamide N-(4-bromo3-methyl5-isoxazolyl)3(4-1so 0. 155' 3.31' pentylphenyI)thiophene-2-sulfonamide N-(4-bromo-3-methy-5-isoxazoly).3[(2,4,6. 0.0457' 4.68' trimethylphenoxy)methyllthiophene2sulfonamide N-(4-bromo-3methyI5isoxazoyi)-2[(24,6 0.0562' 3.39' trimethylphenoxy)methylthiophene-3sulfonamide N-(4-bromo-3-methylBisoxazolyl)3(2,4,6. 0.0490' 1 .86' trimethylcinnamyl)thiophene-2sulfonamide

N-(

4 -bromo- 3-m ethyl- 5isoxazoy)3(2-methyl-4- 0.0468' 3.63' pro pylphenyl) thiop hene-2-sulfo namide N-4boo3mty--sxzll--4iobtl2 0.0468' 1 .66T methylphenyl)thiophene-2-suffonamide

N-(

4 -bromo3methy;5isoxazoyi)3(4-iso-pentyl2- 0.107' 2.40' rnethylphenyl)thiophene2sufonamide N-(4-bromo-3-methyl-5-isoxazolyl)2{t3,4- 0.302' 6.6 1' (methylenedioxy)phenoxylmethytlthiophene3.

sulfonamide N-(4-bromo-3-methy-i.Isoxazolyi)..2..I4,5 0.107' 0.407' (methylenedioxy)-2-propylphenoxylmethy~thio.

phene-3-sulfonamide N-(4-bromo-3-methyl-5-isoxazoi)..2.(24,6 0.0417' 1.23' trimethyfphenethyl)thiophene-3sulfonamide N-(4-bromo-3-methyl5isoxazolyi)3(24,6 0.055' 1.62' trimethylphenethyl)thiophene-2sulfonamide N-(3,4-dimethyl-5-isoxazolyl)2..(2,4,6 0.537' 8 100' trimethylphenoxy)carbonyllthiophene-3sulfonamide N-(4-chloro-3-methyl-5isoxazoyi)2[(2,4, 6 0.0776' 30.2' trimethylphenoxy)carbonyllthiophene-3sufonamide N-(4-bromo3methy5isoxazoyi)-2(2 4 6- 0.479' 24.5' trimethylphenoxy)carbonyllthiophene-3sulfonamide -54results are generally the average of 2 to 5 experiments S preliminary results or results in which one or more data points were only determined approximately assay performed with incubation at 240 C. As described in the Examples, incubation at the higher temperature reduces the activity by a factor of 2- to about 10-compared to the activity at 40 C data not available or measured as inhibition 100 pM inhibition 100 pM It is understood that 4-bromo or 4-chloro groups can be replaced by other 10 4-halo substituents or other suitable substituents for such as alkyl.

2. Ar 2 is a substituted 4-biphenyl group Compounds of formulae I in which Ar 1 is N-(5-isoxazolyl) or N-(3isoxazolyl) in which Ar 2 is selected from biphenyl derivatives. These compounds can be represented by the following formulae (VII): R' R R R 20 so- or so,- N SH

H

R"

0* 25 in which R 26 and R" are each independently selected from H, OH, OHNH, NH, 2

NO

2 halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heterolaryl, alkoxy, alkylamino, dialkylamino, alkylthio, haloalkoxy, haloalkyl, alkylsufinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsufinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, carbonyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, in which the alkyl, alkenyl and alkynyl portions contain from 1 up to about 14 carbon atoms, preferably from 1 to 6 atoms, and are either straight or branched chains or cyclic, and the aryl portions contain from about 4 to about 16 carbons, preferably 4 to 10 carbons. R' 3 and R 26 are preferably each selected from from H, loweralkyl, haloalkyl and halide. Again, it is understood that Ar 2 may be substituted with more than one substituent, each of which is selected independently from the selections set forth for R 26 and R 1 3 and R 2 and R' are as defined above.

In the embodiments herein, the biphenylsulfonamides are substituted 4biphenylsulfonamides,

R

1 3 is preferably at the para position and R 26 if it is not hydrogen, is at any position except the 2-position.

In more preferred embodiments, R' is halide or methyl or higher (Cs-C, 3 alkyl. R 1 is selected from halide, CH,, C 2 5

CF

3

C

2 F, n-C 3 H, and cyclo-C 3

H,,

preferably halide or CH 3 and R 2 is selected from H, CH 3

C

2

H

5 CF3, C 2

F

5 n-

C

3 H, and cyclo-C 3 more preferably R 1 is halide or CH 3 and R 2 are selected from H, CH 3

C

2 Hs, or CF3.

In more preferred embodiments, R' is Cl or Br, or if greater ET, activity is 10 preferred a higher alkyl (C 9

H

19 to C 13

H

27

R

2 is selected from H, CH, C 2 H CF 3

C

2 Fs, n-C 3

H

7 cyclo-C 3 nC 1 3

H

27 and nCH,, In yet more preferred embodiments, R 1 is Br, Cl or C 9

H,

1 to C 13

H

27

R

2 is H, CH 3

C

2 Hs, or CF 3 The biphenyl compounds provided herein are generally ET, active or ET, selective (see, Table i.e. the compounds provided herein inhibit binding 15 of endothelin to ET e receptors at concentrations about 10- to about 30-fold less than they inhibit binding of endothelin to ETA receptors. In particular the 4biphenylsulfonamides are ET, selective.

In general in all embodiments herein, 4-haloisoxazolyl sulfonamides exhibit substantially enhanced activity with respect to at least one of theET receptors (about two-fold to twenty-fold greater activity), as assessed by assays, such as those provided herein, that measure binding to ETA and/or ET, receptors, compared to corresponding sulfonamides in which the substituent at the 4 position in the isoxazolyl is other than halo, such as alkyl. For example: the ICso of N-(3,4-dimethyl-5-isoxazolyl)-2-biphenylsulfonamide for ETA receptors is about 0.008 pM, whereas, the ICso of N-( 4 -bromo-3-methyl-5-isoxazolyl)-2biphenylsulfonamide is about 0.0016 pM (see, Table below); and the ICso of N-(3,4-dimethyl-5-isoxazolyl)-3-biphenylsulfonamide for ET receptors is about 3.48 pM; whereas, the ICso of N-( 4 -bromo- 3 -methyl-5-isoxazolyl)-3-biphenylsulfonamide for ET, receptors is about 0.76 pM and the ICso of N-(4-chloro-3methyl-5-isoxazolyl)-3-biphenylsulfonamide for ET, receptors is about 0.793 pM (see, Table below).

Exemplary biphenyl sulfonamides are the following and those set forth in Table 2, and include, but are not limited to: N- 3 -m ethyl- 5 isoxazolyl)4'-methyl phenyl-4biphenylsulf onamide, N(4-bromo-3methyl-5-isoxazoly)4'..methylphenyl-4biphenysulfonamide, N-(4-chloro-3m ethyl- 5 isoxazolyl) 4'methylp henyl-4bip he nylsu fonamide, isoxazolyl)-4'-trifluorophenyl-4biphenylsulfonamide, 4 -brom o-3-m ethyl- isoxazolyI)-4'-trifluorophenyl4biphenylsulfonamide, 3 -methyl-5-isoxazolyl)4' methyoxyphenyl-4-biphenylsulfonamide, (4-bromo-3-methyl-5-isoxazolyl)-4'.

methoxypheny-4-biphenylsulfonamide, 4 -bromo-3-methyI-5-isoxazolyl)-3 rnethoxyphenyl-4-biphenylsulfonamide, 4 -bromo-3-methyl-5-isoxazolyl)2 Smethoxyphenyl-4-biphenylsulfonamide, N- (4-brom o-3-m ethyl- 5isox azolyl)-.3 :methylenedioxyphenyl-4biphenylsulfonamide and (4-bromo-3-methyl-5- *:*isoxazolyl) -3 ethyl phe nyl-4-biphe nylsuIf on amide. Corresponding 4-chloro and 4-fluoro isoxazolyl compounds are also encompassed herein.

Exemplary biphenyl compounds were tested using the exemplified assays (see, EXAMPLES) and the results, which are intended to be exemplary or provided for comparison with compounds provided herein, and are not limiting, are as set forth in the following table (Table 2): :TABLE 2 COMPOUND EA ETR (jPM)*

N-(

4 -bromo-3-methyk5-isoxazoly)-4biphenylsu.. 3.3 -0.17 fonamide 49t 1.23?

N-(

4 -bromo-5-methy.3.isoxazolyl).4biphenylsu.. 6.4t± 2 0.29 0.02 fonamide 491 1.78T N-4clr--ehl5ioaoy)4bpeysl 4.93:t 3 0.29 ±0.1 fonamide N-(3,4-dimethyl-5-isoxazolyl).4- 9.9±t1.4 0.77 .32 biphenyisufonamide 6.3? 0.

N-(

4 -chloro-5-methy;-3-isoxazolyl)-4biphenylsul- 3 .7 0.23t±0.01 fonamide 18.6? 1.29?

N-(

4 -Methy-3trifluoromethyl5isoxazolyl)-4 19.0 1 .7 biphenyisulfonamide 5.621

N-(

4 .Tr decyl-3..xrifluoromethyl..sisoxazoly).4 34.0 9 0.99 0.2 biphenylsulfonamide 33.0? 0.95? a.

a a a 5 10 COMPOUND ETA N-(3,4-dimethyl-5-isoxazolyl).2-biphenylsulfon- 0-0083:±0.0014 12.8 amide

N-(

4 -bromo-3-methy(-5-isoxazolyl)-2biphenyisul 0.001 27- 8.54** fonamide

N-(

4 -chloro-3-methyi-5-isoxazolyl)-2-biphenlsuffon. 0.001 23* 1 4 amide

N-(

3 4 -dimethyl-5-isoxazolyl)..3.biphenylsulfon. >0.03- 3.48~ amide

N-(

4 -bro mo-3-meth.y-5..isoxazo lyl) -3biphenylsul O.03** 0.76* fonamide

N-(

4 -chloro-3-methyl-5-isoxazolyl)-3biphenyisul 0.793fonamide N-(4-bromo-3-methyl-5-isoxazolyl)-4'. 14.53 ±9.6 0.046 0.044 methylphenyi.4-biphenylsulfonamide 22.17 3.771 0.168:i±0.0032' N-(4-bromo-3-methy..5isoxazolyi).4' 5.4 0.3 0.083 ±0.02 trifluorophenyl-4-biphenyisulfonamide 25.9t± 13.71 0.71 ±0.431 N-(4-bromo-3-methy-5.isoxazolyi).4'. 14.7 ±5.6 1.15 0.44 methoxyphenyl.4-biphenylsulfonamide 121.5 ±2.121 3.94 ±0.891 N-(4-bromo-3-methyl.5.isoxazoiyl)-3'. 4.97 ±3.4 0.66 0.25 methoxyphenyl-4-biphenylsulfonamide 162.6 ±t7.14? 2.08 0.23? N-(4-bromo-3-methyl-5.isoxazolyl).2'. 3.3 3.5 0.41 0.14 methoxyphenyl-4-biphenylsulfonamide N..(4-bromo-3-methy-5..isoxazolyi)-3',4'. 38.2 4.951 3.0 0.78? methylenedioxyphenyl.4-biphenysulfonamide N-(4-bromo-3-methyl-5-isoxazolyl).3'- methylphenyl-4-biphenylsulfonamide *results generally from 1, 2 or 3 experiments with the same preparation **preliminary results Preferred compounds are those in which Ar 2 is a 4-biphenyl in which, referring to formula VII and at least one substitutent R1 3 is at the para position. Preferred substitutents are loweralkyl, halo loweralkyl and lower alkoxy. Such compounds are ET, active.

The preparation of the above and other compounds that possess the requisite acitivities are set forth in the Examples.

-58- B. Preparation of the compounds The preparation of some of the above and other compounds that possess the requisite acitivities are set forth in the Examples. Compounds whose synthesis is not explicitly exemplified can be synthesized by routine modification of one or more methods described in detail in the Examples by substituting appropriate readily available reagents.

The preparation of the above compounds are described in detail in the examples. Any such compound or similar compound may be synthesized according to a method discussed in general below and set 10 forth in the Examples by selecting appropriate starting materials as exemplified.

In general, most of the syntheses involve the condensation of a sulfonyl chloride with an aminoisoxazole in dry pryidine or in tetrahydrofuran (THF) and sodium hydride. The sulfonyl chorides and 15 aminoisoxazoles either can be obtained commercially or synthesized according to methods described in the Examples or using other methods available to those of skill in this art (see, U.S. Patent Nos.

4,659,369, 4,861,366 and 4,753,672).

The N-(alkylisoxazolyl)sulfonamides can be prepared by condensing an aminoisoxazole with a sulfonyl chloride in dry pyridine with or without the catalyst 4 -(dimethylamino)pyridine. The N-(3,4-dimethyl-5isoxazolyl)sulfonamides and N-(4,5-dimethyl-5-isoxazolyl)sulfonamides can be prepared from the corresponding aminodimethylisoxazole, such as 5-amino-3,4-dimethylisoxazole. For example, N-( 3 4 zolyl)- 2 -(carbomethoxy)thiophene-3-sulfonamide was prepared from 2methoxycarbonylthiophene-3-sulfonyl chloride and 5-amino-3,4-dimethylisoxazole in dry pyridine.

The N-(4-haloisoxazolyl)sulfonamides can be prepared by condensation of amino-4-haloisoxazole with a sulfonyl chloride in THF with sodium -59hydride as a base. For example, N-(4-bromo-3-methyl-5-isoxazolyl)thiophene-2-sulfonamide was prepared from 5-amino-4-bromo-3-methylisoxazole and thiophene-2-sulfonyl chloride in THF and sodium hydride. N-(4bromo-3-methyl-5-isoxazolyl)-5-(3-isoxazolyl)thiophene-2-sulfonamide was prepared from 5-amino- 4 -bromo-3-methylisoxazole and 5-(3-isoxazolyl)thiophene-2-sulphonyl chloride.

Alternatively, compounds, such as those in which Ar 2 is thienyl, furyl and pyrrolyl herein, may be prepared by reacting an appropriate sulfonyl chloride with a 5-aminoisoxazole substituted at the 3 and 4 10 positions, such as 5-amino- 4 -bromo-3-methylisoxazole, in tetrahydrofuran (THF) solution containing a base, such as sodium hydride. Following the reaction, the THF is removed under reduced pressure, the residue dissolved in water, acidified and extracted with methylene chloride. The organic layer is washed and then dried over anhydrous magnesium sulfate, the solvents are evaporated and the residue is purified by recrystallization using hexanes/ethylacetate to yield pure product.

These sulfonamides also can be prepared from the corresponding sulfonyl chloride and the aminoisoxazole in pyridine with or without-a catalytic amount of 4 -dimethylaminopyridine (DMAP). In some cases, the bis-sulfonyl compound is obtained as the major or exclusive product. The bis-sulfonated products can be readily hydrolyzed to the sulfonamide using aqueous sodium hydroxide and a suitable co-solvent, such as methanol or tetrahydrofuran, generally at room temperature. For example: N-(4-bromo-3-methyl-5-isoxazolyl)-2-(N-phenylaminocarbonyl)thiophene-3-sulfonamide was prepared from N-(4-bromo-3methyl-5-isoxazolyl)-2-carboxylthiophene-3-sulfonamide and aniline and 1ethyl- 3 3 -dimethylaminopropyl]carbodiimide (EDCl). N-(4-bromo-3methyl-5-isoxazolyl)-2-[(4-methoxyphenyl)aminocarbonyl]thiophene-3-sulfonamide was prepared from 4-methoxyaniline, N,N'-diisopropylethylamine and N-( 4 -bromo- 3 -methyl-5-isoxazolyl)-2-carboxylthiophene.3-sulfonamide. N-( 4 -bromo- 3 -methyl-5-isoxazolyl)-2-(benzylaminocarbonyl).

thiophene-3-sulfonamide was prepared from N-(4-bromo-3-methyl-5isoxazolyl)- 2 -carboxylthiophene.3-sulfonamide and benzylamine as described above.

mieN-( 4 -bromo-3-methyl5isoxazolyl)2.carboxylthiophene3sufona- *e was prepared from N-(4-bromo-3-methyl- S-isoxazolyl)-2-(carbomethoxy)thiophene-3-sulfonamic, which was prepared from the condensation of 5-a min o-4-bro mo-3-methylisoxazol e and 2-croehx~ho phene-3-sulfonyl chloride.

1-( 4 1 -isopropylphenyl) pyrro le- 2-sulfon amid e and N-(4-bromo-3-methyl-5-isoxazolyl). 1 4 '-isopropylphenyl)pyrrole-3-sulfonamide were prepared from 5-amino-4bromo-3-methylisoxazole and a mixture of 1-( 41 -isopropylphenyl)pyrrole- 2-sulf onyl chloride and 1 4 '-isopropylphenyl)pyrrole-3-sulfonyl chloride.

These sulfonyl chlorides were prepared from l-( 4 '-isopropylphenyl)pyr- .9.*role-2-sulfonic acid in phosphorus oxychioride and phosphorus pentachloride. 1 -W 4 -iso pro pylphenyl) pyrrole-2-sulfo nic acid was prepared from l-( 4 '-isopropylphenyl)pyrrole and chiorosulfonic acid. l-( 4 '-isopropylphenyl)pyrrole was prepared from 4-isopropyl aniline and tetrahydrof uran.

Prodrugs and other derivatives of the compounds suitable for administration to humans may also be designed and prepared by methods known to those of skill in the art (see, Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392).

Compounds described herein have been synthesized and tested for activity in in vitro assays and, in some cases, in in vivo animal models.

-61- Nuclear magnetic resonance spectroscopic (NMR), mass spectrometric, infrared spectroscopic and high performance liquid chromatographic analyses indicated that the synthesized compounds have structures consistent with those expected for such compounds and are generally at least about 98% pure. All of the compounds exemplified or described herein exhibited activity as endothelin antagonists.

C. Evaluation of the bioactivity of the compounds Standard physiological, pharmacological and biochemical procedures are available for testing the compounds to identify those that 10 possess any biological activities of an endothelin peptide or the ability to interfere with or inhibit endothelin peptides. Compounds that exhibit in vitro activities, such as the ability to bind to endothelin receptors or to compete with one or more of the endothelin peptides for binding to endothelin receptors can be used in the methods for isolation of endothelin 15 receptors and the methods for distinguishing the specificities of endothelin receptors, and are candidates for use in the methods of treating *endothelin-mediated disorders.

Thus, other preferred compounds of formulas I and II, in addition to those specifically identified herein, that are endothelin antagonists or agonists may be identified using such screening assays.

1. Identifying compounds that modulate the activity of an endothelin peptide The compounds are tested for the ability to modulate the activity of endothelin-1. Numerous assays are known to those of skill in the art for evaluating the ability of compounds to modulate the activity of endothelin (see, eq., U.S. Patent No. 5,114,918 to Ishikawa et al; EP Al 0 436 189 to BANYU PHARMACEUTICAL CO., LTD. (October 7, 1991); Borges et al. (1989) Eur. J. Pharm. 165: 223-230; Filep et al.

(1991) Biochem. Biophys. Res. Commun. 177: 171-176). In vitro studies may be corroborated with in ivo studies (see, U.S. Patent No.

-62- 5,114,918 to Ishikawa et aL; EP Al 0 436 189 to BANYU PHARMACEU- TICAL CO., LTD. (October 7, 1991)) and pharmaceutical activity thereby evaluated. Such assays are described in the Examples herein and include the ability to compete for binding to ETA and ET B receptors present on membranes isolated from cell lines that have been genetically engineered to express either ETA or ET 8 receptors on their cell surfaces.

The properties of a potential antagonist may be assessed as a function of its ability to inhibit an endothelin induced activity in vitro using a particular tissue, such as rat portal vein and aorta as well as rat uterus, 10 trachea and vas deferens (see Borges, Von Grafenstein, H. and Knight, "Tissue selectivity of endothelin," Eur. J. Pharmacol 165:223-230, (1989)). The ability to act as an endothelin antagonist in vivo can be tested in hypertensive rats, ddy mice or other recognized animal models (see, Kaltenbronn et al. (1990) J. Med. Chem. 33:838- 15 845, see, also, U.S. Patent No. 5,114,918 to Ishikawa et al.; and EP Al 0 436 189 to BANYU PHARMACEUTICAL CO., LTD (October 7, 1991); see, also Bolger et al. (1983) J. Pharmacol. Exp. Ther. 225291-309).

Using the results of such animal studies, pharmaceutical effectiveness may be evaluated and pharmaceutically effective dosages determined.

A

potential agonist may also be evaluated using in vitro and in vivo assays known to those of skill in the art.

Endothelin activity can be identified by the ability of a test compound to stimulate constriction of isolated rat thoracic aorta (Borges et l. (1989) "Tissue selectivity of endothelin" Eur. J. Pharmacol. 165: 223-230). To perform the assay, the endothelium is abraded and ring segments mounted under tension in a tissue bath and treated with endothelin in the presence of the test compound. Changes in endothelin induced tension are recorded. Dose response curves may be generated and used to provide information regarding the relative inhibitory potency -63of the test compound. Other tissues, including heart, skeletal muscle, kidney, uterus, trachea and vas deferens, may be used for evaluating the effects of a particular test compound on tissue contraction.

Endothelin isotype specific antagonists may be identified by the ability of a test compound to interfere with endothelin binding to different tissues or cells expressing different endothelin-receptor subtypes, or to interfere with the biological effects of endothelin or an endothelin isotype (Takayanagi et al. (1991) Reg. PeD. 32: 23-37, Panek et al. (1992) Biochem. Biophys. Res. Commun. 183: 566-571). For example, ET, receptors are expressed in vascular endothelial cells, possibly mediating the release of prostacyclin and endothelium-derived relaxing factor (De Nucci al. (1988) Proc. Natl. Acad. Sci. USA 85:9797). ETA receptors are not detected in cultured endothelial cells, which express ET, receptors.

The binding of compounds or inhibition of binding of endothelin to 15 ET, receptors can be assessed by measuring the inhibition of endothelin- 1-mediated release of prostacyclin, as measured by its major stable metabolite, 6-keto PGFi,, from cultured bovine aortic endothelial cells (see, Filep et al. (1991) Biochem. and Biophys Res. Commun. 177: 171-176). Thus, the relative affinity of the compounds for different endothelin receptors may be evaluated by determining the inhibitory dose response curves using tissues that differ in receptor subtype.

Using such assays, the relative affinities of the compounds for ETA receptors and ET 8 receptors have been and can be assessed.

Those that possess the desired properties, such as specific inhibition of binding of endothelin-1, are selected. The selected compounds that exhibit desirable activities may be therapeutically useful and are tested for such uses using the above-described assays from which in vivo effectiveness may be evaluated (see, U.S. Patent No. 5,248,807;

U.S.

Patent No. 5,240,910; U.S. Patent No. 5,198,548; U.S. Patent No.

-64- 5,187,195; U.S. Patent No. 5,082,838; U.S. Patent No. 5,230,999; published Canadian Application Nos. 2,067,288 and 2071193; published Great Britain Application No. 2,259,450; Published International PCT Application No. WO 93/08799; Benigi et al. (1993) Kidney International 44:440-444; and Nirei et al. (1993) Life Sciences 52:1869-1874). Compounds that exhibit in vitro activities that correlate with in vivo effectiveness will then be formulated in suitable pharmaceutical compositions and used as therapeutics.

The compounds also may be used in methods for identifying and 10 isolating endothelin-specific receptors and aiding in the design of compounds that are more potent endothelin antagonists or agonists or that are more specific for a particular endothelin receptor.

2. Isolation of endothelin receptors A method for identifying endothelin receptors is provided. In 15 practicing this method, one or more of the compounds is linked to a support and used in methods of affinity purification of receptors. By selecting compounds with particular specificities, distinct subclasses of -ET receptors may be identified.

One or more of the compounds may be linked to an appropriate resin, such as Affi-gel, covalently or by other linkage, by methods known to those of skill in the art for linking endothelin to such resins (see, Schvartz et al. (1990) Endocrinology 126: 3218-3222). The linked compounds can be those that are specific for ETA or ETB receptors or other subclass of receptors.

The resin is pre-equilibrated with a suitable buffer generally at a physiological pH (7 to A composition containing solubilized receptors from a selected tissue are mixed with the resin to which the compound is linked and the receptors are selectively eluted. The receptors can be identified by testing them for binding to an endothelin isopeptide or analog or by other methods by which proteins are identified and characterized. Preparation of the receptors, the resin and the elution method may be performed by modification of standard protocols known to those of skill in the art (see, Schvartz t al (1990) Endocrinology 126: 3218-3222).

Other methods for distinguishing receptor type based on differential affinity to any of the compounds herein are provided. Any of the assays described herein for measuring the affinity of selected compounds for endothelin receptors may also be used to distinguish receptor subtypes based on affinity for particular compounds provided herein. In particular, an unknown receptor may be identified as an ETA or ETB receptor by measuring the binding affinity of the unknown receptor for a compound provided herein that has a known affinity for one receptor over the other.

Such preferential interaction is useful for determining the particular 15 disease that may be treated with a compound prepared as described herein. For example, compounds with high affinity for ETA receptors and t little or no affinity for ET8 receptors are candidates for use as hypertensive agents; whereas, compounds that preferentially interact with ET 8 receptors are candidates for use as anti-asthma agents.

D. Formulation and administration of the compositions Effective concentrations of one or more of the sulfonamide compounds of formula I or II or pharmaceutically acceptable salts, esters or other derivatives thereof are mixed with a suitable pharmaceutical carrier or vehicle. In instances in which the compounds exhibit insufficient solubility, methods for solubilizing compounds may be used.

Such methods are known to those of skill in this art, and include, but are not limited to, using cosolvents, such as dimethylsulfoxide (DMSO), using surfactants, such as tween, or dissolution in aqueous sodium bicarbonate. Derivatives of the compounds, such as salts of the -66compounds or prodrugs of the compounds may also be used in formulating effective pharmaceutical compositions.

The concentrations or the compounds are effective for delivery of an amount, upon administration, that ameliorates the symptoms of the endothelin-mediated disease. Typically, the compositions are formulated for single dosage administration.

~Upon mixing or addition of the sulfonamide compound(s), the resulting mixture may be a solution, suspension, emulsion or the like.

The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the symptoms of the disease, disorder or condition treated and may be empirically determined.

Pharmaceutical carriers or vehicles suitable for administration of the 15 compounds provided herein include any such carriers known to those i skilled in the art to be suitable for the particular mode of administration.

.i In addition, the compounds may be formulated as the sole pharmaceutically active ingredient in the composition or may be combined with other active ingredients.

The active compounds can be administered by any appropriate route, for example, orally, parenterally, intravenously, intradermally, subcutaneously, or topically, in liquid, semi-liquid or solid form and are formulated in a manner suitable for each route of administration.

Preferred modes of administration include oral and parenteral modes of administration.

The active compound is included in the pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect in the absence of undesirable side effects on the patient treated.

The therapeutically effective concentration may be determined -67empirically by testing the compounds in known in vitro and in vivo systems (see, U.S. Patent No. 5,114,918 to Ishikawa et al.; EP Al 0 436 189 to BANYU PHARMACEUTICAL CO., LTD (October 7, 1991); Borges et a. (1989) Eur. J. Pharm. 165: 223-230; Filep et al. (1991) Biochem. Biophys. Res. Commun. 177: 171-176) and then extrapolated therefrom for dosages for humans.

The concentration of active compound in the drug composition will depend on absorption, inactivation and excretion rates of the active 9compound, the dosage schedule, and amount administered as well as other factors known to those of skill in the art. For example, the amount that is delivered is sufficient to treat the symptoms of hypertension. The effective amounts for treating endothelin-mediated disorders are expected to be higher than the amount of the sulfonamide compound that would be administered for treating bacterial infections.

15 Typically a therapeutically effective dosage should produce a serum concentration of active ingredient of from about 0.1 ng/ml to about 100 /ig/ml. The pharmaceutical compositions typically should provide a dosage of from about 0.01 mg to about 2000 mg of compound per kilogram of body weight per day. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at intervals of time. It is understood that the precise dosage and duration of treatment is a function of the disease being treated and may be determined empirically using known testing protocols or by extrapolation from i vivo or in vitro test data. It is to be noted that concentrations and dosage values may also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and -68that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions.

If oral administration is desired, the compound should be provided in a composition that protects it from the acidic environment of the stomach. For example, the composition can be formulated in an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine. The composition may also be formulated in combination with an antacid or other such ingredient.

999: Oral compositions will generally include an inert diluent or an edible 9: 10 carrier and may be compressed into tablets or enclosed in gelatin capsules. For the purpose of oral therapeutic administration, the active compound or compounds can be incorporated with excipients and used in the form of tablets, capsules or troches. Pharmaceutically compatible binding agents and adjuvant materials can be included as part of the 15 composition.

The tablets, pills, capsules, troches and the like can contain any of 9 the following ingredients, or compounds of a similar nature: a binder, such as microcrystalline cellulose, gum tragacanth and gelatin; an excipient such as starch and lactose, a disintegrating agent such as, but not limited to, alginic acid and corn starch; a lubricant such as, but not limited to, magnesium stearate; a glidant, such as, but not limited to, colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; and a flavoring agent such as peppermint, methyl salicylate, and fruit flavoring.

When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar and other enteric agents. The compounds can also be administered -69as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.

The active materials can also be mixed with other active materials which do not impair the desired action, or with materials that supplement the desired action, such as antacids, H2 blockers, and diuretics. For example, if the compound is used for treating asthma or hypertension, it may be used with other bronchodilators and antihypertensive agents, 10 respectively.

Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include any of the following components: a sterile diluent, such as water for injection, saline solution, fixed oil, polyethylene glycol, glycerine, propylene glycol or other 15 synthetic solvent; antimicrobial agents, such as benzyl alcohol and methyl parabens; antioxidants, such as ascorbic acid and sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid (EDTA); 9. buffers, such as acetates, citrates and phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose. Parenteral preparations can be enclosed in ampules, disposable syringes or multiple dose vials made of glass, plastic or other suitable material.

If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol and mixtures thereof. Liposomal suspensions, including tissue-targeted liposomes, may also be suitable as pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art. For example, liposome formulations may be prepared as described in U.S. Patent No. 4,522,811.

The active compounds may be prepared with carriers that protect the compound against rapid elimination from the body, such as time release formulations or coatings. Such carriers include controlled release formulations, such as, but not limited to, implants and microencapsulated delivery systems, and biodegradable, biocompatible polymers, such as collagen, ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid and others. Methods for preparation of Does such formulations are known to those skilled in the art.

The compounds may be formulated for local or topical application, 10 such as for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, and lotions and for application to the eye or for intracisternal or intraspinal application. Such solutions, particularly those intended for ophthalmic use, may be formulated as 0.01% 10% isotonic solutions, pH about 5-7, with appropriate salts.

15 The compounds may be formulated as aeorsols for topical application, such as by inhalation (see, U.S. Patent Nos. 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for delivery of a steroid useful for treatment inflammatory diseases, particularly asthma).

Finally, the compounds may be packaged as articles of manufacture containing packaging material, a compound provided herein, which is effective for antagonizing the effects of endothelin, ameliorating the symptoms of an endothelin-mediated disorder, or inhibiting binding of an endothelin peptide to an ET receptor with an ICso of less than about pM, within the packaging material, and a label that indicates that the compound or salt thereof is used for antagonizing the effects of endothelin, treating endothelin-mediated disorders or inhibiting the binding of an endothelin peptide to an ET receptor.

The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention.

-71- EXAMPLE 1

N-(

4 -Bromo-3-methyl-5-isoxazolyl)thiophene-2-sulfonamide A solution of 5-amino- 4 -bromo-3-methylisoxazole (177 mg, mmol) in dry tetrahydrofuran (THF, 2 ml) was added to a suspension of sodium hydride (60% dispersion in mineral oil, 90 mg, 2.2 mmol) in dry THF (1 ml) at 0 50 C. After stirring at 0 50 C for 5 min., the reaction was stirred at room temperature for 10 min to complete the reaction.

The reaction mixture was re-cooled to 00 C and thiophene-2-sulfonyl chloride (200 mg, 1.1 mmol) dissolved in dry THF (2 ml) was added 10 dropwise. Stirring was continued for 1 h; during this period the reaction mixture slowly attained ambient temperature. THF was removed under reduced pressure. The residue was dissolved in water (10 ml), the pH was adjusted to 10 11 by adding 5 N sodium hydroxide solution, and was extracted with ethyl acetate (3 X 10 ml) to remove the neutral 15 impurities. The aqueous layer was acidified with concentrated

HCI

(pH 2 3) and extracted with methylene chloride (3 X 10 ml). The combined organic layers was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give N-(4-bromo-3-methyT-5isoxazolyl)thiophene-2-sulfonamide. The pure material was obtained by recrystallization using hexanes/ethyl acetate (110 mg, 34 yield), m.p.

125 1270 C.

EXAMPLE 2

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-5-(3-isoxazolyl)thiophene-2-sulfona mide A solution of 5-amino-4-bromo-3-methylisoxazole (177 mg, mmol) in dry THF (2 ml) was added to a suspension of sodium hydride dispersion in mineral oil, 90 mg, 2.2 mmol) in dry THF (1 ml) at 0 50 C. After stirring at 0 50 C for 5 min, the reaction was warmed to room temperature for 10 min to complete the reaction. The reaction mixture was re-cooled to 00 C, and 5-( 3 -isoxazolyl)thiophene-2-sulphonyl -72chloride (273 mg, 1.1 mmol), which had been dissolved in dry THF (2 ml), was added slowly. Stirring was continued for 1 h; during this period the reaction mixture slowly attained ambient temperature. THF was removed under reduced pressure. The residue was dissolved in water mi), the pH was adjusted to 2 3 by adding concentrated HCI, and was extracted with methylene chloride (3 X 10 ml). The combined organic layers was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give N-(4-bromo-3-methyl-5-isoxazolyl)-5-(- S Isoxazolyl)thiophene-2-sulfonamide. The pure material was obtained by 10 recrystallization using hexanes/ethyl acetate (160 mg, 41% yield), m.p.

120 1230 C.

EXAMPLE 3 N-(4-Bromo-3-methyl-5-isoxazolyl)-5-(2-pyridyl)thiophene-2-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-5-(2-pyridyl)thiophene-2-suffona- *15 mide was prepared in the same manner as described in Example 2 from amino-4-bromo-3-methylisoxazole and 5-( 2 -pyridyl)thiophene-2-sulphonyl chloride in 40% yield. Purification was achieved by recrystallization from ethyl acetate to give a crystalline solid, m.p. 186 1880 C.

EXAMPLE 4

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-4,5-dibromothiophene-2-suffonamide N-(4-bromo-3-methyl-5-isoxazolyl)-4,5-dibromothiophene-2-sulfonamide was prepared in the same manner as described in Example 2 from amino-4-bromo-3-methylisoxazole and 4 ,5-dibromothiophene-2-sulphonyl chloride in 45% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 153 1550 C.

-73- EXAMPLE

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-.5chloro.3methylbenzo (blthiophene-2sulfonamide

N-(

4 -bromo-3-methyl.5.isoxazolyl).5chloro3methylbenzofbjtho phene-2-sulfonamide was prepared in the same manner as described in Example 2 from 5-amino-4-bromo-3-methyhisoxazole and 5-chloro-3methylbenzofblthiophene-2-sulphonyI chloride in 18% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 153 1550 C.

EXAMPLE 6 N 4 Br mo3methyi 5isoxazolyl)5(4chlo robenza mid thl)thio phene-2-sulf onamide N-4Boo3mty--sxzll)5(-hooezmdmty) thiophene-2-sulfonamide was prepared in the same manner as described in Example 2 from 5-amino-4-bromo-3-methylisoxazole and 5-(4-chlorobenzamidomethyl)thiophene.2sulphonyI chloride in 27% yield. The crude product was passed through a silica gel column using hexanes/ethyl acetate as eluent. Purification was effected by recrystallization from-..

*a ethyl acetate/hexanes to give a crystalline solid, m.p. 2100 C (dec).

EXAMPLE 7 N-4Boo3mty--sxzli--(eznsfoy~hohn--ufn amide N-(-Bam--ehl -sxzll--bne sloy~hopee2 sulfonamide was prepared in the same manner as described in Example 2 from 5-amino-4-bromo-3-methylisoxazole and 4 -benzenesulfonylthiophene-2-sulphonyl chloride in 26% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 181 1840 C.

-74- EXAMPLE 8 N-4Boo3mty--sxzll brm--hootipee2sloa mide

N-(

4 Bromo3 methy-soalyl)-4-i o oyl)4brotohtheh 2 sulfonamide was prepared in the same manner as described in Example 2 from 5-.amino-4-bromo-3-tmethylisoxazole and 4 2-upoy chloride in 25% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 143 1450 C.

EXAMPLE 9 N-4Bom--ety--ioao Si-,-dichlorothiophene-3.sulfonamide was prepared in the same manner as described in Example 2 from amino- 4 -bromo3methylisoxazole and 2 ,S-dichlorolthiophene3sulphonyF 15 chloride in 47% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 135 1380 C.

EXAMPLE N* r o -3 m t y -5 i o a o y 2 5 i e h l h o e -3 s l o n fi I N- (4-Brom o -3-methyl5..isoxaz ol -imethyltthiop he..sulfonaod amide was prepared in the same manner as described in Example 1 from 5-amino-4bromo3methylisoxazole and 2 ,5-dimethylthiophene-3 sulphonyl chloride in 55% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 77 800 C.

EXAMPLE 11

N-(

4 -Bromo-3..methyI..5.isoxazolyl)-4,5-dichlorothiophene-2sulfonamide N-+4Bromo-3..methyl5isoxazolyl)-4,5-dichlorothiophene2sulfonamide was prepared in the same manner as described in Example 1 from amino- 4 -bromo.3-methylisoxazole and 4 ,5-dichlorothiophene2sulphonyI WO 96/31492 PCr1US96/04759 chloride in 42% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 135 1380 C.

EXAMPLE 12

N-(

4 -Bromo-3-methyl..5.isoxazolyl)-2,5-dichloro-4-bromothjophene.3-su..

fonamide

N-(

4 -Bromo-3-methyl.s..isoxazoJyl)..2 S-dichloro-4-bromothiophene- 3 -sulfonamide was prepared in the same manner as described in Example 1 from 5-amino-4-bromo-3-methylisoxazole and 4-bromo-2, 10phene-3-sulfonyl chloride in 58% yield. Purification was achieved by from ethyl acetate/hexanes to give a crystalline solid, m.p. 146 1490 C.

EXAMPLE 13 4 Bromo3methy5isoxazoly)2{ 3

N-(

4 Bromo3methylisoxazolyl)2{3.[ 1 -methyl- 5 ulfofl 5 mjde was prepared in the same manner as described in Example 1 from 5-amino- 4 -bromo-3-methylisoxa- .zole and sulphonyl chloride in 30% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 121 1230 C.

EXAMPLE 14 Thiophene-2-sulfonyl chloride (183 mg, 1 mmol) was added to a solution of 3 -amino- 4 -bromo-5-methylisoxazole (177 mg, 1 mmol) in dry pyridine (0.5 ml). The reaction mixture was stirred at room temperature for 3 h. Pyridine was removed under reduced pressure and the residue was partitioned between water and ethyl acetate. The organic layer was washed with 1iN HCI (3 X 10 ml), brine (10 ml) and dried over anhydrous magnesium sulfate. Evaporation of the solvents left an oily residue which -76was solidified at -2Oo C and then purified by recrystallization from ethyl acetate/hexanes, to give the pure product (51 O/o yield) as a tan solid, m.p. 156 1580 C.

EXAMPLE

N-(

4 -Bromo- 3 -methyl-5-isoxazoly)5(benzenesulfonyl)thiophene.2-sulfonamide N-4Boo3mty--sxzll--bneeufnltipee2 sulfonamide was prepared in the same manner as described in Example 2 from 5-amino-4-bromo-3-methylisoxazole and X. 1o phene-2-sulfonyl chloride in 59% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 139 -1420 C.

EXAMPLE 16

N-(

4 -Bromo- 3 -methyl-5-isoxazoly)2(carbomethoxy)thiophene3sulfonamide N- 4 -Bro mo- 3 -methyl- 5-isoxazo lyl)..2.(carbomethoxy)thi p henl .3sulfonamide was prepared in the same manner as described in Example 2 from 5-amino-4-bromo-3-methylisoxazole and 2-(carbomethoxy)thiophene-3-sulfonyl chloride in 73% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 198 -2000 C.

EXAMPLE 17 N ehl -sxzll 2(abxltiphn--ufn i N-(4-Bromo-3-methyl- S-isoxazolyl)-2-(carbomethoxy)thiophene-3 sulfonamide (Example 16) (1.5 g, 3.95 mmol) was dissolved in methanol ml). Sodium hydroxide pellets (1 g, 25 mmol) and a few drops of water were then added. The resultant solution was stirred for 1 6 h at ambient temperature. Methanol was removed under reduced pressure.

The residue was diluted with water and was extracted with ethyl acetate (2 X 10 ml). The aqueous layer was acidified (pH 2) with -77concentrated hydrochloric acid and was extracted with ethyl acetate (2 X ml). The combined organic layers was dried over anhydrous magnesium sulfate and filtered. Removal of the solvent gave N-(4-bromo- 3-methyl-5-isoxazolyl)-2-(carbomethoxy)thiophene-3-sulfonamide (1.2 g, 82% yield), which was purified by silica gel column chromatography using ethyl acetate as eluent, m.p. 188 1940 C.

EXAMPLE 18 N-(4-Bromo-3-methyl-5-isoxazolyl)-2-(N-phenylaminocarbonyl)thiophene-3sulfonamide 10 Aniline (0.093 g, 1 mmol) and 1-ethyl- 3 '[3-dimethylaminopropyl]carbodiimide (EDCI) (0.191 g, immol) were added to N-(4-bromo-3methyl-5-isoxazolyl)-2-(carboxyl)thiophene--sulfonamide (0.368 g, 1 mmol) that had been suspended in methylene chloride (5 ml) to produce a clear solution. Stirring was continued for 1 h at ambient temperature.

15 The reaction mixture was diluted with methylene chloride (50 ml) and washed with 3 N hydrochloric acid solution (3 X 50 ml). The combined organic layers was dried over anhydrous magnesium sulfate and filtered.

Removal of the solvent under reduced pressure gave N-(4-bromo-3methyl-5-isoxazolyl)-2-(N-phenylaminocarbonyl)thiophene-3-sulfonamide.

The crude product thus obtained was purified by column chromatography using ethyl acetate as eluent to yield the product (0.32 g, 72% yield, m.p. 168 170° C.

EXAMPLE 19

N-(

4 -Bromo-3-methyl-5-isoxazolyl) 1-(4'-isopropylphenyl)pyrrole-2-sulfonamide and N-(4-bromo-3-methyl-5-isoxazolyl) isopropylphenyl)pyrrole-3-sulfonamide A. 1-( 4 '-isopropylphenyl)pyrrole Glacial acetic acid (100 ml) was added to a mixture of 4isopropylaniline (10 ml, 72.4 mmol) and 2 (9.6 ml, 72.4 mmol) and the resulting mixture was refluxed for 1.5 h.

-78- The reaction mixture was allowed to cool and acetic acid was removed under reduced pressure. The resulting brown syrup was dissolved in ethyl acetate (200 ml) and washed with water (2 X 200 ml). The organic layer was dried over magnesium sulfate and filtered. Removal of the solvent gave 1-(4'-isopropylphenyl)pyrrole (13.28 g, 99% yield) as a brown syrup.

B. 1-( 4 '-isopropylphenyl)pyrrole-2-sulfonic acid Chlorosulfonic acid (1.82 ml, 27.08 mmol) was slowly added to a solution of 1-( 4 '-isopropylphenyl)pyrrole (5.01 g, 27.08 mmol) in 10 chloroform (100 ml) at 0 0 C. The resulting solution was stirred at 0 0

C

for 1 h and for an additional 1 h at room temperature. Chloroform was removed under reduced pressure. The resultant brown liquid was diluted .with ethyl acetate (200 ml) and washed with 1 N sodium hydroxide. The aqueous layer was then acidified with concentrated hydrochloric acid 15 (pH and then extracted with chloroform (2 X 150 ml). The combined organic layers was dried over magnesium sulfate and was filtered. Removal of the solvent gave 1-( 4 '-isopropylphenyl)pyrrole-2- *i sulfonic acid as a brown syrup (3 g, 42 yield).

C. 1-( 4 '-isopropylphenyl)pyrrole-2-sulfonyl chloride and 1-(4'-isopropylphenyl)pyrrole-3-sulfonyl chloride Phosphorus pentachloride (4.7 g, 22.64 mmol) was slowly added to a solution of 1-( 4 '-isopropylphenyl)pyrrole-2-sulfonic acid (3 g, 11.32 mmol)in phosphorus oxychloride (8.4 ml, 90.57 mmol). The resulting mixture was heated at 700 C for 10 h. The reaction mixture was allowed to cool to room temperature, then carefully poured on to crushed ice (500 g) and extracted with chloroform (200 mi). The combined organic layers was dried over anhydrous magnesium sulfate. This was filtered and removal of the solvent yielded a 4:1 mixture of 1-( 4 '-isopropylphenyl)pyrrole-2-sulfonyl chloride and 1-[ 4 '-isopropylphenyl)pyrrole-3-sulfonyl chloride (2.5 g, 78%) as a brown oil.

-79- D. N-( 4 -bromo-3-methyl..5.isoxazolyl) 1 4 1 -isopropylphenyl)pyrrole-2 sulfonamnide and N-(4-bromo-3-methyl-5-isoxazolyl) 1 isopropylphenyl)pyrrole.3-sulfonamide

N-(

4 -bromo-3-methyl..sisoxazolyl) 1 4 1 -isopropylphenyl)pyrrole-2 sulfonamide and N-(4-bromo-3-methyl-5-isoxazolyl) isopropylphenyl)pyrrole3sufonamide were prepared in the same manner as described in Example 2 from 5-amino-4-bromo-3-methylisoxazole and a mixture of l-( 41 -isopropylphenyl)pyrrole2sulfonyl chloride and 1 isopropylphenyl)pyrrole.3-sulfonyI chloride in 65% combined yield. The 10 mixture was subjected to preparative HPLC to give N-(4-bromo-3-methy[.

l-( 41 isopropylpheny)pyrrole..2.sulfonamide (retention time .22.85 min, 5% to 95% acetonitrile in water with 0. 1% TFA over 30 min period, 018, analytical column) and N-(4-bromo-3-methyl-5-isoxazolyl) 1- 4 '-isopropylphenyl)pyrrole-3sulfonamide (retention time 24.56 min, to 95 acetonitrile in water with 0. 1% TFA over 30 min period, C, :9analytical column) as oils.

EXAMPLE

N-(

4 Bromo3methyl.5isoxazoly).5 ohohnensuf mde 9 20 mide was prepared in the same manner as described in Example 2 from amino- 4 -bromo-3-methylisoxazole and 5-bromothiophene-2-sulfonyl chloride in 30% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 2400 C (dec).

EXAMPLE 21

N-(

4 Bromo3mtythyl..5zoli)-2-azolyl)..2[N(4ethoyphemnc)bnl thiophene-3-sulfonamide 4-Methoxyaniline (0.246 g, 2 mmol), bromo-tris-pyrro lid inophosphonium hexafluorophosphate (PyBrop) (0.466 g, 1 mmol) and N,N'diisopropylethylamine 15 ml) were added to N-(4-bromo-3-methyl-5isoxazolyl)-2-(carboxyl)thiophene3sulfonamide (0.368 g, 1 mmol), which had been suspended in methylene chloride (3 ml), resulting in a clear solution. Stirring was continued for 24 h at ambient temperature. The reaction mixture was diluted with methylene chloride (50 ml) and washed with 3 N hydrochloric acid solution (3 X 50 ml) followed by 5% sodium carbonate solution (2 X 50 ml). The combined organic layers was dried over anhydrous magnesium sulfate and filtered. Removal of the solvent under reduced pressure gave N-(4-bromo-3-methyl-5isoxazolyl)2[N-(4 .methoxyp hen yl) amin ocarbonyl~th iophene-3suf on amid e The crude *6 product thus obtained was purified by column chromatography using 10 ethyl acetate as eluent. This was recrystallized from ethyl acetate/hexanes to give a crystalline solid, m.p. 202 2050 C (0.08 g, 17% yield).

*6 EXAMPLE 22

N-(

4 -Bromo-3-methyl.5-isoxazoly).2[N..(3.methoxyp henyl) amino carbonyllthiophene-3-sulfonamide 666nocarbonyllthiophene3sulfonamide was prepared in the same manner as *described in Example 21 from N-4boo3mtyl5ioaoy)2 6 (carboxyl)thiophene-3-sulfonamide and 3-methoxyaniline in 23% yield.

620 The crude product was p urified by column chromatography using ethyl acetate as eluent. This was recrystallized from ethyl acetate/hexanes to give a crystalline solid, m.p. 200 2020 C.

EXAMPLE 23 N-4Boo3mty--sxzll--N(-ehxpey~mncr bonyllthiophene-3-sulfonamide N-4boo3mty--sxzoy)2[-2mtoyhnlaminocarbonyllthiophene-3sufonamide was prepared in the same manner as described in Example 21 from N-4boo3mtyl5ioaoy)2 (carboxyl) th ioph ene-3..sulfon amid e and 2-methoxyaniline in 26% yield.

The crude product was purified by column chromatography using ethyl -81acetate as eluent. This was recrystallized from ethyl acetate/hexanes to give a crystalline solid, m.p. 74 800 C.

EXAMPLE 24 NuonBaomoe-3-methyl-5-isoxazoly )-2-(N-benzylaminocarbonyl)thiophene-3- ~Benzylamine (0.214 g, 2 mmol), benzotriazole-1-yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate (Bop) (0.442 g, 1 mmol) and N,N'-diisopropylethylamine (0.15 ml) were added to N-(4-bromo-3methyl-5-isoxazolyl)-2-(carboxyl)thiophene-3-sufonamid (0.368 g, 1 mmol), which had been suspended in methylene chloride (3 mi). The resultant solution was stirred for 14 h at ambient temperature. This was diluted with methylene chloride (50 mi) and washed with 3 N hydrochloric acid (3 X 50 mi) followed by 5% sodium carbonate solution (2 X 50 ml). The combined organic layers was dried over anhydrous S 15 magnesium sulfate and filtered. Removal of the solvent under reduced pressure gave N-(4-bromo-3-methyl-5-isoxazolyl)-2-(N-benzylaminocarbonyl)thiophene-3-sulfonamide. The crude product was purified by column chromatography using ethyl acetate as eluent. Recrystallization from ethyl acetate/hexanes gave a crystalline solid, m.p. 186 1900 C (0.14 g, 30% yield).

EXAMPLE N-(4-Bromo-3-methyl-5-isoxazolyl)-2-N-(4-ethylphenyl)aminocarbonyl thiophene-3-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-2- N-(4-ethylpheny)aminocarbonyl]thiophene-3-suffonamide was prepared in the same manner as described in Example 24 from N-(4-bromo-3-methyl-5-isoxazoly)-2- (carboxyl)thiophene-3-sulfonamide and 4-ethylaniline in 31% yield. The crude product was purified by column chromatography using ethyl acetate as eluent. This was recrystallized from ethyl acetate/hexanes to give a crystalline solid, m.p. 187 1900 C.

-82- EXAMPLE 26 thiophene- 3 -suffonamideoll[N bihnlancrbnl

N-(

4 -bromo-3-methyl. Sisoxazolyl)-2-[N-(4..biphenyl) aminocarbonyl]..

thiophene.3.sulfonamide compound was prepared in the same manner as described in Example 24 from N-(4-bromo-3..methyl. -isoxazolyl)-2- (caboxl~tiopen-3-uffnamde nd 4 -phenylaniline in 26% yield. The *crude product was purified by column chromatography using ety .acetate as eluent. This was recrystallized from ethyl acetate/hexanes to 10 give a crystalline solid, m.p. 205 2120 C (dec).

EXAMPLE 27 2-~Methoxycarbonylthophene-3-su~ffyI chloride (2.50 g, 10.05 mmol) was added to a solution of 5-amino-3,4..dimethylisoxazole(0.

9 8 g, 8.75 mmol) in dry pyridine (5.0 ml). The reaction mixture was stirred at room temperature for 16 h. Pyridine was removed under reduced pressure and the residue was partitioned between water and a.dichlo ro methane. The organic layer was washed with 1 N HCI (2_X ml) and dried over anhydrous magnesium sulfate. Evaporation of the solvents left an oily residue, which, after purification by column chromatography over silica gel (1:1 hexanes/ethyl acetate as eluent), yielded 2.20 mg of a brown solid. Further purification was achieved by recrystallization from ethyl acetate/hexanes, giving the pure product as a white solid, m.p. 113..1160

C.

EXAMPLE 28 N 3 4 dimethyI.5.isoxazo ly 2(carb l)thihen--ufn i N- 4-d imeth yl- 5-s aoll 2 croy)tipe -3sl nai was prepared in the same manner as described in Example 1 7 from

N-

(34dmty--sxzli--croehx~hohn--ufnmd by basic hydrolysis in 94% yield. Purification was achieved by -83recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 202 2030 C.

EXAMPLE 29

N-(

3 ,4-dimethyl-5-isoxazolyl)-2-(N-phenylaminocarbonyl)thiophene-3-sulfonamide N-(3,4-dimethyl-5-isoxazolyl)-2-(N-phenylaminocarbonyl)thiophene- 3-sulfonamide was prepared in the same manner as described in Example 18 from N-(3,4-dimethyl-5-isoxazolyl)-2-(carboxyl)thiophene-3sulfonamide in 40% yield. Purification was achieved by recrystallization 10 from ethyl methanol/water to give a crystalline solid, m.p. 176 178 C.

EXAMPLE N. N-( 4 -Bromo-3-methyl-5-isoxazolyl)-5-(2-thienyl)thiophene-2-sulfonamide A. 5-bromo-2,2'-bithiophene N-Bromosuccinimide (NBS, 1.12 g, 6.3 mmol) was added in small 15 portions to a stirred solution of 1.0 g (6.01 mmol) of 2,2'-bithiophene in ml of glacial acetic acid and 10 ml of chloroform. After stirring for 1 h at room temperature, the mixture was poured into ice-water and "extracted into chloroform (75 ml). The organic layer was washed with aqueous sodium bicarbonate solution, water, and then dried overmagnesium sulfate and evaporated. The residue was subjected to flash chromatography on silica gel using hexane to give 1.3 g of a light green solid, m.p. 55 560 C.

B. 5-Chlorosulfonyl-2,2'-bithiophene A stirred solution of 5-bromo-2,2'-bithiophene (1.5 g, 6.1 mmol) in 10 ml of dry ether was placed under an argon atmosphere, cooled to -780 C and 4.3 ml of a 1.7 M solution of t-butyllithium was added over min. Stirring was continued at this temperature for an additional min. Sulfur dioxide gas was then bubbled in at -780 C until a yellow precipitate formed. Bubbling of the sulfur dioxide gas was continued for an additional 3 min and was immediately followed by a dropwise addition -84of N-chlorosuccinimide (NCS, 902 mg, 6.76 mmol) that had been dissolved in THF. The mixture was warmed to room temperature and stirring was continued for an additional 1.5 h. The mixture was then concentrated and the residue dissolved in ether. The organic layer was washed with water, brine solution and dried over magnesium sulfate.

Evaporation of solvent left a pale yellow solid, which was recrystallized from hexane to give 700 mg of a yellow solid, m.p. 63-640 C.

SC. N-(4-Bromo-3-methyl-5-isoxazolyl)-5-(2-thienylthiophene-2sulfonamide 10 N-(4-bromo-3-methyl-5-isoxazolyl)-5-(2-thieny)thiophene-sufona mide was prepared in the same manner as described in Example 2.

Reaction of 2 -chlorosulfonyl-5,2'-bithiophene (300 mg, 1.14 mmol) with 5-amino-4-bromo-3-methylisoxazole (183 mg, 1.03 mmol) yielded, after flash chromatography using 10% MeOH/CHCI 3 430 mg of a pale 15 brown solid, m.p. 2100 C.

C

EXAMPLE 31 N-(4-Bromo-3-methyl-5-isoxazolyl)thiophene- 3sulfonamide A. Thiophene-3-sulfonyl chloride A stirred solution of 3 -bromothiophene (1.5 g, 9.2 mmol) in 10 ml of dry ether was placed under an argon atmosphere and cooled to -780 C. Over the course of 20 min, a solution of t-butyllithium (5.6 ml of a 1.7 M) was added and stirring was continued at this temperature for an additional 20 min. Sulfur dioxide gas was then bubbled in at -780 C and the solution was warmed to 0° C, whereupon NCS (1.47 g, 12 mmol) in 8 ml of THF, was added dropwise. After warming to room temperature, stirring was continued for an additional 1 hour, after which, evaporation of solvents left 1.55 g of a brown oil. Flash chromatography over silica gel using hexanes yielded 1.24 g of a yellow oil which solidified on standing to give a yellow crystalline solid, m.p. 38-390 C.

B. N-( 4 -Bromo-3-methyl-5-isoxazolyl)thiophene-3-sulfonamide

N-(

4 -bromo-3-methyl-5-isoxazolyl)thiophene-3-sufonamide was prepared in the same manner as described in Example 2 from thiophene- 3-sulfonyl chloride with 5-amino-4-bromo-3-methylisoxazole in 22% yield.

Purification by column chromatography using 10% MeOH/CHCI 3 as eluent gave a pale brown oil.

EXAMPLE 32 N-(3,4-dimethyl-5-isoxazoly)-5-phenylthiophene-2-sulfonamide A. N-( 3 ,4-dimethyl-5-isoxazolyl)-5-bromothiophene-2-sulfonamide A solution of 5-bromothiophene-2-sulfonyl chloride (2.75 g, 10 mmol) and 5-amino-3,4-dimethylisoxazole (1.07 g, 9.57 mmol) in pyridine containing a catalytic amount of 4 -dimethylaminopyridine (DMAP, mg) was stirred at room temperature for a period of 3 h. The solution was heated at 500 C for an additional 1.5 h to drive the reaction to completion as judged by TLC. The pyridine was removed under reduced pressure and the residue, after extraction into ethyl acetate, was washed with 1 N HCI (2 x 25 ml), water (1 x 25), brine solution, (1 x 25 ml) and dried over magnesium sulfate. Evaporation of solvent left a viscous brown gum, which was subjected to flash chromatography. Elution with 3% methanol hexanes gave 246 mg of pure sulfonamide.

B. N-(methoxyethoxymethyl)-N-(3,4-dimethyl-5-isoxazolyl)-5bromothiophene-2-sulfonamide N-(3,4-dimethyl-5-isoxazolyl)-5-bromothiophene-2-sulfonamide (680 mg, 2 mmol) in dry THF (2 ml) was added to sodium hydride (121 mg of a 60% oil dispersion, 3 mmol) in dry THF (1 ml). The resulting suspension was cooled to 00 C and methoxyethoxymethyl chloride (334 mg, 2.68 mmol) was added dropwise via syringe. The solution was warmed to room temperature, and stirring continued overnight.

Evaporation of solvent left an oil that was extracted into ethyl acetate, washed with brine, dried over magnesium sulfate and evaporated. Flash -86chromatography of the residue on silica gel using 10-15% ethylacetate/hexanes yielded 480 mg of a colorless oil.

C. N-(methoxyethoxymethyl)-N-(3,4-dimethyl-5-isoxazolyl)-5phenylthiophene-2-sulfonamide Sodium carbonate (2 ml of a 2 M aqueous solution) followed by a. phenyl boronic acid (86 mg, 0.71 mmol) in 2 ml of 95% ethanol were Sadded to a solution of N-(methoxyethoxymethyl)-N-(3,4-dimethyl-5- Sisoxazolyl)-5-bromothiophene-2-sulfonamide (200 mg, 0.47 mmol) and tetrakis (triphenylphosphine) palladium (23 mg, 0.02 mmol) in dry 10 benzene (4 ml) under argon. The mixture was refluxed for 12 h, diluted with 5 ml of water and extracted into ethyl acetate (3 X 25 ml). The combined organic extracts was washed with brine (1 x 25 ml), dried and evaporated. The residue was flash chromatographed on silica gel using 25% ethylacetate/hexanes to afford 123 mg of the sulfonamide as 15 a colorless gum.

D. N-( 3 ,4-dimethyl-5-isoxazolyl)-5-phenylthiophene-2-sulfonamide HCI (3 ml of a 3 N aqueous solution) was added to a solution of N- (methoxyethoxymethyl)-N-(3,4-dimethyl-5-isoxazolyl)-5-phenylthiophene- 2 -sulfonamide (100 mg, 0.24 mmol) in 3 ml of 95% ethanol and-the resulting mixture was refluxed for 6 h. The mixture was then concentrated, diluted with 5 ml of water, neutralized with saturated aqueous sodium bicarbonate solution and acidified to pH 4 using glacial acetic acid. The mixture was extracted with ethyl acetate (2 x 25 ml) and the combined organic extract was washed with brine (1 x 5 ml), dried and evaporated. Flash chromatography of the residue on silica gel using 2% MeOH/CHCI, and further purification by reverse phase HPLC yielded 33.4 mg of the pure sulfonamide as a white powder, m.p. 176-1780

C.

EXAMPLE 33

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-5-(4-ethylphenylthiophene-2-sulfonamide A. N-(5-bromothiophene-2-sulfonyl)-pyrrole Sodium hydride (60% oil dispersion, 191 4.78 mmol) was suspended in dry tetrahydrofuran (2 ml) and the resulting cloudy suspension was cooled to 00 C in an ice bath. Pyrrole (385 mg, 5.75 mmol) in dry tetrahydrofuran (2 ml) was added dropwise over a period of.

10 min. The ice bath was removed and the solution was stirred at room 10 temperature until gas evolution ceased (15 minutes), whereupon bromothiophene-2-sulfonyl chloride (1.0 g, 3.82 mmol) previously dissolved in tetrahydrofuran (4.0 ml) was added dropwise through a steel cannula. After stirring for 1 h at room temperature, the mixture was filtered through Celite. The filter pad was rinsed with tetrahydrofuran, 15 and the filtrate was evaporated, which left a light brown solid that was recrystallized from methanol to produce the sulfonamide (821 mg, 74% yield) as a white powder.

B. 4 -Ethylphenylboronic acid A solution of 1-bromo-4-ethyl benzene (2.0 g, 11 mmol) indry ether (5 ml) was added to magnesium turnings (311 mg, 13 mmol), which had been suspended in dry ether, by dropwise addition. After addition was complete, the suspension was refluxed for a period of min, by which time nearly all of the magnesium had reacted. The solution was then added to trimethy borate (1.12 g, 11 mmol), previously dissolved in ether (5 ml) at -780 C, warmed to room temperature and stirred for 90 min. The reaction was quenched by the addition of aqueous HCI (2 ml) and the solution was extracted with ether. The combined ether extract was extracted with 1 M NaOH (2 X 20 ml), the aqueous extracts were acidified with dilute HCI to pH 2 and extracted with ether (2 X 25 ml). The resulting combined ether extract was -88washed once with water (10 ml), dried and evaporated to produce a white solid (676 mg, 38% yield), m.p. 138-1400 C.

C. 4 -ethylphenyl)thiophene-2-sulfonyl]pyrrole 4 -ethylphenyl)thiophene-2-sulfonyl]pyrrole was prepared, in the same manner as described in Example 32C, from 4-ethylphenylboronic acid and N-(5-bromothiophenesulfonyl)pyrrole. Purification by column chromatography using 10% ethyl acetate/hexanes gave the pure sulfonamide as a tan solid in 81% yield.

D. 5-Chlorosulfonyl-2-(4-ethylphenyl)thiophene A solution of 4 -ethylphenyl)thiophene-2-sulfonyl]pyrrole (100 mg, 0.32 mmol) and 6 N sodium hydroxide (1 ml) in methanol (1.5 ml) was refluxed for approximately 6 h. Evaporation of solvents and drying in vacuo resulted in an oil. Phosphorus oxychloride (258 ml, 2.52 mmol) and phosphorus pentachloride (131 mg, 0.63 mmol) were added to the oil and the resulting brown suspension was heated at 500 C for 3 h.

The resulting clear brown solution was carefully added to about 20 ml of crushed ice and then extracted with ethyl acetate (3x25 ml). The combined organic layers was washed with brine (2x5 ml), dried LMgSO 4 and evaporated to leave an oily residue. Flash chromatography over silica gel using 2% ethyl acetate/hexanes yielded (53 mg, 59%) of the pure sulfonyl chloride as a pale yellow oil.

E. N-(4-bromo-3-methyl-5-isoxazolyl)-5-(4-ethylphenyl)thiophene-2sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-5-(4-ethylphenyl) thiophene-2sulfonamide was prepared in the same manner as described in Example 2.

Reaction of 5-chlorosulfonyl-2-(4-ethylphenyl) thiophene (47.1 mg, 11.16 mmol) with 5-amino-4-bromo-3-methyl isoxazole (29 mg, 0.16 mmol) yielded, after flash chromatography using 10% MeOH/CHCI 3 a pale brown solid (46 mg, 66% yield), m.p. 172-1750 C.

-89- EXAMPLE 34

N-(

3 4 -dimethyl-5-isoxazolyl)benzo[b]thiophene-2-sulfonamide A. Benzo[b]thiophene-2-sulfonyl chloride Benzo[b]thiophene (1.50 g, 11.2 mmol) was stirred at 00 C in 20 ml of THF. t-Butyllithium (1.7 M, 16.8 mmol, 9.9 ml) was slowly added over a 5 minute period. Fifteen minutes later, SO 2 was flushed into the reaction flask and a thick white precepitate formed. The reaction mixture was stirred for 15 minutes at 0° C and then NCS (1.64 g, 12.3 mmol) was added. The reaction was warmed to 250 C and stirred for min. It was then poured into ethyl acetate (150 ml) and washed with brine (3x100 ml). The organic phase was dried with MgSO 4 filtered and concentrated to collect 2.29 g of a brown oil. The brown oil was subjected to flash chromatography ethyl acetate/hexanes), which provided a yellow tan solid (1.39 g, 53% yield).

15 B. N-( 3 4 -Dimethyl-5-isoxazolyl)benzo[b]thiophene-2-sulfonamide S. 3 4 -Dimethyl-5-amino-isoxazole (0.224 g, 2.0 mmol) and 50 mg of DMAP were stirred in 5 ml of pyridine at 250 C. The benzo[b]thiophene- 2-sulfonyl chloride (0.16 g, 2.6 mmol) was added and the dark brownyellow reaction mixture was stirred for 18 h at ambient temperature, poured into 100 ml of ethyl acetate and washed with 2% HCI (3x50 ml).

The organic phase was dried with MgSO 4 filtered and concentrated to collect 0.61 g of a brown oil/solid. The brown oil/solid was subjected to flash chromatography (30% ethyl acetate/hexanes) to provide 0.37 g of a light brown solid. This was stirred in 10 ml of methanol and 0.5 g of NaOH. The methanolic solution was heated for reflux for 1 h, then cooled to 250 C and the methanol was removed in vacuo. The resulting residue was acidified to pH 1 with 2% HCI (100 ml) and extracted with ethyl acetate (2x50 ml) The organic phase was dried with MgSO 4 filtered and concentrated to collect 0.225 g of a yellow-orange solid.

This was recrystallized from CHCI 3 /hexanes to produce a light tan-yellow solid 1949g, 31 yield), m.p. 157-160O

C.

EXAMPLE N-34Dmty--sxzlf~ezfjua--ufnmd A. Benzo~blfuran.2.suffonyl chloride Benzo[blfuran.2.sulfonyl chloride was prepared as in Example 34A from benzo[blfuran (1.61 g, 13.6 mmol), t-BuLi (1.7 M, 17.7 mmol, 10.4 ml) and NCS (2.09g, 15.0 mmol). Flash chromatography ethyl aceatate/hexanes) yielded a brown solid (1.84 g, 62% yield).

10 B. N-34Dmty--ioaoy ez[blfuran-2-sulfonamide N- 3 4Dimethyl....isoxazolyl nz beIf u ranf 2 ffnai wspe pared, in the same manner as described in Example 348, from 3,4-dimethyl-s-amino isoxazole (78 mg, 0.70 mmol) and benzofblfuran-2sul.

afonyl chloride (0.46 g, 2.1 mmol) Flash chromatography (30% ethyl acea15 tate/hexanes) provided 0. 186 g of a fight yellow solid, which was treated with 31 mg of NaOH in 10 ml of methanol at 250 C for 30 minutes. Re- **crystallization from CHCl 3 fhexanes yielded a light tan solid (90 mg, 44% yield), m.p. 160.5-1630

C.

EXAMPLE 36 N- 3 ,4dim ethyl5iso x yfazolyI-fulf amde A. Furan-2-sulfonyl chloride Furan-2-sulfonyl chloride was prepared as in Example 34A from furan (0.96 g, 14.2 mmol), t-BuLi (1.7 M, 17 mmol, 10 ml) and NCS (2.27 g, 17 mmol) using ether (30 ml) as the solvent. Flash chromatography ethyl acetate/hexanes) produced a yellow liquid (1.22 g, 52% yield).

B.

N-(

3 4 dimethyl.5-i isaoxy~an-2-sffoafde N-34dmty--sxzoy~ua--ufnmd was prepared as described in Example 34B from 3 4 -dimethyl-5-amino isoxazole (0.122 g, -91mmol), furan-2-sulfonyl chloride (0.50 g, 3.0 mmol) and NaOH (64 mg). Flash chromatography (50% ethyl acetate/hexanes) yielded 70 mg of a yellow solid. Recrystallization from CHCI 3 /hexanes produced an offwhite solid (46 mg, 29% yield), m.p 107 1100 C.

EXAMPLE 37

N-(

3 4 -Dimethyl-5-isoxazolyl)-3-methoxy-2-thiophene sulfonamide A. 3 -methoxy-2-thiophenesulfonyl chloride Chlorosulfonic acid (CISO 3 H, 2.31 g, 19.62 mmol) was slowly added to a solution of 3 -methoxythiophene (2.29 g, 19.62 mmol) in

CHCI

3 (80 ml) at 00 C. The resulting mixture was stirred at 0° C for min. The solvent was evaporated under reduced pressure at room temperature, the residue was suspended in POCI, (15 ml, 156.96 mmol), and PCI, (8.2 g, 39.24 mmol) was added slowly. The reaction was stirred at 600 C for 18 h, then cooled to room temperature and poured 15 onto crushed ice (200 The aqueous mixture was extracted with

CHCI

3 (2x150 ml) and the combined organic layers was dried (MgSO 4 The solid was removed by filtration and the filtrate was concentrated to give 3 -methoxy-2-thiophenesulfonyl chloride as a brown oil (1.81 g, 43% yield).

B. N-(3,4-dimethyl-5-isoxazolyl)-3-methoxy-2-thiophene sulfonamide Sodium hydride (1.02 g, 25.56 mmol, 60% dispersion in mineral oil) was slowly added to a solution of 3 -methoxy-2-thiophenesulfonyl chloride (1.18 g, 8.52 mmol) and 3 4 -dimethyl-5-aminoisoxazole (1.05 g, 9.37 mmol) in THF (20 ml) at room temperature. The resulting mixture was refluxed for 4 h. THF was removed under reduced pressure. The residue was dissolved in water (10 ml), the pH was adjusted to 10 11 by adding 5 N sodium hydroxide solution, and was extracted with ethyl acetate (3 X 10 ml) to remove the neutral impurities. The aqueous layer was acidified with concentrated HCI (pH 2 3) and extracted with -92methylene chloride (3 X 10 ml). The combined organic layers was dried over anhydrous magnesium sulfate to produce a crude oil. Further purification by reverse phase HPLC yielded a yellow oil (retention time 14.94 min, 5% to 95% acetonitrile in H 2 0 with 0.1% TFA over 30 min period, C, 1 analytical column).

EXAMPLE 38 N-(4-Bromo-3-methyl-5-isoxazolyl)-3-phenyl- 2 -thiophene sulfonamide and N-(4-bromo-3-methyl-5-isoxazolyl)-4-phenyl-2-thiophene sulfonamide A. 3-phenyl-2-thiophenesulfonyl chloride and 4-phenyl-2- 10 thiophenesulfonyl chloride n-Butyllithium (2.38 M, 17.2 ml, 41.03 mmol) was slowly added to a solution of 3 -phenylthiophene (5.47 g, 34.2 mmol) in EtO (25 ml) at S" 00 C. The ice bath was removed, the mixture was stirred at room temperature for 2 h, cooled to -300 C (CO,/acetone) and SO 2 gas was 15 bubbled through the reaction mixture for 20 min. A solution of NCS (6.06 g, 44.5 mmol) in THF (20 ml) was then added. The reaction was allowed to warm to room temperature and stirred for 16 h. The crude mixture was filtered, and the solid was washed with Et20. The combined organic layers was concentrated and the residue was chromatographed (hexanes/CHCI 3 to give 3 -phenyl-2-thiophenesulfonyl chloride and 4phenyl-2-thiophenesulfonyl chloride as a 1:1 mixture (1.46 g, 16.5%, while solid).

B. N-(4-Bromo-3-methyl-5-isoxazolyl)- 3 -phenyl- 2 -thiophene sulfonamide and N-(4-bromo-3-methyl-5-isoxazolyl)-4-phenyl-2-thiophene sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-3-phenyl-2-thiophene sulfonamide and N-(4-bromo-3-methyl-5-isoxazolyl)-4-phenyl-2-thiophene sulfonamide were prepared as described in Example 1. A fraction of the crude mixture of products was purified by HPLC to give N-( 4 isoxazolyl)-3-phenyl-2-thiophene sulfonamide (light brown solid, retention time 20.48 min, 5% to 95% acetonitrile in water with 0.1% TFA over min C 1 analytical column, m.p. 105-1070 C) and N-( 4 isoxazolyl)-4-phenyl-2-thiophene sulfonamide (dull yellow solid, m.p. 108- 110 0 C, retention time 21.35 min, same conditions).

EXAMPLE 39 5 4 -tert-Butyl-N-(4-bromo-3-methyl-5-isoxazolyl)benzenesulfonamide S. A solution of 5-amino-4-bromo-3-methylisoxazole (354 mg, mmol) in dry THF (1 ml) was added to a suspension of sodium hydride (60% dispersion in mineral oil, 188 mg, 4.4 mmol) in dry THF (1 ml) at 0-50 C. After stirring at 0-50 C for 10 min., the reaction was warmed to room temperature for 10 min. to complete the reaction. The reaction mixture was re-cooled to 00 C and 4 -tert-butylbenzenesulfonyl chloride S(512 mg, 2.2 mmol) was added slowly. Stirring was continued for min. at 0-5° C. Excess sodium hydride was decomposed by addition of methanol (0.4 ml) followed by water (0.5 ml). The mixture was acidified with hydrochloric acid and extracted with dichloromethane. The extract was dried over anhydrous magnesium sulfate and the solvent was removed under reduced pressure to give a crude product, which was purified by recrystallization from ethyl acetate/hexanes to give a white-solid in 21% yield, m.p. 1700 C (dec.).

EXAMPLE N-(3,4-Dimethyl-5-isoxazolyl)-2-methylbenzo[b]thiophene-3-sulfonamide A. 2 -Methylbenzo[b]thiophene t-BuLi (1.7 M, 26 mmoles, 15 ml) was added to a stirred solution of benzo[blthiophene (17 mmoles, 2.3 g) and THF (30 ml) at -500C. The resulting bright yellow reaction mixture was warmed to -30 0 C, and iodomethane (26 mmoles, 1.6 ml) was added. After 10 min. at -30 0

C,

the solution was warmed to ambient temperature and stirred an additional min., then diluted with ether (100 ml) and washed with brine (2 x 100 ml). The organic phase was dried (MgSO filtered and -94concentrated to collect 2.48 g of 2 -methylbenzo[b]thiophene as a light yellow solid.

B. 2 -Methylbenzo[blthiophene-3-sulfonyl chloride Sulfuryl chloride (9.5 mmoles, 0.76 ml) was added to a stirred solution of dimethylformamide (DMF; 11.2 mmoles, 0.87 ml) at OOC, and the resulting faint yellow solution was stirred for 20 min at 0 0 C. 2- SMethylbenzo[blthiophene (5.6 mmoles, 0.83 g) was then added, the reaction mixture was diluted with 2 ml of DMF, and then heated to 850C.

After 2.5 hrs at 850C, the brown reaction mixture was cooled to ambient S 10 temperature and added to ice 100 ml). The aqueous phase was extracted with ethyl acetate (100 ml), and the organic phase was dried (MgSO 4 filtered and concentrated to collect an orange-brown solid.

Flash chromatography ethyl acetate/hexanes) provided 0.89 g (64%) of 2-methylbenzolbthiophene-3-sulfonyl chloride as a yellow solid.

C.

N(

3 4 -Dimethyl-5-isoxazolyl)-2-methylbenzobthiophene-3-sufonamide 12-Methylbenzo[bthiophene-3-sulfonyl chloride (1.7 mmoles, 0.41 g) was added to a solution of 3,4-dimethyl-5-aminoisoxazole_(0.75 mmoles, 84 mg), 4-dimethylaminopyridine (DMAP; 50 mg) and pyridine (5 ml) at ambient temperature. After 24 h, the reaction mixture was diluted with ethyl acetate (50 ml) and washed with 2% HCI (3 x 50 ml).

The organic phase was dried (MgSO 4 filtered and concentrated to collect a brown-orange solid, which was dissolved in a solution of methanol ml) and NaOH (60 mg). The solution was stirred 1 h at ambient temperature, then the methanol was evaporated and the resulting residue was diluted with 2% HCI (50 ml), and extracted with ethyl acetate ml). The organic phase was dried (MgSO 4 filtered and concentrated to collect a tan solid. Recrystallization from chloroform and hexanes resulted in 93 mg of N-( 3 ,4-dimethyl-5-isoxazolyl)-2- 0 600.

0 Ut 0.

1.: methylbenzo[blthiophene-3-sulfonamide as light yellow crystals, m.p. 174-176 0

C.

EXAMPLE 41 N-4Boo3mty--sxzli--mtybnobtipee3sloa mide NaH (60% oil dispersion, 2.5 mmoles, 100 mg) was added to a solution of 4 -bromo-3-methyI-5-aminoisoxazole (1.0 mmoles, 0. 177 g).

THF (5 ml) at 000C was added, and the resulting reaction mixture was stirred 10 min at 0CC. 2 Methyl benzo [bjthio phene-3-sulf onyl chloride (1.2 mmoles, 0.28 g) was added, and the reaction mixture was stirred for min at 000, then warmed to ambient temperature for 1 hr followed by addition of 2 ml of water. The mixture was diluted with ethyl acetate (100 ml) and washed with 2% HC! (2 x 50 ml), then brine (50 ml). The organic phase was dried (MgSO 4 filtered and concentrated. Recrystallization of the crude reaction mixture resulted in 0.24 g of N-(4br o -ehy-5isxzll 2mtybno[ t opene3sl nai as an off-white solid, m.p. 131-1 3300.

EXAMPLE 42 N-4Boo3mty--sxzli--tybnobtipee3sfoa mide A. 2-Ethylbenzofblthiophene 2-Ethylbenzo[blthiophene was prepared by the method -of Example with benzo[b~thiophene (7.5 mmoles, 1.0 t-BuLi (1.7 M, 8.9 mmoles, 5.3 ml), iodoethane (8.9 mmoles, 0.72 ml) and THF (20 ml).

1 .2 g of a yellow liquid was isolated.

B. 2-Ethylbenzo [blthiophene-3-sulfonyI chloride 2 -Ethylbenzofblthiophene-3-sulfonyl chloride compound was prepared by the method of Example 40B with dimethylformamide

(DMVF;

13.6 mmoles, 1. 1 ml), sulfonyl chloride (11. 5 mmoles, 0. 93 ml). Flash -96chromatography ethyl acetate/hexanes) provided 1.34 g of a light yellow solid.

C. N-( 4 bromo3methyl5isoxazoly2ethylbenbzhopb~hn- 3 slfonamide N-4boo3mth [ioaoyi--tybnobjthiophene-3-sul.

fonamide was prepared by the method of Example 41 with 4-bromo-3- (1.0 mmoles, 0. 177 NaH (2.5 mmoles, C**100 mg), 2 -ethylbenzo[bJthiophene-3sufonyI chloride (1.2 mmoles, 0.31 g) and THF (7 ml). Recrystallization from chloroform and hexanes 10 provided 0.24 g of a tan crystalline solid, m.p. 11 8.5-1201C.

OV, EXAMPLE 43

N-(

4 -Bromo3methyl.5isxazoly)2benzylbenzo [blthiophene-3-sulfona.

C mide A. 2 -Benzylbenzo[blthiophene 15 2 -Benzylbenzo[blthiophene was prepared by the method of Example 40A with benzo[blthiophene (7.5 mmoles, 1.0 t-BuLi (1.7 M, 11.2 mmoles, 6.6 ml), benzyl bromide (10.2 mmoles, 1.3 ml) and THF 0 00 (20 ml). Flash chromatography (hexanes) provided 0.66 g of a yellow solid.

B. 2 -Benzylbenzo[bthiophene-3sufonyI chloride 2 -Benzylbenzo[blthiophene3sulfonyl chloride was prepared by the method of Example 40B with DMF (5.4 mmoles, 0.41 ml), sulfuryl chloride (4.6 mmoles, 0.37 ml) and 2 -benzylbenzo[blthiophene. Flash chromatography ethyl acetate/hexanes) provided 0.55 g of a yellow solid.

C. N-( 4 bromo3methyl.5isoxazolyl)2..ezybenzolenzop~he 3 sulfonamide N-4boo3mty--sxzll)2bnybnobtipee3 sulfonamide was prepared by the method of Example 41 with 4-bromo-3methyl-5-aminoisoxazole (1.0 mmoles, 0. 177 NaH (2.5 mmoles, -97- 100 mg), 2 -benzylbenzo[blthiophene-3-sulfonyI chloride (1.2 mmoles, 0.39 g) and THF (7 ml). Flash chromatography methanol/chloroform) followed by recrystallization from chloroform and hexanes provided 0. 11 g of a tan crystalline solid, m.p. 120-1231C.

:%se EXAMPLE 44 0006,N-( 4 Bromo3m ethyl-5iso azol-2buyl2b lbenzo[bh- 3 sffo mide A. 2 -Butylbenzo[blthiophene 10 2 -n-Butylbenzofblthiophene was prepared by the method of Example 40A with benzo[blthiophene (7.5 mmoles, 1.0 t-BuLi (1.7 M, S *9.7 mmoles, 5.7 ml), l-bromobutane (9.7 mmoles, 1.0 ml) and THE (20 ml). 0.65 g of a yellow liquid was isolated.

B. 2 -n-Butylbenzofblthiophene-3sufonyI chloride 2 -n-Butylbenzofblthiophene-3sulfonyI chloride was prepared by the method of Example 40B with DMVF (6.6 mmoles, 0.51 ml), sulfuryl chloride (5.6 mmoles, 0.45 ml) and 2 -n-butylbenzofblthiophene (3.3 0 55*.mmoles, 0.63 Flash chromatography ethyl acetate/hexanes) provided 0.68 g (71 of an orange solid.

C. N-( 4 bromo3me hyl..5..zoli)-2-n-buylbenzbult~iophe 3 sulfonamide N-4boo3mty--sxzll---uybnobtipee3 sulfonamide was prepared by the method of Example 41 with 4-bromo-3- (1.0 mmoles, 0. 177 NaH (2.5 mmoles, 100 mg), 2 -n-butylbenzo[blthiophene-3-sulfonyI chloride (1.2 mmoles, 0.35 g) and THE (6 ml). Recrystallization from ethyl acetate and hexanes provided 0.24 g of a yellow solid, m.p. 124.5-1260C.

PCTIUS96IO4759 -98- EXAM PIE

N-(

4 Bromo3methyli io)--noylnroylbenobhen 3 slfo mide A. 2 -n-propylbenzofblthiophene 2 -n-Propylbenzo[bjthiophene was prepared by the method of Example 40A with benzo[blthiophene (7.5 mmoles, 1.0 t-BuLi (1 .7 M, 9.7 mmoles, 5.7 ml), n-bromopropane (9.7 mmoles, 0.88 ml) and THE.

(20 ml). 1.11 g of a light yellow liquid was isolated.

B. 2 -propylbenzofblthiophene-3-sulfony chloride 10 2 -Propylbenzofblthiophene-3sulfoflyI chloride was prepared by the method of Example 408 with DMVF (3.6 mmoles, 0.28 ml), sulfuryl chloride (3.1 mmoles, 0.25 ml) and 2 -propylbenzo[blthiophene (1 .8 mmoles, 0.32 Flash chromatography ethyl acetate/hexanes) provided 0.28 g of a yellow solid.

15 C. N-( 4 -bromo3methyl5isoxazolyl)2npropylbenzo[blthiophene-3sulfonamide N-(-rm -3m t l -s aoy)-2npoplez bt o e -3 sulfonamide was prepared by the method of Example 41 with 4-bromo-3methyl-5-aminoisoxazole (0.68 mmoles, 0.12 g) NaH (1.7 mmoles, 20 6.8 mg), 2 -n-propylbenzo[blthiophene.3-sulfoflyI chloride (0.82 mmoles, 0.23 g) and THE (3 ml). Recrystallization from chloroform and hexanes provided 0. 19 g of a yellow crystalline solid, m.p. 136-138 0

C.

EXAMPLE 46

N-(

4 Bromo3methyl.5isoxazolyl)2propylbenz[bthohe-3sloa mide A. 2 -/-propylbenzofbjthiophene 2 -i-Propylbenzo[bjthiophene was prepared by the method of Example 40A with benzolblthiophene (7.5 mmoles, 1.0 t-BuLi (1.7 M, 11 .2 mmoles, 6.6 ml), 2 -iodopropane (11. 2 mmoles, 1. 12 ml) and THE -99ml) with stirring at ambient temperature for 24 hrs. It was isolated as a yellow oil (1.11 g; 85% yield).

B. 2 -i-propylbenzo[bthiophene3sufonyI chloride 2 -i-propylbenzofblthiophene-3sufolyI chloride compound was prepared by the method of Example 408 with DMVF (5.2 mmoles, 0.40 ml), using sulfuryl chloride (4.2 mmoles, 0.34 ml) and 2-i-propylbenzo[blthiophene (2.1 mmoles, 0.37 Flash chromatography (1 ethyl acetate/hexanes) provided 0. 17 g of a yellow solid.

C

N-(

4 bromo3methyl-soa..i)-2-i-pryliproybblhohe-3 sulfonamide sulfonamide was prepared bthmeodfExpl41using 4-bromo- 3 -methyl-5-aminoisoxazole (0.55 mmoles, 9.7 mg), NaH (1.4 mmoles, 5.5 mg), 2 -ipropylbenzo[bjthiophene..3sulff 0 yI chloride (0.60 mmoles, 15 0. 17 g) and THF (2 ml). Recrystallization from chloroform and hexanes provided 89 mg of a tan crystalline solid, m.p. 157.5-1590C.

EXAMPLE 47

N-(

4 Bromo3methyl.5.isoxazol )2(4ethylbzl)benzob]-thiophene-3sulfonamide A a-(2-benzo [blthiophene)-4-ethylbenzyI alcohol a-1&-Benzo[blthiophene)-4ethylbenzyl alcohol was prepared by the method of Example 40A with benzol~blthioph ene (7.5 mmoles, 1.0 g), t-BuLi (9.7 mmoles, 1.7 M, 5.7 ml), 4 -ethylbenzaldehyde (8.9 mmoles, 1.22 ml) and THF (20 ml). Flash chromatography (10% ethyl acetate/hexanes) provided 1.79 g of a yellow solid.

B. 2 4 -ethylbenzyl)benzofblthiophene To a solution of a-( 2 -benzofblthiophene)-4ethylbenzy alcohol mmoles, 1. 1 triethylsilane (4.4 mmoles, 0. 11 ml) and CH 2 CI, (20 ml) at O'C was added TFA (8.1 mmoles, 0.62 ml). The solution was stirred 30 min at 0 0 C, then diluted with ether (100 ml) and washed with sat.

-100- NaHCQ 3 (100 ml). The organic phase was dried (MgSO 4 filtered and concentrated. Flash chromatography ethyl acetate/hexanes) provided 0.69 g of a white solid.

C. 2 4 -Ethylbenzy)benzo[bthiophene-3suffonyI chloride 2 4 -Ethylbenzyl)benzobthiophene3sulfonyI chloride was prepared by the method of Example 408 with DMF (5.4 mmoles, 0.42 ml), sulfuryl chloride (4.6 mmoles, 0.37 ml) and 2-(4ethylbenzyl)benzotblthiophene (2.7 mmoles, 0.69 Flash chromatography ethyl acetate/hexanes) provided 0.43 g of an 10 orange solid.

D. N-( 4 bromo3me hy5isoxzoli)-2-azolyl2(ethyl)bz[b .thiophene-3-sulfonamide 15thiophene-3-sulfonamide was prepared by the method of Example 41 4-rm--ehl5aiosxzl (1.0 mmoles, 0. 177 NaH mmoles, 100 mg), 2 4 -ethylbenzyl)benzo[bthiophene3-suffnyj chloride (1.2 mmoles, 0.42 g) and THF (6 ml). Flash c hromatography ethyl acetate/hexanes) followed by recrystallization from chloroform and hexanes provided 0.21 g of a tan solid, 20 m.p. 128-130 0

C.

EXAMPLE 48

N-(

4 Bromo3methyl.5isoxazoll)2[ 3 4(thyl1ndioybny benzo [blthiophene-3-suffonamide A. a-( 2 -benzobthienyl).34(methylenedioxy)bezy alcohol a-2-BezofbthieyI)-3,-methylenedoxy)benzyl alcohol was prepared by the method of Example 40A using benzo[blthiophene mmoles, 1 .0 t-BuLi (1.7 M, 9.7 mmoles, 5.7 ml), piperonal (8.9 mmoles, 1 .0 g) and THF (20 ml). Flash chromatography (20% ethyl acetate/hexanes) provided 1.6 g of a yellow solid.

-101- B. 2 3 ,4-(methylenedioxy)benzyllbenzo~blthlophene 2 3 4 -(methylenedioxy)benzyllbenzo[bjthiophene was prepared by the method of Example 478 using a-( 2 -benzo[blthienyl)..3,4.(methylenedioxy)benzyl alcohol (6.2 mmoles, 1 .8 triethylsifane (6.8 mmoles, 1. 1 ml) CH 2

CI

2 (50 ml) and TFA (12.4 mmoles, 0.95 ml). Recrystallization from hexanes provided 1.2 g of a light orange solid.

C. 2 3 4 -(methylenedioxy)benzyllbenzo[bjthiophene- 3 -sIfonI chloride 2 3 4 (methylenedioxy)benzylbenzobthiophene 3 sfonI chloride was prepared by the method of Example 408 using DMF (9.1 mmoles, 0.70 ml), sulfuryl chloride (7.7 mmoles, 0.62 ml) and 2-[3,4- (methylenedioxy)benzyllbenzofblthiophene (4.6 mmoles, 1.2 Flash chromatography ethyl acetate/hexanes) provided 0.71 g of a light yellow solid.

D0. N-4boo3mty--sxzli--34(ehlndoybnyj benzo[blthiophene.3suffonamide N-4bom--eh* .soaoy) -34-mtyendoybenzyllbenzo[blthiophene-3-sulfonamide was prepared by the method of Example 41 using 4 -bromo-3-methyl-5-aminoisoxazole (1.0 mmoles, 0. 177 g), NaH (2.5 mmoles, 100 mg), 2 -f 3 4 -(methylenedioxy)benzyllbeflzo[blthiophene-3-sulfonyl chloride (1.1 mmoles, 0.40 g) and THF (7 ml). Flash chromatography (50% ethyl acetate/hexanes) followed by recrystallization from chloroform and hexanes provided 0.23 g of a tan crystalline solid, m.p. 1 64-1 65 0

C.

EXAMPLE 49 N-4Boo3mty--ioaoy ez-,, 3 -thiadiazole-4..sulfonamide N-4boo3mty--ioaoy ez-,, 3 -thiadiazole-4sufon.

amide was prepared from 5-mn--rm--ehlsxzl and 2,1,3thiadiazole-4-sulfonyl chloride according to the procedures described in Example 39. The crude product was purified by recrystallization from -102ethyl acetate/hexanes to give a crystalline solid, m.p. 177-1 790 C, yield 34%.

EXAMPLE N-(4-Bromo-3-methyf-5isoxazolyl). 1 -methylindole-2-sulfonamide A. 2-Methylindole-2-sulfonyl chloride 2 -Methylindole-2-sulfonyl chloride was prepared by the method of Example 34 with 1-methylindole (7.8 mmols, 1.0 ml), t-BuLi (1.7 m, 9.4 mmols, 5.5 ml), sulfur dioxide, NCS (8.6 mmols, 1.2 g) and THF (15 ml).' Flash chromatography ethyl acetate/hexanes) provided 0.66 g (36%) 10 of a yellow solid.

B .b o o 3 m th l 5 i o a o yi e h l n ol u f n m B. N-( 4 -bromo-3-methyl.5isoxazolyl). 1 -methylindole-2-sulfonamide was prepared by the method of Example 41 with 4 aminoisoxazole (1.0 mmols, 0.18 NaH (2.5 mmols, 60 mg), 1i methylindole-2-sulfonyl chloride (1.2 mmols, 0.26 g) and THF (7 ml).

Recrystallization from chloroform and hexane provided 0.28 g of a brown solid, m.p. 132-1340

C.

EXAMPLE 51 20 A. 2-Dibenzofuransulfonyl chloride 2 -Dibenzofuransulfonic acid (12.8 mmol) was heated at 700 C with phosphorus oxychloride (1.30 ml, 14.0 mol) for 2 h. Excess phosphorus oxychloride was removed under reduced pressure. The residue was decomposed with ice water and extracted with ethyl acetate. The extract was washed with 5% sodium bicarbonate solution, dried over anhydrous magnesium sulfate and concentrated to yield 2.9 g crude 2-dibenzofuransulfonyl chloride.

-103- B. N-( 3 ,4-Dimethyl-5-isoxazolyl)-2-dibenzofuransulfonamide The 2-benzofuransulfonyl chloride from step was added to a solution of 5-amino-3,4-dimethylisoxazole (250 mg, 2.2 mmol) and 4- (dimethyl)aminopyridine (5 mg) in dry pyridine (2.0 ml). The reaction mixture was stirred at room temperature for 4 h. Pyridine was removed under reduced pressure and the residue was partitioned between water and ethyl acetate. The organic layer was washed with 1N HCI (2 X ml), brine (25 ml) and dried over anhydrous magnesium sulfate.

S* Evaporation of the solvents left an oily residue that, after purification by column chromatography over silica gel methanol in chloroform as .eluent), yielded white solid (32% yield). Purification was achieved by recrystallization from chloroform/hexanes to give a white "cotton-like" solid, m.p. 173-175° C (dec.).

EXAMPLE 52 15 N-(4-chloro-3-methyl-5-isoxazolyl)-2-[2-(3,4-methylenedioxy)phenyl]ethoxycarbonyl-3-sulfonamide N-(4-chloro-3-methyl-5-isoxazolyl)-2-[2-(3,4-methylenedioxy)phenyl]ethoxycarbonyl-3-sulfonamide was prepared by the same method as described in Example 97 with the exception that 2 3 ,4-methy 7 ne- 20 dioxy)phenylethanol was used instead of sesamol. The final product was obtained by HPLC purification as a yellowish oil, (500mg, 25% yield).

EXAMPLE 53 N-(4-Bromo-3-methyl-5-isoxazolyl)-2-phenylthiophene-3-sulfonamide A. 3 -bromo-2-phenyl-thiophene Tetrakis (triphenylphosphine) palladium (400 mg), Na 2

CO

3 (4 M, ml, 320 mmol) and phenylboric acid (3.81 g, 30.3 mmol) as a solution in ethanol (80 ml) were sequentially added to a solution of 2,3dibromothiophene (7.33 g, 30.3 mmol) in benzene (100 ml). The mixture was heated at reflux for 12 hours. The aqueous layer of the crude mixture was removed and the organic layer was diluted with EtO2 (200 -104ml), washed with 1N NaOH (2 x 150 mi) and was dried (MgSO 2 filtered and the soluent was evaporated. The residue was chromatographed using hexane as the eluent to give 3 -bromo-2-phenylthiophene as a clear oil (3.31 g, 47% yield).

B. 2 -Phenylthiophene-3-sulfonylchloride n-BuLi (2.38 M, 11.5 ml, 27.28 mmol) was slowly added to a solution of 3 -bromo-2-phenyl-thiophene (22.73 mmol) in ether (50 ml).at 00 C. The reaction was stirred at 00 C for 1 h. SO, was bubbled Sthrough the mixture for 15 minutes at 0 OC followed by the addition of NCS (3.95 g, 29.55 mmol) as a suspension in THF (20 ml). The crude products were purified by column chromatography (hexanes) to give 2phenylthiophene-3-sulfonylchloride as a white solid (1.23 g, 34% yield).

C. N 4 -bromo-3-methyl-5-isoxazolyl)-2-phenylthiophene-3-sulfona- S-mide N-(4-bromo-3-methyl-5-isoxazolyl)-2-phenyl-thiophene-3-sulfonamide was prepared from 2 -phenyl-3-thiophene sulfonylchloride using the method described in Example 1. The product was purified by HPLC, 77% yield, reddish solid, mp 86-890

C.

EXAMPLE 54 20 3-Phenoxy-N-(4-bromo-3-methyl-5-isoxazoly)thiophene2sufon a mide A. 3 -Phenoxythiophene.

Cuprous chloride (3.08 g, 31.1 mmol) and phenol (8.78 g, 93.3 mmol) were sequentially added to a solution of 3 -bromothiophene (5.06 g, 31.1 mmol) in pyridine (150 mi). Sodium hydride (3.73 g, 93.3 mmol, 60% dispersion in mineral oil) was then slowly added. The reaction was heated at reflux for 20 hours under Argon. The pyridine was removed under reduced pressure. The residue was diluted with EtO2 (200 ml) and washed with 1 N NaOH (3 x 100 ml), 1 N HCI (2 x 150 ml) and 1 N NaOH (150 ml). The organic layer was dried (MgSO 4 filtered, and the -105solvent was evaporated. The residue was chromatographed using hexanes to give 3 -phenoxy-thiophene as a clear oil (4.0 g, 74% yield).

B. 3 -Phenoxythiophene-2s uffonyl chloride BuLi (2.38 M, 11.5 ml, 27.28 mmol) was slowly added to a solution of 3 -phenoxythiophene (4.0 g, 22.73 mmol) in ether (50 ml) at 00 C. The reaction was stirred at 00 C for 1 h. SO, was bubbled through the mixture for 15 minutes at 0 OC followed by the addition of NCS (3.95 g, 29.55 mmol) as a suspension in THF (20 ml). The mixture was allowed to warm up to 250 C and stirred for 2 more h. The 10 precipitate was filtered, and the filtrate was concentrated and chromatographed (hexanes) to give 3 -phenoxythiophene-2- sulfonyl chloride as a yellowish solid (1.03 g, 17% yield).

C. N-(4-bromo-3-methyl-5-isoxazolyl)-3-phenoxythiophene-2-sulfonamide 15 N-(4-bromo-3-methyl-5-isoxazoly1)-3-phenoxythiophene-2-sulfonamide was prepared from 3 -phenoxythiophene-2-sulfonyl chloride and Samino-4-bromo-3-methylisoxazole using the method described in Example 1. The product was recrystallized from acetonitrile/H20, to give S 2 a solid m.p. 121-1230C, 61% yield.

20 EXAMPLE N-(4-Bromo-3-methyl-5-isoxazolyl)-2-[N-(4-isopropylphenyl)aminocarbonyl]thiophene-3-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-2-[N-(4-isopropylphenyl)aminocarbonyllthiophene-3-sulfonamide was prepared in the same manner as described in Example 24 from

N-(

4 isoxazoly)-2-(carboxyl)thiophene-3-sulfonamideand 4 -isopropylaniline in 19% yield. The crude product was passed through silica gel column using ethyl acetate as eluent. This was further purified by HPLC CHCN to 100% CHCN over 30 min.) to give a solid.

-106- EXAMPLE 56

N-(

4 -Bromo..3.methyl-5..isoxazolyl) 2 -[N-(4-sec-butylphenyI)aminocarbonyljthiophene.3-sulfonamide

N-(

4 -bromo-3..methyl. 5-isoxazolyl)-2-[N(4..sec..butylphenyl) aminocarbonyllthiophene-3sulfonamide was prepared in the same manner as described in Example 24 from

N-(

4 -bromo-3-methyl..5-.isoxazolyl)- 2 (carboxyl)thiophene-3sulfonamide and 4 -sec-butylaniline in 25% yield.

The crude product was passed through silica gel column using ethyl acetate as eluent. This was further purified by HPLC

CH

3 CN to woo* 10 100%

CH

3 CN over 30 mi.) give a solid, m.p. 205 2080C.

EXAMPLE 57

N-(

4 -Bromo-3-methyl..5.isoxazolyl)..2.[N.(4.tet-butylphenyl)amino *carbonyllthiophene.3-sulfonamide amino carbon yfIthio ph en e- 3-sulfonamid was prepared in the same manner as described in Example 24 from N-( 4 -bromo..3.methyls..

*.ll)2(aboy~hopee3sufnmd and 4 -tert-butylaniline in 28% yield. The crude product was passed through silica gel column using ethyl acetate as eluent. This was further purified by

CH

3 CN to 100%

CH

3 ,CN over 30 min.) give a solid, m.p. 76 860C.

EXAMPLE 58 bony1 Ithiophene-3suffonamide was prepared in the same manner as described in Example 24 from N-4boo3mtyl5ioaoy)2 (croy~hohn--ufnmd and 4 -butylaniline in 18% yield. The crude product was passed through silica gel column using ethyl acetate as eluent. This was further Purified by HPLC

CH

3 CN to 100%

CH.

3 CN over 30 min.) give a solid.

-107- EXAMPLE 59

N-(

4 -bromo-3-methyl-5-isoxazolyl)thiazole-2-sulfonamide A. Thiazole-2-sulfonyl chloride Thiazole (0.51 g, 6 mmol) was dissolved in THF (5 ml) and cooled to -78 0 C under argon atmosphere. n-Butyllithium (2.5 M solution in hexane, 2.4 ml, 6 mmol) was added dropwise under constant stirring.

The resultant reaction mixture was stirred at -78 0 C for 40 min. Sulfur dioxide was bubbled through the reaction mixture for 15 min at -78 0

C.

The reaction mixture was allowed to attain ambient temperature slowly 10 and stirred for 30 min. NCS was added and stirring was continued for S:min. The reaction mixture was diluted with water (50 ml), extracted with i' ethyl acetate (2 X 50 ml) and the combined organic layers was dried over anhydrous MgSO 4 Removal of the solvent under reduced pressure gave crude product which was purified by column chromatography, using 15 hexane as eluent, to give thiazole-2-sulfonyl chloride as a liquid(0.6 g, 54% yield).

N-(4-bromo-3-methyl-5-isoxazolyl)thiazole-2-sulfonamide

N-(

4 -bromo-3-methyl-5-isoxazolyl)thiazole2-sulfonamide was prepared in the same manner as described in Example 2 from 5-amino-4- 20 bromo-3-methylisoxazole and thiazole-2-sulfonyl chloride in 57% yield.

This was purified by HPLC

CH

3 CN to 100% CH 3 CN over 30 min.) to give a solid., m.p. 175 177 0

C.

EXAMPLE

N-(

4 -chloro-3-methyl-5-isoxazolyl)thiazole-2-sulfonamide N-(4-chloro-3-methyl-5-isoxazolyl)thiazolesulfonamide was prepared in the same manner as described in Example 2 from 5-amino-4chloro-3-methylisoxazole and thiazole-2-sulfonyl chloride in 33% yield.

This was purified by HPLC

CH

3 CN to 100% CH 3 CN over 30 min.) to give a solid, m.p. 171-173OC.

-108- EXAMPLE 61

N-(

3 ,4-dimethyl-5-isoxazolyl)thiazole-2-sulfonamide N-(3, 4 -methyl-5-isoxazolyl)thioazole-2-sulfonamide was prepared in the same manner as described in Example 14 from 5-amino-3,4dimethylisoxazole and thiazole-2-sulfonyl chloride in 37% yield. This was purified by HPLC

CH

3 CN to 100%

CH

3 CN over 30 min.) give a solid, m.p. 118 120 0

C.

EXAMPLE 62 :*to 5-benzyl-N-(4-

B

romo-3-methyl-5-isoxazolyl)thiophene-2-sulfonamide 10 A. 1-( 2 -Thienyl)benzyl alcohol Si:: Sodium borohydride (0.37 g, 10 mmol) was added to 2benzoylthiophene (1.88 g, 10 mmol) dissolved in methanol/THF mixture (1:10 ratio, 11 mi). This was stirred at room temperature for 10 h. The reaction mixture was decomposed by addition of saturated ammonium 15 chloride solution (50 ml) and was extracted with ethyl acetate (2 X ml). The combined organic layers was dried over anhydrous MgSO,.

Removal of the solvent gave 1-( 2 -thienyl)benzyl alcohol as a solid (1.75 g, 92% yield).

B. 2 -Benzylthiophene 20 Acetic anhydride (5 ml) was added to a solution of 1-(2thienyl)benzyl alcohol in pyridine. The resultant solution was stirred at 0 C for 3h. Water (50 ml) was added and the reaction mixture was stirred at room temperature for 2h. This was extracted with ethyl acetate (2 X 50 ml) and the combined organic layers dried over anhydrous MgSO 4 Removal of the solvent gave crude product, which was purified by passing through silica gel using 3:1 hexane/ethyl acetate mixture to give 1-( 2 -thienyl)benzyl acetate.

A solution of 1-( 2 -thienyl)benzyl acetate in THF (5 ml) was added carefully to dry liquid ammonia (100 ml). Lithium metal was added in -109small portions until the blue color persisted. The resulting reaction mixture was stirred for 30 min, and the reaction was quenched by addition of solid ammonium chloride. The residue, after complete evaporation of liquid ammonia, was dissolved in water (50 ml) and was extracted with methylene chloride (2 X 50 ml). The combined organic layers was dried over MgSO, and filtered. Removal of the solvent gave crude product, which was purified by column chromatography using hexane as eluent to give 2 -benzylthiophene (1.2 g, 68 yield).

5-Benzylthiophene-2-sulfonyl chloride 10 To a solution of 2 -benzylthiophene (0.875 g, 5 mmol) in chloroform (2 ml) at 0°C was added chlorosulfonic acid dropwise and the reaction was stirred at 0°C for 30 min. The reaction mixture was decomposed by pouring onto crushed ice (20 The mixture was extracted a with ethyl acetate, dried over MgSO, and filtered. The solvent was re- 15 moved under reduced pressure to give 5-benzylthiophene-2-sulfonic acid.

Phosphorous pentachloride (2.08 g, 40 mmol) was added to a solution of 5-benzyithiophene-2-sulfonic acid in phosphorous oxychloride g, 40 mmol) at 00C. The reaction mixture was kept at 500C for 1 h, cooled to room temperature, then poured onto crushed ice (50 g) and 20 extracted with ethyl acetate (2 X 30 ml). Removal of the solvent under reduced pressure gave a crude product, which was purified by column chromatography using 3% ethyl acetate in hexane to give 2-benzylthiochloride (0.6 g, 39 yield).

SD. N-(4-bromo-3-methyl-5-isoxazolyl)-5-benzylthiophene-2-sulfonamide N-(4-Bromo-3-methyl-5-isoxazolyl)-5-benzylthiophene-2-sulfonamide was prepared in the same manner as described in Example 2 from amino-4-bromo-3-methylisoxazole and 5-benzyl-2-thiophenesulfonyl chloride in 22% yield. The product was purified by HPLC

CH

3 CN to 100%

CH

3 CN over 30 min.) to give a solid, m.p. 49 500C.

-110- EXAMPLE 63

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-3-phene thliophene-2sulfonamide A. 1 3 -Thienyl)phenethyl alcohol Benzyl bromide (25.65 g, 150 mmol) was added dropwise over 8 h to a suspension of magnesium (3.6 g, 150 mmol) in ether (75 ml) dissolved in ether (30 ml). The resulting mixture was cooled to -100C.

3 -thiophenecarboxaldehyde in ether (45 ml) over 30 min was then added and the resultant reaction mixture was stirred at room temperature for 6 S 10 h. This was cooled to 00C and the reaction mixture was decomposed by addition of 0.1 N HCI. The ether layer was separated and the aqueous phase was extracted with ethyl acetate (2 X 50 ml). The combined organic layers was dried over MgSO 4 and filtered. Removal of the solvent gave 1-( 3 -thienyl)phenethyl alcohol (16 g, 78% yield).

B. 1-( 3 -Thienyl)phenethyl acetate 1-( 3 -Thienyl)phenethyl alcohol (10 g, 49 mmol) was dissolved in a 2:1 pyridine and acetic anhydride mixture (50 ml). This was stirred at 800C for 4 h. Excess of pyridine and acetic anhydride mixture was S: removed under reduced pressure and the residue was dissolved in-water 20 (100 ml). This was extracted with methylene chloride (3 X 75 ml) and the combined organic layers was dried over MgSO, and filtered. Removal of the solvent gave 1-( 3 -thienyl)phenethyl acetate (10.2 g, 84% yield).

C. 3 -Phenethylthiophene 1-( 3 -thienyl)phenylethyl acetate dissolved in THF (20 ml) was added carefully to dry liquid ammonia (300 ml). Lithium metal was added in small portions until the blue color persisted. The resulting reaction mixture was stirred for 30 min and the reaction was quenched by addition of solid ammonium chloride. The residue, after the complete evaporation of liquid ammonia, was dissolved in water (100 ml) and was extracted with methylene chloride (4 X 50 ml). The combined organic -111layers was dried over MgSO 4 and filtered. Removal of the solvent gave a crude product, which was purified by column chromatography using hexane followed by mixture of ethyl acetate in hexane as eluent to give 3 -phenethylthiophene (3.2 g, 34 yield) and 1-( 3 -thienyl)phenethyl acetate (starting material, 7g).

D. 3 -Phenethylthiophene-2-sulfonyl chloride and 4phenethylthiophene-2-sulfonyl chloride 3 -Phenethylthiophene (0.94 g, 5 mmol) was dissolved in THF (12 10 ml) and cooled to -780C under argon atmosphere. n-Butyllithium (2.5 M solution in hexane, 4.4 ml, 5.5 mmol) was added dropwise with constant stirring under an argon atmosphere. The resultant reaction mixture was stirred at -100C to 00C for 3 h, cooled to -780C and sulfur dioxide was bubbled through the reaction mixture for 15 min. The reaction mixture 15 was allowed to attain ambient temperature slowly and stirring continued for 30 min. NCS (1 g) was added and stirring was continued for 1 h. The ::reaction mixture was diluted with water (50 ml), extracted with methylene chloride (2 X 50 ml) and the combined organic layers was dried over anhydrous MgSO 4 Removal of the solvent under redueed 20 pressure gave a crude product which was purified by column chromatography, using 0.2% ethyl acetate in hexane as eluent, to give 3phenethyl-2-thiophenesulfonyl chloride (0.06 g, 4% yield) and 4phenethyl-2-thiophenesulfonyl chloride (0.72 g, 45% yield).

2 E. N-( 4 -bromo-3-methyl-5-isoxazolyl)-3-pheneththtiophene- 2 -sufonamide N-(4-bromo-3-methyl-5-isoxazolyl)-3-phenethylthiophene-2-sulfonamide was prepared in the same manner as described in Example 2 from amino-4-bromo-3-methylisoxazole and 3-phenethyl-2-thiophenesulfonyl chloride in 48% yield. This was purified by HPLC CHCN to 100%

CH

3 CN over 30 min.) to give a solid.

-112- EXAMPLE 64

N-(

4 2 -ufnmd

N-(

4 bromo3.methyl5.isoxazoly4phenthlthihe-2sfoa mide was prepared in the same manner as described in Example 2 from amino- 4 -bromo3methylisoxazole and 4 -phenethyl-2-thiophenesulfonyl chloride in 32% yield. This was purified by HPLC

CH

3 CN to 100%

CH

3 CN over 30 min.) to give a gum.

EXAMPLE 4 -Bromo3methyl5.isoxazolyl).5(3methoxypheny,)thiophene 2 l- A. 5-bromothiophene.2-sulfonyl chloride Chlorosulfonic acid was added dropwise over 20 min. to a cold solution (-78 0 C) of 2 -bro moth io phene (16.3 g, 100 mmol) in methylene chloride (50 ml) was added After addition of chlorosulfonic acid was complete, the cold bath was removed. The reaction mixture was allowed to attain room temperature slowly (2 was added dropwise onto the crushed ice (1000 g) and was extracted with methylene chloride (4 X .20 100 ml). The combined organic layers was dried over MgSO 4 filtered and the solvent was removed under reduced pressure to give a crude product. This was purified by column chromatography using hexane as eluent to give 5-bromothiophene.2-sulfonyl chloride (22 g, 75% yield).

B. N-(5-bromothiophene..2.sulfonyl)pyrrole N-(5-bromothiophene..z-sulfonyl)pyrrole was prepared in the same manner as described in Example 33A from 5-bromothiophene-2-.sulfonyl chloride and pyrrole in 88% yield. This was purified by recrystallization using hexane/ethyl acetate as a solvent.

C. 3 -Methoxyphenylboronic acid 3 -Methoxyphenylboronic acid was prepared in the same manner as described in Example 338 from 3-bromoanisole and triisopropyl borate in -113- 82% yield. This was used in the next step without any further purification.

D. N-[5-(3-methoxyphenyl)thiophene-2-sulfonyl]pyrrole 3 -methoxyphenyl)thiophene-2-sulfonyl]pyrrole was prepared in the same manner as described in Example 32C from 3methoxyphenylboronic acid and N-(5-bromothiophene-2-sulfonyl)pyrrole in 93% yield. This was purified by recrystallization using hexane/ethyl acetate as solvent.

E. 5-( 3 -Methoxyphenyl)thiophene-2-sulfonyl chloride To a suspension of N-[5-(3-methoxyphenyl)thiophene-2-sulfonyllpyrrole (1.4 g, 4.5 mmol) in ethanol (15 ml) was added 6 N sodium hydroxide solution (15 ml) and the resultant reaction mixture refluxed for 14 h. The reaction mixture was cooled to room temperature. Ethanol was removed under reduced pressure and the resultant precipitate was filtered and dried under vacuum (1.1 g, 91% yield).

Phosphorous pentachloride (2.08 g, 10 mmol) was added to the suspension of sodium slat of sulfonic acid (0.62 g, 2.5 mmol) (obtained from above step) in phosphorousoxy chloride (0.93 ml, 10 mmol) and the resultant reaction mixture stirred at room temperature for 3 h. This was decomposed by adding on to crushed ice and the product was extracted with methylene chloride (2 X 50 ml). The combined organic layers dried over MgSO, and filtered. Removal of the solvent gave the crude product which was purified by column chromatography using 2% ethyl acetate in hexane to give 5-( 3 -methoxyphenyl)thiophene-2-sulfonyl chloride (0.51 g, -114-

C

C.

C C C.

C

C.

F. N-( 4 bromo3.methyl5isoxazoly)5(3-methoxyhi)tipe- 2-sulfonamide N- 4 -bromo-3methyl 5isoxazo lyl)5(3-meth oyhiny)tiphne 2 -sulfonamide was prepared in the same manner as described in Example 2 from 5-amino-4-bromo.3methylisoxazole and 5- 3 -methoxyp hen yl)thiophene-2-sulfonyl chloride in 48% yield. This was purified by HPLC

CH

3 CN to 100% CH 3 CN over 30 min.) give a solid.

EXAMPLE 66

N-(

4 Bromo.3methyl5.isoxazol )5(2thylfur Iy)thohn--uf amide A. N-(pyrrole)5(2methylfuyraiony-thiopenaifde t-BuLi (1.7 mn solution in hexane, 7.9 ml, 14.6 mmol) was added dropwjse under constant stirring under a nitrogen atmosphere to a 15 solution of 2-methyl furan 1 .0 g, 12 mmol) in THIF (20 ml) at -78 0 C.

The solution was then warmed to -100 C and stirring was continued for min. The solution was then added to a solution of zinc chloride (27 ml of a 0.5 M solution in THF) at -300 C and then warmed to room temperature where stirring continued for 1 hr. resulting in a pale yellow 20 clear solution. The solution was then transferred via a steel canula under nitrogen to a solution of N-(pyrrole)-5bromothiophene.2sulfonamide (Example 33A, 3.5 g, 12 mmol) and tetrakis(triphenylphosphine)palladium (693 mg, 0.6 mmol) in THF (15 ml) at -78 0 C. The solution was then warmed to room temperature and stirred for a period of 2 hours. Purification by column chromatography using 2% ethyl acetate gave 680 mg of N-(yr e-5(-ehf rny)ti hn-2sf nai as a pale yellow powder (19% yield).

B. 2 2 -methylfuranyl)thiophene5sulfonyI chloride 2 2 -methyfuranyl)thiophene5sulfonyI chloride was prepared in the same manner as described in Example 33D from N-(pyrrole)-5-(2mehfuay~hohn--ufnmd (300 mg, 1.02 mmol).

-115-

C

C

C

Purification by column chromatography using 2% ethyl acetate/hexanes gave 145 mg of the sulfonyl chloride as a pale yellow solid.

C. N-( 4 -bromo-3-methyl-5-isoxazolyl)-5-(2-methyl-furanyl)thiophene-2sulfonamide N-(4bromo-3-methyl-5-isoxazolyl)-5-(2-methyl-furanythiophene sulfonamide was prepared in the same manner as described in Example 2.

Reaction of 2 2 -methylfuranyl)thiophene-5-sulfonyl chloride (55 mg, 0.21 mmol) with 5-amino-4-bromo-3-methyl isoxazole (41 mg, 0.21 mmol), after purification by column chromatography using MeOH/CHCI 3 gave 45 mg of the pure sulfonamide as a brown semisolid, 54% yield, m.p. 123-124o

C.

EXAMPLE 67 N-(4-Bromo-3-methyl-5-isoxazolyl)-5-(4-methoxyphenyl)thiophene-2-sulfonamide A. 4 -methoxyphenyl)thiophene-2-sulfonyl]pyrrole N-[5-(4-methoxyphenyl)thiophene-2-sulfonyl]pyrrole was prepared, in the same manner as described in Example 32C, from 4 -methoxyphenylboronic acid and N-(5-bromothiophene-2-sulfonyl)pyrrole.

Recrystallization using hexane/ethyl acetate gave a solid in quantative yield.

B. 5-chlrnom rlfnn IL y. i- I I oxuAyphenyl/thiophene A solution of N-[5-(4-methoxyphenyl)thiophene-2-sulfonyl]pyrrole (1.4 g, 4.5 mmol) was suspended in ethanol (15 ml). A 6N sodium hydroxide solution was added, and the resulting suspension was refluxed for 14 hr. to give a clear solution. This was cooled to room temperature.

Ethanol was removed under reduced pressure. A precipitate was formed on standing at room temperature which was filtered and washed with methylene chloride and dried under vacuum giving a solid (1.2 g, 91%).

The solid (0.67 g, 2.5 mmol) was suspended in phosphorous oxychloride (0.92 ml, 10 mmole) and phosphorous pentachloride (2.08 g, -116mmole) was added. The resulting mixture was stirred at room temperature for 3 hr. The reaction mixture was decomposed by pouring onto crushed ice (50 The mixture was extracted with methylene chloride (2 x 50 ml) and the combined organic layers was dried over MgSO 4 Removal of solvent under reduced pressure gave a crude product which was purified by column chromatography using 2% EtOAc in hexane as solvent to give 5-chlorosulfonyl-2-(4-methoxyphenyl)thiophene (530 mg, 86%).

1 C. N-(4-brom '-3-methyl-5-isoxazolyl)-5-(4-methoxyphenyl)thiophene- 10 2 -sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-5-(4-methoxyphenyl)thiophene-

S

2 -sulfonamide was prepared in the same manner as described in Example S 1. Reaction of 5-chlorosulfonyl-2-(4-methoxyphenyl)thiophene with amino-4-bromo-3-methylisoxazole gave N-( 4 isxazolyl)-5-(4-methoxyphenyl)thiophene-2-sulfonamde in 50% yield, m.p. 128-1300 C.

EXAMPLE 68

N-(

4 -Bromo-3-methyl-5-isoxazolyl)5(3thienyl)thiophene-2sufonamide 20 A. 3 -Thiopheneboric acid n-Butyllithium (2.5 M solution in hexane, 20 ml, 50 mmol) was added dropwise to a solution of 3 -bromothiophene (8.15 g, 50 mmol) in THF (20 ml) at -78 0 C under an argon atmosphere. The resulting solution was stirred at -780C for 45 min, and then added to a solution of triisopropyl borate (9.4 g, 50 mmol) in THF at -780C over 30 min through a steel cannula. The resulting reaction mixture was stirred at room temperature for 12 h and was decomposed by the addition of 100 ml 1N HCI. The aqueous layer was extracted with ether (2 X 100 ml) and the combined organic layers was extracted with 1 M NaOH (3 X 30 ml), the aqueous extract was acidified with concentrated HCI to pH 2 and .117extracted with ether (3 X 50 ml). The combined ether extract was washed once with water, dried over MgSO 4 and filtered. Removal of the solvent gave 3 -thenylboronic acid as a solid (5.2 g, 80% yield).

B. 3 -thienyl)thiophene.2.sulfonyllpyrrole N-L.-(.-thienyI)thiophene2.sulfonyllpyrrole was prepared in the same manner as described in Example 32C from 3 -thienylboronic acid and N- (5-bromothiop hen e.2-sufonyl) pyrrole in quantative yield. This was.

purified by recrystallization using hexane/ethyl acetate as solvent.

C 5-( 3 -Thienyl)thiophene-2sulfonyI chloride 5-( 3 -thienyl)thiophene-2sufonyI chloride was prepared in the same manner as described in Example 64E from 4 -methoxyphenyl)thi 0 phene-2-sulfonyllpyrrole in 74% yield.

N-4boo3mty--sxzll--3tinltipee2sfoa mide *.mide was prepared in the same manner as described in Example 2 from amino- 4 -bromo.3-methylisoxazdle and 5-( 3 -thienyl)thiophene.2-sulfonyl a~a. chloride in 40% yield. This was purified by HPLC

CH

3 CN to 100% 20 CH 3 CN over 30 min.) give a solid.

EXAMPLE 69 N (-Bom-3mehl -sxaoy)f a--uf ai N-4boo3mty--sxaoy~ua--ufnmd was prepared by the method of Example 1 with 5-amino-4-bromo-3-methylisoxazole (0.266 g, 1.5 mmol), NaH (60% oil dispersion) (0.15 g, 3.8 mmol) and furan-2-sulfonyl chloride (Example 36A) (0.30 mg, 1.8 mmol).

Flash chromatography (50% EtOAc/hexane) and recrystallization from

CHCI

3 and hexane provided 90 mg (20% yield) of light yellow crystals 117-119 0

C).

-118- EXAMPLE N-4br o3mty-5ioaoy)-5(hnlhofrn--ufn i A. 2-(phenylthio)furan t-BuLi (1.7 m, 10 ml, 1.7 mmol) was added to a solution of furan (1 .24 ml, 17 mmol) in 20 ml of THF at -601C. Thirty minutes later diphenyldisulfide (3.7 g, 17 mmcl) was added via cannula in 8 ml of THF.

The reaction was warmed to ambient temperature for 30 minutes, then diluted with 150 ml of ether and washed with 3% NaOH (3 x 100 ml).

The organic was dried (MgSO 4 filtered and concentrated to collect 2.92 g (97% yield) of a light yellow liquid.

B. 5 -phenylthiofuran-2-suffonyl chloride a S-phenylthiofuran-2-sulfonyl chloride was prepared by the method of Example 34A with 5-phenylthiofuran (1.5 g, 8.5 mmol), t-BuLi (1.2 m, 8.9 mmcl, 5.3 ml) and NOS (1.14 g, 8.5 mmol). Flash chromatography EtOAc/hexane) provided 1.61 g (69% yield) of a yellow-orange liquid.

C. N-( 4 bromo3methyl5isoxazoly)5(phenylthio)furan2sffo mide 20 N-4boo3mty--sxzll)5(hnlhofrn2sloa mide was prepared by the method of Example 1 with 4 -bromo-3-methyl- 2 -aminoisoxazole (0.354 g, 2.0 mmol), NaH (60% oil dispersion) (0.20 g, mmol) and 5-phenylthiofuran-2-sulfonyl chloride (0.66 g, 2.4 mmol).

Flash chromatography (50% EtOAc/hexane) and recrystallization from CHC1 3 /hexane provided 82 mg (10% yield) of a tan solid 91.5 0

C).

-119- EXAMPLE 71

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-5-phenylfuran-2-suffonamide A. 2-Phenylfuran 2-phenylfuran was prepared by the method of Example 32C from 2-bromofuran (0.93 g, 6.3 mmol), sodium carbonate (18 ml of 2 M aqueous solution), phenyl boric acid (0.93 g, 7.6 mmol) and tetrakis (triphenylphosphine) palladium (0.36 g, 0.32 mmol). Flash chroma- Stography with hexane provided 0.79 g (87% yield) of a colorless liquid.

B. 5-phenylfuran-2-sulfonyl chloride 5-phenylfuran-2-sulfonyl chloride was prepared by the method of Example 34A with 2-phenylfuran (0.79 g, 5.5 mmol), t-BuLi (1.7 m, mmol, 3.6 ml) and NCS (0.73 g, 5.5 mmol). Flash chromatography EtOAc/hexane) provided 0.84 g (63% yield) of a light red solid.

C. N-(4-bromo-3-methyl-5-isoxazolyl)-5-phenyfuran-2-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-5-phenylfuran-2-sufonaide was prepared by the method of Example 1 with 4-bromo-3-methyl-2amino isoxazole (0.354 g, 2.0 mmol), NaH (60% oil dispersion) (0.20 g, :5.0 mmol) and 5-phenylfuran-2-sulfonyl chloride (0.58 g, 2.4 mmol).

Flash chromatography (50% EtOAc/hexane) and recrystallization from CHCI,/hexane provided 0.23 g (29% yield) of light yellow crystals (m.p.

124-1260C).

EXAMPLE 72 N-(4-Bromo-3-methyl-5-isoxazolyl)-5-(4-isopropylphenyl)thiophene-2-sulfonamide A. 4 -sopropylphenyl boronic acid 4 -sopropylphenyl boronic acid was prepared in the same manner as described in Example 33B from 1-bromo-4-ethyl benzene. The boronic acid was isolated as a white powder in 63% yield, m.p. 133-135 0

C.

B. N-5-(4-isopropylphenyl)thiophene-2-sulfonylpyrrole -120- 4 -isopropylphenyl)thiophene-2-sulfonylpyrrole was prepared in the same manner as described in Example 33C, from 4 -isopropylphenyl boronic acid and N-(5-bromothiophene sulfonyl)-pyrrole. Purification by column chromatography using 10% ethyl acetate/hexanes gave the pure sulfonamide as an off white colored solid in 84% yield, m.p. 112-114 0

C.

C. 5-chlorosulfonyl-2-(4-ethylphenyl)thiophene 5-chlorosulfonyl-2-(4-ethylphenyl)thiophene was prepared in the same manner as described in Example 33D. Hydrolysis of 526 mg (1.59 mmol) of N-[5-(4-isopropylthiophene)-2-sulfonyl]pyrrole with 6N sodium hydroxide followed by chlorination using phosphorous oxychloride and phosphorous pentachloride gave the crude sulfonyl chloride as dark oil.

Flash column chromatography over silica gel using 2% ethyl acetate/hexanes yielded 262 mg of the pure sulfonyl chloride as a light brown oil.

D. N-( 4 -bromo-3-methyl-5-isoxazolyl)-5-(4-isopropylphenyl)thiophene- S. 2 -sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-5-(4-isopropylphenyl)thiophene- 5: 2 -sulfonamide was prepared in the same manner as described in Example .20 2. Reaction of 5-chlorosulfonyl-2-(4-isopropyl)thiophene (260 mg, 0.87 mmol) with 5-amino-4-bromo-3-methylisoxazole (161 mg, 0.91 mmol) yielded after flash chromatography using 10% MeOH/CHCI 3 a pale brown solid (265 mg) which was further purified using preparative HPLC to give the pure sulfonamide as a light tan colored solid, m.p. 114-116 0

C.

EXAMPLE 73 -idBromo-3-methyl-5-isoxazolyl)-5-(4-propylphenyl)thiophene-2-sulfona- A. 1-bromo-4-propylbenzene A solution of l-bromopropane (1.32 g, 0.6 mmol) was added dropwise at room temperature at a rate such that a gentle reflux was -121maintained to a suspension of magnesium (258 mg, 12 mmol) in dry tetrahydrofuran. The cloudy suspension was stored at room temperature for an additional 30 minutes to produce a gray solution that was then added dropwise over 15 minutes to a mixture of 1-iodo-4-bromobenzene (3.0 g, 10.6 mmol) and tetrakis (triphenylphosphine) palladium in mL of dry benzene at room temperature. The mixture was stirred for 2 hours, diluted with 50 mL of water, the organic layer was separated and the aqueous layer was extracted with ether (2 x 50 mL). The combined organic extracts was dried and evaporated to yield 1.69 g of a 10 light brown oil, which was used in the next step without further purification.

B. 4 -propylphenyl boronic acid 99 To a suspension of magnesium shavings (217 mg, 8.9 mmol) in 3 mL of dry tetrahydrofuran under argon, a crystal of iodine along with a solution of 4 -bromopropylbenzene (1.69 g, 8.5 mmol) dissolved in 6 mL of tetrahydrofuran was added at such a rate that a gentle reflux was maintained. The solution was refluxed for an additional 0.5 h, cooled to room temperature and added in portions over 10 min to a solution-_f trimethylborate (924 mg, 8.9 mmol) previously dissolved in 4 mL of dry ether at -78 0 C. After 30 minutes, the solution was warmed to room temperature; and stirring was continued for 90 min. The reaction was then quenched by the addition of 2 mL of a 10% hydrochloric acid solution. The tetrahydrofuran was removed under reduced pressure and the remaining residue was extracted into diethyl ether (3 x 25 mL). The combined ether extracts was extracted with 1 M NaOH (3 x 25 mL) and the resulting aqueous layer was acidified to pH 2.0 using 6N HCI, then reextracted back into diethyl ether (3 x 25 mL). The combined organic layers was washed with water (1 x 25 mL), brine (1 x 25 mL) and dried over magnesium sulfate. Evaporation of solvent left a brown solid which -122pe Ct

I

C

C.

C

C,

9 9

S

C. CC 9 C 9 was filtered through a small plug of silica gel using 10% MeOH/CHCI,.

Evaporation left 448 mg of a brown solid, m.p. 90-93 0

C.

C. 4 propylphenyl)thiophene2sulfonylpyrrol 4 -propylphenyl)thiophene..2.sulfonyllpyrrole was prepared in the same manner as described in Example 33C, from 4 -propylphenyl boronic acid and N-(5-bromothiophenesulfonyl)pyrrole. Purification by column chromatography using 10% ethyl acetate/hexanes gave the pure sulfonamide as a white solid in 55% yield, m.p. 106-1080C.

D. 5-chlorosulfonyl..2.(4..propylphenyl)thiophen 10 5-hooufnl2(-rplhnltipeewas prepared in the same manner as described in Example 33D. Hydrolysis of 240 mg (0.73 mmol) of N-5(-rplhnlhohn)2sloylyrl with 6N NaOH followed by chlorination using phosphorous oxychloride and phosphorous pentachloride gave the crude sulfonyl chloride as a greenish-brown oil.

15 Flash chromatography over silica gel using 2% ethyl acetate/hexanes yielded 83 mg (81 of the pure sulfonyl chloride as a pale yellow oil.

E.

N-(

4 bromo3.methy-sxll)-5.isoa opyl)(phl)thioh. e sulf onamide 20 N-4boo3mty--sxzli)5(-rplhnltipee2 sulfonamide was prepared in the same manner as described in Ex ample 2.

Reaction of 5-hooufnl2(-spoy~hohn (260 mg, 0.87 mmol) with 5-amino-4-bromo..3.methylisoxazole (161 mg, 0. 91 mmol) yielded after flash chromatography using 10% MeOH/CHCI 3 a brown solid (76.1 mg) which was further purified using preparative HPLC to give the pure sulfonamide as a tan colored oil.

CC

9

S

CCC.

5* 9 C 94

SC

*C C C C C CC -123- EXAMPLE 74 N-4Boo3mty--sxzli--(,,-rmtoyezi-ez~l thiophene-3-sulfonamide A. a-( 2 -benzo[blthienyl)..3,4,5..trimethoxybenzyI alcohol a-{ 2 -benzofblthienyl)..3,4,5-trimethoxybenzyl alcohol was prepared in the same manner as described in Example 40A. Reaction of benzofblthiophene (7.5 mmoles, 1 .0 t-BuLi (1 .7m, 9.7 mmoles, 5.7, 00 mls) and 3 4 5 -trimethoxybenzaldehyde (8.9 mmoles, 1.8 g) in THF S* ml) yielded, after flash chromatography using 50% ethyl acetate/hexanes, 2.4 g of a yellow-white solid.

B. S-trimethoxybenzly)-benzofblthiophene 2 3 4 ,5-trimethoxybenzly)..benzo[bjthiophene was prepared in the 5* same manner as described in Example 47B. Reaction of a-(2-benzo[bI- *0*15 thienyl)-3,4,5-trimethoxybenzyl alcohol (4.5 mmoles, 1.5 triethylsilane mmoles, 0.80 mls), CH 2

CI

2 (50 ml) and TFA (9.1 mmoles, 0.70 mls) too&% yielded, after flash chromatography using 20% ethyl acetate/hexanes, 0.77 g of a white solid.

0 C. 2-(3,4,5-rmtoyezl-ez~jhohn--ufnlhod 2- (3,,trimethoxybenzyl) be nzo [bthiophen e3sulf onylch lrde was prepared in the same manner as described in Example 40B. Reaction of dimethylformamide (DMVF; 4.8 mmoles, 0.40 mls), sulfuryl chloride (4.1 mmoles, 0.33 mls) and 2 3 4 thiophene (2.4 mmoles, 0.75 g) yielded, after flash chromatography using 20% ethyl acetate/hexanes, 0.29 g of a yellow-orange oil.

D. N-( 4 bromo3.methyisoxazioxazolyl)2..(345..mtrimbl)benzo[blthiophene.3.sulfonamide N-4boo--ehl5-sxzSl)2(,,-trimethoxybenzyl).

benzo[blthiophene-3sulfonamide was prepared in the same manner as described in Example 41. Reaction of 4 -bromo-3-methyl-s..aminoisoxazole (0.55 mmoles, 97 mg), NaH (1.4 mmoles, 55 mg), and 2- -124- S-trimethoxylbenzyl).benzo[blthiophene3sulf onyl chloride (0.66 mmoles, 0.27 g) in THF (2 ml) yielded, after flash chromatography using ethyl acetate/haxanes and recrystallization from chloroform and hexanes, 94 mg of a tan solid, m.p. 154-1561C.

EXAMPLE

N-(

4 -bromo-3-methyl.5.isoxazoly) 2 -ethyl-5-methybenzofbthiophene- 3 sulfonamide A. 2-ty--ehlez~jhohn 2-ty--ehlezflhohn was prepared in the same manner as described in Example 40A. Reaction of methylbenzo[bjthiophene (3.4 mmoles, 0.50 t-BuLi (1.7 m, 5.1 9 mmoles, 3.0 ml) and ethyl iodide (6.8 mmoles, 0.54 ml) in THF (10 ml) yielded 0.58g of a light yellow liquid.

B 2 -ethyI-5-methylbenzorblthiophene.3suffonylchoride 2-ty--ehlezflhipee3sloyclrd was prepared in the same manner as described in Example 40B. Reaction of DMVF 9 mmoles, 0.50 ml), sulfurylchloride (5.5 mmoles, 0.44 ml) and methylbenzofbjthiophene (3.2 mmoles, 0.57 g) yielded, after flash__ chromatography, using 2% ethyl acetate/hexanes, 0.58 g of an orange solid.

C.

N-(

4 bromo3.methyl-sox..i)-2-etlf)2ethylbezbtho phene-3-sulfonamide mehlez~lhohn--ufnmd was prepared in the same manner as described in Example 41. Reaction of 4 aminoisoxazole (1.0 mmole, 0. 18 NaH (2.5 mmoles, 0.l1Og), and 2ety-5mtybno[lhipee3sloyclrd (1.3 mmoles, 0.36 g) in THE (6 ml) yielded, after re crystallization from chloroform and hexanes, 0.25g of a light brown crystalline solid, m.p. 176-1780

C.

-125- EXAMPLE 76

N-(

4 chloro3m ethyl-5iso zol- l)2[4(methylened)bzl] benzo [blthiophene-3-sulfonamide

N-(

4 chloro3methyl5isoxazolyl)2[34(methylendiox)benyj benzo[blthiophene.3.sulfonamide was prepared in the same manner as described in Example 41. Reaction of 4 -chloro- 3 (0.61 mmoles, 81 mg), NaH (1.5 mmoles), 61 mg), and 2-[3,4- (methylenedioxy)benzylbenzoblthiophene3sulfony chloride (0.74 10 mmoles, 0.27 g) in THF (4ml) yielded, after flash chromatography using ethyl acetate/hexanes followed by recrystallization from ethyl acetate and hexanes, 0.23 g (81 of a light brown solid, m.p. 178- 181 0 C.

EXAMPLE 77 15 N-( 4 Bromo3 nethylio zol- 2(4-dimethoxbl)benzo [blthiophene-3-suffonamide A. 2 -benzo[blthienyl)..3,4..dimethylbenzyl alcohol a-( 2 -benzofbthienyl)34dimthylbelzyI alcohol was prepared in the same manner as described in Example 40A. Reaction of benzo[bjthiophene (7.5 mmoles, 1.0 t-BuLi (1.7 m, 97 mmoles, 5.7 ml) and 3 4 -dimethoxybenzaldehyde (8.9 mmoles, 1.5 g) in THE (20 ml) yielded, after flash chromatography using 30% ethyl aceteate/hexanes, 2.25 g 100%) of a white gummy solid.

B. 2 3 4 -di meth oxyb enzyl) benzo thiop hen e 2 3 4 -dimethoxybenzyl)benzo[bjthiophene was prepared in the same manner as described in Example 47B. Reaction of a-(2benzo[bjthienyl)-3..4.dimethoxybenzyl alcohol (7.5 mmoles, 2.25 g), triethylsilane (8.2 mmoles, 1.3 ml) and CH 2 Cl 2 (20 ml) in TEA mmoles, 1 .2 m~s) yielded, after flash chromatography using 1 0% ethyl acetate/hexanes, 1 .77g of a colorless oil.

-126- C. 2 3 4 dimethoxybenzyl)benzobihioe--slfnlchlor 2 3 4 dimethoxybenzyl)..zobnziopbhioe-sfnlhloid was prepared in the same manner as described in Example 40B. Reaction of DMVF (90 mmoles, 7 ml) and 2 3 4 -dimethoxybenzyl)..benzo[bjthiophen (6.0 mmoles, 1.7 g) yielded, after flash chromatography using 15% ethyl acetate/hexanes, 1 .24g of a green oil.

D.

N-(

4 -bromo-3..methyl..5.isoxazolyl)..2.(3,4..dimethoxybenzyl)benzo [blthiophene-3..sulfonamide benzo[blthiophene-3sulfonamide was prepared in the same manner as described in Example 41. Reaction of 4 -bromo-3-methyl-.s.

**aminoisoxazole (1.0 mmoles, 0. 18 NaH (2.5 mmoles, 60 mg), and 2- (3,-dimehoyb nzl)bezo blhiphee-3-ulfony~h l rde(1.2 mmoles, O.44g) in THF (6m1), after recrystallization from chloroform and hexanes, yielded 0.42 g of a brown solid, m.p. 151-153o

C.

EXAMPLE 78

N-(

3 4 dimethyl.5isoxaz lyl2( 3 4 -thylndoybnobtipee3 sulfonamide N 4 -d met yl-5 iso azo yl) 4 -m thyl ned oxybenzo[57jthio.

phene-3-sulfonamide was prepared in the same manner as described in Example 41. Reaction of 3,-iehl5aiosxzl (1 .0 mmoles, 0.11 Nail (2.5 mmoles, 60 mg) and 2 3 4 -(methylenedioxy)benzyl..

b no[]t pee--uIoyc ord (1.1 mmoles, 0.40 g) in THF (6 ml) yielded, after flash chromatography using 50% ethyl acetate/hexanes followed by recrystallization from chloroform and hexanes, 0.35 g (79%) of a tan solid, m.p. 135-137o

C.

-127- EXAMPLE 79

N-(

4 Bromo3methylsox..li)-2-(4-lyl)2(4ehbl)bezbthiophene-3-sulfonamide A. a-( 2 -benzofbjthienyl)..4.methyoxybenzol alcohol a-( 2 -benzofbjthienyl).4-methyoxybenzol alcohol was prepared by the method of Example 40A with benzo~blthiophene (7.5 mmoles. 1.0. g), t-BuLi (1.7 m, 10.4 mmoles, 6.1 ml), 4 -methoxy-benzaldehyde (8.9 mmoles, 1.1 ml) and THF (20 ml). Flash chromatography (20% ethyl 10 acetate/hexanes) provided 1.75 g of a yellow solid.

B. 2 4 methyoxybenzyl)-benz 0 [bithiophene 4mtyxbny)-ezflhohn was prepared by the method of Example 478 with 2 benzo[blthienyl)..4.methoxybenzyI alcohol (1.9 mmoles) 0.50 triethysilane (2.0 mmoles, 0.32 mls), 15 CH 2

CI

2 (20 ml) and TFA (3.7 mmoles, 0.30 ml). Recrystallization with hexanes and chloroform provided 0.40 of a pink solid.

C. 2 4 methoxybenzyl)benzobthiophene3sulfolhlrd 20prepared by the method of Example 40B with DMVF (3.2 mmoles,.fl.24 sulfurylchloride (2.7 mmoles, 0.22 ml) and 2 4 -methoyxybenzyl).

benzofblthiophene (1.6 mmoles. 0.4 Flash chromatography using 2% ethyl acetate/hexanes provided 0. 19 g (33 of a light yellow solid.

D.

N-(

4 bromo3.methyl-sox..i)xazo(4-2(thxbeny benzo[blthiophene.3.sulfonamide N-4boo3mty--sxzoy)2(-ehxbny) benzofblthiophene.3.sulfonamide was prepared by the method of Example 41 with 4-rm--ehl5aiosxzl (0.48 mmoles, mg), NaH (1.2 mmoles, 48 mg), 2 4 -methoxybenzyl)..benzo[blthiophene- 3 -su Ifo nylc hlo ride 53 mmoles, 0. 19 g) and THF (3 ml). Flash chromatography (50% ethyl acetate/hexanes) followed by -128- C.

C

C

recrystallization from methanol 'and water provided 46mg of a white crystalline solid, m.p. 120-1220

C.

EXAMPLE

N-(

4 -Bromo- 3 -methyI-5-isoxozolyi).2(2..methoxybenzyl)benzofblthiophene-3-suffonamide A. a-2bnobtipee--ehxbny alcohol a-( 2 -Benzo[blthiophene)..2.methoxybenzyl alcohol was prepared -by the method of Example 47A with benzo[bjthiophene (7.5 mmoles, 1 .C g) t-BuLi (1.7 m, 9.7 mmoles, 5.7 ml), 2 -methoxybenzaldehyde (8.9 mmoles, 1.1 ml) and THF (20 ml). Flash chromatography (20% ethyl acetate/hexanes) provided 1 .9 g of a yellow oil.

B. 2 2 -Methoxybenzyl)-benzo[blthiophene 2 2 -Methoxybenzyl)-benzofbjthiophene was prepared by the method of Example 478 a-( 2 -benzo[blthiophene)-2methoxybenzyl alcohol (7.1 mmoles, 1.9 triethylsilane (7.9 mmoles, 1.3 ml) and CH 2

CI

2 (30 ml) at 0 0 C was added TFA (14.3 mmoles, 1.1 Flash chromatography ethyl acetate/hexanes) provided 1 .31g of a yellow solid.

20 C. 2 2 -methoxybenzyI)-benzo[bthiophene3sufonyI chloride 2 2 -methoxybenzyl)benzotblthiophene-3sulfonyI chloride was prepared by the method of Example 40B with sulfuryl chloride (8.4 mmoles, 0.7 ml), DMVF (9.8 mmoles, 0.8 ml) and 2 2 -methoxybenzyl)benzolthiophene (4.9 mmoles, 1.25 Flash chromatography (2% ethyl acetate/hexanes) provided 0.94 g of a yellow solid.

D. N-( 4 -bromo- 3 -methyI.5-isoxozoly)..2.(2..methoxybenzyl)benzoI~blthiophene-3-sulfonamide N-4boo3mty--sxzll--2mtoyezl-ez~l thiophene-3-sulfonamide was prepared by the method of Example 41 with 5-amino- 4 -bromo-3-methylisoxazole (1.0 mmoles, 0.18 NaH mmoles, 100 mg), 2 2 -methoxybenzyl)-benzofbthiophene-3-suffonyI -129chloride (1.4 mmoles, 0.49 g) and THF (7m1). Flash chromatography ethyl acetate/hexanes) followed by recrystallization from chloroform and hexanes provided 0.30 g (61 of a brown solid, m.p.

80-840

C.

EXAMPLE 81 fofla-d meV)( 4 -chsoaly)(-horoen Ibenzyl) be nzo btp he e3s ul.

A a-( 2 -benzolblthienyl)-4chlorobenzyI alcohol ~10 a-( 2 -benzofblthienyl)-4chlorobelzyI alcohol was prprdbyth method of Example 40A with benzofblthiophene (7.5 mmoles, 1.0 t- BuLi (1 .7 mn, 9.7 mmoles, 5.7 ml), 4 -chlorobenzaldehyde (9.7 mmoles, 1.4 g) and THF (20 ml). The crude material (2.45 g) was taken forward without further purification.

B. 2 4 -chlorobenzyl)benzofblthiophene 2 4 -chlorobenzyl)benzofblthiophene was prepared by the method of Example 47B with u-( 2 -benzofblthiophene)..4.chlorobenzyl alcohol (8.

mmoles, 2.45 triethylsilane (9.8 mmoles, 1 .6 ml), CH 2

CI

2 (40 ml) and TFA (13.4 mmoles, 1.0 ml). Flash chromatography (1 ethyl ace- :20 tate/hexanes) provided 1.3 g (67% 2 steps) of an off-white solid.

C. 2 4 -chlo robe nzyD benzo [b1thiop hene 3sulfonylchl rde 2-4clrbny~ezfjhipee3sfoyclrd was prepared by the method of Example 40B with DMVF (70 mmoles, 5.4 ml), sulfurylchloride (2.3 mmoles, 1.9 ml) and 2 4 -chlorobenzyl)-benzo[bjthj 0 phene (4.6 mmoles, 1 .2 Flash chromatography ethyl acetate/hexanes) provided 0.51 g (31 of an orange-yellow oil.

D. N 3 4 d imethyl 5.iazoxalyl)2(4chllobeny) bezo[ t1ophne- 3 -sulfonamide bhope- 3 -sulfonamide was prepared by the method of Example 41 with 3,4- (1.2 mmoles, 1.4 NaH (3.0 mmoles, 73 -130mg), 2 4 -chlo robenzyl) benzo [bthi oph en e3-sulfo nylcho ride (1.4 mmoles, 0.50 g) and THF (8 ml). Flash chromatography (50% ethyl acetate/hexanes) followed by recrystallization from methanol and water provided 1.04 g of a yellow solid, m.p. 100-1020

C.

EXAMPLE 82

N-(

4 .chlo ro3meth yl5iso xazolyl) 2( 4 .diehl mi ny)b b]thio A a-2bnobti.enJlu.4.imtylaminobenzyI alcohol 10 u-( 2 -benzofbthienyl)4dimethylaminobenzyl alcohol was prepared by the method of Example 40A with benzofblthiophene (7.5 mmoles, t-BuLi (1.7 M, 8.9 mmoles, 5.3 ml), 4 -dimethylaminobenzaldehyde (8.9 mmoles, 1.3 g) and THF (20 ml). The crude product (2.4 g) was carried forward without further purification.

B. 2 4 -dimethya min obe nzyl) benzo [b Ithio phen 2 -d i eth a m i o b e z y~ b e z of l th o p h n e w a s p re p a re d b y th e method of Example 47B with a-2bnobtinl-4dmtyaioez alcohol (7.5 mmoles, 2.1 triethylsilane (2.8 mmoles, 1.3 ml), CH 2

CI

2 (50 ml) and TFA (11.2 mmoles, 0.9 ml). Flash chromatography .20 ethyl acetate/hexanes) provided 1.5 g (73% -for two steps) of a white solid.

C. 2 4 -dimethylaminobenzyl)benzobthiophene3sulfofylch,.rde Chlorosulfonic acid (9.4 mmoles, 0.6ml) was added to 2-(4dimethylaminobenzyl)-benzo[blthiophene (3.7 mmoles, 1 .0 g) in CH 2 C1 2 (100 ml) at -780 C. The solution was stirred 20 min. at -780 C.

Phosphorous oxychloride (11 .2 mmoles, 1.0 ml) and phosphorous pentachloride (11.2 mmoles, 2.3 g) were added to the reaction mixture.

The reaction mixture was warmed to ambient temperature and stirring was continued an additional 1.5 hr. followed by dilution with ice 200 ml) and extraction with ethyl acetate (200 ml). The organic layer was -131washed carefully with sat. NaHC0 3 (3 x 100 ml), then dried (MgSO 4 filtered and concentrated. Flash chromatography ethyl acetate/hexanes) provided 0.61 g of a yellow solid.

D. N-( 4 chloro3.methyl.5.isoxazolyl)..24dimtyimethylm l)benzofblthiophene-3sulfonamide o -ehy-5ioaoy)--4diehlamnoezl benzo[blthiophene.3-sulfonamide was prepared by the method of Exampie 41 with 4-hoo3mty-5aiosxzl (0.52 mmoles, 69 mg), NaH (1.3 mmoles, 31 mg), 2 4 -d imethyl amin obenzyl) benzo [b~thioph e n e 3 -sulfonylchloride (0.58 mmoles, 0.21g) and THF (4 mJ). Flash chromatography methanol/chloroform) provided 0. 16 g of a yellow gummy solid, m.p. 105-1100

C.

EXAMPLE 83 A. 2 ,5-dimethyl-furan-3..sulfonylchloride meh l- ura- 3suf nychordewas prepared by the method of Example 40B with IDMVF (28 mmoles, 2.2 ml), sulfurylchloride (24 mmoles, 1.9 ml) and 2 ,5-dimethyl-furan (14 mmoles, 1.5 ml). Flash ,20 chromatography ethyl acetate/hexanes) provided 0.61 g (2 of a yellow liquid.

B. N-( 4 -bromo-3-methyl..s.isoxazolyl)..2,5-dimethyI-furan-3..sulf 0 on..

mide

N-(

4 -bromo-3-methyl..s-jsoxazoly)-25diehy-urn3-ufoa mide was prepared by the method of Example 41 with 4 -bromo-3-methyl- (2.0 mmoles, 0.35 NaH (5.0 mmoles, 200 mg), 2,-iehlfrn3sloyclrd (2.4 mmoles, 0.47 g) and THF (9 ml). Flash chromatography methanol/chloroform) followed by recrystallization from chloroform and hexanes provided 0.21 g (31 of a light brown solid, m.p. 85.5-870

C.

-132- EXAMPLE 84

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-2,5-dimethyl-4-phenylhiophene-3-sulfonamide A. 2 5 -dimethyl-3,4-dibromothiophene NBS (13.2 mmoles, 2.4 g) was added to 2 5 -dimethylthiopene (5.3 mmoles, 0.59 g) in CHCI 3 (30 ml). The reaction mixture was stirred at ambient temperature for 1.5 hr., then diluted with ether (50 ml) and washed with water (3 x 50 ml). The organic was dried (MgSO 4 filtered 10 and concentrated. Flash chromatography (hexanes) provided 1.2 g of a white solid.

B. 2,5-dimethyl-3-bromo-4-phenylthiophene Phenyl boronic acid (5.0 mmoles, 0.61 g) was added to dimethyl-3,4-bromothiophene (4.5 mmoles, 1.2 tetrakis (triphenyl- 15 phosphine) palladium(0) (0.23 mmoles, 0.26 g) and Na 2 CO, (2 M, 26 mmoles, 13 ml) in benzene (20 ml). The biphasic reaction mixture was heated to reflux for 24 hr. then cooled to ambient temperature and diluted with ether (100 ml) and washed with water (100 ml). The organic later was dried (MgSO 4 filtered and concentreated. Flash 20 chromatography (hexanes) provided 0.60 g of a yellow solid.

C. 2 ,5-dimethyl-4-phenyl-3-sulfonylchloridethiophene t-BuLi (1.7 M, 2.7 mmoles, 1.6 ml) was added to 2 ,5-dimethyl-3bromo-4-phenyl thiophene (2.2 mmoles, 0.59 g) in THF (8 mis.) at -300 C. The solution was stirred 20 min. at -30 0 C, then the flask was evacuated with sulfur dioxide, and warmed to -20° C upon which NCS (2.2 mmoles, 0.30 g) was added. The reaction mixture was warmed to ambient temperature for 30 min., then diluted with ethyl acetae (50 ml) and washed with water (2 x 50 ml). The organic layer was dried (MgSO 4 filtered and concentrated. Flash chromatography ethyl acetate/hexanes) provided 0.26 g of a light yellow solid.

-133- D. N-(4-bromo-3-methyl-5-isoxazole)-2, 5-dimethyl-4-phenylthiophene- 3-sulfonamide N-(4-bromo-3-methyl-5-isoxazole)-2, 5-dimethyl-4-phenylthiophene.

3-sulfonamide was prepared by the method of Example 41 with 4-bromo- 3-methyl-5-amino isoxazole (0.79 mmoles, 0.14 NaH (2.0 mmoles, mg), 2.5S-d imethyl-4-p henylthiophene-3-sulfo nylchlo ride (0.91 mmoles, 0.26 g) and THF (3 ml). Recrystallization twice from chloroform and hexanes provided 0. 15g (45 of a white crystalline solid, m.p.

16~6a.EXAMPLE

N-(

4 -Bromo- 3 -methyl5isoxazoly)2.(hydroxymethyl)thiophene3sulfonamide

BH

3 -THF (3.62 ml, 1 M in THF) was added to a solution of N-(4br o -ehy-5isxz y)-2cabxlhiop ne -ufnai (1 .0g, 2.72 mmol) in dry THF (1 5 ml) at room temperature. After stirring at room temperature for 10 minutes, the mixture was refluxed for 1 hour.

The reaction was cooled with an ice-bath and 1N HCI (10 ml) was added.

The resulting mixture was concentr ated. The aqueous residue was then partitioned between 1 N HCI and EtOAc. The organic layer was dried (MgSO 4 The solid was filtered and the filtrate concentrated. The residue was treated with MeGH and concentrated again. This process was repeated 3 more times to give N-(4-bromo-3-methyl-5-isoxazolyl)-2(hy.

droxymethyl)thiophene-3-sulfonamide (680 mg, 71 yield) as a yellow oil.

EXAMPLE 86

N-(

4 -Bromo-3-methyl-5-isoxazolyl)..2.[(3..methoxyphenyl)aminomethyl] thiophene-3-sulf onamide

BH

3 -THF (15 ml, 1 M in THF) was added to a solution of N-(4br o -ehl -sxz y)-2 -(3mtoyhnl i cr y hio phene-3-sulfonamide (Example 22) (1.0 g, 2.12 mmol) in dry THF -134- Ml). The mixture was refluxed for 8 hours and cooled. THF was evaporated on a rotavap and MeOH was added to the residue. The resulting solution was concentrated. The final residue was purified by HPLC to give N-( 4 -bromo3methyl.5.isoxazoly)2(3-methoxyhiy)aminomethyulthiophene.3-sulfonamide (113 mg., 12% yield) as a grey Powder, m.p. 70-73 0

C.

EXAMPLE 87 0

N-(

4 bromo3.methyl5isoxazoly2[N( 3 aboyhnlmocrnlthiophene-3-sulfonamide Et 3 N (2.27 ml. 1 6. mmol), ethyl 3 -aminobenzoate (836 ml, 5.44 mmol) and phosphonitrilic chloride trimer (1.89 g, 5.44 mmol) were sequentially added to a solution of N-( 4 -bromo-3-methyl.s..ioxazolyl).

2 (carbonyl)thiophene-3 5 ulfoflamjde (Example 17) (1g, 2.27 mmol) in dry THE (20 ml). The reaction was stirred at room temperature for 1 hour and cooled. Water (5 ml) was added to quench the reaction. The resulting solution was concentrated on a rotavap. The residue was 5005 diluted with EtOAc and washed with 2N HCI (2 x 150 mi). The organic layer was dried (MgSO 4 The solid was filtered off and the filtrate-was .20 concentrated. The residue was treated with 1iN NaOH (200 mi) and stirred at 00) C for 1 5 minutes. The mixture was then acidified with conc. HCI to pH The resulting yellow precipitate was filtered off and recrystalized from

CHCN/H

2 0 to give N-4boo3mtyl5ioaoy) 2-[N 3c bxpe l i cabnltipee l nmd (153 mg., 11.6%) as a yellowish powder, m.p. 183-1850

C.

EXAMPLE 88

N-(

4 -Bromo3.methyl5-isoxazoy) 2 2 carboxylphenyl) amino arbonl iohiop 3 lfonmd N-4boo3mty--sxzll)2[-2croypey)aio carbonyllthiophene.3.ulfoflamjde was prepared by the same method as -135described in Example 87, with the exception of using ethyl-2aminobenzoate instead of ethyl-3-aminobenzoate.

EXAMPLE 89

N-(

4 -bromo- 3 -methyl-5-isoxazolyl)-2-(aminocarbonyl)thiophene-3-sulfonamide Carbonydiimidazole (485 mg, 2.99 mmol) was added to a solution of N-( 4 -bromo-3-methyl-5-isoxazolyl)-2-carboxylthiophene-3-sulfonamid (1 g, 2.72 mmol) in THF (10 mi) at room temperature. The mixture was 10 stirred for 15 minutes. Aqueous NH, (5 mi) was then added, and the mixture was stirred at room temperature for 30 minutes. The solvent S was evaporated and the residue was partitioned between EtOAc and 1N SHCI. The organic layer was dried (MgSO 4 The solid was filtered and the filtrate concentrated. The oily residue was recrystalized from EtOAc 15 to give N-(4-bromo-3-methyl-5-isoxazolyl)-2-(aminocarbonyl)thiophene-3sulfonamide (946 mg, 95% yield) as a white solid, m.p. 168-1700 C.

S EXAMPLE

SN-(

4 -Bromo-3-methyl-5-isoxazolyl)-2-[(5-dimethylamino-1 -naphthyl)sulfonylaminocarbonyl]thiophene-3-sulfonamide Dansylchloride (90.2 mg, 0.328 mmol) was added to a solution of N-(4-bromo-3-methyl-5-isoxazolyl)-2-(aminocarbonyl)thiophene-3-sulfonamide (Example 89) (100 mg, 0.273 mmol) and NaH (43.7 mg, dispersion in mineral oil, 1.10 mmol). The reaction was stirred at room temperature for 1 hour. Water was added to quench the reaction and THF was stripped off on a rotavap. The aqueous residue was partitioned between 1N HCI and EtOAc. The organic layer was dried (MgSO 4 The solid was filtered and the filtrate concentrated. The residue was recrystalized from EtOAc to give N-( 4 -bromo-3-methyl-5-isoxazolyl)-2-[(5dimethylamino- 1 -naphthyl)sulfonylaminocarbonylthiophene-3-sulfonamide mg., 34% yield) as a white powder 184-1860 C).

-136- EXAMPLE 91 N-4Boo3mtyl5ioaoy)2[(3, 4 -methylenedioxyphenyl)aminocarbonyllthiophene-3-sulfonamide

N-(

4 -bromo- 3 methyl5isoxazoly)2[(34methylenedioxyphenyl) aminocarbonylthiophene-3-sulfonamide was prepared by the same method as described in Example 89 with the exception that 3,4methylenedioxyaniline was used in place of ammonium hydroxide. N-(4b romo- 3 -meth yl- 5isoxazo lyl) 2 (34methylened ioxyph enyl) amin ocarbonylIthiophene-3-sulfonamide was purified by HPLC to give 1 5% yield of the desired product as a dark grey powder, m.p. 138-1400 C.

EXAMPLE 92

N-(

4 -Bromo- 3 -methyl..5.isoxazoly).2.[(34.methylenedioxy)phenoxy Carbonyldiimidazole (530 mg., 3.26 mmol.) was added to a solution of N-4boo3mty--sxzly)2croytipee3sl fonamide (Example 17) (1.0g, 2.72 mmol) in dry THF (10 ml). The mixture was stirred at room temperature for 1 5 minutes. Sesamol (767 mg., 5.44 mmol) and imidazole (185 mg, 2.72 mmol) were added simultaneously. The resulting mixture was refluxed for 1 hour and allowed to cool to room temperature. The solvent was evaporated. The residue was partitioned between 1 N HCI and EtOAc. The organic layer was dried (MgSO 4 The solid was filtered and the filtrate concentrated to give a yellow oil which was recrystalized from EtOAc/Et 2 O/Hexane.

N-

(4boo3mty-5ioaoy)2[(3, 4 methylenedioxy)phenoxycarbo nyll thioph ene-3-sulf on amid e was obtained as a white powder (494 mg, 37% yield), m.p. 174-1760

C.

-137- EXAMPLE 93

N-(

4 Bromo3methyl5isoxazoly).2.[(34methylenedioxy)bol] thiophene-3-sulfonamide A. N-( 4 -bromo3methyl5isoxazolyl).2[(N-methoxy-N methyl) amino ca rbonyllthop hene.3sulf onamide

N-(

4 -bromo-3-methyl..5-isoxazolyl)..2-.[(Nmethoxy-N methyl)carboxamidelthiophene-3sulfonamide was prepared by the same method as described in Example 89 with the exception that N,0dimethylhydroxylamine was used in place of ammonium hydroxide. The yield was

N-(

4 Bromo3methyl.5..isoazoly yl)..2..[(34mhyetdix)benzoyllthiophene.3.sulfonamide Freshly prepared 3 4 -methylenedioxy)phenyl magnesium bromide 15 (1.28 g of 3 4 -methylenedioxy)bromobenzene and 172 mg Mg turnings) was added to a solution of N-( 4 -bromo3methyl5-isoxazolyl)-2-[(Nmehx--ehlaioabnylhohn--ufnmd (652 mg, 1. 59 mmol) in THF (10 ml) at room temperature. The resulting mixture was refluxed for 30 minutes. To workup, the mixture was allowed to cool to room temperature and was quenced with 1iN HCl (10 ml). I-F was then evaporated. The aqueous residue was partitioned between 1 N HCI and EtOAc. The organic layer was concentrated and the residue was purified by HPLC to give N-4boo3mty--ioaoy)2[34 methylenedioxy)benzoyllthiophene-3 5 suffnamid (90 mg, 12% yield) as a dark yellow powder, m.p. 47-490

C.

EXAMPLE 94 N 4 Bromo 3 metythyis xl..5..isoxazoyd roxyph eyrxapmin o a b ll thiophene-3-sulfonamide N-4boo3mty--sxzll--(-yrxpey)aio carbonyllthiophene-3..sulfofl 5 mjde was prepared by the same method as described in Example 89 with the exception that 3 -aminophenol was used -138in place of ammonium hydroxide. The product was purified by HPLC to give N-(4-bromo-3-methyl-5-isoxazolyl)-2-l(2-hydroxyphenyl)aminocarbonyl]thiophene-3-sulfonamide (50 mg, 18% yield) as a dull yellow solid, m.p. 42-440

C.

EXAMPLE N-(3,4-dimethy 5-isoxa zolyI)-2[(3,4-methylenedioxy)phenoxycarbonyllthiophene-3-suffonamide N-(3,4-dimethyl-5-isoxazolyl)-213,4-(methylenedioxy)phenoxycarbonyl]thiophene-3-sufonamide was preparedby the same method as was prepared by the same method as described in Example 92 with the exception that N-( 3 i.soxazolyl)-2-carboxylthiophene-3-sulfonamide was used instead of N-(3bromo-4-methyl-5-isoxazolyl)-2-carboxylthiophene-3-sulfonamide. N-(3,4dimethyl-5-isoxazolyl)-21(3,4-methylenedioxy)phenoxycarbonyllthiophene- 15 3 -sulfonamide was purified by HPLC and was obtained as an orange oil S.(200 mg., 15% yield).

~EXAMPLE 96 N-(4-Bromo-3-methyl-5-isoxazolyl)a2- (e3,4-methylenedioxy)benzoylla min ocarbonyl~thiophene-3-sulf onamide 20 Carbonyldiimidazole (213 mg. 1.31 mmol) was added to a solution of piperonylic acid (181.5 mg., 1.09 mmol) in dry THF (10 ml). The resulting mixture was stirred for 15 minutes.

N-(

4 isoxazolyl)-2-aminocarbonylthiophene-3-sulfonamide (Example 89) (400 mg, 1.09 mmol) and NaH (175 mg, 60% in mineral oil, 4.37 mmol) were added sequentially. The mixture was stirred at room temperature for 8 hours. Water was added to destroy the excess NaH. The solvent was then evaporated and the residue was partitioned between 1N HCI and EtOAc. The organic layer was dried (MgSO 4 the solid filtered and the filtrate concentrated. The residue was recrystalized from EtOAc to give N-(4-bromo-3-methyl-5-isoxazolyl)-2-{[(3,4-methylenedioxy)benzoyl]- -139aminocarbonyllthiophene-3-sufonamide (20 mg, 3.6% yield) as a yellowish powder 90-930 C).

EXAMPLE 97

N-(

4 -chloro-3methyl.5isoxazolyl).2..(34methylenedioxy)pheoy carbonyllthiophene-3-sulfonamide i--(,-ehlndoyphenoxycarbonyllthiophene-3..sulfoflamjd 5 was prepared by the same method as described in Example 93 with the exception that N-(4-chloro-3-methyl.5isxzli--abxlhohn--ufnmd was used instead of N-(4br mo -mty-5ioaoll -abxlhohe ufo mde N-(4thiophene-3-sulfonamide was recrystalized from EtOAc (49% mg, yield) as a white solid, m.p. 189-191O0

C.

EXAMPLE 98

N-(

4 -Bromo-3-methyl..5.isoxazolyl)..2.[3,4..methylenedioxy)phenyl.

N-(

4 -bromo-3-methyl.s..soxazolyl)..2.j(3,4-methylene dix~hnlctltiohn--ufnmd was prepared by the same method as described in Example 93 with the exception that piperonylmagnesium chloride was used instead of 3 4 -methylenedioxy)phenylmagnesium bromide and the reaction mixture was stirred overnight at room temperature instead of refluxing for 30 minutes. The crude mixture was purified by HPLC to give N-( 4 2-34(ehlndoypeyaeylhohn--ufnmd mg, yield) as a yellow oil.

-140- EXAMPLE 99

N-(

4 Bromo3.methyl..5..xzoi)-2-[(3yI)2[(34methedi)pheoy carbonylaminolthiophene.3sulfonamide Triethylamine (2.28 ml, 16.35 mmol) and diphenylphosphorylazide (773 mg., 2.72 mmol) were sequentially added to a solution of N-(4bromo- 3 methyl5.isoxazolyl)..2..carblthoxyhnpheneulfomde (Example 17) (1.0 g, 2.72 mmol) in dry THF (40 ml). The mixture was stirred for 8 hours. Sesamol (1.54 g, 10.9 mmol) was added and the *10 mixture was refluxed for 2 hr. The mixture was allowed to cool to room temperature. The solvent was stripped off on a rotavap and the residue was partitioned between EtOAc and 1iN HCI. The organic layer was dried (MgSO 4 The solid was filtered and the filtrate concentrated. The *~*residue was purified by HPLC to afford N-(4-bromo-3-methyl5 15 isxzll--34(ehlndoy~hnxcroyaiotipee3 sulfonamide (400 mg, 29% yield) as a beige powder, m.p. 39-430

C.

EXAMPLE 100

N-(

4 -Bromo3methyI.5isoxazolyl).2[(3 4 mthylendx)peyuedthiophene-3-sulfonamide ureidolthiophene-3sulfonamide was prepared by the same method as described in Example 99 with the exception that 3 4 -methylenedioxy.

aniline was used instead of sesamol.

N-(

4 2 3 4 (meth yl enedioxy) ph eny ureidothio phene-3-uIfo n md e(157 mg, 12% yield) was obtained via HPLC purification as a brownish-greyish powder, m.p. 62-650

C.

-141- EXAMPLE 101

N-(

4 chloro3methyl5.isoxazolyl) .2..{(34..mleethoyeniylo carbonyllthiophene.3.sulfonamide

N-(

4 chloro3methy5o ioxalyl)2[{344methylenedo)bzloy carbonyulthiophene.3-sulfonamide was prepared by the same method as described in Example 97 with the exception that piperonyl alcohol was **used instead of sesamol.

N-(

4 -chloro-3-methyl5soxazolyl)2[( 3 4 (methylen edioxy) benzyloxycarbo nyllth ihhenene-s dlfaide (210 mg, 15% yield) was obtained via HPLC purification as a yellowish powder, m.p. 35-380

C.

EXAMPLE 102 **acetyllthiophene-3sulfonamide was prepared by the same method as described in Example 98 with the exception that N-(4-chloro-3-methyl-..

isxzli--abxlhohn--ufnmd was used instead of N-(4phene-3-sulfonamide (3g, 50% yield) was obtained via HPLC purification as a yellow solid, m.p. 35-380) C.

EXAMPLE 103

N-(

4 chloro3ty--ioaoyl-.,4-meoll)[(4ethdox~penihyl carbonyllthiophene.3.sulfonamide N-4clr--ehl5ioaoy)2[34mtyeeix) phntyoyabnlthohn--ufnmd was prepared by the same method as described in Example 97 with the exception that (3,4methylenedioxy)phenethyl alcohol was used instead of sesamol. N-(4choo3mty--sxzli--3,-mtyeeix~hntyoy -142carbon yllthiophene-3sulf onamide (500 mg, 34% yield) was obtained via HPLC purification as a yellowish oil.

EXAMPLE 104

N-(

4 -chloro3.methyl.5isoxazoly)2{ 4 3 4 mehyedixbny pipe razin -1 -yll ca rbonyllthiophene.3-su fona mide

N-(

4 -chloro-3-methyl..5.isoxazolyl)- 2 4 3 4 -methylenedioxyben.

zyl)piperazin- 1l-yfl carbonyl~thiophene- 3-suIf on amide was prepared by the same method as described in Example 89 with the exception that N-(4c o -ehl -ioaoy)2croyt io phen-3- ulfonaie was 1 used instead of N-4boo3mty--sxzoy)2croytipee3 **sulfonamide, and 1-piperonylpiperazine was used instead of ammonium hydroxide. N-4clr--ehl5ioaoy)2f4(,-ehlndoy .benzyl)piperazin- 1 yjcronlt oh n-3s fnai (872 mg, 54% yield) was obtained via HPLC purification as a white powder, m.p.

221-2230

C.

EXAMPLE 105

N(

4 -chloro3methyl.5isoxazoly)2amnthiohen--uf 3 md S. was prepared by the same method as described in Example 99 except that the mixture was refluxed without the addition of sesamol. N-(4 choo3mty--sxzll-2aiohohn--ufnmd (298 mg, 31 yield) was obtained via HPLC as a yellow solid, m.p. 39-420

C.

EXAMPLE 106 N 4 -chl oro-3..methyl- 5.isoxazolyl)2{ 1 -cya no 1- 3 4 -meth ylenedi oxy) phenyllacetyllthlophene.3-sulfonamide Carbonyldiimidazole (603 mg, 3.72 mmol) was added to a solution of (Example 17) (1.0g, 3.1 mmol) in dry THF (40 ml). The mixture was stirred at room temperature for 1 5 minutes.

-143- NaH (868 mg, 60% in mineral oil, 21.7 mmol) was added to a solution of 3 4 -methylenedioxy)phenylacetonitrile in THF (100 ml). The mixture (II) was refluxed for 30 minutes and then allowed to warm to room temperature.

The mixture was then cannulated into mixture (ill) while cooled by ice-bath and the resulting mixture was allowed to warm to room temperature. Water was added to quench excess NaH. THF was then stripped off on a rotavap. The residue was partitioned between 1N NaO-f and Et20. The aqueous layer was acidified with concentrated HCI with 10 cooling to pH 1 and extracted with EtOAc. The organic layer was dried (MgSO 4 the solid filtered and the filtrate concentrated. The residue was purified by HPLC to give N-( 4 2-{1 -cyano-1 -[3,4-(methylenedioxy)phenyllacetylthiophene-3-sulfonamide (277 mg, 19% yield) as a yellowish powder, m.p. 142-142 0

C.

15 EXAMPLE 107

N-(

4 -chloro-3-methyl-5-isoxazolyl)-2-13-(dimethylaminophenoxycarbonyl]thiophene-3-sulfonamide N-(4-chloro-3-methyl-5-isoxazolyl)-2-13-(dimethylamino)pheaoxy- S 20 carbonyl]thiophene-3-sulfonamide was prepared by the same method as described in Example 97 with the exception that 3 -dimethylaminophenol was used instead of sesamol. N-(4-chloro-3-methyl-5-isoxazolyl)-2-13- (dimethylamino)phenoxycarbonyl]thiophene-3-sulfonamide mg, 7.3% yield) was obtained via HPLC purification as a dark brown oil.

EXAMPLE 108 N-(4-chloro3-methyl-5-isoxazolyl)-2-(cyclohexyloxycarbonyl)thiophene-3- N-(4-chloro-3-methyl-5-isoxazolyl)-2-(cyclohexyloxycarbonyl)thiophene-3-sulfonamide was prepared by the same method as described in Example 97 with the exception that cyclohexanol was used instead of -144sesamol.

N-(

4 -chloro-3-methyl-5-isoxazolyl)-2-(cyclohexyloxycarbonyl)thiophene-3-sulfonamide (29 mg, 5% yield) was obtained via HPLC purification as an off-white solid, m.p. 134-1370

C.

EXAMPLE 109 N-(4-chloro-3-methyl-5-isoxazolyi)-2--hydroxy(3,4-methylenedioxy)phenylethyllthiophene-3-sufonamide LiBH 4 (36.6 mg, 1.68 mmol) was added slowly to a solution of N- 4 -chloro-3-methyl-5-isoxazolyl)-2-[3,4methylenedioxy)phenylacetyl]thiophene-3-sufonamide (Example 102) (74 Smg, 0.168 mmol) in the THF (10 mi). The resulting mixture was stirred for 8 hours. Saturated

NH

4 CI (aq) was added to quench the excess LiBH 4 The resulting mixture was concentrated on a rotavap. The residue was partitioned between EtOAc and 1N HCI. The organic layer was dried (MgSO 4 and the solid was filtered.

EXAMPLE 110 N,N'-bis {3-(3,4-dimethyl-5-isoxazolyi)aminosufonylthien-2-yurea Triethylamine (1.4 ml, 9.93 mmol) and diphenylphosphorylazide (9.39 mg., 3.31 mmol) were sequentially added to a solution of N-3,4dimethyl-5-isoxazolyl)-2-(carboxyl)thiophene-3-su If onamid (Example 17) g, 3.31 mmol) in THF (50 mi). The resulting mixture was stirred for minutes at room temperature and then refluxed for 1 hour. The mixture was allowed to cool to room temperature. THF was stripped off by use of a rotavap. The residue was partitioned between EtOAc and 1N HCI. The organic layer was dried (MgSO 4 the solid filtered, and the filtrate concentrated. NN'-bis3-(3,4-dimethy-5-isoxazolyl)aminosulfonylIthien-2-yljurea (140 mg, 14% yield) was obtained via HPLC purification as a pale powder, m.p. 112-114C.

-145- EXAMPLE 111 N,N '-bis{ 3 -[(4-bromo-3methyJ.5.isoxazolyl)aminosulfonylthien2yllurea N, N'-b is 3 [(4-bro mo-3-.methy- 5-isoxazo lyl) amin osulf onyllthien- 2 yllurea was prepared by the same method as described in Example 110 with the exception that N-( 4 -bromo-3-methyl-5-isoxazolyl)-2..carboxythio.

phene-3-sulfonamide was used instead of N-(3,4-dimethyl-5-isoxazolyl).2 carboxylthiopehene-3-sulfonamide. N,N'-bis{3-[(4-bromo-3-methyl-5isoxazolyl)aminosulfonyllthien2yllurea (80 mg, 1 5.5% yield) was obtained via HPLC purification as an off-white solid, m.p. 127-129O

C.

EXAMPLE 112 fonamide OV A. 2 -(benzyloxymethyl)thiophene Sodium hydride (0.41 mg, 20 mmol) was added to a solution of 2thiophene methanol (2.0g, 0.l18mmol) in THF (20 ml) at -40 1C. The reaction was stirred at -400 C for 25 min., then neat benzylbromide (3.6 g, 20 mmol) was added by syringe. The solution was stirred at -400 C for 0.5 hr, then at room temperature for 1 hr. The THF was evaporated off and the remaining residue was taken up in ether 50 ml). The organic solution was washed with water (1 x 10 ml), brine (1 x 10 ml) and dried over MgSO 4 Evaporation of solvents left an oil which was purified by column chromatography using 1 ether-hexanes to give 2.6 g of the thiophene as a pale yellow oil (78% yield).

B. 2 2 -chlorosulfonyl-5-(benzyloxymethyl)thiophene was prepared in the same manner as described in Example 1 32A from 2 -(benzyloxymethyl)thiophene (1.0 g, 5.25 mmol). Purification by column chromatography using 2.5% ethyl acetate/hexanes gave 520 mg of the pure thiophene as a brown oil (32% yield).

-146- C. N 4 -b romo 3 -m ethyl -5-isoxazolyl) 5.(be nzyloxymethyl)thop hen e 2 sulf onamide

N-(

4 -bromo- 3 -methyl-5-isoxazolyl)-5.(benzyloxymethyl)thiophene-2 sulfonamide was prepared as described in Example 2 from 2chlorosulfonyl-5-(benzyloxymethyl)thiophene (520 mg, 1 .72 mmol) and amino-4-bromo-3..methyl isoxazole (319 mg, 1.8 mmol). Purification by column chromatography using 10% MeOH/CHC 3 gave 238 mg of pure N-4bo o3-eh 5isxzll--(ezlxm.hltiph n ufonamide as brown semisolid (31 yield, m.p. 920 C).

EXAMPLE 113 4 -Chloro-3-methyl-5-isoxazoly)..2.ethylbenzo [blfuran-3-sulfonamide A. 2-Ethylbenzo[blfuran 2-Ethylbenzo[blfuran was prepared by the method of example *with benzo[b~furan (7.3 mmols), 0.86 g) t-BuLi (1.7 m, 9.4 mmols) iodoethane (4 mmols, 0.9 mls) and THF (15 ml). 1.0 g of a yellow liquid was isolated.

B. 2 -Ethylbenzo[blfuran-3-sulfonyl chloride 2 -Ethylbenzofbjfuran-3-sulfonyl chloride was prepared by the method of Example 82C with 2-ethylbenzofblfuran (6.9 mmols, 1.0 g), chlorosulfonic acid (8.9 mmols, 0.6 ml) phosphorous oxychloride (21 mmols, 1.9 ml), phosphorous pentachloride (6.9 mmols, 1 .4 g) and

CH

2

CI

2 (10 ml). Flash chromatography ethyl acetate/hexanes) provided 0.71 g of an orange solid.

C. N-4clr--ehl5ioaoy)--tybnobfrn3sloa mide N-4clr--ehl5ioaoy)--tybnobfrn3sloa mide compound was prepared by the method of Example 41 with 4choo3mty--mn-sxzl (1.0 mmols, 0. 13 NaH (2.5 mmols, mg), 2 -ethylbenzo[blfuran-3-sulfonyl chloride (1.2 mmols, 0.28 g) and THE (7 ml). Flash chromatography (20% ethyl acetate/hexanes) followed -147by recrystallization from chloroform and hexane provided 97 mg of a white solid, m.p. 132-133.50 C.

EXAMPLE 114

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-2-[(3,4-methylenedioxy)benzyl])thiophene-3-sulfonamide A. N-( 2 -carbomethoxythiophene-3-sulfonyl)pyrrole

N-(

2 -carbomethoxythiophene-3-sulfonyl)pyrrole was prepared in the same manner as described in Example 33A by reacting 2 -carbomethoxy- 10 thiophene-3-sulfonyl chloride with pyrrole, 50% yield.

B. 2 -hydroxymethyl)thiophene-3-sulfonyl]pyrrole To a stirred solution of N-( 2 -carbomethoxythiophene-3sulfonyl)pyrrole (3.0 g, 11.02 mmole) in a mixture of THF and methanol (3:1 mixture, 40 ml) was added sodium borohydride (1.2 g) in six lots over a 30 min. period (exolhermic reaction) with constant stirring. The solvent was removed under reduced pressure and the residue was dissolved in saturated ammonium chloride solution (50 ml). The crude product was extracted with ethyl acetate, and the combined organic *layers was dried over MgSO 4 and evaporated to give a crude product S 20 (2.4 g, which was used directly in the next step.

C. N-[ 2 -(bromomethyl)thiophene-3-sulfonyl]pyrrole Bromine was added to a stirred mixture of triphenylphosphine (3.1 g, 12 mmole) in methylene chloride (50 ml) at 00 C under nitrogen atmosphere with stirring until the yellow color persisted. A few crystals of triphenylphosphine were added to consume excess bromine followed by pyridine (1.21 ml), and 2 -hydroxymethyl)-3-sulfonyl]pyrrole dissolved in methylene chloride (10 ml) was added. The reaction was stirred at 00 C for 1 hr and concentrated to give a crude product. The crude product was purified by flash chromatography on silica gel using -148- 1 hexane in ethyl acetate to give N-[ 2 -(bromomethyl)thiophene-3 sulfonyllpyrrole (2.7 g, 80% yield).

D. N-{ 2 [(34methylenedioxy)benzylthiophene3ulfnlyrro}j

N-{

2 -[(3,4-methylenedioxy) benzyljthiophene-3-sulfonylJ~pyrrole was prepared in the same manner as described in Example 320 using N- 2 -(bromomethyl)thiophene-3sulfonyllpyrrole and 3 4 -methylenedioxy.

phenylboronic acid, 53% yield.

E. 3clrsloy--(,-ehlndoybnylhohn 10 prepared in the same manner as described in Example 65E from N-{2f( 3 4 -methylenedioxy)benzylI.3-sufonylpyrrole by basic hydrolysis of the 0 OV.sulfonamide to the sodium salt of sulfonic acid and by conversion of this salt to the corresponding sulfonyl chloride, 54% yield.

thiophene-3-sulfonamide N-(4broo-3-ethl-5isoxzoll)-2[(34-mthylnedoxybenzyllthiophene-3-sulfonamide was prepared in the same manner as described in Example 2 by reacting 5-amino- 4 -bromo-3-methylisoxazole and 3- 0.:chI orosu f ny-2[ (,4metyln eioy) enyllh op ene.The crude product was purified by H PLC, 37% yield.

EXAMPLE 115 f-(4-romo--metyl)( 2 methoyl)5(-hflylphniophene-2sul- 2 -methoxyphenl)thne2sffnlpyrl N-5-(2methoypeyltopene--suonylpyrrole was prepared in the same manner as described in Example 32C using 2 -methoxyphenylboronic acid and N-(5-biomothiophenesulfonyl)pyrrole, 74% yield.

B. 5-hooufnl2(-ehxpey~hohn 5-hooufnl2(-ehxpey~hohn was prepared in the same manner as described in Example 65E from 2 -methoxyphenyl)- -149thiophene-2-sulfonyl]pyrrole by hydrolysis of the sulfonamide to the sodium salt of sulfonic acid followed by conversion of the salt to the corresponding sulfonyl chloride, resulting in a 24% yield.

C. N-(4-bromo-3-methyl-5-isoxazolyl)-5-(2-methoxyphenyl)thiophene- 2-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-5-(2-methoxyphenyl)thiophene- 2-sulfonamide was prepared in the same manner as described in Example 1. Reaction of 2 -chlorosulfonyl-5-(2-methoxyphenyl)thiophene with amino-4-bromo-3-methylisoxazole gave N-(4-bromo-3-methyl-5- 10 isoxazolyl)-5-(2-methyoxyphenyl)thiophene-2-sulfonamidein 32% yield, m.p. 114-1170 C.

EXAMPLE 116 N-(4-Bromo-3-methyl-5-isoxazolyl)-5-(2-tolyl)thiophene-2-sulfonamide A. 2 2 -tolyl)thiophene 15 Sodium carbonate (5 ml., 2M aqueous solution) followed by 2methylphenylboronic acid (0.294 g, 2.4 mmol) were added to a solution of 2-bromothiophene (0.542 g, 2 mmol) and tetrakis(triphenylphosphine)palladium(O) (100 mg) in toluene (5 ml) and ethanol (5 ml) under nitrogen.

The mixture was refluxed for 2 hours, cooled to room temperature and extracted with ethyl acetate (2 x 50 ml). The combined organic layers was dried over MgSO 4 and evaporated. The residue was flash chromatographed on silica gel using hexane as eluent to afford 1.2 g of 2 2 -tolyl)thiophene as a colorless gum.

B. 2 -chlorosulfonyl-5-(2-tolyl)thiophene To the cold to 00 C) solution of 2 2 -tolyl)thiophene (0.87 g, mmole) was added chlorosulfonic acid (0.33 ml, 5 mmole) over a 15 min.

period with constant stirring. After 10 min., phosphorous oxychloride (2 ml) and phosphorous pentachloride were added. The reaction mixture was slowly allowed to attain ambient temperature and stirred for 3 hours.

The mixture was then poured onto crushed ice (50 g) and was extracted -150- .4 @4 4 4 4 4 0ebO 4* 4 4

S.

4.

5 4

S

.4 4.

4 4 4*4* 44 4 44* .4 .4 4 .4 44 4

S

4* 4 .4 44 04 4 .4 44 with ethyl acetate (2 x 50 ml). The combined organic layers was dried over MgSO 4 and evaporated. The residue was purified by flash column chromatography on silica gel using 2% ethyl acetate in hexane to give 2chlooufcy -2-oy~hipee (1 .1 g, 72% yield).

C. N-( 4 bromo3methyl..5..xzoli)-5-(2olyl)5(tlthiohee 2 sffo mide N-4boo3mty--sxzll--2tlltipee2sfoa mide was prepared in the same manner as described in, Example 2.

Reaction of 5-chlorosulfonyl-2(2tolyi)thiophefle with 5-amino-4-brorno.3 10 methylisoxazole gave the crude product which was purified by column chromatography giving the pure product (gum). This gum was dissolved in 5 ml of NH 4 OH, concentrated and dried under high vacuum to get the ammonium salt of N-4boo3mty--sxzli--2tlltipee 2 -sulfonamide in 67% yield, m.p. 180-184O C (NH 3 salt).

EXAMPLE 117

N-(

4 Bromo3.methyl..5..xzoli)-5-xaoly)5(tiohnh 2 -sf nm A. 2 3 -tolyl)thiophene 2 3 -tolyl)thiophene was prepared in the same manner as described 20 in Example 11 7A using 2 -bromothiophene and 3 -methylphenylboronic acid. The crude product was purified by flash chromatography on silica gel using hexane as the eluent (86% yield).

B. 2 -chlorosulfonyl-5..(3-tolyl)thiophene 2 -chlorosulfonyl.-5-(3-tolyl)thiophefle was prepared in the same manner as described in Example 11 7B from 2 3 -tolyl)thiophene, 22% yield.

C. N-( 4 bromo3.methy..5-ioazi)-5-xazolyl)(thoyphnh 2 -sfo mide mide was prepared in the same manner as described in the Example 11 7C using 2 -chlorosulfonyl-5-(3-tolyl)thiophene and 5-amino-4-bromo-3 -151methylisoxazole. To obtain the ammonium isalt of the final product, aqueous

NH

4 0H was used (31 yield; hygroscopic).

EXAMPLE 118

N-(

4 Bromo3methyl5oxoy)3-zoylthiobene-h~ 2 -ufnm A. 3 -benzylthiophene 3 -benzylthiophene was prepared in the same manner as described in Example 32C using 3 -thienylboronic acid and benzyl bromide, 74% yield.

B. 2 -chlorosulf onyl.3-benzylthiophene 0 2 -chlorosulfonyl3benzylthiophene was prepared in the same manner as described in Example 1178 using 3 -benzylthiophene, 78% yield.

*SO

.00.* 6* a i 6605 5@O* 0 55 5S *0 C. N-( 4 bromo3methyl5.isoxazolyI)33belthohe-2sl nmd

N-(

4 bromo.3methyl5isoxazolyl)3bzlthioh.--ul nm 15 was prepared in the same manner as described in Example 2 by reacting 2 -chlorosulfonyl.3-benzylthiophene with 5-amino-4-bromo-3.

methylisoxazole resulting in a 24% yield, m.p. 180-183o

C.

EXAMPLE 119 amideY)S(m ethl5isxayluranmehyfuny)tioph ene--lfofl t-BuLi (1.7 m solution in hexane, 7.9 ml, 14.6 mmol) was added dropwise under constant stirring under a -nitrogen atmosphere to a solution of 2 -methylfuran (1.0 g, 12 mmol) in THF (20 ml) at -780 C.

The solution was then warmed to -100 C and stirring continued for min. The solution was then added to a solution of zinc chloride (27 ml of a 0.5 M solution in THE) at -301 C and then warmed to room temperature where stirring continued for 1 hr. resulting in a pale yellow clear solution. The solution was then transferred via a steel canula under nitrogen to a solution of N-5boohohn-2sloy~yrl (Example -152- 33A, 3.5 g, 12 mmol) and tetrakis(triphenylphosphine)-palldiu (693 mg, 0. 6 mmol) in THF (15 ml) at -78O0 C. The solution was then warmed to room temperature and stirred for a period of 2 hours. Purification by column chromatography using 2% ethyl acetate gave 680 mg of 5-(2- I mety-5~-ury)thiophene-2..sulfony II Iol as a pale yellow powder (19% yield).

B. 2 2 chord 2-(2metyl--fuyl~hiohen-5-ulfnyIchloride was prepared in the same manner as described in Example 33D from N-f S-( 2 -methyl-s..

:10 furyl)thiophene-2sulfonylpyrrole (300 mg, 1.02 mmol). Purification by colum chromatography usn %ethyl acetateIhexaes av 145 m of the sulfonyl chloride as a pale yellow solid.

C. N 4 brom o- 3 .m ethyl 5 .is xaz y) 5 2 -m t l -urlt i ph ne 2 *sulfonamide sulfonamide was prepared in the same manner as described in Example 2.

Reaction of (55 mg, 0.21 mmol) with 5-amino-4-bromo-3.methylisoxazIl (41 mg, 0.21 **mmol), after purification by column chromatography using 10% MeOH/CHCI 3 gave 45 mg of the pure sulfonamide as a brown semisolid 54% yield, m.p. 123-124o

C).

EXAMPLE 120 amide was prepared in the same manner as described in Example 2 using 5-amino-4bromo3methylisoxaol (132.75 mg, 0. 75 mmole) and 4- (phenethyl)thiophene-2-sulffn chloride (Example 119D); 225.62 m g, 0.75 mmol). The product was purified by HPLC (5-95% acetonitrile with 0. 1 TFA over 30 min.) to give N-(4bromo3methyl5ioxazoy)- 4 (henthyitiphene-2-sulfonamide as a brownish oil 7 2 .3mg, 32% yield.

-153- EXAMPLE 121 N-(3,4-dimethyl-5isoxazolyl)-2-(4-methylphenyl)aminocarbonylthiophene-3-sulfonamide Phosphonitrilic chloride trimer dissolved in THF (5 ml) was added to a suspension of N-(3,4-dimethyl-5-isoxazolyl)-2-(carboxyl)thiophene-3-sulfonamide (2.0 g, 6.6 mmol) (Example 28) in THF (5 mi) and Et 3 N at 00 C.

The cold bath was removed and the reaction mixture stirred at room temperature for 2 hours. The mixture was diluted with water (150 ml) and acidified to pH 2 using concentrated hydrochloric acid. The reaction mixture was then extracted with methylene chloride (2 x 100 ml), and the combined organic layers was washed with 2N hydrochloric acid (3 x 100 ml), dried over MgSO 4 and concentrated be get crude product. The crude product was dissolved in ether and allowed to stand at room temperature to give a precipitate which was filtered and washed with cold ether giving N-(3,4-dimethyl-5-isoxazolyl)-2-(4-methylphenyl)aminocabnItin:r...*.~~a carbonyl]thiophene-3-sulfonamide (1.6 g, 61% yield).

EXAMPLE 122

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-5-(4-tolyl)aminocarbonylthiophene-2sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-5-(4-tolyl) aminocarbonyl]thiophene-2-sulfonamide was prepared in the same manner as described in Example 24 from N-(4-bromo-3-methyl-5-isoxazolyl)-5-(4-carboxybenzene)thiophene-2-sulfonamide (Example 148), 110 mg, 0.25 mmol) and 4 -methylaniline (53 mg, 0.49 mmol). Purification by recrystallization from methanol/water gave 91 mg of the pure sulfonamide as a light tan powder (61% yield, m.p. 1880 C).

-154- EXAMPLE 123 sulfonamide A.

N-(

4 chloro3methyi..5..ioly)2(abo )tipee3sfo mide Yrx~hohn.3ufn

N-(

4 chloro3.methyl5iso azlyl) )2(caby)thioh.--ufo mide was prepared in the same manner as described in Example 1 7 using

N-(

4 chloro3.methy--ioxly.iox-aboml e(hoytipe- 10 sulfomnanide (Example 123), 78% yield.

B. N 4 -chlo ro 3.methyl.5is oxazolyl) 2 4 -tll in) ab nl thiophene-3..sulffnamide

N-(

4 chloro.3.methyl5isoxazoly) 2 f( 4 -toll)amncroyl thiophene-3-sulfonamide was prepared in the same manner as described 15 in Example 122 using

N-(

4 chloro3..methyl.5.isoxazolyl)- 2 (carboxyl)thiophene-3sulfonamide. The crude product was dissolved in a small quantity of EtOAc (2 ml) and ether (15 ml) was added. The resulting precipitate was filtered and washed with cold ether (50 ml) giving

N-(

4 chloro3methyl.5isoxazolyl) 2 f( 4 -toll)amncroy* thiophene-3-sulfonamide in 53% yield 177-1790).

.:EXAMPLE 124

N-(

4 -Bromo-3methyl..5.isoxazolyl) 2 [(Nmehl).N-hnlmocrny A solution of N-4boo3mty--sxzli--croy) thiophene-3-sulfonamide (183.6 mg, 0.5 mmol) in dry THF (I ml) was added to a solution of N-methyl-4-methylaniline (0.066 ml, 0.6 mmol) in THF (0.5 ml). Triethylamine (0.63 ml, 4.2 mmol) was added to the mixture and, after 1 0 min., a solution of phosphonitrilic chloride trimer (210.7 mg, 0.6 mmol) was added. The mixture was stirred for 1 hr. at 5000C, cooled, neutralized with 10 ml 1 N HCI (pH 3) and extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate and the solvent was removed under reduced pressure to give a crude -155product which was purified by HPLC (5-95 acetonitrile with 0. 1 TFA over 30 min.) resulting in N-( 4 -bromo.3-.methyl.5.isoxazoly) 2 methyl)Nphenylaminocarbonylihnh 3 -ufnmd as a white solid (92.1 mg, 39.4% yield, m.p. 51-55 0

C).

EXAMPLE 125

N-

4 -om3-methyl.5..isoxazolyl)3[3,4.(methylenedioxy)phelI- A. 3 -bromothiophene.2-sulfonyl chloride Chlorosulfonic acid (20 ml, 300 mmol) was added to a solution of 3 -bromothiophene (8.15 g, 50 mmol) in methylene chloride (50 ml) at 9e* 9 -780 C over a 20 min. period. After the completion of addition, the cold bath was removed and stirring continued at ambient temperature for 1 hr.

The reaction mixture was carefully added, dropwise, to crushed ice *15 (100 The mixture was extracted with methylene chloride (2 x 100 ml). The combined organic layers was dried over MgSO 4 and evaporated.

The crude product was purified by flash chromatography on silica gel using hexane as the eluent resulting in 3 -bromothiophene2sulfonyl chloride (4 g, 30% yield) and 4 -bro moth ioph ene2-sulf onyl chloride 2 0 0 mg, :5 1 B. N-3boohohn-2sfoy~yrl

N-(

3 -bromothiophene..2.sulfonyl)pyrrole was prepared in the same manner as described in Example 33A by reacting 3 -bromothiophene.2sulfonylchloride with pyrrole (for 16

N-(

3 -bromothiophene-2 sulfonyl)pyrrole was obtained in 54% yield.

C. 3 -(3,4methylenedioxy)phenyI hiohe-2sfoyprol N-[-34mtyeeix~hnltipee2sloy~yrl was prepared in the same manner as described in Example 320 using 3,4mehlndoyhnlooi acid and N-( 3 -bromothiophene-2 sulfonyl)pyrrole. The crude product was purified by flash column -156chromatography on silica gel using 2% EtOAc in hexane as the eluent resulting in 3 (3,4.methylenedioxy)phenylthohe-2sfoypyrrole in a 90% yield.

D. 2 -chlorosufonyl3[3(meth edx)pheythoe 2 -chlorosulfony..3j3,4(methndy.)phnlhoh was prepared in the same manner as described in Example 1 12B using 3 4 -methylenedioxy)phenylthiophen 2 5 flfonlpyrrebybai hydrolysis of the sulf onamide to the sodium sulfonate 0100% yield) followed by conversion of the salt to the corresponding sulfonyl chloride 10 resulting in a 34% yield of the final product.

N-(

4 3 3 4 (mehlndoype thiophene..2-sulfonamide was prepared in the same manner as described in Example 1 by reaction of 2-hooufnl3[,-mtyeeix) pheny1Ithiophene with resulting in a yield, m.p. 183-1860

C.

EXAMPLE 126

N-(

4 Bromo.3ehy-ethoalil)- 2 -[(2clr-,-ehlndoy ~20 phenoxymethyllthiophene.3sulfonamide A N 2 ,4 m e th y le n e d io x y )p h e x m t yl th o e n s u f y l pyrrole Sodium hydride (100 mg, 5 mmole) was added to a stirred solution of 3 4 -methylenedioxyphenoJ (0.607 g, 4.5 mmol) in DMF (dry, 5 ml) at 00 C under a nitrogen atmosphere with stirring. The reaction mixture was permitted to attain room temp and stirring continued for 1 hr. The reaction mixture was cooled to 00 C and 2 bromomethyl)thiophen 3 sulfonyllpyrrole was added. Stirring was continued at ambient temperature for 1 6 hr. The reaction mixture was diluted with water (1 00 ml), extracted with ethyl acetate (2 x 50 ml) and washed with 1IN NaOH -157- (2 x 25 ml) to remove phenol derivative. The mixture was dried over MgSQ 4 and concentrated resulting in N-{ 2 3 ,4-methylenedioxy)phenoxy.

methyllthiophene-3-sufonyllpyrrole, which was recrystallized using hexane/EtOAc (1.0 g, 92% yield).

B. 3-h rs fnl2 tyendoy e ynehl thiophene 3-hooufnl2[2clr-,4mtyeeix~hnxmtyl thiophene was prepared in the same manner as described in Example 64E using N{-(,-ehlndoypeoyehltipee3sloyl pyrrole by conducting a basic hydrolysis (using potassium hydroxide in a iso-propanol) to the potassium sulfonate followed by conversion of the salt to the corresponding sulfonyl chloride in an overall yield of C. N-( 4 -bromo3methyl5isoxazoly).2[(2chloro34methylephnx*ety hopee3sufnmd was prepared in the same manner as described in Example 1 by reaction of 3-chlorosulfonyl-2-[(2choo-,-mtylndixpenx:mty tiohn with 5-amino-4- S bromo-3-methylisoxazole, 47% yield, m.p. 152-154 0

C.

EXAMPLE 127 A. Diethyl 2 3 -[(N-pyrrolyl)sulfonyllthienylmethyllphosphofl 81 e

N-[

2 -bromomethyl)thiophene.3sulfonyllpyrrole (0.91 5 g, 3 mmol) was suspended in tri ethyl phosph ite (5 ml) and was heated to 1400 C for 1 hr. with stirring under nitrogen atmosphere. Excess triethylphosphate was removed under reduced pressure and the residue was dried under vacuum resulting in 0.9g, 83% yield of diethyl 2 3 -f(N-pyrrolyl)sulfonyljthienylmethyl~phosphonate.

-158pyrrole rfeSUI lyJ Sodium hydride (200 mg, 60% dispersion) was added in two lots to the stirred solution of diethyl 2 3 -[(N-pyrroly)sufonyllthienylmethyl}phosphonate (900 mg, 2.48 mmol) in dry THF (10 ml) at 00 C. The mixture was stirred at room temperature for 1 hr. then piperonal (600 mg) was added. Stirring was continued for 12 hours. The mixture was diluted with water (100 ml) and extracted with methylene chloride (2 ml). The combined organic layers was dried over MgSO 4 evaporated, and the residue was flash chromatographed on silica gel using 0.5% ethyl *acetate in hexane to give N-(-[tas 34mtye doy innaml thiophene-3-suffonyl~pyrrole (750 mg, 84% yield).

C. 3 -chl orosulfony2[trans3,4-(methylenedioxy) cinna myl th phee 3-hooufoy-- trns 3,4 (mtyeeix)cnay~hoh was prepared in the same manner as described in Example 64E from N- {2 rn-34(ehlndoy inaylhohn--ufnl yrl by basic hydrolysis (using isopropanol and potassium hydroxide) to the corresponding potassium sulfonate (100%) followed by conversion of the salt to the corresponding sulfonyl chloride in a 31 overall yield.- D. N-( 4 bromo3methy.5..isoxazol 2[tr ras3, 4 -(thledioxy)cinnamyllthiophene-3-sulfonamide cin n amyl~th iophene3-sulf on amide was prepared in the same manner as described in Example 1 by reaction of 3 -chlo rosulf onyl-2- [tras- 3 4 (methylenedioxy)cinnamyIlthiophene with 5-amino-4-bromo.3-methylisoxazole. The crude product was purified by HPLC resulting in a 33% yield, m.p. 147-149 0

C.

-159- EXAM PLE 128 N-h-oo-hn 3 -suefofl--isxazokl)2[34(methylenedioxy)phelethyl]- A.

N-{

2 [34(methylenedioxy)phenethyl jthio h- 3 lffoylyrl An ethyl acetate (15 ml) solution of N-{ 2 -ftrans-3,4-(methylenedioxy)cinnamythiophene3sulfonylrle (Example 1278, 0.6 g, 1.67 a. mmol) was subjected to catalytic hydrogenation using 10% Pd-C (100 at 55 psi for 14 hr. The catalyst was filtered~jdd the filtrate lo*::1 concentrated to resulting in N-{ 2 -[3,4(methylenedioxy)phenethyllthio a phene- 3 -suffonyl~pyrrole (0.55 g, 91 yield).

3 -chlorosulfonyl.2[3,4(methylenedioxy)phefylth~ihe a prepared in the same manner as described in the Example 64E using

N-

{2[,-mehlndoxIhntyltipen--ufnIlyrl by conducting basic hydrolysis (iso-propanol and potassium hydroxide) of the sulfonamide to the potassium salt of sulfonic acid followed by conversion of the salt to the corresponding sulfonyl chloride in a 42% yield.

2 0 C N r m e h l 5 i o a o y 2 m t y e e i x

N-(

4 -bromo-3-.methyl..s-isoxazoly) 2 f[ 3 4 -(methylenedioxy).

phnty~hohee -ufnai was prepared in the same manner as described in Example 2. By reacting 3 -chlorosufonyl.2f3 (mtyeeix~hntylhohn with 5-amino-4-bromo-3 methylisoxazole and purifying the crude product by HPLC, N-(4-bromo-3.

l--34(mtyeeix~hnthltipee3sl fonamide was obtained in a 30% yield, m.p. 1800 (dec.).

-160- EXAMPLE 129

N-(

4 -Bromo-3-methyl-5-isoxazolyl)-3-(phenylthio)thiophene-2-suffonamide A. 3 -phenylthiothiophene-2-sulfonyl chloride A stirred solution of 3 -(phenylthio)thiophene (1.0 g, 5.2 mmol) in ml of dry THF was placed under an argon atmosphere and cooled to -78 C. n-butyl lithium (2.78 ml of 2.3M solution) was added over 20 min.

and stirring was continued at this temperature for an additional 20 min.

Sulfur dioxide gas was then bubbled in at -780 C for a period of 30 min., resulting in the formation of a yellow percipitate. This was immediately 10 followed by dropwise addition of N-chlorosuccinimide (764 mg, 5.72 mmol) dissolved in THF. The mixture was warmed to room temperature and stirring continued for an additional 1.5 hr. The mixture was then concentrated and the residue dissolved in ether. The organic layer was washed with water, brine solution and dried over magnesium sulfate.

15 Evaporation of solvents left a light brown oil which was subjected to S flash chromatography. Elution with 2% ethylacetate-hexanes gave 840 mg of a pale yellow solid.

S* B. N'( 4 "bromo-3-methyl-6-isoxazolyl)-3-(phenylthio)thiophene2sul fonamide N-(4-bromo-3-methyl-6-isoxazolyl)-3-phenythio)thiophene--sulfonamide was prepared in the same manner as described in Example 2 using 5-amino-4-bromo-3-methylisoxazole (192 mg, 1.1 mmol) and 3phenylthiothiophene-2-sulfonyl chloride (300 mg, 1.0 mmol). Purification by column chromatography using 10% MeOH/CHCI 3 yielded 358 mg of the pure sulfonamide as a brown oil.

-161- EXAMPLE 130 N-(3,4 dimethyl-5-isoxazolyl)3(phenlaminocarbIh.ohn--ufn amide n topee2sfn.

A. N 3 4 dimeth yl 5is oxazolyl)thi oph ene2sufo mde -34dmty--sxzly~hohn--ufnmd was prepared in the same manner as described in Example 14 using thiophene-2sulfonyl chloride and 3 4 -dimethylaminoisoxazole. Purification by column 0% chromatography using 3% MeOH/CHC 3 yielded 48% N-(3,4-dimethyl-5.

isoxazo lyl) thiop hene- 2-sulf on amide.

B. N-(methyoxyethoxymethyl)..N.(3,4dimethyl-5-isoxazolyl)thiophe- 4 2 -sulfonamide .N-(methyoxyethoxymethyf)..N.(3,4dmty--sxzlltipee 2 -sulfonamide was prepared in the same manner as described in Example 32BusngN- 3 4 dimeth yl.5isoxaz yl t opy~hnehenulfnaidean methoxyethoxymethyl chloride resulting in 34% yield. HPLC analysis of :.the crude oil obtained after workup showed that the oil was approximately 96% pure and so was used in the next step without further purification.

C. N-(methyoxyethoxymethyl).N.(3,4-dimethyl-5-isxaIl)-5 :20 trim ethylsilyl)thio phen e2-sulf onamide A stirred solution of isoxazolyl)thiophene.2.sulfonamide (300 mg, 0.87 mmol) in 5.0 ml of dry THF was placed under an argon atmosphere and cooled to -780 C. Over the course of 20 min, a solution of t-BuLi in hexanes (461 ml of a 2.25 M solution) was added dropwise and stirring was continued at this temperature for about 25 min. Then neat trimethylsilych lo ride (135 ml, 1 mmol) was added dropwise and the solution was stirred at -781 C for min. then at room temp. for 1 .5 hr. TLC (1 CHCI 3 in MeOH) showed complete reaction of starting material at-this time thus, the reaction was quenched by addition of 2.0 ml of water. After evaporation of solvents -162the remaining residue was extracted into ethyl acetate, washed with brine solution and dried over magnesium sulfate. Purification by column chromatography using 20% ethyl acetate/hexanes gave the pure sulfonamide as a clear oil (52% yield).

D. N-(methyoxyethoxymethyl)-N-(3,4-dimethyliisoxazolyl)-3-(phenylaminocarbonyl)-5-(trimethylsily)thliophene-2-sufonamide A stirred solution of N-(methyoxyethoxymethyl)-N-(3,4-dimethyl-5isoxazolyi)-5-trimethylsilyl)thiophene-2-sulfonamide (180 mg, 0.43 mmol) in 4 ml of dry THF was placed under an argon atmosphere and cooled to- 780 C. At this temperature a solution of t-BuLi in hexanes (215 p1 at a 2.55 M solution) was added dropwise and stirring was continued at -780 C for 0.5 hr. resulting in a clear yellow solution. Phenylisocyanate (77 pl, S 0.65 mmol) was added dropwise at -780 C and the solution allowed to reach room temperature. The solution was then worked up as above in part C. Purification of the final product was achieved by column chromatography using 30% ethyl acetate/hexanes to give 108 mg of the sulfonamide a 54% yield.

E. N-(3,4dimethyl-5-isoxazolyl)-3-(phenylaminocarbonyl)thiophene-2- C sulfonamide N-(3,4 dimethyl-5-isoxazolyl)-3-(phenylaminocarbonyl)thiophene-2sulfonamide was prepared in the same manner as described in Example 32D using N-(methyoxyethoxymethyl)-N-(3,4-dimethyliisoxazoly)-3-(Nphenylcarboxamide-5-trimethylsilyl)thiophene-2-sufonamide (108 mg, 0.23 mmol). Purification was achieved by recrystallization from acetonitrile/water to give 62 mg (71% yield) N-(3,4 isoxazolyl)-3-(phenylaminocarbonyr)thiophene-2-sulfonamide as a brown powder, m.p. 152 0

C.

-163- EXAMPLE 131 N-(3,4-dimethyl-5-isoxazolyl)-2-(a-hydroxybenzyl)thiophene-3-sulfonamide A. Thiophene-3-sulfonyl chloride n-BuLi (2.38M, 17ml) was slowly added to a solution of 3-bromothiophene (6.5 g, 40 mmol) in ether (30 ml) at -780 C. The reaction was stirred at -780 C for 45 min. SO was bubbled through the mixture for minutes at -780 C followed by the addition of NCS (6.4 g, 48 mmol) as a suspension in THF (40 ml). The crude product was purified by column chromatography using 5% ethyl acetate/hexanes to give 3.92 g 10 thiophene-3-sulfonyl chloride as a pale yellow solid (54% yield).

B. N-( 3 ,4-dimethyl-5-isoxazolyl)thiophene-3-sulfonamide N-(3,4-dimethyl-5-isoxazolyl)thiophene-sulfonamide was prepared in the same manner as described in Example 14 using thiophene-3sulfonyl chloride and 3 4 -dimethylaminoisoxazole resulting in 66% yield S 15 as a pale brown solid.

C. 2-[2-(trimethylsilyl)ethoxymethyll-N-(34-dimethyl-5 isoxazolyl)thiophene-3-sulfonamide N,N-diisoproplyethylamine (222 pl, 128 mmol) was added to a solution of N-(3,4-dimethyl-5-isoxazolyl)thiophene-3-sulfonamide (300 mg, 1.16 mmol) in methylene chloride (5 ml), and the mixture was stirred at room temperature for 15 min. The mixture was then cooled to 0° C and 2 -(trimethysilyl)ethoxy methyl chloride (SEM chloride) (226 p1, 1.28 mmol) was added dropwise via syringe, and the resultant yellow solution was stirred at room temperature for 5 hours. Evaporation of solvents left an oil which was extracted into ethyl acetate, washed with water and brine solution and dried over magnesium sulfate. Flash chromatography of the residue using 10% ethyl acetate/hexanes yielded 321 mg 2-[2- (trimethylsilyl)ethoxymethyl]-N-(3,4-dimethyl-5-isoxazolyl)thiophene-3-sulfonamide as a clear colorless oil which solidified into a white solid upon standing (71% yield).

-164- D. 2 2 (trimethylsilyl) ethoxym ethyl] -N -(34-dimthyI-5-sxzl) (a-hydroxybenzy)thiophene3suffonande n-BuLi (2.39 M, 177 p1) was slowly added to a solution of (2trimethylsilylethoxymethyl)N(34dimethyl15-ioaoy*hipee3s fonamide (156 mg, 0.38 mmol) in THF at -78 0 C under nitrogen. The reaction was stirred at -781 C for 45 min., then benzaldehyde (45 p1l, 0.42 mmol) was added in one lot at -780 C and the solution was allowed to come to room temperature. Stirring was continued for 1 hr., Purification was achieved by column chromatography using 10% ethyl 10 acetate/hexanes to give 179 mg 2

-L

2 -(trimethylsilyl)ethoxymethyJJN( 3 4 diehl5ioaoy)2(-yrxbny~hohn--ufnmd as a yellow viscous oil (90% yield).

E. N-( 3 4 dimethyl5.isaoyl)-2 l)2(adrxbezltipn-3ua a fonamide .15 To a solution of 2-2(rmtysllehxmtylN(,-iehl mmol) in DMVF (2 ml) was added cesium fluoride (26 mg, 0. 17 mmole) in one portion. The resulting mixture was heated to 10001 C for 10 hours.

The solvents were removed by evaporation under vacuum and the remaining residue was extracted into ethyl acetate, washed with water, brine, and dried over MgSO 4 The product was then purified by chromatography using 50-70% ethyl acetate/hexanes to give 26.2 mg N- (3,4-iehl5ioaoy ahdoyezlthohn--ufnmd as a pale white semisolid (51 yield).

EXAMPLE 132 amideoo--etyl-soaol~mtylphen(4-mthlphentipe e2sulfon- A. N-lIS-( 4 methylphenyl)thiophene2sulfonyI] Iol N-5(-ehlhnitipee2sloylyrl was prepared in the same manner as described in Example 32C using 4 -methyl-phenyl- -165.

boronic acid and N- (5-bro moth ioph enesulf onyl) pyrro le. Purification by column chromatography using 2% ethyl acetateihexanes gave methylphenyl)thiophene.2-sulfonyulpyrrole as a pale yellow solid in 77% yield.

B. 2 -chlorosulfonyl..5-(4..methylphenyl)thiophen was prepared in the same manner as described in Example 33D using N-[5-(4-methylphenyl) thiophene-2-sufonyllpyrrole. Purification by column chromatography using 2% ethyl acetate/hexanes gave 2 -chlorosulfonyls5(4methylphenyl)thiophene as a pale yellow powder (61 yield).

sulfonamide sulfonamide was prepared in the same manner as described in Example 2.

Reaction of (100 mg, 0.37 *mmol) with 5-mn--rm--ehlsxzl (65 mg, 0.37 mmol) yielded, after column chromatography using 10% Me0H/CHCI 3 96 mg final product as a pale yellow solid, (63% yield, m.p. 1750

C).

EXAMPLE 133 4 Bromo3.meth ;.isoxazolyl( 4 hnl)tipe-2sfoade A. N-(pyrrole)5.(4phenViophoe-2lfnmde 4phnl~hophn--ufnai was prepared in the same manner as described in Example 32C using phenyl boronic acid and ohip nsloy)proe Purification by column chromatography using 2% ethyl acetate/hexanes gave the pure sulfonamide as a yellow powder in 67% yield.

B. 2 -chlorosulf onyl..5.(4..phenyl)thiophen 2 -chlorosulfonyl..s(4-phenyl)thiophefle was prepared in the same manner as in Example 33D from N-proe--4penltipee2sl fonamide. Purification by column chromatography using 2% ethyl -166acetate/hexanes gave the pure thiophene in 77% yield as a yellow power.

C. N-( 4 bromo3methyl-i ioxazol(4-hyl)thphenlehi 2 lffo mide N-(4-bromo- 3 -methyl-5-isoxazolyl).

S.(

4 -phenyl)thiophene-2..sulfona.

mide was prepared in the same manner as described in Example 2.

Reaction of 2 -chlorosulfonyl-5-(4-phenyl)thiophene (94 mg, 0.36 mnmol) with 5-amino-4-bromo-3-methyl isoxazole (64 mg, 0.36 mmol) yielded.

after column chromatography using 10% MeOH/CHCI 3 85 mg of N-(4bromo 3 methyl5isoxazlyl)5(4py~tihen-thshlfonmd as a light brown solid, (59% yield, m.p. 132 0 C).

EXAMPLE 134

N-(

4 2 -sulfonamide 9 prepared in the same manner as described in Example 320 using 4trifluoromethylbenzene boronic acid and N-(5-bromothiophene sulfonyl)pyrrole. Purification by column chromatography using 2% ethyl am~tate/hexanes gave the pure sulfonamide as a white powder in 75% yield.

B. was prepared in the same manner as Example 33D from N-{5-[4-(trifluoro.

methyl)phenyllthiophene.2suffnylpyrrole. Purification by column chromatography using 2% ethyl acetate/hexanes gave the pure thiophene as a white powder in 41 yield.

thiophene-2..suffonamide thiophene-2-sulfonamide was prepared in the same manner as described -167in Example 2. Reaction of 2 -chlorosulfonyl-5-14-(trifluoromethyl)phenyl]thiophene (100 mg, 0.31 mmol) with 5-amino-4-bromo-3-methyl isoxazole (54 mg, 0.31 mmol) yielded, after column chromatography, 39 mg of N-( 4 -bromo-3-methyl-5-isoxazolyl)-5-14-(trifluoromethyl)phenyllthiophene-2-sulfonamide as a pale yellow powder, (27% yield, m.p.

1320

C).

EXAMPLE 135 N- 4-Bromo-3-methyl-5-isoxazolyl)-5-(2-formylphenyl)thiophene-2-sufona- A. N-(3-methyl-5-isoxazoly)-5-bromothiophene-2-suffonamide

N-(

3 -methyl-5-isoxazolyl)-5-bromothiophene-2-sufonamide was prepared in the same manner as described in Example 14 from bromothiophene-2-sulfonyl chloride and 5-amino-3-methyl isoxazole.

Purification was achieved by extraction of the crude sulfonamide into aqueous 2N NaOH, washing of the aqueous layer with ethyl acetate and i acidification, using concentrated HCI, to pH Re-extraction into ethyl acetate was followed by washing of the organic material with water, brine and drying over magnesium sulfate. After evaporation of solvents, a brownish solid remained which was sufficiently pure to use in the next step.

B. N- (3-methyl-5-isoxazolyl)-5-(2-formylphenyl)thiophene-2-sulfonamide N-(3-methyl-5-isoxazolyl)-5-(2-formylphenyl)thiophene-2-sulfonamide was prepared in the same manner as described in Example 32C from 2 -formylbenzeneboronic acid (281 mg, 1.87 mmol) and N-(3-methyl- 5-isoxazolyl)-5-bromothiophene-2-sulfonamide (550 mg, 1.7 mmol).

Purification by column chromatography using 15% MeOH/CHCI 3 gave 163 mg of the pure sulfonamide as a brown oil.

30C. N-(4-bromo-3-methyl-5-isoxazoly)-5-(2-formylphenyl)thiophene-2sulfonarnide -168- N-bromosuccinamide (81 mg, 0.45 mmol) was added to a solution of N-( 3 -methyl-5-isoxazolyl)-5-(2-formylphenythiophene-2-sulfonamide (155 mg, 0.45 mmol) in CHCI 3 (5 ml). The resulting brownish solution was stirred at room temperature for 3 hours. The solvent was stripped off and the material extracted into ethyl acetate and washed with brine solution. Evaporation of solvents gave 85 mg of the product (45% yield).

A portion of this was further purified by preparative HPLC. N-(4-bromo-3ethyl-5-isoxazoly)-5-(2-formylphenyl )thiophene-2-sulfonamide was isolated as a pale brown oil.

EXAMPLE 136 SN-(4-Bromo-3-methyl-5-isoxazolyl)-5-(3-aminophenyl)thiophene-2-sulfonamide A. N-(3-methyl-5-isoxazolyl)-5-(3-aminophenyl)thiophene-2-sulfonamide 15 N-(3-methyl-5-isoxazolyl)-5-(3-aminophenyl)thiophene-2-sulfonamide was prepared in the same manner as described in Example 32C from 3 -amino benzene boronic acid (256 mg, 1.87 mmol) and N-(3methyl-5-isoxazolyl)-5-bromothiophene-2-sufonamide mg, 1.7 mmol).

Purification by column chromatography using 15% MeOH/CHCI 3 gave 20 318 mg of the product.

B.

N-(

4 -bromoe3-methyl- 5-isoxazolyl)- 5-(3-aminophenyl)thiophene-2- N-(4-bromo-3-methyl-5-isoxazolyl)-5-(3-aminophenyl)thiophene-2- Sulfonamide rpe i h sulfonamide was prepared in the same manner as described in Example 30A (without acetic acid) using

N-(

3 -methyl-5-isoxazolyl)-5-( 3 aminophenyl)thiophene-2-sulfonamide resulting in a 33% yield. Further purification was achieved using preparative HPLC giving pure N-(4-bromo- 3-methyl-5-isoxazoly)-5-(3-aminophenyl)thiophene-2-sufonamide as a clear colorless oil.

-169- EXAMPLE 137

N-(

4 Bromo.3methyl5isoxall)- 3 3 -dmty y-y)thiophene.2 A. N-f 3 -dimethylbutyn. 1-yl)thiophene-2sulfonyllpyrrole A mixture of N-(5-bromothiophene2sulfonyl)pyrol (600 mg, 2.05 mmol), 3 3 -dimethyl- 1 butyne (338 mg, 4.1 mmol), copper iodide (39 mg, 0.21 mmol), tetra kistriph enylph osph in e palladium LPd(PPh 3 4

J

(118 mg, 0.1 mmol) and piperidine (5 ml) was stirred at room temperature for a period of 24 hours under a nitrogen. atmosphere. The mixture was then diluted with water (10 ml) and extracted with 3 x ml portions of ether. The combined ether extracts are washed with brine and dried over MgSO 4 The solvent was removed under reduced pressure and crude product purified by column chromatography using 2% ethyl acetate/hexanes to give 423 mg N-[5-(3,3-dimethylbutyn- 1 -yl)thiophene.

2 -sulfonyllpyrrole as a yellow powder (70% yield).

B 2 -chl oTosulfonyl 5(3,3dimeth ylbutynl -yl)thiop hene 2 was prepared in the same manner as described in Example 33D from 1-ltipee2sfoaie Purification by column chromatography using 2% ethyl acetate/hexanes gave the pure sulfonamide in 33% yield.

C. N-4brorno.3.methyl5isoxazly)-5(3,3.dimty y-

N-(

4 -bromo..3.methyl-5-.isoxazoyf) 3 3 -dimethylbutyn. 1 yltipee2-ufnmd was prepared in the same manner as described in Example 2. Reaction of 2 -chlorosulfonyi.5-(3, 3 dimethylbutyn.1.yl)thiophefle (120 mg. 0.46 mmol) with 5-amino-4brm--ehlsxzl (85 mg, 0.48 mmol) yielded, after column chromatography using 10% MeOH/CHCI 3 116 mg N-( 4 -bromo-3methyl.

5-isoxazolyl).5.(33dimethybutyn- 1 yl)thiophene-2-sulf onamide as a viscous clear oil -170- EXAMPLE 138

N-(

4 -Bromo-3-methyl..5.isoxazolyl..

3 ,S-bis(trifluoromethyl)phenyll- A. 2 -sfoylyrl N-{5-[3,5bis(trifluoromethpeylplthohe-2sloyproe was prepared in the same manner as described in Example 32C from bis(trifluoromethyl)benzeneboronic acid (619 mg, 2.26 mmol) and bromothiophene-.2..sulfonyllpyrrole (60 mg, 2.05 mmol). Purification bycolumn chromatography using 2% ethyl acetate/hexanes gave the pure sulfonamide as a white solid in 93% yield.

B. 2 -chlorosufonyl.5[5bis(tri lothl)pheythoen prepared in the same manner as described in Example 33D from *[3,-bstilooehlpeythohn--ufnlpro. Purification .15 by column chromatography using 2% ethyl acetate/hexanes gave the pure thiophene in 73% yield as a brownish clear oil.

C. N-4boo3mty--sxzll--35 uistriTluooety) phenyllthiophene.2.suffonamide 4 -bromo.3methyl5isoxazoly) 3 5 bis(trifluoromethyl).

a. 20 phnltipee--ufnmd was prepared in the same manner as in Example 2. Reaction of phenyllthiophene (250 mg, 0.63 mmoi) with 5-amino-4-bromo3methyl.

isoxazole (118 mg, 0.67 mmol) yielded, after column chromatography using 5% MeOH/CHCI 3 115.2 mg N-4boo3mtyl5ioaoy)5 [3,5 bis(trifluoromethl)peyhoh en-.oamdeas a white powder (34% yield). A portion of this sample was further purified by preparative HPLC, m.p. 1400

C.

-171- EXAMPLE 139 fo(4-Bromo-3-methyl-5-isoxazolyl)-5-(5-methyl-2-thienyl)thiophene-2-sul- A. 2 -methylthiophene-5-boronic acid n-BuLi (2.38 M, 16 ml) was slowly added to a solution of 2 -methyl thiophene (3.0 g, 31 mmol) in THF (20 ml) at -78° C. The solution was kept at -78o C for 10 min. then warmed to 00 C for an additional 0.5 hr.

SThe solution was then transferred by steel canula under nitrogen into a 10 vessel containing triisopropylborate (6.3 g, 33 mmol) in ether (15 ml) at 780 C. The resulting milky white solution was stirred at -78° C for min. then at room temperature for 2 hours. The reaction was quenched by the addition of 10% aqueous HCI (5.0 ml) and the solution was extracted into ether. The combined ether extracts were extracted with 15 10 M NaOH (2 x 30 ml), the aqueous extracts were acidified with dilute HCI to pH 2 and extracted into ether (3 x 25 ml). The combined ether extracts were washed once with water (10 ml), once with brine (10 ml) and dried and evaporated to give 3.91 g 2 acid as a light brown solid. This was used in the next step with no 20 further purification.

B. N-[5-(5-methyl-2-thienyl)thiophene-2-sulfonyl]pyrrole N-[5-(5-methyl-2-thienyl)thiophene-2-sulfonyl]pyrrole was prepared in the same manner as described in Example 32C from 2-methylacid and N-(5-bromothiophene-2-sulfonyl)pyrrole.

Purification by column chromatography using 2% ether/hexanes gave the pure sulfonamide in 72% yield as a white solid.

C. 2 -chlorosulfonyl-5-(5-methyl-2-thienyl)thiophene 2-chlorosulfonyl-5-(5-methyl-2-thienyl)thiophene was prepared in the same manner as described in Example 33D from N-[5-(5-methyl-2thienyl)thiophene-2-sulfonyl]pyrrole (570 mg, 1.84 mmol). Purification by -172column chromatography using 2% ethyl acetate/hexanes gave 258 mg of the sulfonyl chloride as a light green solid.

D. N-4boo3mty--sxzll-5(-ehl2tinltipee 2 -sulfonamide

N-(

4 bromo3.methyl.5oxoxaly)5-mthl 2 hiey. hipee 2 -sulfonamide was prepared in the same manner as described in Example 2. Reaction of 2 -chlorosulfonyl5(5..methyl-2thienyl)thiophen (200 mg, S0.72 mmol) with 5-a min o-4-bro mo3methyliso xazo le (127 mg, 0.72 mmol) yielded 273 mg of the crude sulfonamide. After passing through a small plug of silica gel, a portion of the product was further purified by preparative HPLC to give the pure sulfonamide as a white powder, m.p. 161-162O

C.

EXAMPLE 140

N-(

4 -Bromo-3-methyl..5.isoxazolyl) 5-(5-ethyl12-heytioen2suf amide A 2 -ethylthiophene.5boronic acid 2 -ethylthiophene-5-boronic acid was prepared in the same manner as described in Example 140A from 2 -ethylthiophene (2.0 g, 18 mmol).

.20 Evaporation of solvents after workup gave 2.1 6g of 2acid as a white sold which was used on the next step with no further purification.

B. N-[5-(5-ethyl-2thienyl)thophene2sulff II Irol N-5(-ty--hey~hipee2sloylyrl was prepared in the same manner as described in Example 32C from 2 boronic acid (411 mg, 2.64 mmol) and N-{-bromothiophene..2.sulfonyl)pyrrole (700 mg, 2.39 mmol). Purification by column chromatography using 2% ethyl acetate/hexanes gave 6.30 mg of the pure product as a dark brown solid (90% yield).

C. 2 -chlorosulf onyl.5-(ethyl2thienyl)thihen -173- 2 -chlorosulfonyl-5-(5-ethyl-2-thienyl)thiophene was prepared in the same manner as described in Example 33D from N-(pyrrole)-5-(5-ethyl-2thienyl)thiophene.-2-sulfonamide (630mg, 2.16 mmol). Purification by column chromatography using 1% ethyl acetate/hexanes gave 400.3 mg of the pure sulfonyl chloride as a bright yellow solid (57% yield).

D. N-(4o-bromomo-3-methyl-5-isoxazolyl)-5-(5-ethyl-2-thienyl)thiophene-2sulfonamide SN-(4-bromo-3-methyl-5-isoxazolyl)-5-(5-ethyl-2-thienyl)thiophene-2sulfonamide was prepared in the same manner as described in Example 2.

10 Reaction of 2-chlorosulfonyl-5-(5-ethyl-2-thienyl)thiophene (200 mg, 0.68 mmol) with 5-amino-4-bromo-3-methyl isoxazole (121 mg, 0.68 mmol) yielded 174 mg N-(4-bromo-3-methyl-5-isoxazolyl)-5-(5-ethyl-2-thienyl- 0 thiophene-2-sulfonamide (59% yield). After passing through a small plug of silica gel with elution using 10% MeOH/CHCI 3 a small fraction of the 15 product was further purified using preparative HPLC to give the sulfona- ,00. mide as a light tan colored powder, m.p. 1260C.

EXAMPLE 141 N-(4chloro-3-methyl-5-isoxazolyl)-5-(5-ethyl-2-thienyl)thiophene-2-sulfonamide

N-(

4 -chloro-3-methyl-5-isoxazoly)-5-(5-ethyl-2-thienyl)thiophene-2sulfonamide was prepared in the same manner as in Example 2. Reaction of 2-chlorosulfonyl-5-(5-ethyl-2-thienyl)thiophene (Example 141C, 200 mg, 0.68 mmol) with 5-amino-4-chloro-3-methylisoxazole (91 mg, 0.68 mmol) yielded 188 mg of the final product (71% yield). A small portion of the product was further purified by preparative HPLC to give the pure sulfonamide as a tan colored solid.

-174- EXAMPLE 142 fN-(-ro3mid hazlY5i)soxzoyen nzoth bnYthin2h iophee2-sul A. Benzofbjthiophene-2boronic acid Benzo[blthiophene-2boronic acid was prepared in the same manner as described in Example 140A from benzo[blthiophene except that t-BuLi was used as the base in place of n-BuLi resulting in a tan solid in 78% yield.

B. N-[(benzobthien2yl)thiophen 2 lfon IIro N-fS-(benzo[bthien2yl)thiophene 2 sufoyprol wa prpa 99 a.in the same manner as described in Example 32C from benzo[bIthiophene-2-boronic acid (426 mg, 2.39 mmol) and 2 -sulfonyl)pyrrole (700 mg, 2.39 mmol). Purification by column chromatography using 2% ethyl acetate/hexanes gave the pure sulfonamide in 68% yield as a brownish-red solid.

C. 2 -chlorosulfonyl.5-(benzo [blthien-2-yl)thiophene 2-hooufnl5(ezobtin2y~hohn was prepared in 9* the same manner as in Example 33D from N-f 5-(benzofb thien-2-yl)thiophene-2-sulfonyllpyrrole (520 mg, 1.5 mmol). Purification by column chromatography using 2% ethyl acetate/hexanes gave 153 mg (32% yield) of the pure sulfonyl chloride as a white solid.

D.

N-(

4 bromo3.methyl.5isoxazoly5(boblthie--ltipe 2 -sulfonamide N-4boo3mty--sxzll-5(ez~lhe--ltipee 2 -sulfonamide was prepared in the same manner as in Example 2.

Reaction of 2-hooufnl5(ezobtin2y~hohn (150 mg, 0.48 mmol) with 5-amino-4-bromo.3-methyl isoxazole (84 mg, 0.48 mmol) resulted in 97 mg of pure N-4boo3mtyl5ioaoy)5 (bnobtin2y~hohn--ufnmd as a light tan powder, (45 yield, m.p. 1640

C).

-175- EXAMPLE 143 N-(Bromo-3-methyl-5..isoxazolyl)..5-( -pentynyl)thiophene-2sulfonamide A. l-pentynyl)thiophene-2-sufonyl)pyrrole -pentynyl)thiophene.2-sulfonyl)pyrrole was prepared as in Example 1 38A from N-(5-bromothiophene.2.sulfonyl)pyrrole (600 mg, 2.05 mmol) and 1-pentynel (280 mg, 4.1 mmol). Purification by column *0 chromatography using 2% ethyl acetate/hexanes gave 424 mg of the pure sulfonamide as a brown oil (74% yield).

B. 2-chlorosulfonyl-5( 1 -pentynyl)thiophene :10 2-hooufnl5(lpnyy~hohn was prepared in the same 9 9 manner as described in Example 33D from N-[5-(l-pentynyl)thiophene-2 sulfonyllpyrrole (420 mg, 1.5 mmol). Purification by column chromatography using 1 ethyl acetate/hexanes gave 55 mg of the pure thiophene as a brown oil (15 yield).

mide N-(bromo-3-methy1..sisoxazolyi).5.( 1 -pentynyl)thiophene-2suffona- 9~9: mide was prepared in the same manner as described in Example 2.

Reaction of 2 -chlorosuffonyl-5-(l -pentynyl)thiophefl 8 (55 mg. 0.22 mmol) with 5-a min o-4-bromo3methylisoxazol e (43 mg, 0.22 mmol) yielded mg of N-(bromo-3-methyl..sisoxazolyl).5-(l1-pentynyl)thiophene-2-suj.

fonamide (87% yield). A portion of the product was further purified by preparative HPLC to give the pure sulfonamide as a light brown oil.

EXAMPLE 144

N-(

4 -Bromo-3methyl5isoxazolyl)..5( 1 -n t ~hohne2s fnai A. -napthyl)thiophene-2sufonyllpyrrole N-50-napthyl)thiophene-2sulfonylpyrrole was prepared in the same manner as described in Example 32C from 1 -napthaleneboronic acid (353 mg, 2.05 mmol) and N-(5-bromothiophene2sulfonyl)pyrrole (600 mg, 2.05 mmol). Purification by column chromatography using 2% ethyl -176acetate/hexanes gave the pure sulfonamide in 87% yield as a pale yellow clear oil.

B. 2-chlorosulfonyl-5-(l -napthyf)thiophene was prepared as described in Example 33D from -napthyl)thiophene-2-suffonyllpyrrole (604 mg, 1.28 mmol). Purification by column chromatography using 2% ethyl acetate/hexanes gave 376 mg of the pure thiophene in 68% yield.

C. N-(4-bromo-3-methyl-5-isoxazolyl)-5-(1 -napthyl)thiophene-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyi)-5-(1 -napthyl)thiophene-sulfonamide was prepared in the same manner as described in Example 2.

Reaction of 2-chlarosulfonyl-5-(l-napthyl)thiophene (200 mg, 0.65 mmol) with 5-amino-4-bromo-3-methylisoxazole (0.65 mmol), after purification by column chromatography using 1 MeOH/CHC13, gave 65.3 mg of pure N-(4-bromo-3-methyl-5-isoxazolyi)-5-(l-napthyl)thiophene-suffonabeet mide as a brown solid, (22% yield, m.p. 1180 C).

too* EXAMPLE 145 N-(4-Bromo-3-methyl-5-isoxazolyl)-5-(3-nitrophenyl)thiophene-2-suffonamide A. N-(3-methyl-5-isoxazolyl)-5-(3-nitrophenyl)thiophene-2-sulfonamide N-(3-methyl-5-isoxazolyi)-5-(3-nitrophenyl)thiophene-2-suffonamide was prepared in the same manner as described in Example 32C from 3nitrobenzene boronic acid (362 mg, 2.17 mmol) and isoxazo lyl) 5-bromoth iop hene-2-sulf on amid e (700 mg, 2.17 mmol).

Purification by column chromatography using 10% MeOH/CHCI, gave 166 mg of the pure suffonamide (21 yield).

B. N-(4-bromo-3-methyl-5-isoxazolyl)-5-(3-nitrophenyl)thiophene-2-sulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)'-5-(3-nitrophenyl)thiophene-2-sulfonamide was prepared in the same manner as described in Example -177- 136C. Reaction of N-( 3 -methyl-5-isoxazolyl)-5-(3-nitrophenyl)thiophene- 2-sulfonamide (328 g, 0.90 mmol) with N-bromosuccinamide (160 mg, 0.90 mmol) gave the final product. A fraction of this material was further purified by preparative HPLC to give the pure sulfonamide as a brown solid, m.p. 132 0

C.

EXAMPLE 146 N-(4-Bromo-3-methyl-5-isoxazolyl)-5-(4-methyoxycarbonylphenyl)thiophene-2-sulfonamide A. 2 4 -methoxycarbonylphenyl)thiophene 10 2 4 -methoxycarbonylphenyl)thiophene was prepared in the same manner as described in Example 32C from thiophene-2-boronic acid g, 7.81 mmol) and methyl-4-bromobenzoate (1.68 g, 7.81 mmol).

Purification by column chromatography using 2% ethyl acetate/hexanes gave 1.1 g of 2 4 -methoxycarbonylphenyl)thiophene as a white solid (65% yield).

B

2-chlorosulfonyl-5-(4-methoxycarbonylphenylthiophene Chlorosulfonic acid (1.06 g, 9.16 mmol) was slowly added to a solution of 2 4 -methoxycarbonylphenyl)thiophene (500 mg, 2.29 mmol) in CH 2

CI

2 (10 ml) at -780 C. The resulting solution was stirred at -780 C S 20 for 1 hr. by which time the sulfonic acid had completely formed as judged by TLC using 10% ethyl acetate/hexanes. Phosphorous oxychloride (2 ml) was then added at -780 C followed immediately by addition of phosphorous pentachloride (954 mg, 4.58 mmol). The resulting solution was stirred at -780 C for 0.5 hr. and then at room temperature for min. The solution was then carefully poured onto crushed ice (100 g) and extracted into diethyl ether (100 ml). The combined organic layers was washed with brine (1 x 25 ml) and dried over MgSO,. After filtration, removal of solvents left a light.greenish solid which was further purified by column chromatography using 2% ethyl acetate/hexanes to -178give 620 mg of pure 2 -chlorosulfonyl5(4methoxycarbonhnl) thiophene as a pale yellow solid (85% yield).

N-(

4 4 ehoxcroypey thiophene-2-sulfonamide was prepared in the same manner as described in Example 2 from phene (646 mg, 2.04 mmQl) and (361 mg, 2.04 mmol). Purification by column chromatography using 10%/ MeOH/CHCI3 gave 384 mg of 4 -methoxyca rbonylph en yl) th iph ne-2 5 uf o md sabrw i (41 yield).

EXAMPLE 147 15 4 -Brrno.3.methyl..5.isoxazolyl)S( 4 -c r o y h nltl p e e2 sl Lithium hydroxide (13.3 mg, 0. 32 mmol) in methanol (2 ml) was added to a solution of N-( 4 bromo3methyl5-isoxazo 1 yf) 5 4 amethoxycarbonylphenyl)thiphn-flfonmd (Example 1470, 121 20 mg, 0.27 mmol) previously dissolved in methanol (5 ml). The solw-tion stirred at room temperature for a period of 18 hours. The methanol was removed in vacuo and the remaining residue dissolved in water. 4N HCl was added until pH 2.0 was reached, and then the aqueous solution was extracted with ethyl acetate (3 x 25m1). The combined organic layers was washed with water (1 x 10 ml), brine (1 x 1lOmI) and dried over MgSO 4 Evaporation left 50 mg (43% yield) of a pale yellow residue which was further purified by preparative HPLC to give N-(4-bromo.3as a white solid, m.p. 219-228'

C.

-179- EXAMPLE 148 Carbonyldiimidazole (553 mg, 3.41 mmcl) was added to a solution of N-( 4 chloro3mehyl.5isoxazol2crylbhioh.--ufnmd g, 3.1 mmol) in dry DMF (10 inl). The mixture was stirred at room temperature for 1 5 minutes to give mixture NaH (60% dispersion in mineral oil, 521 mg, 13.02 mmol) was added to a solution of 2 1 (1.13 g, 6.2 inmol) in dry DMF (10 ml) at 0 0 C. Th -e mixture was stirred at 0 0 C for 15 minutes to give mixture Mixture was slowly cannulated into mixture (11) at 0 0 1C. The resulting mixture was stirred at 0 0 C for 4 hours. The reaction mixture was poured into 2 N HCI faq., 200 ml) and the resulting precipitate was filtered. The solid was washed 15 with water (2x10 ml) and ethyl ether (2x10 ml) to give N-(4-chloro-3.

2 -{[2-acetyl-4,5-mtyeeix~peylmncr bonyllthiophene.3sulfonamide (730 mg, 49% yield) as a dull yellow powder, m.p. 191-193'

C.

:EXAMPLE 149

N-(

4 chloro..3.methyl..5.isoxazoly) 2 4 ,5dmtoy2mtoyabnl phenyl)aminocarbonyllthiophene.3-sufonainide and N-( 4 .chloro-3-methyl- 5-isoxazolyl)2{45mthx-2 5-dYimethy 2 -ehxcr The title compounds were prepared by the method set forth for N- (4clr--mty -sxaoyS-[2aey-,-(methylenedioxy)phenylj..

aminocarbonyl~thiophene-3-suffonl ide (EXAMPLE 148) except that mehl2aio45-iehxbnot was used instead of 2 '-amino- '(ehlndox~ctpeoe The crude product was purified via HPLC to give N-4clr--ehl5ioaoy)2[45dmtoy2 methoxyca rbo lnylphemnyj)bnl~hope e -sl o mi .a yellow powder (13% yield, in.p. 167-168' C) and N-( 4 -180isoxazolyl) 5-dimethoxy-2,4, S-dimethoxy-2-methoxycarbonyl).

phenyllphenylaminocarbonyllthiophene.3-sulfonamide as a dull yellow solid (1 yield, m.p. 228-230'

C).

EXAMPLE 150

N-(

4 -chloro-3-methyl.5.isoxazoly)2[(2-methyl-1 3 yl)a min oca rbo nyll thi ophene.3-suffo n amide Carbonyldiimidazole (553 mg, 3.4lmmol) was added to a solution of N-4clr--ehl5ioaoy)2croy-hohn--ufnmd g, 3.1 mmol) in dry DMF (10 ml). The mixture was stirred at room 10 temperature for 15 minutes before the sequential addition of m ehyl 1 3,-thadiazle(736 mg., 6.2 mmol) and pyridine (10 ml). The resulting mixture was stirred at room temperature overnight. To work up, the reaction mixture was poured into 1 N HCI (150 ml) and extracted with EtOAc. The organic layer was dried (MgSO4), the solid was filtered 15 and the filtrate was concentrated. The residue was purified via HPLC to :give N-4c lr--ehl5isxz i--(-ehl1 3 yl) aminocarbonyllthiophene.3-sufonamide as a white powder (15% yield, m.p. 192-194'

C).

EXAMPLE 151 20 N(-hoo3mty--s~aoy)-[-abxl45 *mtyendoy phe.l i cr n#tipen-3sloai NaOH (1.5 N, 250 ml) was added to N-( 4 -chloro-3-methyl.s.

isoxazolyl)-2-[(4, S-dimethoxy-2-methoxycarbonylpheny) aminocarbo ltiphee3sloai (EXAMPLE 149); 410 mg). The resulting suspension was stirred at room temperature overnight to give a clear solution. The mixture was acidified using concentrated HCl with cooling.

The resulting precipitate was filtered, washed with water (3x50 ml) and dried on a lyophilyzer to give N-( 4 -chloro.-3-methyl-5-.isoxazolyl) 2 -{f 2 carboxyl-4, S-(methylenedioxy)phenyl] aminocarbonyl~thiophene.3.sulfonamide as a yellow powder (87% yield, m.p. 192-1 950 C).

-181- EXAMPLE 152

N-(

3 4-dimethy5isoxazyl)-}{ 2 acey 1 4 5 -mtye- 4 -dimethyl-5-isoxazolyl)- 2 {l2-acetyl-4, S-(methylened ioxy) phenyll amin ocarbon ltiohnh 3 -slo md was prepared by the method set forth for N-( 4 -chloro-3..methyl. -isoxazolyl) 2 -{[2-acetyl.

4 ,S-(methylenedioxy)phenyfinaincabhohen--ufnm (EXAMPLE 148) except that N-( 3 ,4-dimethyl.5.isoxazolyl)- 2 Scarboxylthiophene3sulfonaide was used instead of N-(4-chloro-3 meth yl 5 iso xazol yl 2carblth ihe- sufoamd.

N-(

3 ,4-dimethyf- 5 -isoxazolyi)2{2acetyl-4,S(methylenedioxy)phenyaminobnlJti phene-3-sulfonamide was obtained as a yellow powder yield, m.p. 228-2310

C).

EXAMPLE 153

N-(

4 -chloro3..methyl5-isoxazoly,).

2 4 -mtoy2mtypey) .5*aminoarbonylthiophele3-sufonai was prepared by the method set fot o N-( 4 -chloro3-methyl..

5 ool~ 0 y) 2 2 -mty-, 3 ,4-thii-iazoI- 5-yl) amino carbonyllthiophene-3..suffonamide (XML 5) xetta 4mtoy2mtyaiiewas used instead of 2 -amino5methyl1 3 4 thiadiazole and that pyridine was not used. The title compound- was obtained via HPLC purification as a dull yellow powder (66% yield, m.p.

58-620

C).

EXAMPLE 154 N-(4-chlo ro-3-methyl.

5 -isoxazolyl)-2-[(2-.cyano 0 phenyl) amino ca rbo nylthiophene3-sulf onamid ewas prepared by the method set forth for N-4clr--ehy--sxz l 2 -{[2-acetyr..4,5- -182- (methyen edioxy) phen yll amionblltipee3su md (EXAMPLE 148) except that 2 -amino-4, S-dimethoxybenzonitrile was used in stead of 2 1 -amino4'5'(methylenedio)aetpno. N-(4chloro.3- 2 2 -c.co 4 ,5-dimethoxyphenyl) aminocarbonyl]..

thiophene-3..sulfonamide was obtained via HPLC purification as a light brown powder (36% yield, m.p. 53-56'

C).

EXAMPLE 155

N-(

4 chloro 3thmethoxoy ,-dmtoxpeyamn c *bonyllthiophene3sulfonamide

~N(

4 chloro3methy--ioxlyl)- 2 [2,-ithxpe )ano carbonyllthiphene3sulfonaide was prepared by the method set forth a. minocafbo n ylIthiophen e3suf nami (EXAMPLE 153), except that 2 4 dimethoxyaniline was used in stead of 4-ehx--etyaiie

N-

4 -chloro-3-m ethyl5 isoxazl) 2 2 -i e h x p ny am oc r n l] thiophene-3sufonamide was obtained via recrystalization (CH3CN/H2o) as yellow crystals 016% yield, m.p. 162-1640

C).

EXAMPLE 156 :20 boylhohn--ufnmd N-4clr--mty -sxzoy)2' mthl6prdlamjnocarbonyl~h iphee--s lfo amdewas prepared by the method set forth for N-( 4 chloro3.methyl5soxzoy) 2 2 -acetyl-4, S-(methyvenedioxy)phenyllaminocarbonylthiophene.

3 sulfonfl 8 (EXAMPLE 148), except that 2 amino- 5-pico line was used in stead of 2'aio4,'-mtyee dioxy)acetophenone,

N-(

4 2 3 tyl6 pyridyl) amin ocarbo nylthioph e- lfon amid e was obtained, via HPLC purification of the crude reaction mixture, as a bright yellow powder (17% yield, m.p. 158-1600

C).

WO 96131492 PC~fUS96/04759 -183- EXAMPLE 157 N-(4-fomo-3-~ l~ 2 (methyl5ioa ylph-[4-ethylhl)acetlthiophele-3acetyllthiophene.3-sulfonamide was prepared in the same manner as described in Example 938 by reacting 4 -methylbenzylmagnesium chloride with N-( 4 bromo3.methyl...5..iszoxazolyl)2[thx(mtyai carbonyl)Ithiophene.3-sulfonamide in THF (see Example 93A) at -78' C to, romtemperature, rsligin 78% yield. 146150C.

10 EXAMPLE 158

N-(

4 -bromo-3..methyl5isoxazolyi) 2 4 -mehIl)(inmlItopesulfonamide

N-[

2 (4methtasylrn)-sulfon)3sulfl *-ety-rnssyy)--ufny~yrl was prepared inthe 15 same manner as described in example 1278 using diethy113-[(Npyrllufnltin2flmtyhshnt and 4 -methylbenzaldehyde 30% yield.

B. 2(-ehltassyrltipee3sfoy chloride 2 4 -methyl-trans-styryl)thiophene-.3.

ufflfychriewsppad in the same manner as described in Example 64E from

N-[

2 -(4-methyl- 5. yyl--slonlpyrl by basic hydrolysis (using ethanol and sodium hydroxide) to the corresponding sodium sulfonate followed by conversion to the corresponding sulfonyl chloride in 13% yield.

C.

N-(

4 bromo3.methyl.5.isoxazol)- 2( 4 mthyl-ta styryI)thiophene.3.sulfonamide phene-3-sulfonamide was prepared in the same manner as described in Example 2 by reaction of 2 -(4-methyl-trans.styryl)thiophene-3-sulfonyI chloride with 5-mn--rm--mtyioaoe The crude product -184.

was purified by HPLC followed by crystallization resulting in a 34%Z yield, m.p. 101-1051C.

EXAM PLE 159 sulfonamide A. N-2[4mty~hntylhipee3sfoylyrl

N-{

2 f(4methyl)phenethylthiophene3ufflpyrop was prepared as described in Example 1 28A by the catalytic hydrogenation of

N-[

2 -(4-methyl.trans..styryl)-.3..

5 ufyl)pyrl in 80% yield.

B 2 4 -methyl) phen ethyl] thioph ene- 3 -sulfonylchloride 2 -m e hyl phe ethylth oph ne- 3 -sulfonylchlorid e w as prepared, as described in Example 64E, using N{-(-ehlpeehltipee 3 -sulfonyl~pyrrole by basic hydrolysis (KOH/ethanol) of the sulfonamide to *this potassium salt followed by conversion of salt to the corresponding 15 sulfonyl chloride in 51 yield.

C. N-4boo3mty--sxzli)2[4mty~hntylho S Sphene-3-sulfonamide 555N-( 4 bromo3.methylisoxazoy 2 f( 4 ehy)phntylh phene-3-sulfonamide was prepared, as described in Example 2, usinrg 2- 20 (4mt yl Seehy~hipen-3sloy ord and 5-amino-4-bromo- U 3 -methylisoxazole in 52% yield.

EXAMPLE 160 A.

N-

2 [(4methylphenoxy)methyuhe- 3 sffoylyrl N-2[4mtypeoymtyltipee3sfoy~yrl was prepared, as described in Example 126A, by reacting N-f2--bromomethyl)thiophene-3sulfonylpyrrole with 4 -methylphenol, in 81 yield.

-185- B. 2 4 -methylphenoxy~mthl.ipee3sff Iclrd 2 -[(4methylphenoxy)methylthiophee- 3 lfonl chloride was prepared, as described in Example 64E, using

N-{

2 -[(4-methylphenoxymethyllthiophene.3-sulfonyllpyrrole by basic hydrolysis (NaOH/EtOH) followed by conversion to the corresponding sulfonyl chloride, in 46% yield.

*ope.e-methyl]-id

N-(

4 bromo3.methyl5isoxazol) 2 4 -mtypeoymtyl thiophene-3-sulfonamide was prepared, as described in Example 2, by rectng 3 -chlorosulfonyl2.(4thmpehypmehltipe a. mi:--r o -ehyioaoe eutn in a 64% yield, m.p. 128- 1300 C.

4* EXAMPLE 161 sulfonamide thiophene-3..sulfonamide a. 20 thiophene-3..sulfonamide was prepared, as described in Example 93A, by reacting N-34dmty--sxzll--croy~hohn-3 sulfonamide with N--iehlyrxlmn hydrochloride using triethylanine as base and carbonyldiimidazole, resulting in a 23% yield.

Crude product purified by column chromatography using 1:1 hexanes/EtOAc as the eluent.

sulfonamide sulfonamide was prepared, as described in. Example 93B, by reacting

N-

(34dmty--sxzll--Nmtoymtyaioabnljho -186phene-3-sulfonamjde with 4 -tolylmagnesium chloride, resulting in a yield, m.p. 9.5-100o C.

EXAM PLE 162

N-(

3 4 dimethyl-5isoxazolyi)2.[34-(methylenedioxy)phenylcty,,] thiophene-3-sulfonamide

N-(

3 4 -dimethyl-5isoxazoly).2[34(methylenedioxy)phenyIct]thiophene-3-sulfonamide was prepared, as described in Example 93B, by reacting

N-(

3 4 -dimethyl-5-isoxazolyl)2[N-methoxy(methylaminocarbonyl)thiophene-3sulfonamide with 3 4 -methylenedioxy)p[,eny,.

magnesium chloride, resulting in a 65% yield, m.p. 95-1000

C.

EXAMPLE 163

N-(

4 -chloro-3-metliyl..5.isoxazolyi)..2.{[2.cyano- 4 ph enyl a min ocarb onyllthi ophe ne.3-sulfo namide 9e oo3-ehy--soaoyi--{2can-,-(methylene- 15 dioxy)phenyllaminocarbonyl~thiophene-3.

5 ufonamide was prepared as described in EXAMPLE 148, except that 2 dioxy)benzonitrile was used instead of 2 I,...acetophenone. It was obtained via HPLC purification as a yellowish solid in -40% yield, m.p. 167-168*

C.

20 EXAMPLE 164

N-(

4 -chloro-3methy[-5.isoxazolyi)2(3-meth 4 ,xycarbony1246trimethyl)phenylaminocarbonyl.3-thiophenesuffonamide A. Methyl 3 -a min o-2,4,6-trimethylbe nzoate Methyl 3 -amino-2,4,6-trimethylbenzoate was synthesized in the same manner as 3 4 -methylenedioxy)-6-methylaniline (see Example 177, below).

B. N-4clr--ehl5ioazll--3mtoyabnl 2 4 ,6-tri methyl) phenylamin rocarb-tonyptihenenlfonaide i--3mtoyabnl246trimethyl) ph enylamin ocarbon yl.3-th iophenesu If o namid was synthesized in the same manner as for Example 94 except that DMF was used instead -187of THF and the reaction was heated at 801C for 5 hours. The crude product was purified via preparative HPLC to give N-(4-chloro-3-methyl-5 isoxazolyl) -2-(3metho xycarbony-,4,-trimethyl) phenyla min oc arb on yl 3 thiophenesulfonamide as an off-white powder (48 mg, 1 yield, m.p. 66- 70 0

C).

EXAMPLE 165 N 4 -chloro3rethyl 5isoxoazolyI)2(246trimth) h lact-

N-(

4 -chloro-3-methyl-5..isoxazolyl)..2.( 2 4 ,S-trimethyl)phenylacetyl.

10 3 -thiophenesulfonamide was synthesized in the same manner as for Example 102 using 2 4 6 -trimethylbenzyl chloride and N-(4-chloro-3thiophenesulfonamide. The crude product was purified by flash column chromatography (eluent 1 methanol in CH 2

CI

2 to give N-(4-chloro-3thiophenesulfonamide as a solid (31 yield, m.p. 42-46 0

C).

EXAMPLE 166

N-(

4 ch o ro 3metyth-io a lyl) 2-azolyl)2(24Gth)h nla i :carbonyl- 3 -thiophenesulfonamide cabnl3tipeeufnmd was synthesized in the same manner as Example 94. The crude product was purified via preparative HPLC to give N-( 4 -chloro-3-methyl-.s.isoxazolyf).

2 2 4 6 trmt l e lmncronl -ho eeuf md as a yellowishbrownish powder (410 mg, 30% yield, m.p. 45-48'C).

-188- EXAMPLE 167

N-(

3 4 dimethyl5..isoxzoylyl)..2..(24dml~pethylphtl 3 hohesulfonamide 4 -dimethyl-5isoxazoly)2(2,4dimethyl)phenylacetyl- 3 thiophenesulfonamide was synthesized by the same method as described for Example 102 using 2 ,4-dimethylbenzyl chloride and N-(3,4-dimethyl- 4* 44phenesulfonamide. The crude product was purified by flash column 4**4chromatography (eluent 1 methanol in CH 2

CI

2 and further by prepara-- 10 tive HPLC to give N-( 3 4 -dimethy5isoxazolyl)2(2,4-di meh4*hnyaetl3-hopeeslonmd as a semi-solid (34% yield).

44EXAMPLE 168

N-(

4 chloro3.methyl..5..isaolyl)-2 yl)2(24dmthl)hlactl 4* 4 thiophenesulfonamide thiophenesulfonamide was synthesized in the same manner as for Example 102 using 2 4 -dimethylbenzyl chloride and N-(4-chloro-3-methyl.

2 N-ehy-N-ehoy amino carbo n yI 3-th iophe ne- :sulfonamide. The crude product was purified by flash column 4 44 20 chromatography (eluent 1 methanol in CH 2

CI

2 to give N-(4chforo-3as a solid (52% yield, m.p. 48-54 0

C).

EXAMPLE 169 thiophenesulfonamide

N-(

4 -bromo-3-methyl-5-..isoxazolyl)-2-.( 2 4 -dimethyl)phenylacetyl.3 thiophenesulfonamide was synthesized in the same manner as for Example 102 using 2 ,4-dimethylbenzyl chloride and N-(4-bromo-3-methyl.

5-isoxazoly).2(N..methyl.N '-methoxy)aminocarbonyl.3 thiophenesulfonamide. The crude product was purified by flash column -189chromatography (eluent 1 methanol in CH 2

CI

2 and further by preparative HPLC to give N-( 4 -bromo-3methyl5isoxazolyl)2.( 2 4 dimethyl)phenylacetyl-3thiophenesulfolamide as a solid (28% yield, m.p.

58-63 0C).

EXAMPLE 170

N-(

4 -chloro-3-methyl.5-.isoxazolyl).2( 3 ,S-dimethyl)phenylacetl3thiophenesulfonamide N-(4chlro--mehyl5-ioxaoly)-2(3,-dimethyl)phenylacetyl.3 thlophenesulfonamide was synthesized in the same manner as for 10 Example 102 using 3 ,5-dimethylbenzyl bromide and N-( 4 -chloro-3-methyl.

9thiophenesulfonamide. The crude product was purified by flash column chromatography (eluent 2% methanol in CH 2

CI

2 to give N-(4-chloro-3- *methyl-5-isoxazolyl)2(3,5-dimethyl)phnlct hohneufnmd as a solid (57% yield, m.p. 45-501C).

.:..EXAMPLE 171

N-(

4 -chloro-3-methyl..s-jsoxazolyl).2.( 2 ,5-dimethyl)phenylacetl3.

thiophenesulfonamide

N-(

4 -chloro 3methyl.5.isoxazoly).

2 2 ,5d m t y~ h nl ctl3 20 thiophenesulfonamide was synthesized in the same manner as for Example 102 using 2 ,5-dimethylbenzyl chloride and N-( 4 -chloro-3-methyl.

2 Nmty-'-ehx)amnocroy-3 thiophenesulfonamide. The crude product was purified by flash column chromatography (eluent 2% methanol in CH 2

CI

2 to give N-(4-chloro.3 methyl- 5-isoxazolyl) -2-(2,-iehl pheyaey-3t o ee l nai as a solid (33% yield, m.p. 72-761C).

-190- EXAMPLE 172

N-(

4 -chloro- 3 -methyl-5-isoxazolyl)-2-[3,4-(methylenedioxy)-6-( 2 acetoxyethyl)]phenylaminocarbonyl-3-thiophenesulfonamide A. 2 3 4 -methylenedioxy)phenyl- 1-ethanol To a solution of 2 3 4 -methylenedioxy)phenylacetic acid (5 g, 25.75 mmol) in anhydrous THF (20 ml) at 0°C was added BH 3 THF mi, 1.0 M in THF). The mixture was stirred at room temperature for 1h.

To work up, THF was evaporated on a rotavap. The residue was treated with water (100 ml) Acidified and extracted with ether (2 x 100ml).

10 Removal of the solvent under reduced pressure gave 2-(3,4methylenedioxy)phenyl- -ethanol as an oil (4.7g, 98% yield).

B. 1 -acetoxy-2-[(3,4-methylenedioxy)phenyl ethane To a stirred solution of 2 3 4 -methylenedioxy)phenyl-1-ethanol (1.68 g, 10 mmol) in dry pyridine was added acetic anhydride and the resultant reaction mixture was stirred at 80 0 C for 1h. The reaction mixture was poured into ice-water and was extracted with ether (2 x ml). The combined ether extract was washed with water (2 x 50 ml), Hcl (2 x 50 ml) and then with 5% NaHCO 3 (2 x 50 ml). The organic layer was dried over magnesium sulfate and the solvent was removed 20 under reduced pressure to give 1-acetoxy-2-[(3,4methylenedioxy)phenyl]ethane as a solid (1.7 g, 81% yield).

C. 1 -acetoxy-2-[(3,4-methylenedioxy)-6-nitrophenyl]ethane To a stirred solution of 1-acetoxy-2-[(3,4-methylenedioxy)phenyl]ethane (1.7 g, 8.09 mmol) in acetic acid (10 ml) was added, dropwise, concentrated

HNO

3 (4.5 ml). This was stirred at room temperature for 30 min. The reaction mixture was poured into water (100 ml). The precipeated solid was filtered, washed with water and dried under high vacuum to get 1-acetoxy-2-[(3,4-methylenedioxy)-6nitrophenyl]ethane (1.8 g, 88% yield).

-191- D. 1l-acetoxy2[(34meth lened.x)- 6 -aiohnIehn The solution of 1 -acetoxy-2-[(3, 4 -methylenedioxy).6nitrophenlethan (0.8 g, 3.13 mmol) in ethyl acetate (25 ml) was subjected to catalytic hydrogenation using 10% palladium on carbon (100 mg) at 50 psi for min. The catalyst was filtered and the solvent was removed under reduced pressure to give 1-ctx--(,-mtyeeix)6 aminophenyllethane as a solid (0.69 g, 98% yield).

E. N-( 4 2 f[ 3 ,-mtyedix 2 -acetoxyethyl)phenylaminbnl- 3 thiophenesulfonamide was synthesized in the same manner as Example 87. The crude product was purified by preparative HPLC to give -4clr3mehl5 Nsxzli--34(mtyeeix)6(-etxyhll..etylmno cabnl3tipeeufnmd as a dull yellow powder (12% yield, m.p. 7 8-821C).

EXAMPLE 173 a.aN-( 4 chloro.3ehy-ethyalysl)- 2 -[,-mtyeeix)6( hydroxyeth yl) I phenyla min crocl 3 hioheeufn To a stirred solution of N-( 4 -chloro-3methyl-. -isoxazolyl).24[3,4- (mtye dox)6 2aeoyt l hnl i cronl3t o phenesulfonamide (35 mg, 0.066 mmol) in methanol was added NaOH powder (40 mg) and stirred at room temperature for 30 min.

HPLC

analysis showed complete consumption of starting material. The reaction mixture was diluted with water and acidified to pH 2 This was extracted with ethyl acetate (2 x 25 ml). The combined organic layer was dried over magnesium sulfate and the solvent removed under reduced pressure to give

N-(

4 chloro3methyr..s..soxazoly) 2 3 4 -192- (methylenedioxy).6(2hydroxthyilheyaiorbn-3topnsulfonamide as a solid (84% yield, m.p. 4 7-520C).

EXAMPLE 174

N-(

44 .chloro.3.methyl5isoxaoly)-2[3-(ehlndoy6a cetoxyeth oxy) Ip hen y lnoami nbl- 3 -hoensl am A. Methyl 2 3 ,4-(methylenedioxy)phenoxyI ctt and 2-[3,4- (meth ylenedioxy)p hen oxyla cetic acid The mixture of sesamol (13.8 g, 100 mmol, methyl bromoacetate g,100mmol) and potassium carbonate in acetone (200 m) wa refluxed for 24h with stirring. Acetone was removed under reduced pressure. The residue was dissolved in water 20 0ml) and was extracted with ether (2 x 100' ml). the combined ognclayer was~~ dried oe Ott: 0.magnesium sulfate and the solvent removed under reduced pressure to g i v m e h y l 2 4 m e h y l n e d o x y p e n o y l c e t t e a s a n o il 1 2 g 5 7 yield). The aqueous phase was neutralized to pH 2 3 with concentrated HCl and the precipitated solid was filtered to get 2-[3,4hnxlaei acid as a solid (6 g, 31 yield).

B 2 3 4 m e t y l e e d i o y p e n o x i e t h a n o l 2-34mtyeeix~hnx- -ethanol was synthesized iZ3 the saemnnra for Example i172(A) using 2 3 ,4-(methylenedioxy)phenoxylacetic acid and

BH

3 *THF complex. The reaction was carried out for 12 hours at room temperature (98% yield).

C. 1 -acetoxy-2.f34(methyledi)phnxleh l-Acetoxy234(methyldx)phnxle n was synthesized in the same manner as for Example 1 72(B) by acetylation of 2-34mtyeeix~hnx- -ethanol using acetic anhydride and pyridine (92% yield).

D. 1 aeoy2[,-mtyeedoy--irpeoyehn 1 aeoy2[,-mtyeedoy--irpeoyehn was synthesized in the same manner as for Example 1 72(C) by nitration of 1- -193acetoxy 2 34methylendiy~heoxyhleta The reaction was carried out between OOC and 5 0 C for 30 min (78 yield).

E. 1 -acetoxy-2[34(methylenedi)- 6 iohnoyeh 1 aeoy2[,-Mtyeedoy--mnpeoyehn was synthesized in the same manner as for Example 1 72(D) by reduction of I1a c e t x y 3 4 e t h l e n d i o x -6 nit o p h n o x l e t a n e 1 0 0 y ie ld 2 -a cetoxyethoxy) p henyl inoa b l-~ 3 thlophenesulfonamide p urfied b p hep rive H Pm eto yg ie l or e hy -s xa o y 2 syntphensized nathe s annul elas Examper 871 usid acety1[- N(methlo- dioy)6-ioai)-2-helh and ylne( 4 choro..-..mthyl hydrxyeth xy I 'h en yl mi ocrb nyl3-tiodmhenTeuf crudeprdtwa purified by rparatieHCth5ioo ive N 4 -(met.3.mhyleiox)-6 2 4. tox~phnlmioaroyl3tiophenyslfamarbnwas 3 oyjheyaincronl3thiophenesufonamide asadllylowpwer(1 yield, m.p. 17 .*:18100O).

-194- EXAMPLE 176

N-(

4 2 4 thoxcron26dimethyl)phenylaminocarbony 3 thohesufnmd A. -4-chlro3-methyV[5.,soxazolyN(ehoxehxmty To a mixture of N- 4 -chloro-3-methyl S-isoxazolyl)-2carboxylthiophene-3..sulfonamide (3.23 g, 100 mmol) and diisopropylethyl amine 9(3 ml) in ethyl acetate (20 ml) was added methoxyethoxymethyl chi-r)ide and the resultant reaction mixture was stirred at room temperature or 1 2 10 hours. This was diluted with ethyl acetate (100 ml) and washed with 1 N HCI (2 x 50 ml). The organic layer was dried over magnesium sulfate 9* *9and the solvent removed under reduced pressure to give N-(4-chloro-3 methylS.isoxazoy)N(mhxethxmty)2(croehx) thiophene-3..sulfonamide as light brown oil.

The crude

N-(

4 chloro3.methylisxaoJf)-N(ehxyto met yl)-2 -car ome h o y) hi ph n e 3 ul o nami e asdissolved in methanol (50 ml) and potassium hydroxide (5 g) and water (5 ml) were added. The reaction mixture was stirred at room temperature for 1 2 *9 9.hours and extracted with ethyl acetate (2 x 50 ml). The aqueous-phasewas neutralized to pH 2 3 and was extracted with ethyl acetate (2 x ml). The combined organic layer was dried over magnesim sulfate and solvent removed to give

N-(

4 -chloro..3.methyl S-isoxazolyl).N..* (mtoytoyehl--abxlhohn--ufnmd as light brown solid (3.5 g, 85% yield).

B. 4 -Carbomethoxy26dimethyanli.

To a warm solution of 3 ,5-dimethyl benzoic acid (5 g, 33.33 mmol) in acetic acid (30 ml) was added fuming nitric acid (30 ml), dropwise.

After completion of the addition the reaction mixture was warmed with a heat gun. This was stirred for an additional 2 hours during which period -195a solid was precipitated. The reaction mixture was diluted with water (200 ml) and filtered. The solid was dried under reduced pressure.

To the above solid was added 20 ml of oxalyl chloride and a catalytic amount of DMF (2 drops). This was stirred at room temperature for 3 hours during which period a clear solution was formed. Excess oxalyl chloride was removed under reduced pressure to give yellow solid.

To the yellow solid was added dry methanol (100 ml) and the mixture stirred at room temperature for 1 hour. Excess methanol was removed under reduced pressure and the residue dissolved in ether (200 10 ml). This was washed with water (100 ml) followed by saturated NaHCO3 solution (100 ml). The oganic layer was dried over magnesium sulfate and the solvent removed giving 4-carbomethoxy-2,6-dimethylnitrobenzene as a yellow solid (5.8 g, 83% yield).

4-Carbomethoxy-2,6-dimethylnitrobenzene (2 g, 9.5 mmol) was 15 dissolved in in ethyl acetate (20 ml) and subjected catalytic hydrogenation using 10% palladium on carbon (300 mg) at 55 psi for mmin. The catalyst was filtered and solvent removed to give 4-carbomethoxy-2,6-dimethylaniline as a solid (1.7 g, 100% yield).

C. N-(4-chloro-3-methyl-5-isoxazolyl)- 2 -(4-methoxycarbonyl- 2,6-dimethyl)phenylaminocarbonyl-3-thiophenesulfonamide N-(4-chloro-3-methyl-5-isoxazolyl)-N-(methoxyethoxymethyl)-2carboxylthiophene-3-sulfonamide (3.5 g, 8.5 mmol) (from step A) was dissolved in oxalyl chloride (5 mi) and a drop of DMF was added. This was stirred at room temperature for 6 hours. Excess oxalyl chloride was removed under reduced pressure and the mixture dried under high vacuum.

To a solution of 4-Carbomethoxy-2,6-dimethylaniline (0.9 g) (from step B) and triethyl amine (2 ml) in methylene chloride (20 mi) at O°C was added the acid chloride in 10 ml of methylene chloride (2.4 g, 4.98 mmol) prepared in the above step. The reaction mixture was permitted to -196warm to room temperture, was diluted with methylene chloride (50 ml) and was washed with 1 N HCI followed by saturated NaHCO, solution The organic layer was dried over magnesium sulfate and the solvent removed giving the crude product. This was purified by column chromatography using 4:6 ethyl acetate and hexane as a eluent to give N-(4-chloro-3-methyl-5-isoxazolyl)-N-(methoxyethoxymethyl)2-(4methoxycarbonyl-2,6-dimethyl)phenylaminocarbonyl-3thiophenesulfonamide an oil (0.6 g, 20% yield).

N-(4-chloro-3-methyl-5-isoxazolyl)-N-(methoxyethoxymethyl) 2(4- S 10 methoxycarbonyl-2,6-dimethyl)phenylaminocarbonyl3thiophenesulfonamide (0.6g) was dissolved in a mixture of methanol (8 ml) and concentrated HCI (1.5 ml) and the reultant reaction mixture was refluxed under stirring for 8 hours. Excess methanol was removed under reduced pressure and the residue dissolved in ethyl acetate (50 ml) This was 15 washed with saturated sodium chloride solution. The organic layer was dried over magnesium sulfate and the solvent removed giving N-(4-chloro- 2EXAMPLE 177 N-(4-chloro-3-methyl-5-isoxazolyl).2-[3,4-(methylenedioxy)-6methyl]phenylaminocarbonyi-3-thiophenesufonamide A. (3,4-methylenedioxy)-6-methylaniline To a solution of (3,4-methylenedioxy)toluene (5 ml) in acetic acid (20 ml) cooled with a cold water bath was added, dropwise, nitric acid 5 ml). The mixture was stirred for 45 min. To work up, water (100 ml) was added and the resulting yellow precipitate was filtered and washed with water until the aqueous filtrate was colorless. The yellow solid was dissolved in EtOAc (250 ml) and dried (MgS0 4 and the solid was fltered off. The filtrate was subjected to catalytic hydrogenation -197- Pd/C, 1 atm) for 12 hours. The reaction mixture was then filtered off the catalyst and the filtrate was concentrated on a rotavap to give 3 4 -methylenedioxy)-.6-methylaniline as a brownish grey solid (5.49g, 87% yield).

B. N-( 4 choro3methyisoxa5..ioxazoyI)23(mehyetdix)-6 methyllphenylaminocarbonyl3thiophenesulfnaide N-(4-chloro-3-methyl 5-isoxazolyf)-2-[3, 4 -(methylenedioxy).6 methyl] eyaioaroy--hoheeufnmd was synthesized in :9::the same manner as Example 94 using 3 4 -methylenedioxy)-6 10 methylaniline. The crude product was purified by preparative HPLC to 9 9 nlmioaboy--tipensfonmd as a yellow solid (45% yield, m.p. 60-621C).

9 EXAMPLE 178 N (-hIor--ehl -sxz y) -(,-iehoy6amno 9N-(Wc h loro methyl.55- .isoxaolyl) 3,4d 4 dmeh th- 6 -amn cabnl~hnlaioaroy-3tiphnslfnmd was synthesized in the same manner as for Example 94 using N-(4chloro-3-methL5.

20 isoxazolyl)2-{[2-carboxyl-4,5-(methylenedioxy)phenylamin 0 o 5 a..

bonyllthiophene.3-sulfonamide (Example 151) and ammonium hydroxide.

The crude product was purified by preparative HPLC to give N-(4-chloro- 3-ehl5ioaoy)2(,4dmtoy6aioabn phenylaminocarbony[.3-thiophenesulfonamide as yellow powder (66% yield, m.p. 189-1921C).

-198- EXAMPLE 179

N-(

4 -chloro-3-methyl..5.isoxazolyl).2[3,4-(methylenedix)-6-methyII phnlctl3tipeeufnmd A. 3 ,A-methylenedioxy)-6methylbenzy chloride To a 1: 1 mixture of ethyl ether (100 ml) and conc. HCI (100 ml) at 0OC was added 3 4 -methylenedioxy)toluene (10 ml). Formaldehyde ml, 37% in water) was then added dropwise. The reaction was stirred at 0C for 2 hours and at room temperature for an additional 10 hours. The reaction mixture was then diluted with ethyl ether (100 ml) and the two layers were separated. The organic layer was dried (MgSO 4 the solid was filtered and the filtrate was concentrated. The residue was then heated with hexane (200 ml) and the insolubles were filtered off the hot solution. The filtrate was concentrated to give a mixture of (3,4methylenedioxy)-6methylbenzyl chloride (9.4 g, 63% yield) and bis[(3,4mehlndoy--etylhnlehn (3.6 g) as a white solid. This 4e mixture was carried on to the next step without further purification.

B. N-( 4 chloro3methyl5isoxazoly)2[34(methylndix)-6 a .20 mehlpeyaey--tipeeufnmd was synthesized in the same manner as for Example 102 using 3 4 -methylenedioxy)-6methylbenzyI chloride instead of 3 4 -methylenedioxy)benzyl chloride. The crude product was purified by preparative HPLC to give N-( 4 isoxazolyl)-2-[3, 4 -(methylenedioxy)6methyllphenylacetyj 3 thiophenesulfonamide as a yellow powder (71 yield, m.p. 42-45 IC) -199- EXAMPLE 180

N-(

4 .cloro.3methyl-5...zly-23,.mtledix)6 Sulfonamide A.

N-(

3 4methylenedioxy)benlehaeufnmd To a solution of piperonylamine (6.07 g, 38.95 mmol) and trlethylamine (5.37 g, 53.12 mmol) in dichloromethane (100 ml) at 000 S Swas added methanesulfonyl chloride (4.14 g, 35.41 mmol). The reaction was stirred at 0 0 C for 1 hour. The Mixture was then diluted with *5 10 dichloromethane (100 ml) and washed with 1 N HCl (2 x 100 Ml). The organic layer was dried (MgSO 4 the solid was filtered and the filtrate Swas concentrated to give

N-(

3 ,4methylenedioxy)bezl-mta sulfonamide as a grey solid (8.4 g, 92% yield).

B N m t yl n d o y mi o b ny..ha e uf n m d C. N-([3hl4 o-3methyln ioxaz) 6 2[ ,-mehlndix)6 N- 4cl oro4 3m ethyl-diox zly-2[,4 ehye eioy -i sufnmd was synthesized in the same manner as forpl 94 h cr d proaniline (E a p e cr s ai e 1ro 77).ti n at rt iv -4 clr eh -5io a oy)2[ -m tye eix)6methanesulfonyl amnoe. oyl3thiophenesulfonamidaeanoff wh t soiS1 y ed.m p 4 5 0 -200- EXAMPLE 181

N-(

4 chloro3methy..5-is ioxazo-yI)2[4(mt thlediox)- 6 -ca methyllphenylaminocarbonyl.3.thiophenesulfonamide A. 3 4 -(methylenedioxy)-6-aminolphenylacetonitrile 3 4 -(methylenedioxy) -6-a min olphenylaceton itrile was synthesized in the same manner as for 3 4 -methylenedioxy).6-methylaniuine (Example 177).

B. N-( 4 chloro3methyl.5.isoxazoyl) [34..(myethedi)- 6 cyan omethyll phenyla min ocarb onyl 3thio phenesuf mde

N-(

4 chloro3methyl5isoxazolyi234(methylendi)- 6 cyanomethyllphenylaminocarbonyl3thiopheneslfaide was synthesized in the same manner as in Example 94. The crude product was recrystalized from acetonitrile/water to give N-(4-chloro-3-methyl-5.

*isoxazo lyl)- 2 4 3 4(meth ylenedioxy)6cyan omethll h enl ioabo 3 -thiophenesulfonamide as a red-brownish powder (15 yield, m. p. 190- 1 93 0

C).

EXAMPLE 182

N-(

4 chloro3methyl.5.isoxazolyl 2..[34u.(mleedhyd) 6 3 hydroxypropyl)phenylaminbca banyl3thiopheesufnelfde A. 3 3 4 -methylenedioxy) phenyl-. 1 -pro panol- To a solution of 3 3 4 -methylenedioxy)phenylpropanoic acid (5 g, 25.75 mmol) in anhydrous THF (20 ml) at 0 0 C was added BH 3 *THF (51 ml, 1.0 M in THF, 51.5 mmol). The mixture was refluxed for 1 hour.

Then the THF was evaporated on a rotavap. The residue was treated with methanol (20 ml) and the solution was concentrated. This process was repeated 6 times to give 3 3 4 -methylenedioxy)phenyl1.-propanoI as an oil (4.7g, 100 yield).

-201- B. 3 [1 3 4 -(methylenedioxy)6aminophenl -propanol 3 3 4 (methylenedioxy) 6aminolphenyl rpnl was synthesized in the same manner as for 3 4 -methylenedioxy).6methylaniline (Example 177).

C.

N-(

4 chloro-methyl.sisoxazolyl) 2 2[ 3 4(ehylndox) (3uhyfoamjepyl~phyaminocarbonyl-3thiophene- N- 4 -chloro- 3-methyl..5.isoxazo lyl) 2 3 4 (methylenedioxy) 6- 3 *S.hydroxypropyl)phenylaminocarbol 3 thohesufnmdwa synthesized in the same manner as Example 94. The crude product was purified by preparative HPLC to give N-( 4 2 4 -(methylenedioxy)6(3hydroxyppl)Ipheyaiorbnl thiophenesulfonamide asa dull yellow powder (18% yield, m.p. 66- 69 0

C).

EXAMPLE 183

N-(

4 chloro.3methyl5isoxal) 2 3 4 -mtyedix) cyanophenylacetl3thiophelesufmde A. Methyl 3 4 -methylenedioxy)phenylacetate *Methyl (,-ehlndoypeyatt was prepared as 20 described by known methods (see, Rachele (1963) Journal of Oranic Chemistry 28:2898).

B. Methyl G-bromo(3,4methylenedioxy)phenylatat To a solution of methyl (34mtyeeix~hnlctt (5 g, 25.8 mmol) in acetic acid (1 5 ml) was added, dropwise, bromine until a red-brown color persisted. After stirring at RT for 30 minutes, the reaction mixture was partitioned between water (200 ml) and ether (200 ml). The organic layer was washed with water (3 x 200 ml), dried (MgSO 4 the solid was filtered off and the filtrate was concentrated to giv mthy 6bromo (34-ehe xy) ph enyl acetate as an oil (5.9 g, 84% yield).

-202- C. Methyl 3 ,4-methylenedioxy)-6-cyanophenylacetate Methyl (3,4-methylenedioxy)-6-cyanophenylacetate was prepared as described by L. Friedman and H. Shechter in the Journal of Organic Chemistry 26:2522 (1961).

D. t-Butyl 3 ,4-methylenedioxy)-6-cyanophenylacetate To a solution of methyl 3 ,4-methylenedioxy)-6-cyanophenylacetate (5 g, 18.32 mmol) in methanol (100 ml) was added 1 N NaOH (50 ml).

S. The reaction was stirred at room temperature for 1.5 hours. Methanol was then stripped off on a rotavap. The aqueous residue was acidified 10 with conc. HCI to pH- 1 and extracted with ethyl acetate. The organic layer was dried (MgSO,), the solid was filtered and the filtrate was concentrated to give a solid. The solid was treated with thionyl chloride (50 ml) and the mixture was refluxed for 10 min before the volatiles were stripped off on a rotavap. The residue was dissolved in dichloromethane 15 (15 ml) and the solution was added dropwise to a solution of 2-methyl-2propanol (6.8 g, 91.6 mmol) and triethylamine (9.3 g, 91.6 mmol) in dichloromethane (100 ml) at 0 0 C. The mixture was stirred at OC for 1 hour and at room temperature for 2 hours. The mixture was then washed with water (3x150 ml). The organic layer was dried (MgSO 4 the solid was filtered and the filtrate was concentrated to give t-butyl (3,4methylenedioxy)-6-cyanophenylacetate as a solid (335 mg, 7% yield).

E.

N-(

4 -chloro-3-methyl-5-isoxazolyl)-2-[ 3 ,4-(methylenedioxy)-6cyanolphenylacetyl-3-thiophenesulfonamide To a solution of N-(4-chloro-3-methyl-5-isoxazolyl)-2-carboxyl-3thiophenesulfonamide (2.78 g, 8.63 mmol) in anhydrous DMF (30 ml) was added carbonyldiimidazole (1.40 g, 8.63 mmol). The mixture was stirred at room temperature for 20 min to give mixture

I.

To a solution of t-butyl 3 ,4-methylenedioxy)-6-cyanophenylacetate g, 5.75 mmol) in anhydrous DMF (15 ml) was added NaH (1.2 g, 60% dispersion in mineral oil, 29.9 mmol) at 0°C. The mixture was -203stirred at room temperature for 30 min to give mixture 11. Mixture

I

was syringed to mixture 11 at 0 0 C and the resulting mixture was stirred at 0 0 C for 1 hour and at RT for 10 hours. The crude mixture was poured to a 2:2:1 mixture of acetonitrile/water/conc. HCI and the resulting mixture was heated at 40 0 C for 12 hours. Acetonitrile was then removed on a rotavap and the aqueous residue was partitioned between ethyl acetate (200 ml) and 1 N HCI (150 The organic layer was washed with 1 N HCI (3 x 10ml) addried (MgSO 4 the solid was filtered off and the filtrate was concentrated. The residue was purified by preparative

HPLC

10 to give

N-(

4 chloro.3methyl5isoxazol) 2 f 3 4 (mtyedix)6 cynlhnlctl3thohnsloaiea a lgtdull yellow powder (450 mg, 17% yield, m.p. 105-1080C).

EXAMPLE 184 dimet yla min ocarb o nmylm ehyla iocabnhl.tiohee sulfonamide e .SA. N,N-dimethyl (,-ehlndox~hnlctmd N, N-dimethyl 3 4 -methylenedioxy)phenyl 8 cetamde was synthesized in the same manner as described in Example 94.

20 B. N ,N -dim ethyl 3 4 m t y e e i x 6 a i o h n l c a id N,N-dimethyl 3 4 -methylenedioxy)ohnyact..e a synthesized in the same manner as 3 4 -methylenedioxy)-6-mtylanilin (see Example 177).

N-(

4 -chloro..3.methyl.5.isoxazoll)..

2 3 4 -mtyedix ~dimethylaminocarbofylmethylhnlaiorbnl sulfonamide was synthesized in the same manner as Example 94. The crude product was recrystalized from acetonitrile/water to give N-(4chloro3.methyl...isoxazol l)...2f34(mhyeeioyd)-6dmtyaio -204carbonylmethylphenylaminocarbonyl-3-thihnamide as a greyish powder 400 mg, 19% yield, m.p. 190-193 0

C).

EXAMPLE 185

N-(

4 -chloro-3-methyl-5-isoxazolyl)-2-3,4-(methylenedioxy)-6methyllphenylhydroxyimino-3-thiophenesulfonamide To N-( 4 -chloro-3-methyl-5-isoxazolyl)-2-3,4-(methylenedioxy)-6methyllphenylacetyl-3-thiophenesulfonamid (100 mg) was added

NH

2 OH*HCI (300 mg) and water (15 ml). After stirring for 5 min, NaOH pellet (300 mg) and methanol (2 ml) were added. The warm mixture was heated at 80 0 C for 20 min and was cooled to OOC. It was then poured into a dilute HCI solution 30 ml). The resulting white precipitate was filtered to give N-(4-chloro-3-methyl-5-isoxazoly,)-2-[3,4- (methylenedioxy) meth yllphlhydroxyimino-3-thiophenesulfonamide as a white solid 72 mg, 70% yield, m.p. 154-1560C).

EXAMPLE 186

N-(

4 -chloro-3-methyl-5-isoxazolyl)-2-{l-acetoxy-2- cis-[3,4- (methylenedioxy)-6-methyllphe n lvinyl-3-thiophenesuffonamide To a solution of N-( 4 -chloro-3-methyl-5.isoxazol).-2-[ 3 4 (methylenedioxy)-6-methyllphenylacetyl-3-thiophenesufonamide50 mg, 0.11 mmol) in anhydrous DMF (1 ml) was added NaH (11 mg, dispersion in mineral oil, 0.275 mmol). After stirring at room temperature for 5 min, acetic anhydride (16.8 mg, 0.165 mmol) was added. After stirring at room temperature for an additional 10 min., the mixture was poured into dilute HCI solution and the resulting precipitate was filtered to give N-( 4 -chloro-3-mmethyl5-isoxazolyl)-2-{l -acetoxy-2- cis-[3,4- (methylenedioxy)-6methyllphenyl)vinyl-3hnamide as a yellowish Powder (40 mg, 73%, m.p. 55-58 0

C).

-205- EXAMPLE 187

N-(

4 chloro3.methyl5isoxazll) 2 2 3 -trimethoxy-6cyano)phenylaminocarbonyl thio hensffnmd A. 2 -amino-3,4,5..trimethoxybenzonitrile 2 -amino-3,4,5-trimethoxybenzonitrile was synthesized in the same mane a (,4meYn nedixy) 6 eth ylaniine (see Example 1 77), and the crude product was recrystalized from methanol/water to give a yellow powder (13 yield).

N-(

4 chloro.3.methyl5isxazl) 2 2 3 -trimethoxy-6 ~cyano)phenylaminocarbony3thihesufnmd i--ety-5ioxzly) 2 3 -trimethoxy-6.

cyano)phenyaminocarby-topl3 nesu~ffnmd a synthesized in the same manner as Example 148. The crude product was purified via *preparative HPLC to give N-4clro3mty-5ioao.i-12, 3-tnimethoxy6.cyano)phenym micabonl- 3 -tipeeufnm as a yellow powder (180 mg, 10% yield, m.p. 88-901C).

EXAMPLE 188 mehylpenyactl3-hiphneufo md was synthesized in the same manner as Example 179. The crude product was purified via preparative HPLC to give N-34dmty--sxzll)2[,-mtyeeix)6 methylIpheny eyllhtohnsonajf. as a yellow powder (41 7 mg, 14% yield, m.p. 4 5-501C).

-206- EXAMPLE 189 N-(4-ch ho o romehy. 3io5zoy..methyf 3 yl neio y -6 mehy eiphenylacetyl hihsfonmd mehylpheylaety-3-hiohensulonaidewas synthesized in the same manner as Example 179. The crude product was purified via preparative HPLC to give N-(4-chloro-5.methy..3.isoa i)-2[,-mtyeeix) 6- Smethylphenylacety3thohesufnmd as a yellowish powoer (330 mg, 16% yield, m.p. 46-50 0

C).

EXAMPLE 190 Other compounds that have been prepared by the above methods or routine modifications thereof, include, but are not limited to: N- 4-chlro-3-metyi S-isoxazolyl)2A(4methoxyphenoxy) carLbuIyllthio.

phene-3-suffonamide,

N-(

4 bromo.3methy

J..

5 .jsjji-2-(4 15methylphenoxy)carbonlyjJhipee3sfoaie

N-(

4 bromo3methyl- 1 5 -s x z 4 -m t l he x m t y thi phe uf nam de -4 br m e hy -s x zo y) 2 4 e h l h no y m ty*t i p e e 3 n-2ufoamide,

N-(

4 3 4 -ehl 20 phnty~hohne2sloaie

N-(

4 [(-ehlhny 4 toly)thiophene2sufmde N-4boo3mty--isoxazofyl) m eth l-tran -st ryl -5 -4 -t lyl~hio hen -2 uff nam d ,N (4-chloro 3 mdN( 4 chloro3methyisxlyl--(-ety-isxzoy)aio carbonylthiophene3uff 0 .ide

N-(

4 bromo.3methyl.5o szoj)- 2 3 -hy dro xy l 6pyrid azi i car on l t i ene fo m d e -4 -207braomo- 3methy[ 5isoxazoy) 3{[34(th yeeix)phnoy ehl thiophene-2-sulfonamide,

N-(

4 -bromo-3methyl5isoxazolyl) 2 4 methyl) (cinnamyl)] thiophene-3-sulfonamide, N-(4-bromo-3-methyl.5 isoxazolyl)- 3 4 -(methylenedox)henehylltipee2sloai

N-

4 -bromo 3methy 5isoxazoly) 3 3 4 (ehylnd )-tas styryllthiophene-2sulfonamide,

N-

4 bromo3.methyl5isoxazoly) 2 4 methyl)phenethyllthiophene3sulfnmde

N-(

3 2 4 -tolyacetylphenyl)thiophe3sulfnmde N-(3,4-dimethyl-5 isoxazo yl) 2 .[34-(methylenedi) hnlctI t ohne-3 fo mde

F~

4 ~horo xamehyl..5isoxaolmyl[ 2 hd oy)--[-ety-4 blthiophene 3sulfonamid aa d -4boothers, inl i aolIL3-2 dimehylinna yl~hio hene2-s lfon mi ha e be nthos ae d etfothei Foemanr ampleN- 4 brmo-3 3..methy-i isoxazll)- 3 2 (methylenedioxy)cnnamtyyllthioph- 2 -slfoa e N-4-bromo3 15methyl..Sisoxazoly).3[2(hydroxy(mthl 4 doypentylh phn--ufnmd and N-4boo3mty--methyleio)-2(.4 cineamyllthiophene..2.sufonide (seebExample 159 thyl-isoxazlI) 3 2 -tahy ro4H..i-3{-rpylran..2hyloen 14S(eihyleeioycnnml phnx*ety .opee2sufnmd hav been prepared in the m 2 samanner a s 4 bro-mo..3...methazyli)-3xazolyl) 2 f( 4 oxy -208methyllthiophene-2sulfonamide and N-( 4 3 3 4(methyenedioxy)phenoxymelthyiophene 2 sufoaide N-(4bromo-3-methyI5isoxazoly)2[2-methyl-4,5- (methylenedioxy)phenethyllthiophene3sulfnmde has been prepared in the same manner as N-( 4 -bromo-3-methyl..5..soxazoly)..-[( 3 4 methylenedioxy)phenelthyliophee3suf Compounds, such asN-( 4 bromo3.methyl-isoxazl 3(2toil)hohe-2sloai, 4 rromo.metl thsxayl5.i)-3-azolyl)3(3..tl)hiohe2 sua (lbomide N(4rmethyl methyazo -iso3(tlyl)-3h(h mehxyhn a ohee2sulfonamide,

N-(

4 bromo-3.me-metjoxhylJ).

3 isxzl--2methoxyphenylthopenhslonamid -ulomie N-( 4 -bromo-3met. yl 5-oxa ol) lyl)2-3-methyphenyl) th ohe 2 fnaide -4 N bro o-3 met yl- -is xaz lyl -3-4-i o-p opy phe yf~ Nio he4-br m -3 methflnSmideoxaz-bolyl)3m(ethylioaoy)(-uyphenyl)thh a .hene15 brom o naethyl.5..isoxa olyl3-4proplhf5ioaoy)--2 4 .hipe-2slon soamide, N-(4-bromo-3-methylis-ioxalyl)-(3-i 4.butyl2m yphenyl) .thiophene-2-sulfonamide a d N -r m eh,- -s x z li- -4i o sae ane as N-4brm--bmo3-5isethoyl)-35.jsoxaz eyl( 2 ~dimeyphenylthiophene2sufonamid se Exa-bme 125). th-op-ene mo-sulfonamid -iandaNoly-bromo2-methyl.5..uisoxazlI) 3 4 dioxy)phenethyllthiophene-3sulfonamide has been prepared in the same -209m anner as N (4-rom o 3-m ethyl- is oxz.i -M t y e e dioxy)phenethylthiophe 3 -ufnmd (Example 1 28). N-(4-bromo.3- methyl-- isoxazoly)-.2 [2-meth 1-4,5-(methylenedioxy) cifnamyI]thjophene- 3 -SUffonamide has been prepared in the same manner as N-(4boo methyl-5-isoxazolyI) cna y) thop en.. .ulomoaid (Example 158). 2[4mty)cnayl hohn-sufaid N- 4 -bromo..3.methy,-soxzl.-- [3 4 -(methylenedioxy) phenoxYlmethyhlthiophee- 3 -sfoai N-(4bromo3methl-5 isoazS. [2,,6trimethylphenoxy)mthltipee3sloaie 1 hn-4 oyl me lth y5iohen 3 ,5(m ethylenedioxy).

2 propyl phe oxy m et yl~ hio h en -3 ulf nam dehave been prepared in the sam e m a ne as N (4 -brom o .3 m eth y 5is. x z l l e h l h n x 3 lfoaie(xml16) Any corresponding

N-'

4 methYI-3..isoxazolyl)

N-(

3 3 t .i s o x a z o l y l 4 h l e h l 5 i o a o y 4 5 d m t y s x z l l derivative of any of these compounds or any compound dislosed herein S may also be prepared and used as described herein.

EXAMPLE 191 Other compounds that can be prepared by the above methods or routine modifications thereof, include, but are not limited to: N chloro..3.m ethyl.. 5 Qso 8 1 y 2 -2 3 4 ti e h x 6 m th l en aminocarbonyl)thiopen--s fnmie N-4clr--ehl--sxzl 2 3 4 -trimethoxys6 acetylp hen yla minoc bo ~hip ne -unaie N c h lo ro .3 m eth y is o xz.l -2 3 4 tr m t o y 6 m t o y N -(4chloro 3 m eth y 5is o a o y) 2 4-r m t xy 6 -210-

N-(

4 -chloro-3-methyI.5.isoxazoy)2(2,3,4-trimethoxy-6methanesulfonylphenylaminocarbonyl)thiophene 3 sufaide N-(4-chloro-3-methyI-5.isoxazoy) 2 2 3 ,4-trimethoxy-6- (cyanomethylphen ylaminocarbo n y lltohioe- n3ifd e

N-(

4 -chloro-3-methy..5..isoxazolyi).2[ 2 3 ,4-trimethoxy-6-(2hydroxyethyl) phenylaminocarbonyllthiophene-3-sulfo'namide N- 4 choro3m ethy- 5isoxazoazo2yI)2f34(mthlendiox)- 2 -ehx methylphenylaminocarbonyllthiophene3sulf oide

N-(

4 choro3 mehy y;.5...solyl) oly[,4-mehl34(tdix)-2mtoy 10 acetylphenylaminocarbonylthiophene3sufaide

N-(

4 -chloro-3-methy..5.]soxazoly)- 2 3 4 -(methylenedioxy)..2.methoxy-6 **methoxycarbonylphenylaminocarbony ulthihne3-ufohmide

N-(

4 -chloro-3-methyl-5-isoxazoly).2[3, 4 -(methylenedioxy)2methoxy-6 carboxylphenylaminocarbonylthiophene3sufnmde **.methanesulfonyphenylioaminycarbophn,]thifohe ide 4 choro-me hyJ.5Isxaoyazo2-[,2.[34..(thdix)-2mtoy cyanophenylaminocarbonylthiophene3sufaide

N(

4 choro3methy5i.5..soaoly yl)2[34(mmtyetdi)- 2 cyanomlpehylphnyamiontoarbny--lfthophn id 4 choro3methy5isoxazioyzo)- I)2.4-(e mehyenedioxy)ct1 2 -h dmethylphenylaminocarbonyllthiophene3sufonid 4 choro3methy..5..isoxzolyI).2..[34..(myetdix)methoxycarbonyl2methylphenylaminocarybhopylhne 3 -sfoaie -211-

N-(

4 choro3.methy..5-ioazixazo3,4-[ 3 4 (hylndoy--abx,2 methylphenyaminocarbonyllthnpe-3esuslfaide

N-(

4 -chloro-3-methyl..5.isoxazoy) 2 3 4 -(methylenedioxy) -6-methoxy-2methylphenylaminocarbonyllthiophenef ide

N-(

4 choro3methy5isoxazoy)2[3 4 (thylneioy)6 metha nesu fonyk-2methylphenylamin robfylth iop n--uf mde

N-(

4 choro3methy.5isoxazoi u 2 [3, 4 -(hylndoy-6cao meth ylp hen ylaminocarbo nylthiophen e- 3 -sufo amde too. N- 4 cho ro 3methy5isoxazoy 2 3 4 (hylndix)to 10 (cyanomethyl)2methylphenylaminocbythihe-3sfoai, 0. too

N-(

4 choro3methy5isoxazoy) 2 3 4 (hylndoyh ydroxyethyl) 2meth heylhenyloaminocbfh i hene -u mde

N-(

4 choro3methy5isoxaz yI2[3, 4 -(hylndoy-2cao *.*meth ylp hen yla min oca rbonyllthophne 3 -sulf o mde *to.*

N-(

4 -choro3.methy.5isoxazo yI 2 3 (ehylndoy--ctl cetanophenylaminocarbonyllthiophe-sufaid 0 to

N-(

4 -choro3 .methy5isoxazol) 2 3 4 (mtye dix)6 eho 3 -suffoaie

N(

4 choro.3methy5-isoxazoi)- 2 3 croyl24 acetylphenylaminocarbonyllthiophen 3 ulf aie 4 -choro3methyI.5-.isoxazoi)2(-yf) 2 3 2 4 -imthytrimethylphenylammcabnylcarbopnl-thoplfaide

N-(

4 choro3methy5isoxazoy).

2 3 -mhans onl246trimethylphenylaminocarbonyllthiophlufnmde -2 12-

N-(

4 chro3methy5isoxazlI)- 2 3 -caoehl246trimethylphenylaminocarbolthihe-3sloai,

N-(

4 choro3methyI5isoxazoly).

2 -f 3 2 hdoyty)246trimethylphenylaminocarbonyfltiohnh 3 -sfoaie

N-(

4 chloro3methy5isoxazly) 2 3 f(abxlety)246 trimethylphenyfaminocarbonyuthihe-3sfoai,

N-(

4 choro3.methy5isoxazliI)- 2 4 -cao26dimethylphenyaminocarbonyf~ihnh 3 -sfoaie

N-(

4 choro3methy5isozy)- 2 -(4-abx 6dimethylphenylaminocarbonthihe-3sloai, 4 choro3methy5isozly)- 2 -[4-2hdoyty)2 *adimethylphenylaminocarbonythhe-3sfoai,

N-(

4 choro3methy5isoxly) 2 4 (abxlmty26dimethylphenylaminocarbonyitheh..

3 -sfoaie

N-(

4 -choro-3.methy5isoxzli)-2f(-ehnsufnl2 dimethyl ph enyainoca rbo nyl th ohen- foamd, aN-( 4 choro3methyI5isoxazoy)2-(,,4t etoy6- *dmeth ylphen yamy)tiorheny3-uffhih de r-3mty--ioaoy)--234trmtoy6-

N-(

4 choro.3methy5isoxIy) 2 .(2,,-rmtoy dmethycarhenylaminolcarbol)thihe-3sloai,

N(

4 chloro3methy5isoxazly)..

2 2 3 4 -trimethoxy.6carboxylp he nyacetyl)th io phe 3 suffoai -213-

N-(

4 -chloro-3-methy...S.isoao 4 -trimethoxy.6methanesulfonylphenylatf)thioh. uloaie N-(4choro3methy.

5 -ioxzoy.

1 [2 4 -trimethoxy.6 (cyanomethyl)phenyacetthohe-3sfoai,

N-(

4 -choro3-methYl-5-isoxazolyl).

2 2 4 -trimethoxy-6-( 2 hydroxyethyl)phenyactyIlhioh. uloai N (4 c h lo ro 3 m e th y l.

5 -i o a o y 2 [3 4 e h l n d ox.2 m t o y 6 N -ch or -3 eth l -is xaz ly -2 [3 4 eth y en ed io x y m eth o y 6 methylphen aceylcty i phn e- 3 so 6-de N (4-ch loro..3 m ethyl.

5 iso a o y) 2 m t y e e i x 2 m t o y N -c h o r o 3 m t h y -5 s o x z o l l) 3 4 m e t h y l e n e d i o x y -2 m t o y 6 10 N-4clr--ehl5ioaoy)2[,-mtyeei 2 -methoxy..6.

(aet~phenylacetyllthioh.e 3 -sloai dmethoylhncrboyltphenlij lonmie

N(

4 choro--memethy5isoaz.

5 i 34siehleedoy) mecaroxabnlmtylpheny laceyIttlh(thYioene-3-suY).2methxy6 -214-

N-(

4 choro3me hy5isoxazoyl)o2- I)2f3(thylndioy-cabxl2 methylphenylacetyllthiophene-3sulfonamide

N-(

4 choro3methyI.5-isoxaoyl I)2f 3 3 4 -(mhylndoy-6mtoy methylphenylacetyllthiophene-3-sufonamide

N-(

4 -chloro-3-.methyl.5isoxazoy) 2 f[ 3 4 -(methylenedioxy).6methanesulfony2.methylphenyactllthihe-3sloai,

N-(

4 -chloro-3-methy.. -isoxazofyl)2[34(methyenedioxy) 6 cyao 2methylphenylacetylthiophene.3sulfaide

N-(

4 chloro3methyI5.isoxazoy).

2 f 3 4 -(hyedix)6 10 (cyanomethyl).2..methylphenylactllthihe-3sloai, 4 -c h Ioro- 3-.meth yl.-isxaoll -f34 m t diox)-6 2 N-(4chloo-3methl-5isoxazolyl)-23,4-(methyenedioxy) 62 ao h ldroxyethytylmetiypphen-3-setylnahihe, 4 -chloro3methy5isoxazoy 2 3 4 (hyedix)6mto- *.*methylphenylaceylthiophene3sulfoide

N(

4 chloro3methy5isoxazoy) 2 3 can( ,4 cyanohyphenylacetylthiophene3sulf amde

N(

4 -chloro3methy5isoxazo) 2 3 croy( 24 rmethyph en ylacetylthio0phen e- 3 -sulfonamide, 4 chloro3methyl5isoxazoyi) 2 3 -hdoymty-24 20 aetylphenylacetylthiophene3sulfoaide

N-(

4 -choro3methyl5isoxazoly).

2 3 2 4 6ufon-24 trimeth ylphen ylacetyl) thio ph ene3..sf o ide -2 NJ(4 chloro .3 m eth l 5is. x z l l -3 y n m t y) 2 4 6 trimethylphenylacelthohe-3ufoai, N-(4-chloro-.3..methyl-5-isoxazolyl)..243.(2.hydroyty)4,6trimethylphenylaceylthohe-3sloai, N-(4-chloro..3.meth y--isoxazolyl)23-(cbxlmeh )2 6trimethylphenylacetyllthihen--ufnmd, N-(4-chloro..3-methyl- .isoaoy)2-4cao6dimethylphenylacetl)thohe-3sloai, -chloro3m eth yl5 -i o a oy) 2 -a b x l2 **.dimethylphenylacetyl)thihe-3sfoai, N- 4 -chloro..3.methy 5ioaoy)2[-2hdoyty)26- N-4-choro--met Yl-5-isoxazoly )2 4 .hy roxt yl (dimethylphenylacetlthihe-3sfoai, N(4-chloro..3-methyl -isoxazolyl)-2 [4-(2hroxymethyl) -2,6dimethylphenylaceyulthipee3sfoai n dimethyl)phenylacetlthiophene 3 sfoai

N(

4 ~chlrEXAMPLEt 192 containeshylheeoylctngl shasthinlfrl and proesloa N- 4 y f,- 2 4 -x c lo e y o y ar ny th -2 16phene-3-sulfonamide, 2 -[3,4(methyenedioxy)phenytIhihe-3 sulfonamide,

N-(

4 -chloro..3-methyl- 5 -isoxazolyl)..{2r[3, 4 -(methylenedioxy)phenyll acety~thiophesufn amid e oxime,

N-(

4 isox8zoly)2.phenybenzobjthihen sulfonamide,

N-(

4 -chloro-3-methyJ- 65-Sisoxazoyi)2.[(4tolaiyIcambnI 1 -methyindoe.3-sulfonamide

N-

4 -chloro-3methyJ5isoxazoyl) 2 4 -ehxpnoycrnlti- Phene-3-suffonamide,

N-(

4 -bromo-.3methy- 5 -isoxa i)l[34(ehbenylbezofltiohen-3sufonmieN-( 4 mie N-4boo3mty--sxzli--4mtyrns-(4-cl~h oo_ 2 4 ehoxpey~ctltipee3sloaie N-(4 ch or me m ethy5isoxa oj)2-l h d o y l (m t le di x .benzyllel)zhopbh n enlf amdN-(bromo..3..methl -ioaoy) ZOI4 l)eth mthyl4tlhiphen e--uloamdhI-4-r mo3 m e th N- 54- soao-3 m t y-5iso2[(4th ih 4 m ty-rn -trl--4 mo ylie 2 s l o am d ,N -h o o- e h l 5 i o az l i -f~ N- N-h or- 3-broml-5-soo.. ety2 9 (i eh lam no -3 4 mehye phenetysuhfoamideN-(-broonmie, met-hlorio 3 mehyl) pheethydoy-[3thiophyedxpheny 1 f

N-(

4 -bromop3.ethy-su -217benyljenzfblhiohen-3-ulfnamde;N-( 4 -bromo..3..methyI isoxazoly)3styrythiophen 2 slfoaie

N-(

4 isoxazoIyi)-2-st...tyrylhnp 3 loai

N-(

4 isoxazolYl) 2 (benzoyfa min o)thihe- sufnai; N-(4-bromo-3 m ethy-5-isoxazo 2[(Phl~ eIthyai no ab nlti p e e3 s fo md;N-( 4 -bromo3methyl- 5 -isxzj)- 5 -(hnlhofrn2slo am nocar b rom~thophenethysl ona iI) N (h-r m ehy- -s x z l rom2-ufoname; -hyIom-3mehysjisxao) 5 *1 0 s t r l h o h n -ufo n a m id e N h o o 3 m t y s x z l i 3 mie;N-4choN-( brm o.hy 3...methzoyl)2[34(mtyendoxf)nyl *yezfjtip en--ufoa ie N dimethyffuran..hy u- fon 20 -ufnaie N

N(

4 4bromoro--methysoxzoy5)2-[o4(mehyl thio phene yi3. 5.su tf o xybie z f j h o h n uf n m d ;N h o o 3 meth l-5-sox zoly)-2-3 N4(etbromeo3... e yl5-iso I 1) 2 aminocarzobnlthiophee3eslfoaie

N(

4 ro3mothyethylsiso 25 lsSsyyluan2ufonamide;NN(-hoo3mty-(-roo...-2-3,4methylenedo 1

J)

phnxybno .pee3-ufnaie

N-(

4 -chloro.3..methyl5 isiis o(elyl)2 en,-(etylenzyuIS (d hxl5dmtya ioezfjh ph ne 4 lfo cioro3.met-h yIo--ety-5ioxjjy)--(3 methndoxypeno labtophen ethlmnbezIfhipee3-uf -2 18amide; N-( 4 choro3methyI.5.isoxazoyi)2.[ 3 4 -(hyndix benzylcarbonyllN-methylindole-3slfaide

N-(

4 isoxazoy)2.[34(methyenxyeoxycphboylnoe3sfnai;

N-(

4 -chloro3.methy;.5-isoxazoyi)-2.[ 3 4 (hylneio phenoxycarbonyII-N-methyindoe3sulfonamide

N-(

4 2 3 4 -(methylenedioxy)phenoxycarbonyi de 3 ufonmd; chloro 3 methy I5isoaoyazo2-I)2[3(thldioybny]Nmtl aindole-3-sulfonamide;

N-(

4 -chloro-3methy5isoxazoy) 2 f[ 3 4 (methylenecioxy)benzyindoe3sufoamde

N-(

4 isoxazolyI)-2..[3,4..(methyI enedioxy)benzyoxycarbony7NNdiehy .amino) benzo blthiophene3sufonamide

N-(

4 zolyl) 2 [3,4 (methyen edioxy) benulj7(NN-d iehl i )bnof.h a. *phene-3-sulfonamide;

N-(

4 -chloro-3-methy-5-isoxazo~lyl)..

2 3 4 (mehyeneioy)benzoyJJ-7(N, NdimethyI) amino z bnziopb~thih 3 15 fonamide; -4clr--eh-5ioaoy)7Ndmtyamino) bnzo( 4 lhiohene3.metyJ..5.ioxao y7.(Nhor--edi--io amnobenziofbhee- eneson aid

N-(

4 -chloro-3methyisoxzo5iox Y 7 (methoxy carbfbthonyIeesufnzoamtih N-(4-crm-3-e hlioo- N-*-bo-3methI-5-isoxazoiyIy..).34methylhd.)bhipen--sl fonamde; -(4-bomo--Net 4 l-chso roly33methyph.5isoxazo (methe-ufnzoai;Nhenerm3slfo Nxazbomoi)32methy5iox fonmidthpee; N-sbro onamie;yI..5isomao-y,3mehl-5ioaoy Pheanethylhy~in m tiohee siop n--uon amidee NN(-bom 43mehyrooazoety) (trsoasly4)-metyan3mthinmyoh 2 ifpee2sfo

N-(

4 -boo3ety brIoo-el)-isotansl)3(tylcn-2-thylinml~hhhne2sl -219fonamide;

N-(

4 -bromo.3.methyl5isoxazliI)- 3 f1(4-ehlhnx methylithiophene.2-sulfonamide

N-(

4 bromo3methyJ5-isoxazoy)-3- 3 4 -(methylenedioxy)phenethylthihe 2 -sloaie N-(4-ch10r-3methyl 5 isoxazo yl)...2{4(34(deth)phenlaey)tipee3sf namide;

N-(

4 choro3methyl5isoxazlif) 2 3 5 -dimethoxyphenyl)acetyllthiophene-3suffonamide

N-(

4 -choro3.methyl5isoxazoly)-2 S-trimeth oxyphenyl) acetyllthioh- 3 -sl mde N-(4-chloro-3 methyJ-5-isoxazoy2, 34.(thndioybnysloylhohns u lfo n a m id e 4 c l r e h l 5 i o a o y 2 3 m t y n d o x 1 0 b n z yl uff ny 4 hi h n eo r lo am h .i e o x4 l).

2 3 e h l 5 i o a bzo ylsui-[3 n-yuthln oen e nzul ufnltip ee -ufdna i ;N (4clr--ehl5ioaoy)2* 0 0 Ni( 4 thlioo..3,4-(ethyI en zolyi) 2 -f[34ethy(methylenediox lneioy~heylehy~tibIee 3clro3amiethyl..5sloxomy{l-5ioaoy)2f-(ehxlmn) dioxY)Pehyenylethoylthenopehylftipee3sloi

N-(

4 -chloro...mthti zoleyl-- smetolai)2.f(c eyle di)thlndix~heyl 3 -slftoamide N-(schfoi; -(4thlioxazetl-5isxaoy 2 *croy) [34 mt le doy bez0 viyl thohn-methfoxnyamino)2

N-I

4 -(mhlor en-edioyl--sphen i--[-4-mtyleethyIxthph n-(4-oo 5-methzy-2isoxaz,yl)..

2 {l(croY)234.etyenedioxy)pezl-124oxdao--y~hio ethyl-thiophene.3.loai 3-eh**ioxzll-- [2(ehne( 4 phenll amin c rboymthyl penei3-suffenzyImi;n-(4th 20isoxaol-Chlro.3..mty..isl.

2 4 -(methylenedioxy)-6phenyl]-ylamnoar bl 2 4-xaaophn 5..lylfo tioe; l n

N-(

4 -chloro-3mmethylsxaoll)2 S[,-isoxazoyle'ned3 4 3 4 -(methlnoxyrbnyllhe1,2, 4 -oaizlr-~ t -220byaoflvllthiohenedixlf {lamnoaboY12-ohne3 sulonaide N(4-hloo--methytS..isoxazolyi) 2 4 ,-(methylenedioxy)- (meylthnhes3fonyI) 4 namie; -(4oro-3-meth5ioxazolyi)- 2 2 (mthnsuf Sulfonamide;laincabnyltiohne3- Soam e N(4-choro3mthyl5isoaoy)212croy--eh ph0 h en y am in o carb o n y l th io h e e 3 s l o a i e -4 c l r e h l phn--u*oaie N-4cloo3mthl5iNxzi-2ch(oroyathI .0 i eto x a z oY )2 ho x m t y 4 e h l h n y l m n c b o n yS P h n h3 si o n e 3 s l o n a iemN hod- e hye ;s x a o y phehyflmnhcanyyl) hiop n-3sbfnaie N-4clr-4mchyro 3 3 -s lfo am de; N -4 N (4o-3 mc h l o xazoth i 5 -im t o y ch'or -3met hY[5..isoxa ou,) 2 (f -2 cy n ,5 d m th x p en l a in c S Sb o fyl l th io p e e 3 lfo a i e 4 c l r 3 m t y i s x z y i 2 d. meL(oxydimhoxyatNhoomethYI5peyaiocroytioxazonyl) 4thycheiooy.3ethI5carbthohene3-shionamie; N-(4-choflam3d Phenboylla 5meocarb nyujtih)2p e ya eytip e e3sfo a i e 00 N(4lchooro3..m ethI.S dimethoxy.2h,-( etylneixy -2 mehdy -221carbonylphenyllacetylthiophe 3 lfoaie

N-(

4 -chloro..3.methylisoxazolyi)-2.{2%cyano[4,S-(methyenedioxy).phenyacetylthohen-- Sulfonamide;

N-(

4 2 2 (methylenedoxy)phenyacetlthohe-3sfoai; N-(4-chloro-3 m ethyl-5-isoxazoy 2 2 4 dm h ox h n la i o a b n lt i p e 3 sulfonamide; N- 4 -c h oro 3methy-5-isxz lyI)-2[(mehoy2-et pheny)aminocarbonyllthiophene- 3 -sufoaie

N-(

4 *ie N. (oxzoii).2..o(2 3 -methylphenyxal) ehyp ey~mn c incaronamtiocarny-slfoth id e;Ne (4 mienyl 4 hi 2 2 4 d.lyi- 0bonyIlthiophene3sufonmde

N-(

4 -chloro3methyl 5isoxazly)-23mehyd5imethzoypeni)2[2mtoy6ehlpnl)aminocarb n l t i p e e 3 s f o a i e -4chloro-3-m ethyl-5 i ox.ly 2 chlro3-mth isoxazoy)2[4 (,-dihxymethylpheny minocar bonylthiophene3sulfonide N-(t 4 -chloro 3 methyf5isoxazoyf) 2 -2 ethyl 4-dmethoy-pheny) aminocarbonltipee3sfoaie

II

5choro-3methy-5isoxazol)- 2 2 -iorpl4mtoypey~mncr bonyllthiophene3.sufonamd

N-(

4 2 dpro thyl l)Pt xphenyl oaino. cro i-hpen3suIon amide;ro.3 chIor-3mt metylsouxoxa o )2-t [-4mtoy -bp nyamnocrolI bhophnllhiohen..uonamd N-4clr- 4 mechloismethyi)-o II)4 (ethylen~eioxy)- mehylny )a ceytiph n ufnbie -4 -222chloro-3-methyl-5-isoxazolyl)-2{[3,4-(methylenedioxy)-6-ethylphenyl)acetyl}thiophene-3-sulfonamide;

N-(

4 -chloro-3-methyl-5-isoxazolyl)-2- {[3,4-(methylenedioxy)-6-methoxyphenyllacetyllthiophene-3-sufonamide EXAMPLE 193

N-(

3 ,4-Dimethyl-5-isoxazolyl)-4-biphenylsuffonamide 4 -Biphenylsulfonyl chloride 4 -Biphenylsulfonic acid (3.0 g, 12.8 mmol) was heated at 700 C with phosphorus oxychloride (1.30 ml, 14.0 mol) for 2 h. Excess phosphorus oxychloride was removed under reduced pressure. The residue was decomposed with ice water and extracted with ethyl acetate.

The extract was washed with 5% sodium bicarbinate solution, dried over anhydrous magnesium sulfate and concentrated to yield 2.9 g of crude 4biphenylsulfonyl chloride.

N-(3,4-Dimethyl-5-isoxazolyl)-4-biphenylsulfonamide 15 The 4 -biphenylsulfonyl chloride from step was added to a solution of 5-amino-3,4-dimethylisoxazole (250 mg, 2.2 mmol) and 4- (dimethyl)aminopyridine (5 mg) in dry pyridine (2.0 ml). The reaction mixture was stirred at room temperature for 4 h. Pyridine was removed under reduced pressure and the residue was partitioned between water and ethyl acetate. The organic layer was washed with 1N HCI (2 X ml), brine (25 ml) and dried over anhydrous magnesium sulfate.

Evaporation of the solvents left an oily residue that, after purification by column chromatography over silica gel (1 methanol in chloroform as eluent), yielded 337 mg of a white solid. Recrystallization from ethyl acetate/hexanes gave white crystals, m.p. 154-155'

C.

-223- EXAMPLE 194

N-(

4 -Bromo.3-methyl-5-isoxazoly)-4-biphenysufonamide 5-Amino-4-bromo-3-methylisoxazole 5-Amino-3-methylisoxazole (0.98 g, 10 mmol) was dissolved in chloroform (15 ml) and cooled to 00 C. N-Bromosuccinimide (1.78 g, mmoles) was added in small portions over a period of 10 min. The stirring was continued for another 10 minutes at 00 C. The reaction mixture w diluted with chooom(50 ml),wahdit with chloroform (50 ml), washed with water (2 X 50 ml) and 10.. the organic layer was dried over magnesium sulfate. Removal of the sol- S. vent under reduced pressure gave the crude product, which was purified by column chromatography using 9: 1, hexanes/ethyl acetate as the eluent, to give 5-amino-4-bromo-3-methylisoxazole (1.55 g, 87% yield).

N-(4-Biphenysulfonyl)-N-(4-bromo-3-methyl-5-isoxazolyl)-4- 15 biphenylsulfonamide mino-4-bromo-3-methylisoxazole (0.179 g, 1.0 mmol) was dissolved in dry pyridine (2 ml). 4-Biphenylsulfonyl chloride (0.509 g, 2.2 mmol) was added wth g, 2.2 *mmol) was added with stirring at ambient temperature. N,N-dimethylaminopyridine (5 mg) was added, and stirring was continued at t5O C for 16 h. The reaction mixture was diluted with dichloromethane (75 ml), washed with 1 N HCI (2 X 50 mi) and the organic phase was dried over magnesium sulfate. The solvent was removed under reduced pressure to yield a crude product, which was purified by column chromatography using 8:2, hexanes/ethyl acetate, to give 0.390 g (60% yield) of N-(4bmiphenylsulfonyl)-N-(4-bromo-3-methyl-5-isoxazolyl)-4-biphenylsulfonamide.

N-(4-Bromo-3-methyl-5-isoxazolyl)-4-biphenylsuffonamide N-(4-biphenylsulfonyl)-N-(4-bromo-3-methyl-5-isoxazoly)-4-bi phenylsulfonamide (0.150 g, 0.233 mmol) was dissolved in tetrahydrofuran (THF). Sodium hydroxide (0.120 g, 3.0 mmol) was added and the -224solution was warmed to 450 C to dissolve the sodium hydroxide. Stirring was continued for 20 min. Tetrahydrofuran was removed under reduced pressure. The residue was dissolved in water, cooled to 00 C and acidified to pH 3-4 with concentrated HCI. The solid precipitate was filtered off and dried in vacuo to give N-( 4 4 -biphenylsulfonamide (94% yield), which was further purified by recrystallization from chloroform/hexanes, m.p. 133-135*

C.

EXAMPLE 195 SN-(3,4-Dimethyl-5-isoxazolyl)-2-dibenzofuransulfonamide N-(3,4-Dimethyl-5-isoxazolyl)-2-dibenzofuransulfonamide was prepared, using the method described in Example 193b, from 3 4 -dimethylisoxazole and 2 -benzofuransulfonyl chloride in 32% yield.

Purification was achieved by recrystallization from chloroform/hexanes to give a white "cotton-like" solid, m.p. 173-175' C (dec.).

15 EXAMPLE 196 N-(4-M ethyl- 3-trifluoromethyl 5-iso xazo lyl)-4-biphenylsulfon amide N-(4-Methyl-3-trifluoromethyl-5-isoxazolyl)-4-biphenylsulfonamide was prepared in the same manner as described in Example 194b from amino-4-methyl-3-trifluoromethyl-isoxazole and 4 -biphenylsulfonyl chloride in 78% yield. Purification was achieved by recrystallization from methanol/water to give a white solid, m.p. 139-1400

C.

EXAMPLE 197

N-(

4 Tridecyl-3-trifluoromethyl-5-isxazolyl)-4-biphenysufonamide

N-(

4 -Tridecyl-3-trifluoromethyl-5-isoxazolyl)-4-biphenysulfonamide was prepared, in the same manner as described in Example 194b, from amino-4-tridecyl-3-trifluoromethyl-isoxazole and 4 -biphenylsulfonyl chloride in 81% yield. Purification was achieved by recrystallization from methanol/water to give an off white solid, m.p. 115-1160

C.

EXAMPLE 198 -225-

N-(

4 Methyl3trifloeomethylsoxaoll) 4 -bipe. sffnm

N-(

4 4 bpheniufnm was prepared, as described in Example 194, from 5-amino-4-methyl-3 trifluoromethylisoxazole and 4 -biphenylsulfonyl chloride in 78% yield.

Purification was achieved by recrystallization from ethyl acetate/hexanes to give a white solid, m.p. 139-140'

C.

EXAMPLE 199

N-(

4 Bromo-methy.3isoxazolyl) 4 bh.hyisufn mn rm ehlsxz e 10 3 -Amino-5-methylisoxazole (1.96 g, 20 mmol) was dissolved in chloroform (10 ml) and cooled to 0* C. N-Bromosuccinimide (3.56 g, mmol) was added in small portions over a period of 10 min. The stirring 9 was continued for another 1 5 minutes at 00 C. The reaction mixture was diluted with chloroform (100 ml), washed with water (2 X 50 ml) and the 15 organic layer was dried over magnesium sulfate. Removal of the solvent under reduced pressure gave the crude product, which was purified by column chromatography, using 9:1 hexanes/ethyl acetate as the eluent, to give 3-mn--rm--ehlsxzl (1.40 g, 40 yield).-

N-(

4 bromo.5.methyl3isoxazoly) 4 bipheniufnm N- 4 bro mo 5methyl 3-isoazoyl) 4 .bp nlufonai a prepared, using the method in Example 193b, from 3 methylisoxazole and 4 -biphenylsulfonyl chloride in 5% yield. The crude product was purified by column chromatography. After recrystallization from ethyl acetate/hexanes,

N-(

4 -bromo5-methyl.3..isoxazolyl)- 4 biphenylsulfonamid 8 154-156' C) was obtained in 51 yield.

-226- EXAMPLE 200

N-(

4 -Chloro-3-methyl-5-isoxazolyl)-4-biphenyIsuffonamide 5-Amino-4-chloro-3-methylisoxazole Using the method in Example 194a, 5-amino-4-chloro-3-methylisoxazole was prepared from 5-amino-3-methylisoxazole and N-chlorosuccinimide in 90% yield.

N-(

4 -Chloro-3-methyl-5-isoxazoly)-4-biphenysufonamide .Sodium hydride (188 mg, 4.4 mmol) was suspended in dry THF S(1 ml) and cooled to 00 C. A solution of 5-amino-4-chloro-3-methylisoxazole (mg, mmol) in dry THF (1 ml) was added with stirring. Once the addition was complete, the reaction mixture was warmed to room *temperature for 10 min. The solution was recooled to O0 C, and 4biphenylsulfonyl chloride (0.283 ml, 2.2 mmol) was added. Stirring was :continued at 250 C for 2 h. Excess sodium hydride was decomposed by the addition of methanol (0.4 mi) followed by water (0.5 ml). The THF was removed under reduced pressure and the residue was dissolved in .I water (20 ml) and basified by addition of sodium hydroxide (pH 9 Neutral impurities were removed by extraction with ethyl acetate (2 X ml). The aqueous layer was acidified to pH 2-3 using concentrated

HCI

and extracted with ethyl acetate (3 X 10 ml). The combined organic layer was dried over magnesium sulfate. Removal of the solvent gave

N-

(4-chloro-3-methyl-5-isoxazolyl)-4-biphenylsulfonamide in 83% yield.

This product was purified by recrystallization from ethyl acetate/hexanes as a white solid, m.p. 129-1320

C.

EXAMPLE 201 2,5-Dimethoxy-N-4-bromo-3-methyl-5-isoxazolyl)benzenesulfonamide 2, 5 -Dimethoxy-N-(4-bromo-3-methyl-5-isoxazolyl)benzenesulfonamide was prepared from 5-amino-4-bromo-3-methylisoxazole and methoxybenzenesulfonyl chloride according to the procedures described -227in Example 200. The crude product was purified by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 118-1200 yield 58%.

EXAMPLE 202 N-(4-bromo-3-methyl-5-isoxazolyl)-2biphenylsulfonamide A. 2 -Biphenylsuffonyl chloride 2-Bromobiphenyl (2.33 g, 10 mmol) was dissolved in ether (10 ml) and cooled to -78 0 C. n-Butyllithium (2.5 M solution in hexane, 4.8 ml, 4 12 mmol) was added dropwise under constant stirring and an argon atmosphere. The resultant reaction mixture was stirred at -700C to 600C for 1h. The reaction mixture was cooled to -78C and sulfuryl chloride (0.88 ml, 11 mmol) was added dropwise. After addition,he tion mxturewas allowed to attain ambient temperature slowly and tirred for 1 h. The reaction ixture was diluted with ethyl acetate 15 ml), washed with water and the organic layer dried over anhydrous MgSO 4 Removal of the solvent under reduced pressure gave a crude Sproduct, which was purified by column chromatography, using hexane followed by 5% ethyl acetate in hexane as eluent, to give 2-biphenylsulfonyl chloride as a solid (1.3 g, 51% yield).

B.

N-(

4 -bromo-3-methyl-5-isoxazolyl)- 2 -biphenylsulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)2.-biphenylsulfonamide was prepared in the same manner as described in Example 200b from amino-4-bromo-3-methylisoxazole and 2 -biphenylsulfonyl chloride in 71% yield. Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 145 147 0

C.

-228- EXAMPLE 203

N-(

4 -Chloro-3-methyl-5-isoxazolyl)-2.biphenylsulfonamide

N-(

4 -Chloro-3-methyl-5-isoxazolyl)-2-biphenylsulfonamide was prepared in the same manner as described in Example 202 from 4 -chloro-3-methylisoxazole and 2 -biphenylsulfonyl chloride in 74% yield.

Purification was achieved by recrystallization from ethyl acetate/hexanes to give a crystalline solid, m.p. 132 134 0

C.

EXAMPLE 204

SN-(

4 -Bromo-3-methyl-5-isoxazolyl)-3-biphenylsulfonamide 10 A. 3 -Biphenylsulfonyl chloride 3 -Bromobiphenyl (1.5 g, 6.4 mmol) was dissolved in ether (15 ml) and cooled to -78 0 C. t-Butyllithium (1.7 M solution in hexane, 3.8 ml, 6.4 mmol) was added dropwise under constant stirring and an argon atmosphere. The resultant reaction mixture was stirred at -100C to -500 for 6h. The reaction mixture was cooled to -78 0 C and sulfuryl chloride (0.64 ml, 6.4 mmol) was added dropwise. After completion of the addition, the reaction mixture was allowed to attain ambient temperature slowly and stirred for 1 h. The reaction mixture was diluted with ethyl acetate (50 ml), washed with water and the organic layer dried over anhydrous MgSO4. Removal of the solvent under reduced pressure gave crude product, which was purified by column chromatography, using hexane followed by 5% ethyl acetate in hexane as eluent, to give 3biphenylsulfonyl chloride as an oil (0.8 g, 49% yield).

B.

N-(

4 -bromo-3-methyl-5-isoxazolyl)-3-biphenylsulfonamide N-(4-bromo-3-methyl-5-isoxazolyl)-3-biphenylsulfonamide was prepared in the same manner as described in Example 200b from amino-4-bromo-3-methylisoxazole and 3 -biphenylsulfonyl chloride in 22% yield. This was purified by HPLC

CH

3 CN to 100%

CH

3 CN over min.) to give a solid., m.p. 78 82° C.

-229-

S

a. a

S

*5 .5 S S 5 EXAMPLE 205

N-(

4 -chloro-3-methyl-5-isoxazolyl)-3-biphenysuffonamide

N-(

4 -chloro-3methyl-5-isoxazolyl)-3-biphenylsuffonamide was prepared in the same manner as described in Example 204 from 4 -chloro-3-methylisoxazole and 3 -biphenylsulfonyl chloride in 63% yield.

This was purified by HPLC

CH

3 CN to 100%

CH

3 CN over 30 min.) to give a solid, m.p. 84 86 0

C.

EXAMPLE 206

N-(

3 -methyl-5-isoxazolyl)-4-(4-methylphenyl)benzenesuffonamide

N-(

3 -methyl-5-isoxazolyl)4-bromobenzenesulfonamide 4 -brombenzenesulfonyl chloride (solid) was added, in five portions, to a solution of 3 -methyl-5-aminoisoxazole (3.82 g, 40 mmol) in dry pyridine (30 ml). This was stirred at room temperature for 3 h and the pyridine was removed under reduced pressure. The residue was dissolved in THF (300 ml) and a 5% NaOH solution (100 ml) was added. Stirring continued for 1 h at room temperature. The THF was removed under reduced pressure and the resultant residue was neutralized to pH 2 using concentrated hydrochloric acid. This was extracted with ethyl aeetate (3 x 200 ml) and the combined organic layer was dried over MgSO 4 and concentrated. The crude product was recrystallized using hexane/ethyl acetate giving N-(3-methyl-5-isoxazoly)-4-bromobenzenesulfonamide (9.2 9, 72% yield).

N-(3demethyl-5-isoxazolyl)--(4-methylphenyl)benzenesulfonamide Nitrogen was bubbled through a biphasic mixture of ethanol ml), toluene (15 mi) and 2M sodium carbonate solution (15 mi). N-(3methyl-5-isoxazolyl)-4-bromobenzene sulfonamide (0.951 g, 3 mmol), 4methylbenzeneboronic acid (0.56 g, 4 mmol) and tetrakistriphenylphosphine palladium (300 mg) were added. The reaction mixture was kept at 800 C, under a N, atmosphere for 24 h, with stirring, and was -230then diluted with water (50 ml) and extracted with ether (50 ml) to remove neutral impurities and excess 4 -methylbenzeneboronic acid. The aqueous phase was neutralized to pH 2 using concentrated hydrochloric acid and the resultant solid was filtered. This was dried under vacuum and recrystallized using hexane/ethyl acetate giving isoxazolyl)-4-(4-methylphenyl)benzenesulfonamide g, 100% yield, m.p. 194-198 0

C).

EXAMPLE 207

SN(

4 bromo-3-methyl-5-isoxazolyl)-4-(4-methylphenyl)benzenesulfonamide S 10 N-bromosuccinimide (NBS) (0.178 g, 1 mmol), in one lot, was added to a stirred suspension of N-(3-methyl-5-isoxazolyl)-4-(4-methylphenyl)benzenesulfonamide (0.

3 27g, 1 mmol, Example 206b) in chloroform (12 ml). The reaction mixture was stirred for 10 min then diluted with dichloromethane (50 ml). This was washed with water (2 x 50 ml).

15 The organic layer was dried over MgSO 4 and concentrated. The crude product was recrystallized using hexane/ethyl acetate giving N-(4-bromo- S3-methyl-5-isoxazolyl)-4-(4-methylphenylbenzenesufonamide (350 mg, 86% yield, m.p. 153-1560

C).

EXAMPLE 208

N-(

4 -chloro-3-methyl-5-isoxazolyl)-4-(4-methylphenyl)benzenesulfonamide N-chlorosuccinimide (0.266 g, 2 mmol) was added, in one lot, to a stirred suspension of N-(3-methyl-5-isoxazolyl)-4-(4-methylphenyl)benzenesulfonamide (0.

3 27g, 1 mmol, Example 206b) in chloroform (10 ml) and stirred at room temperature for 2 h The reaction mixture was diluted with dichloromethane (50 ml) and washed with water (2 x 50ml). The organic layer was dried over MgSO 4 and concentrated. The crude product was purified by column chromatography using ethyl acetate as eluent to give N-(4-chloro-3-methyl-5-isoxazolyl)-4-(4-methylphenyl)benzenesulfonamide [210 mg, 58% yield, m.p. 2600 C (de coup)].

-23 1- EXAMPLE 209

N(

3 methyl5 soxzoly4(4tluorotrl eybenzenesulfon.

amide was prepared in the same manner as described in Example 206b, and 4-trifluoromethylbenzeneboronic acid resulting in the final product in a 78% 9. 0Oyield, m.p. 150-153' C. The product was recrystallized using an acetonirile and water mixture.

9 9EXAMPLE 210 sulfonamide f 0

N-(

4 bromo3methyl5-so 5 xazoly) 4 4 -trflurmehl phenyll benzenesulfonamide was prepared in the same manner as described in Exam ple 207, using N e h l 5 i o a o y) 4 -rf u r m t y p e y) benzenesulfonamide (Example 209) and NBS (reaction time 30 min at room temperature). The crude product was purified by column chromatography on silica gel using ethyl acetate as eluent resulting in the final product in 56% yield, m.p. 113-117'

C.

EXAMPLE 211

N-(

3 methy5isoxazolyl(4ethyhenlbneeufnmd was prepared in the same manner as described in Example 206b, using N- 3 -meth yk 5-iso xazo lyl) 4bro mobeznslonaie (Example 2 06a) and 4-ehxbneeooi acid resulting in an 82% yield of the final product, m.p. 194-196' C. The product was recrystallized using hexane/ethyl acetate.

-232- EXAMPLE 212 N-(4-iebromo-3-meth-yl)-4- oxazoz)4oyphenyl)benzenesulfon- N-(4-bromo-3-methyl-5-isoxazolyl)-4-(4-methoxyphenyl)benzenesulfonamide was prepared in the same manner as described in Example 207 using

N-(

3 -methyl-5-isoxazolyl)-4-(4-methoxyphenyl)benzenesulfonamide (Example 211) and NBS (reaction time 30 min at room temperature). The crude product was purified by column chromatography on silica gel using ethyl acetate as eluent giving the final product in 78% yield, m.p. 2080

C

(dec). The product was recrystallized using hexane/ethyl acetate.

EXAMPLE 213 9N-( 4 bromo-3-methyl-5-isoxazolyl)-4-(3-methoxyphenyl)benzenesulfonamide 15 N-(3-methyl-5-isoxazolyl)-4-(3-methox b fonamide peybeznsl N-(3-methyl-5-isoxazolyl)-4-(3-methoxyphenyl)benzenesulfonamide was prepared in the same manner as described in Example 206b, using

N-

(3-methyl-5-isoxazolyl)-4-bromobenzenesulfonamide (Example 206a) ad 9* 9 0 (znslonmdExample 2 O6a) and 20 3-methoxybenzeneboronic acid resulting in a 77% yield.

zN-uromo-3methyl-5-isoxazolyl)-4-(3-methoxyphenyl)benzenesulfonamide

N-(

4 -bromo-3-methyl-5-isoxazoly)-4-(3-methoxyphenyl)benzenesulfonamide was prepared in the same manner as described in Example 207 using

N(

3 -methyl-5-isoxazoyl)-4-(3-methoxyphenyl)benzenesul and NBS (reaction time 30 min at room temperature). The crude product was purified by column chromatography on silica gel using ethyl acetate as eluent giving the final product, after recrystallization using hexane/ethyl acetate, in 75% yield, m.p. 140-1440

C.

-233- EXAMPLE 214 N-m-bode 3-etyl-isoxazolyl)-4-(2-methoxypheflyl)befzensffn

N-(

3 methyl5isxazoly)4( 2 mthoxpey~eznsl fonamideeoyhnlbzneu azoly) 4 .(2-methoxyphenyl)benzslfonm was prepared in the same manner as described in Example 206 using

N-

3 -methyl5isoxazolyl)4bromobenzsffnmdad -ehxb zeneboronic acid resulting in an 81 yield of the final product.

W N-( 4 bromo.3methyl5isoxozll) 4 2 -ehxpn)b Szenesulfonamide 4 -bromo.3methyl5isoxozl)- 4 2 -ehxpnybnzesl fonamicje was prepared in the same manner as described in Example 207, usn

N-(

3 methyl.5-iso aolyl)4(2-m thyhenlbneeufnm and NBS (reaction time 30 min at room temp.) The crude product was purified by column chromatography on silica gel using ethyl acetate as eluent to give the final product in 68% yield, m.p. 205.2090

C.

S. S EXAMPLE 215

N-(

4 -bromo-3methyl 5-isoxzll)-4(34mtyedixpnl)bznsulfonamide

N.(

3 methyl.5isoxazolyl).

4 3 4 ehylndoyhlb zenesulfonamide yn N-3mty--sxzlt--34mtyeeixpey~eznsl fonamjcje was prepared in the same manner as described in Example 206, sin N(3metyl5-ioxzoyi)4romobenzenesulfonamide and 3,-ehlndoyhnlooi acid resulting in a 67% yield of final product.

N-(

4 bromo3methyl5isoxazolyl)4( 3 4mtyendoy phenyl)benzenesulfoflamjde N-4boo3mehl5ioazl 3 4 -methylenedioxypheny,)benzenesulfonamide was prepared in the same manner as described in -234- Example 207, using

N-(

3 -methyl-5-isoxazolyl)-4-(3,4-methylenedioxyphenyl)benzenesulfonamide and NBS in THF as solvent resulting in a yield. The crude product was purified by HPLC, m.p. 172-1740

C.

EXAMPLE 216

N-(

4 -bromo-3-methyl-5-isoxazolyl)-4-(3-methylphenyl)benzenesulfonamide

N

(3methyl-5-isoxazolyl)-4-(3-methylpheny)benzenesulfonamide N-(3-methyl-5-isoxazolyl)-4-(3-methylphenyl)benzenesulfonamide was prepared in the same manner as described in Example 206b, using

N-

10 (3-methyl-5-isoxazolyl)-4-bromobenzenesulfonamide (Example 2 06a) and 3-methylbenzeneboronic acid resulting in an 82% yield.

N-(

4 -bromo-3-methyl-5-isoxaz o lyl)-4-( 3 -methylphenyl)ben- zenesulfonamide "N-(4-bromo-3-methyl-5-isoxazolyl)-4-(3-methylphenylbenzenesul- S 15 fonamide was prepared in the same manner as described in Example 207, using N-(3-methyl-5-isoxazolyl)-4-(3-methylphenylbenzenulf mide and NBS in THF as solvent (reaction time 30 min at room temperature).

The crude product was purified by HPLC resulting in a 31% yield of the inal product, m.p. 186-189°

C.

20 EXAMPLE 217 Assays for identifying compounds that exhibit endothelin antagonistic and/or agonist activity Compounds that are potential endothelin antagonists are identified by testing their ability to compete with 25 I-labeled ET-1 for binding to human ET, receptors or ETB receptors present on isolated cell membranes. The effectiveness of the test compound as an antagonist or agonist of the biological tissue response of endothelin can also be assessed by measuring the effect on endothelin induced contraction of isolated rat thoracic aortic rings. The ability of the compounds to act as antagonists or agonists for ET

B

receptors can be assess by testing the -235ability of the compounds are to inhibit endothelin-1 induced prostacyclin release from cultured bovine aortic endothelial cells.

A. Endothelin binding inhibition Binding Test Inhibition of binding to ETA receptors TE 671 cells (ATCC Accession No. HTB 139) express

ETA

receptors. These cells were grown to confluence in T-175 flasks. Cells from multiple flasks were collected by scraping, pooled and centrifuged for 10 min at 190 X g. The cells were resuspended in phosphate buffered saline (PBS) containing 10 mM EDTA using a Tenbroeck homogenizer.

The suspension was centrifuged at 40 C at 57,800 X g for 15 min, the pellet was resuspended in 5 ml of buffer A (5mM HEPES buffer, pH 7.4 containing aprotinin (100 KIU/ml)) and then frozen and thawed once. 5 ml of Buffer B (5 mM HEPES Buffer, pH 7.4 containing 10 mM MnC 2 I and 0.001% deoxyribonuclease Type 1) was added, the suspension mixed by inversion and then incubated at 370 C for 30 minutes. The mixture was centrifuged at 57,800 X g as described above, the pellet washed twice with buffer A and then resuspended in buffer C (30 mM HEPES buffer, pH 7.4 containing aprotinin (100 KIU/ml) to give a final protein concentration of 2 mg/ml and stored at -70° C until use.

The membrane suspension was diluted with binding buffer (30 mM HEPES buffer, pH 7.4 containing 150 mM NaCI, 5mM MgCI 2 Bacitracin) to a concentration of 8 pg/50 pl. 125 1-endothelin-l (3,000 cpm, 50 mL) was added to 50 pL of either: endothelin-1 (for non specific binding) to give a final concentration 80 nM); binding buffer (for total binding); or a test compound (final concentration 1 nM to 100 pM). The membrane suspension (50 pL), containing up to 8 pg of membrane protein, was added to each of or Mixtures were shaken, and incubated at 40 C for 16-18 hours, and then centrifuged at 4° C for 25 min at 2,500 X g. Alternatively, the incubation was conducted at 240 C. When incubated at 240 C, the IC 50 concentrations -236are 2- to 10-fold higher than when the incubation is conducted at 40 C.

This, must be kept in mind when comparing IC,, concentrations among compounds provided herein.

The supernatant, containing unbound radioactivity, was decanted and the pellet counted on a Genesys multiwell gamma counter. The degree of inhibition of binding was calculated according to the following equation:

(A)

D 100- X 100 10

(A)

Each test was generally performed in triplicate.

b: B. Endothelin binding inhibition Binding Test Inhibition of binding to ET, receptors COS7 cells were transfected with DNA encoding the ET, receptor, 15 The resulting cells, which express the human ET, receptor, were grown to confluence in T-150 flasks. Membrane was prepared as described above. The binding assay was performed as described above using the membrane preparation diluted with binding buffer to a concentration of 1 pg/50/ l.

20 Briefly, the COS7 cells, described above, that had been transfected with DNA encoding the ET, receptor and express the human ET, receptor on their surfaces were grown to confluence in T-175 flasks. Cells from multiple flasks were collected by scraping, pooled and centrifuged for min at 190 X g. The cells were resuspended in phosphate buffered saline (PBS) containing 10 mM EDTA using a Tenbroeck homogenizer. The suspension was centrifuged at 40 C 57,800 X g for 15 min, the pellet was resuspended in 5 ml of buffer A (5mM HEPES buffer, pH 7.4 containing aprotinin (100 KIU/ml)) and then frozen and thawed once.

Five ml of Buffer B (5 mM HEPES Buffer, pH 7.4 containing 10 mM MnCI, and 0.001% deoxyribonuclease Type 1) was added, the -237suspension mixed by inversion and then incubated at 370 C for minutes. The mixture was centrifuged at 57,800 X g as described above, the pellet washed twice with buffer A and then resuspended in buffer C (30 mM HEPES buffer, pH 7.4 containing aprotinin (100 KIU/ml) to give a final protein concentration of 2 mg/ml.

The binding assay was performed as described above using the membrane preparation diluted to give 1 pg/50 p1 of binding buffer.

SC. Test for activity against endothelin-induced contraction of isolated rat thoracic aortic rings 10 The effectiveness of the test compound as an antagonist or agonist of the biological tissue response of endothelin also is assessed by measuring the effect on endothelin induced contraction of isolated rat thoracic aortic rings (see, Borges et al. (1989) Eur. J. Pharmacol.

165:223-230) or by measuring the ability to contract the tissue when added alone.

Compounds to be tested are prepared as 100 pM stocks. If necessary to effect dissolution, the compounds are first dissolved in a minimum amount of DMSO and diluted with 150 mM NaCI. Because DMSO can cause relaxation of the aortic ring, control solutions containing varying concentrations of DMSO were tested.

The thoracic portion of the adult rat aorta is excised, the endothelium abraded by gentle rubbing and then cut into 3 mm ring segments. Segments are suspended under a 2 g preload in a 10 ml organ bath filled with Krebs'- Henseleit solution saturated with a gas mixture of 95% 02 and 5% CO, (118 mM NaCI, 4.7 mM KCI, 1.2 mM MgSO 4 1.2 mM KH 2

PO

4 25 mM NaHC03, 2.5 mM CaCI 2 10 mM D-glucose).

There is a correlation between activity as an antagonist of endothelin-induced thoracic aortic ring contraction and activity as an inhibitor of binding of endothelin to endothelin receptors. The pA 2 is a linear function of the log of the ICso.

-238- D. Assay for identifying compounds that have agonist and/or antagonistic activity against ET receptors 1. Stimulation of prostacyclin release Since endothelin-1 stimulates the release of prostacyclin from bovine aortic endothelial cells, the compounds that have agonist or antagnoist activity are identified by their ability to inhibit endothelin-1 induced prostacyclin release from such endothelial cells by measuring 6keto

PGF

1 substantially as described by (Filepet al. 9 9 1) Biochem.

SBiohs Res. Commun. 177 171-176. Bovine aortic cells are obtained from collagenase-treated bovine aorta, seeded into culture plates, grown in Medium 199 supplemented with heat inactivated 15% fetal calf serum, and L-glutamine (2 mM), penicillin, streptomycin and fungizone, and Ssubcultured at least four times. The cells are then seeded in six-well plates in the same medium. Eight hours before the assay, after the cells reach confluence, the medium is replaced. The cells are then incubated with a) medium alone, b) medium containing endothelin-1 (10 nM), c) test compound alone, and d) test compound endothelin-1 (10 nM).

After a 15 min incubation, the medium is removed from each well and the concentrations of 6-keto PGFare measured by a direct P are measured by a direct immunoassay. Prostacyclin production is calculated as the difference between the amount of 6-keto PGF1a released by the cells challenged with the endothelin-1 minus the amount released by identically treated unchallenged cells. Compounds that stimulate 6 -keto PGFIa release Possess agonist activity and those which inhibit endothelin-1 6-keto

PGF

1 release possess antagonist activity.

2. Inhibition of sarafotoxin 6c induced contraction Sarafotoxin 6c is a specific ET, antagonist that contracts rat fundal stomach strips. The effectiveness of tests compounds to inhibit this sarafotoxin 6 c-induced contraction of rat fundal stomach strips is used as a measure

ET

6 antagonist activity. Two isolated rat fundal stomach strips -239are suspended under a 1 g load in a 10 ml organ bath filled with Krebs'- Henseleit solution containing 10 pM cyclo(D-Asp-Pro-D-Val-Leu-D-Trp) (BQ-123; see, U.S. Patent No. 5,114,918 to Ishikawa et indomethacin, and saturated with a gas mixture of 95% 02/5%

CO

2 Changes in tension are measured isometrically and recorded using a Grass Polygraph coupled to a force transducer. Sarafotoxin 6c is added cumulatively to one strip while the second strip is preincubated for -min with a test compound prior to addition of cumulative doses of sarafotoxin 6c. The effects of the test compounds on the concentrationresponse curve for sarafotoxin 6c are examined.

E. Deoxycorticosterone acetate (DOCA)-salt hypertensive S rat model for assessing in vivo activity of selected compounds- 1 n Selected compounds disclosed herein have been tested for activity 15 in the deoxycorticosterone acetate (DOCA)-salt hypertensive rat model.

To perform these tests, silastic MDX4-4210 elastomer implants containing 47 mg (DOCA) were prepared according to the method of Ornmsbee et a. ((1973) the J. Pharm. Sci. 62:255-257). Briefly,

DOCA

is incorporated into silicon rubber implants for sustained release. To S 20 prepare the implants the DOCA is incorporated into unpolymerized silicone rubber, catalyst is added and the mixture is cast in a hemicylindrical shape.

Sprague Dawley rats (7-8 weeks old) were unilaterally nephrectomized under ketamine anesthesia and a DOCA-implant was placed on the left lateral dorsal abdomen of the animal. The rats were allowed to recover for three weeks. During recovery they were permitted free access to normal rat chow and 0.9% NaCI drinking solution in place of drinking water. The rats develop hypertension within 3 weeks.

-240- All animals were used in the tests between 21 and 30 days post surgery. The mean arterial blood presure in these animals ranged from 165-200 mm Hg.

On the day of experimentation, catheters were inserted under brevital anesthesia into the right femoral artery for measurement of blood pressure, and into the right femoral vein for administration of a selected compound. The animals were placed in a restrainer and allowed to recover for a minimum of 60 min or until a steady mean arterial blood pressure was recorded. At that time, the selected compound or control 10 vehicle was administered either intravenously, as a 60 minute infusion, or orally by oral gavage. Blood pressure was recorded continuously for a fruther 10 hrs.

F. Effect of Intravenous administration on ET-1-induced 15 pressor responses in conscious, autonomically blocked rats; a model for assessing in vivo activity of selected compounds :i Male Sprague Dawley rats (250-450 g) were anesthetized (Brevital 50 mg/kg, IP) and cannulae were placed in the femoral artery to measure mean arterial pressure (MAP) and in the femoral vein for intravenous drug 20 administration. Animals were placed in a restrainer and allowed to regain consciousness. Thirty minutes later autonomic blockade was administered (atropine methyl nitrate, 3 mg/kg, IV, followed by propranalol, 2 mg/kg, IV). An hour later animals received a bolus injection of vehicle (0.5 ml) followed thirty minutes later by intravenous bolus administration of ET-1 (Control, 1 pg/kg). Following recovery from this challenge, test -compounds were administered by intravenous bolus administration (0.5 ml) and then re-challenged with ET-1 thirty minutes later. Results are expressed as the percent inhibition of the ET-1-induced pressor response after administration of the test compound compared to the pressor response induced by the control ET-1 challenge. In some -241cases a third ET-1 challenge was administered ninety minutes after administration of the test compound.

G. Results 1. In~ yjtM The IC 50 for each of the compounds of the preceding Examples for ETA and ETB receptors has been measured. Almost all of the compounds have an IC 50 of less than 10 /pM for either or both of the ETA and

ET,

:0,40,receptors. Many of the compounds have an IC 50 less than about 10 pM, 9 others have an I C 50 less than about 1 uM and some of the compounds 10 have an IC 50 less than about 0. 1 uM. A number of the compounds have :an

IC

50 for ETA receptors that is substantially less (10 to 1 QO-fold or 9 9more) than for ET, receptors, and, thus are selective for ETA receptors.

Others of the compounds are ET, selective.

2. In vivo a. Selected compounds, such as N-(4-chloro.3 *S*methyI 5 isoxazo lyI)2-(N- th lphenl i cab l hohe -3 *sulfonamide,

N-(

4 bromo3.methyl5isoxall).

2 3 ,-mtyedix) 3 -sIfoaie

N-(

4 z4cooly)-~3mty,4 sxzli)2[6hdoy3methylenedioxy)bIenyb ethyllthiophene-3sufonamide,

N-(

4 -chloro.3.methyl5isoxazoly)-2-3 methylenedioxybenzylcarbonyl)thihe-3sloai, have been tested in the hypertensive rat model, and were effective in decreasing blood pressure.

b. Selected compounds, such as N-(4-chloro-3 2 3 4 -(methylenedioxy)phenylactylthiopee3 sulfonamide,

N-(

4 -chloro3.methyl.5isoxall) 2 2 (methylenedioxy)phenyllamicabnblltipee3sfoaie ch lo ro-3.methyl.5isoxazo yl) 2 -f( 4 -mtoy2mt l e l aminocar- -242bonyl~thiophene-3-.sulfonamide and N-(4-chloro-3-methyl..5.isoxazdlyl)- 2 [2-cyano-4, S-dimethoxyphenyl)aminocarbonyl thophen--ufnalide and 4 chloro3m methyl..5isaoyl)olyl)2[2methyl4S(m thlndix phenylacetyllthiophene-3sujffnamid have been tested in the autonomically blocked, normotensive rat model and shown to have substantial activity, reducing pressure about 30% in 30 min at dosages as low as 30 mg/kg, and more than 50% at dosages of 60 mg/kg. On the average dosages of 30-60 mg/kg of the test compound resulted in a 40-60% inhibition of pressor response.

Since modifications will be apparent to those of skill in this art, it is intended that this invention be limited only by the scope of the appended claims.

Claims (23)

1. A compound that has formula I: Ar- SO- N- Ar 1 H or a pharmaceutically acceptable salt, acid or ester thereof, wherein: Ar 1 is a five or six membered aromatic or heteroaromatic ring, preferably isoxazolyl, pyridazinyl, thiazolyl, pyrimidinyl or phenyl or is a bicyclic or tricyclic carbon or heterocyclic ring; and Ar 2 has formula: R 13 R26 5 1 *o wherein: 4* 5 alkoxycarbonyl, carbonyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, formyl, substituted or Sunsubstituted amido, substituted it eor unsubstituted ureido, in which the alkyl, alkenyl and alkynyl portions conntain from 1 up to about 14 carbon atoms, preferably from 1 to 6 atoms, and are either straight or branched chains or cyclic, and the aryl portions contain from about 4 to about 16 carbons, preferably 4 to 10 carbons, with the proviso that if there is only one R 13 it is not hydrogen, and with the proviso that the compound of formula is a 4-biphenylsulfonamide that is unsubstituted at the 2 or 6 position on the sulfonamide-linked phenyl group.

2. A compound of claim 1, or a pharmaceutically acceptable salt, acid or ester thereof, wherein Ar is an isoxazolyl, a thiazolyl, a pyrimidinyl, a pyridazinyl or a phenyl group.

3. A compound of claim 1, or a pharmaceutically acceptable salt, acid or ester thereof, that has formula VII: [RA\LIBA102588 doc TLT K< R R R R R2 SO-- NN or H H R R 2 wherein the biphenyl group is substituted with one or more than one substituent, each of which is selected independently from the selections set forth for R26 and R 13 and R 2 6 and R 13 are each independently selected from H, loweralkyl, haloalkyl and halide with the proviso that if there is only one substituent on the second phenyl ring it is not hydrogen.

4. A compound of claim 3, or a pharmaceutically acceptable salt, acid or ester thereof, in which one of the R 1 3 substituents is in the para position.

5. A compound of claim 3 or 4, or a pharmaceutically acceptable salt, acid or ester thereof, wherein Ri is halide or methyl or (C9-C 13 alkyl.

6. A compound of any of claims 3-5, or a pharmaceutically acceptable salt, acid or ester thereof, wherein R' is selected from halide, CH 3 C 2 H 5 CF 3 C2F 5 n-CsH 7 and cyclo-C 3 H 7 and R 2 is selected from H, CH 3 C2H 5 CF 3 C 2 F 5 n-C3H 7 and cyclo-C 3 H7.

7. A compound of any of claims 3-6, or a pharmaceutically acceptable salt, acid or ester thereof, wherein R 1 is halide or CH 3 and R 2 is H, CH 3 C2H 5 or CF 3 5 8. A compound of claim 2, wherein, the isoxazolyl is a 4-halo-isoxazolyl or a 4-higher alkyl (C8 to

9. The compound of claim 3, or a pharmaceutically acceptable salt, acid or ester thereof, that is selected from among N-( 4 -bromo-3-methyl-5-isoxazolyl)-4'-methylphenyl-4 biphenylsulfonamide, (4-bromo-3-methyl-5-isoxazolyl)-4'-trifluorophenyl-4-biphenylsulfonamide, N- o (4-bromo-3-methyl-5-isoxazolyl)-4'-methoxyphenyl-4-biphenylsulfonamide, N-(4-bromo-3-methyl-5- Isoxazolyl)-3'-methoxyphenyl-4-biphenylsulfonamide, N-(4-bromo-3-methyl-5-isoxazolyl)-2'- methoxyphenyl-4-biphenylsulfonamide and N-( 4 -bromo-3-methyl-5-isoxazolyl)-3',4'- methylenedioxyphenyl-4-biphenylsulfonamide. The compound of claim 3, or a pharmaceutically acceptable salt, acid or ester thereof, that is N-(4-bromo-3-methyl-5-isoxazolyl)-4'-methylphenyl-4-biphenylsulfonamide or N-(4-bromo-3- methyl-5-isoxazolyl)-4'-trifluorophenyl-4-biphenylsulfonamide.

11. A compound of formula I substantially as hereinbefore described with reference to the Examples. .\LIBA]02588 docTLT 245

12. A pharmaceutical composition, comprising a compound of any one of claims 1-11, or a pharmaceutically acceptable salt, acid or ester thereof, in a pharmaceutically acceptable carrier.

13. Use of a compound of any one of claims 1-11, or a pharmaceutically acceptable salt, acid or ester thereof, for treatment of an endothelin-mediated disorder.

14. A method for the treatment of endothelin-mediated diseases, comprising administering an effective amount of one or more compounds of any one of claims 1-11, or pharmaceutically acceptable salts, acids or esters thereof or an effective amount of a composition of claim 12, wherein the effective amount is sufficient to ameliorate one or more of the symptoms of the disease. 1o 15. Use of a compound of any one of claims 1-11 or a pharmaceutically acceptable salt, acid or ester thereof for the manufacture of a medicament for the treatment of endothelin-mediated o: diseases or disorders.

16. A compound according to any one of claims 1-11 or a pharmaceutically acceptable salt, acid or ester thereof, or a composition of claim 12 when used in the treatment of endothelin- I mediated diseases.

17. The method, use or compound of any one of claims 14-16 respectively, wherein the disease is selected from the group consisting of hypertension, cardiovascular disease, asthma, pulmonary hypertension, inflammatory diseases, ophthalmologic disease, menstrual disorders, obstetric conditions, wounds, gastroenteric disease, renal failure, immunosuppressant-mediated renal vasoconstriction, erythropoietin-mediated vasoconstriction endotoxin shock, anaphylactic shock and hemorrhagic shock.

18. The method, use or compound of any one of claims 14-16 respectively, wherein the disease is selected from the group consisting of hypertension, cardiovascular disease, pulmonary hypertension, erythropoietin-mediated vasoconstriction endotoxin shock, anaphylactic shock and hemorrhagic shock.

19. The method, use or compound of any of claims 14-16 respectively wherein the disease is selected from the group consisting of asthma and inflammatory diseases. A method for inhibiting the binding of an endothelin peptide to an endothelinA (ETA) or endothelinB (ETB) receptor, comprising contacting the receptor with one or more compounds of any one of claims 1-11, or pharmaceutically acceptable salts, acids or esters thereof or a pharmaceutical composition according to claim 12, wherein: the contacting is effected prior to, simultaneously with or subsequent to contacting the receptor with the endothelin peptide. [R \LIBA]02588.doc:TLT 246

21. A method for altering endothelin receptor-mediated activity, comprising contacting an endothelin receptor with one or more compounds of any one of claims 1-11, or pharmaceutically acceptable salts, acids or esters thereof or a pharmaceutical composition according to claim 12.

22. A pharmaceutical composition formulated for single dosage administration, comprising an effective amount of one or more compounds of any one of claims 1-11, or pharmaceutically acceptable salts, acids or esters thereof, wherein the amount is effective for ameliorating the symptoms of an endothelin-mediated disease.

23. An article of manufacture, comprising packaging material and one or more compounds of any one of claims 1-11, or pharmaceutically acceptable salts, acids or esters thereof or a o pharmaceutical composition according to claim 12, contained within the packaging material, wherein the compound or composition is effective for antagonizing the effects of endothelin, ameliorating the symptoms of an endothelin-mediated disorder, or inhibiting the binding of an endothelin peptide to an ET receptor with an IC50 of less than about 10 tM, and the packaging material includes a label that indicates that the sulfonamide or salt thereof is used for antagonizing 5 the effects of endothelin, inhibiting the binding of endothelin to an endothelin receptor or treating an endothelin-mediated disorder.

24. A medicament when prepared by the use of any one of claims 15 or 17-19. A compound according to any one of claims 1 to 11 or pharmaceutically acceptable salts, acids or esters thereof, or a composition according to claim 12 when used for inhibiting the 20 binding of an endothelin peptide to an endothelinA (ETA) or endothelinB (ETB) receptor.

26. A compound according to any one of claims 1 to 11 or pharmaceutically acceptable salts, acids or esters thereof, or a composition according to claim 12 when used for altering endothelin receptor-mediated activity.

27. The method of claim 14 wherein the disease is glaucoma. 12 28. The use of claim 15, wherein the disease is glaucoma.

29. The compound when used according to claim 16 wherein the disease is glaucoma Dated 8 September, 2000 Texas Biotechnology Corporation Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON [R \.IBA]02588doc TL I