60/111,025 filed on December 4,1998, the disclosure of which is incorporated herein by reference.

Field of the Invention The present invention relates to novel selectin modulating compounds having the structural Formula 1, as shown below, to methods of their preparation, to compositions comprising the compounds, to their use for treating human or animal disorders, to their use for purification of proteins, and to their use for in diagnostics. These compounds are modulators of selectin (P-, E-and L-selectin) Ligand (e. g. Sialyl Lewis X (sLeX)) interactions for the'management, treatment, control, or as an adjunct of diseases in humans caused by selectins. More particularly, this invention relates to the administration of compounds according to Formula 1 which are selectin/Ligand

antagonists, for the management of diseases and disease states such as 1) acute respiratory distress syndrome (ARDS), 2) diseases that may be controlled via inhibition of angiogenesis, 3) asthma, 4) atherosclerosis, 5) atopic dermatitis, contact dermatitis, and cutaneous inflammation, 6) bowel inflammation, 7) diabetes/diabetes-associated pathologies, 8) Grave's disease and associates conditions, 9) multiple sclerosis (MS), 10) myocardial ischemia/reperfusion injury, 11) organ transplantation, 12) psoriasis, 13) rheumatoid arthritis, 14) stroke and ischemic brain trauma, 15) trauma-induced organ injury, 16) thrombosis, 17) reduction of tumor metastasis and/or tumor growth, and the like.

Background of the Invention The immune response relies on the ability of specialized immune cells--leukocytes and lymphocytes--to migrate to sites of tissue damage, infection, or other insult to the body. Once there, these cells mount a defense against the intruding organism, help to repair the injured tissue, and protect the body from further damage. The immune system is also in constant"surveillance mode". Circulating lymphocytes monitor

the body for pathogens by migrating through lymphoid tissues, where they can be exposed to antigens and become activated.

In order for these processes to occur, various chemoattractants, cytokines, and cell adhesion molecules (CAMs) act in a programmed, sequential manner to form what has been termed the leukocyte-endothelial cascade (Tedder et al., FASEB 9: 866 (1995), Albelda et al., FASEB 8: 1756, (1994)).

Three known families of CAMs participate in this cascade: the selectins, the integrins and the immunoglobulin superfamily.

The first step, rolling of leukocytes and lymphocytes along the blood vessel wall, is mediated by the selectins.

Selectins are a small family of transmembrane glycoproteins that bind to cell surface carbohydrate ligands (for reviews see: Lasky, Science 258: 964 (1992); McEver, Curr.

Opin. Immun. 6: 75 (1994); McEver, J. Biol. Chem. 270: 11025 (1995)). To date, three members have been identified: P-selectin (expressed on platelets and vascular endothelial cells, L-selectin (on leukocytes), and E-selectin (on vascular endothelial cells).

Common structural features include a calcium-dependent (C- type) lectin domain, an epidermal growth factor (EGF)-like domain, and a series of short consensus'complement regulatory protein'repeat sequences. Rodent homologs have

been cloned and they share a high degree of sequence homology with their human counterparts.

Several selectin counter-receptors have been identified (for review see: Lasky et al., in CellularAdhesion : Molecular Definition to Therapeutic Potential, Metcalf et al., Eds. pp. 37-53 (1994) and the like). L-selectin binds to at least three different ligands: Glycam-1, CD34 and MAdCAM-1, each being expressed on different tissues. P-selectin has been found to bind to PSGL- 1, and E-selectin has been found to bind to ESL-1. These cell- surface selectin ligands are capped with clusters of oligosaccharides (for discussion see: Rosen et al., Curr. Opin.

Cell Biol. 6: 663 (1994), and Bertozzi et al., Chemistry. 8s Biology 2: 703 (1995)). The specific carbohydrate moieties necessary for selectin binding have been identified: the sialylated and fucosylated tetrasaccharide sialyl Lewis X (sLeX), and a related structure sialyl Lewis a (sLea), are common motifs recognized by all three selectins.

Although leukocyte recruitment into the tissue is a normal, indeed essential, component of the immune response, excessive and uncontrolled recruitment results in inflammatory disease. As adherence of immune cells to vascular endothelium is a critical event in the pathogenesis of acute inflammation,

modulation of selectin function is indicated in the management of diseases and disease states as described below.

Selectin function can be modulated by altering cell- surface expression, by competitive inhibition, or by shedding/cleavage from the cell surface (Diaz-Gonzalez, et al., J. Clin. Invest. 95: 1756 (1995) ; Whelan, Trends Biochem. Sci.

21 (1996)). While they have been identified as inhibitors of selectin-ligand interactions in vitro, compounds of Formula 1 may reduce inflammation in vivo via any or all of these modes.

Accordingly, the compounds of the present invention, which exhibit inhibitory activity against the selectins, are indicated in the treatment or management of the foregoing diseases (references supporting each indication are noted): 1) acute respiratory distress syndrome (ARDS) (Carraway et al., Am. J. Respir. Crit. Care Med. 157: 938 (1998); Moss et al., Crit. Care Med. 24: 1782 (1996) and others); 2) diseases that may be controlled via inhibition of angiogenesis (Koch et al, Nature 376: 517-519 (1995); Detmar et al., J.

Invest. Dermatol. 111: 1 (1998) ; Nguyen et al, Nature 365 : 267- 269 (1993)); 3) asthma (Gundal et al., J. Clin. Invest. 88: 1407 (1991) ; DeSanctis et al., J. Appl. Physio. 83: 681, (1997); Kogan et

al., J. Med. Chem. 41: 1099 (1998); PRNewswire, Sept. 9, 1998); 4) atherosclerosis (Dong et al., J. Clin. Invest. 102: 145 (1998); Frijns et al., Stroke 28: 2214 (1997); Tenaglia et al., Am. J.

Cardio. 79: 742 (1997); Zeitler et al., Eur. J. Med. Res. 2: 389 (1997), and others); 5) atopic dermatitis, contact dermatitis, and cutaneous inflammation (Teixeira and Hellewell, J. Immunol. 161: 2516 (1998) ; Staite et al., Blood 88: 2973 (1996); Todderud et al., J.

Pharmacol. Exp. Therap. 282: 1298 (1997); Ohnishi et al., Immunopharmacol. 34: 161 (1996), and the like); 6) bowel inflammation (Schurmann et al., Gut 36: 411 (1995); Koizumi et al., Gastroenterology 103 : 840 (1992); Bhatti et al., Gut 43 : 40 (1998); Cellier et al., Eur. J. Gastroenterol. Hepatol.

9: 1197 (1997)); 7) diabetes/diabetes-associated pathologies (Kunt et al., Exp.

Clin. Endocrinol. Diabetes 106: 183 (1998); Kopp et al., Exp.

Clin. Endocrinol. Diabetes 106: 41 (1998); Albertini et al., Diabetes Care 21 : 1008 (1998); Bannan etal., Diabetologica 41: 460 (1998), and others); 8) Grave's disease and associates conditions (Hara et al., Endocr.

J. 43: 709 (1996); Pappa et al., Clin Exp. Immunol. 108: 309 (1997); (Miyazaki et al., Clin. Exp. Immunol. 89: 52 (1992);

Aubert et al., Clin. Immunol. Immunopathol. 76: 170 (1995), and the like); 9) multiple sclerosis (MS) (McDonnell et al., J. Neuroimmunol.

85: 186 (1998)); Washington et al., Ann. Neurol. 35: 89 (1994); Vora et al., Mult. Scler. 3: 171 (1997); Archelos et al., J. Neurol. Sci. 159: 127 (1998)) ; 10) myocardial ischemia/reperfusion injury (reviewed in Lefer, Ann Thorac Surg. 60: 773-777 (1995), also Yamada et al., Eur.

J. Pharmacol. 346 217 (1998), Kilgore et al., J. Pharmacol.

Exp. Ther. 284: 427 (1998); Lefer et al., Circulation 90 : 2390 (1994)); 11) organ transplantation (Naka et al., Proc. Natl. Acad. Sci.

94: 757 (1997); Andreassen et al., Am. J. Cardio. 81: 604 (1998); Koo et al. Am. J. Pathol. 153: 557 (1998); Dulkanchainun et al., Ann. Surg. 227: 832 (1998); Takada et al., Transplantation 64: 1520 (1997); Brandt et al., Eur. J.

Cardiothorac. Surg. 12: 781 (1997) ; Garcia-Criado et al., J.

Surg. Res. 70: 187 (1997)); 12) psoriasis (Veale et al., Br. J. Dermatol. 132: 32 (1995); Bonifati et al., Dermatol. 190: 128 (1995); Danno et al., J.

Dermatol. Sci. 13: 49 (1996)); 13) rheumatoid arthritis (Veale and Maple, Drugs Aging 9 : 87 (1996); Hersmann et al, Cell Adhesion Comm. 6: 69 (1998);

Walter and Issekutz, Eur. J. Immunol. 27: 1498 (1997); Ertenli et al., J. Rheumatol. 25: 1054 (1998) and others); 14) stroke and ischemic brain trauma (Suzuki et al., Neurosci. Lett. 13: 151 (1997); Connolly et al., Circ. Res. 81: 304 (1997); Morikawa et al, Stroke 27: 951 (1996)); 15) trauma-induced organ injury (Simons et al., J. Trauma 41 : 653 (1996), Cocks et al., J. Trauma 45: 1 (1998); Mulligan et al., Nature 359: 843 (1994); Rubio-Avilla et al., J. Trauma 43: 313 (1997) and others); 16) thrombosis (Minamino et al., J. Clin. Invest. 101: 1643 (1998); (Downing et al., J. Vasc. Surg. 25: 816 (1997) and the like); 17) reduction of tumor metastasis and/or tumor growth (Hebbar et al., Proc. Amer. Assoc. Cancer Res. 39: 501, (1998); Khatib et al., Proc. Amer. Assoc. CancerRes. 39: 501, (1998); Kim et al., Proc. Natl. Acad. Sci. USA. 95: 9325-9330 (1998) ; El-Hariry et al., Exp. Opin. Invest. Drugs 6: 1465-1478 (1997), and others).

Comparison with other Selectin-Ligand Inhibitors/Antagonists Sialyl-LewisX analogs/mimetics reported in the literature include :'GSC-150' (Kanebo) which has been

reported to have ICso values of 280 µM, 100 µM, and 3011M against E-, P-, L-selectin respectively when assayed using an ELISA assay (Tsujishita et al., J. Med. Chem. 40: 362 (1997)); TBC-1269 (Texas Biotech) which has been reported to have ICso values of 5001lM, 701lM, and 56011M against E-, P-, and L-selectin respectively, when assayed using a cell adhesion assay (Kogan et al., J. Med. Chem. 41: 1099 (1998)); a macrocyclic derivative, which has an ICso of 390 ru against E-selectin (Kolb, Bioorg. Med. Chem. Lett. 7: 2629 (1997)); and C-mannose derivatives which have ICso values of 100- 160 RM against E-selectin (Marron et al., Tet. Lett. 37: 9037 (1996)). Some of the most potent derivatives that have been reported are multivalent sialyl-LewisX analogs which have ICso values of-1 nM in an L-selectin cell adhesion assay (Renkonen et al., Glycobiology 7 : 453 (1997)).

Some additional sugar based inhibitors of interest include inositol hexakisphosphate (IP-6) and sulfated galactocerebrosides ("sulfatides"). IP-6 has been reported to have ICso values of 1601lM and 2 jj, M, against P-and L- selectin respectively, in competition ELISA assays (Cecconi et al., J. Biol. Chem. 21: 15060 (1994)). Sulfatides have ICso values in the 0.1-1211M range when tested in a P-selectin competition ELISA assay (Marinier et al., J. Med. Chem. 40:

3234 (1997)). BMS-190394, a sulfatide analog, has been reported to have ICso values of 18 zip and 10 pM, in P-, and L-selectin cell adhesion assays respectively (Todderud et al., J. Pharmacol. Exp. Therap. 282: 1298 (1997)). Mannose- containing natural products showed inhibition of P-selectin with an ICso value of 60 p, M (Ikeda et al., Bioorg. Med. Chem.

Lett. 7: 2485 (1997)).

Non-carbohydrate inhibitors include peptides based on a conserved region of the lectin domain of the selectins, which have activity in P-and E-selectin cell adhesion assays with ICso values of ~2011M (Briggs et al., Glycobiology 5: 583 (1995)). Additional peptides, discovered by random screening, have ICso values of 5-10 J. M in an E-selectin cell adhesion assay (Martens et al., J. Biol. Chem. 270: 21129 (1995)).

Summary of the Invention The present invention is based on the discovery that compounds of Formula 1 are inhibitors or modulators of selectins which render them particularly useful for the treatment or management of a large number of disease states in which the role of selectins has directly or indirectly been implicated.

It has been found that the requisite selectin modulating activity can be obtained by employing thiazole template which acts as a framework, to which one can attach the necessary appendages that are required for activity. In order to obtain the desired selectin modulating activity the appendant groups that must be attached to the central thiazole template are 1) a carboxylic acid moiety as defined in Group I, or a carboxylic acid isostere; or other calcium binding moiety which will be apparent to those skilled in the art; and 2) a hydrophobic moiety such as a Cl2H2s alkyl chain. Additional substitution about the central core thiazole is necessary to modify the potency, selectivity and physiological properties, of the compounds claimed herein.

Accordingly, an object of the present invention is to provide a method for inhibiting or modulating selectins in a mammal by the administration of compound according to Formula 1.

Another object of the present invention relates to pharmaceutical compositions containing an effective inhibiting amount of compound according to Formula 1. These compounds have the following general structural formula 1: Where at least one of R1, R2 or R3 = a calcium binding moiety e. g. C02H etc.

Formula 1

Definitions As used herein, the term"attached"signifies a stable covalent bond, certain preferred points of attachment being apparent to those skilled in the art.

The terms"halogen"or"halb"include fluorine, chlorine, bromine, and iodine.

The term"alkyl"includes Cl-Cl6 straight chain saturated, Cl-Cl6 branched saturated, C3-C8 cyclic saturated and Cl-C16 straight chain or branched saturated aliphatic hydrocarbon groups substituted with C3-C8 cyclic saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, this definition shall include but is not limited to methyl (Me), ethyl (Et), propyl (Pr), butyl (Bu), pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, isopropyl (i-Pr),

isobutyl (i-Bu), tert-butyl (t-Bu), sec-butyl (s-Bu), isopentyl, neopentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropylmethyl, and the like.

The term"alkenyl"includes Ca-C16 straight chain unsaturated, Cz-Cn branched unsaturated, Cs-Cs unsaturated cyclic, and Ca-Cie straight chain or branched unsaturated aliphatic hydrocarbon groups substituted with Cs-Cs cyclic saturated and unsaturated aliphatic hydrocarbon groups having the specified number of carbon atoms. Double bonds may occur in any stable point along the chain and the carbon- carbon double bonds may have either the cis or trans configuration. For example, this definition shall include but is not limited to ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, 1,5-octadienyl, 1,4,7-nonatrienyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, ethylcyclohexenyl, butenylcyclopentyl, 1-pentenyl- 3-cyclohexenyl, and the like.

The term"alkyloxy" (e. g. methoxy, ethoxy, propyloxy, allyloxy, cyclohexyloxy) represents an alkyl group as defined above having the indicated number of carbon atoms attached through an oxygen bridge.

The term"alkylthio" (e. g. methylthio, ethylthio, propylthio, cyclohexylthio and the like) represents an alkyl

group as defined above having the indicated number of carbon atoms attached through a sulfur bridge.

The term"alkylamino"represents one or two alkyl groups as defined above having the indicated number of carbon atoms attached through an amine bridge. The two alkyl groups maybe taken together with the nitrogen to which they are attached forming a cyclic system containing 3 to 8 carbon atoms with or without one Cl-Cl6 alkyl, aryl Co-Cl6 alkyl, or Co-Ciealkylaryl substituent.

The term"alkylaminoalkyl"represents an alkylamino group attached through an alkyl group as defined above having the indicated number of carbon atoms.

The term"alkyloxy (alkyl) amino" (e. g. methoxy (methyl) amine, ethoxy (propyl) amine) represents an alkyloxy group as defined above attached through an amino group, the amino group itself having an alkyl substituent.

The term"alkylcarbonyl" (e. g. cyclooctylcarbonyl, pentylcarbonyl, 3-hexylcarbonyl) represents an alkyl group as defined above having the indicated number of carbon atoms attached through a carbonyl group.

The term"alkylcarboxy" (e. g. heptylcarboxy, cyclopropylcarboxy, 3-pentenylcarboxy) represents an alkylcarbonyl group as defined above wherein the carbonyl is in

turn attached through an oxygen.

The term"alkylcarboxyalkyl"represents an alkylcarboxy group attached through an alkyl group as defined above having the indicated number of carbon atoms.

The term"alkylcarbonylamino" (e. g. hexylcarbonylamino, cyclopentylcarbonyl-aminomethyl, methylcarbonylaminophenyl) represents an alkylcarbonyl group as defined above wherein the carbonyl is in turn attached through the nitrogen atom of an amino group. The nitrogen group may itself be substituted with an alkyl or aryl group.

The term"aryl"represents an unsubstituted, mono-, di- or trisubstituted monocyclic, polycyclic, biaryl and heterocyclic aromatic groups covalently attached at any ring position capable of forming a stable covalent bond, certain preferred points of attachment being apparent to those skilled in the art (e. g. 3-indolyl, 4-imidazolyl). The aryl substituents are independently selected from the group consisting of halo, nitro, cyano, trihalomethyl, C1-16 alkyl, arylCl-16 alkyl, Co-16 alkyloxy Co-i6 alkyl, aryl Co-i6 alkyloxy Co-i6 alkyl, Co-i6 alkylthio Co-i6 alkyl, aryl C0-16 alkylthio C0-16 alkyl, C0-16 alkylamino C0-16 alkyl, aryl Co-i6 alkylamino Co-i6 alkyl, di (aryl Cl-16 alkyl) amino Co-i6 alkyl, C1-16 alkylcarbonyl Co-i6 alkyl, aryl Cl-16 alkylcarbonyl Co- 16 alkyl, C1-16 alkylcarboxy Co-i6 alkyl, aryl Cl-16 alkylcarboxy Co-

16 alkyl, Cl-16 alkylcarbonylamino Co l6 alkyl, aryl Cl l6 alkylcarbonylamino Co-m alkyl,-Co-16 alkyl COORl,-Co-i6 alkyl CONR2R3 wherein Ri, R2 and R3 are independently selected from hydrogen, Cl-Cllalkyl, arylCo-Clalkyl, or R2 and R3 are taken together with the nitrogen to which they are attached forming a cyclic system containing 3 to 8 carbon atoms with or without one Cl-Cl6 alkyl, aryl Co-Cl6 alkyl, or Co-Cl6 alkylaryl substituent.

The definition of aryl includes but is not limited to phenyl, biphenyl, naphthyl, dihydronaphthyl, tetrahydronaphthyl, indenyl, indanyl, azulenyl, anthryl, phenanthryl, fluorenyl, pyrenyl, thienyl, benzothienyl, isobenzothienyl, 2,3-dihydrobenzothienyl, furyl, pyranyl, benzofuranyl, isobenzofuranyl, 2,3-dihydrobenzofuranyl, pyrrolyl, indolyl, isoindolyl, indolizinyl, indazolyl, imidazolyl, benzimidazolyl, pyridyl, pyrazinyl, pyradazinyl, pyrimidinyl, triazinyl, quinolyl, isoquinolyl, 4H-quinolizinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, chromanyl, benzodioxolyl, piperonyl, purinyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, benzthiazolyl, oxazolyl, isoxazolyl, benzoxazolyl, oxadiazolyl, thiadiazolyl.

The term"arylalkyl" (e. g. (4-hydroxyphenyl) ethyl, (2- aminonaphthyl) hexyl, pyridylcyclopentyl) represents an aryl group as defined above attached through an alkyl group as defined above having the indicated number of carbon atoms.

The term"carbonyloxy"represents a carbonyl group attached through an oxygen bridge.

In the above definitions, the terms"alkyl"and"alkenyl" maybe used interchangeably in so far as a stable chemical entity is formed, as obvious to those skilled in the art.

The compounds of the present invention also includes racemic mixtures, stereoisomers and mixtures of said compounds, including isotopically-labeled and radio-labeled compounds (Goding ; Monoclonal Antibodies Principles and Practice ; Academic Press, p. 104 (1986)). Such isomers can be isolated by standard resolution techniques, including fractional crystallization and chiral chromatography (Eliel, E. L. and Wilen S. H.; Stereochemistry in Organic Compounds ; John Wiley Sons, New York, (1993)).

The term"therapeutically effective amount"shall mean that amount of drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor or other clinician.

Detailed Description This application relates to compounds having the general Formula 1. Accordingly, an object of the present invention is to provide a method for inhibiting or modulating selectins in a mammal by the administration of a compound according to the general Formula 1 as defined below. In addition, this application relates to the preparation of said compounds, to compositions comprising the compounds, to their use for treating human or animal disorders, to their use for purification of proteins, and to their use in diagnostics or medical devices.

1 3<BR> R Where at least one of Ru, R2 or R3 contains calcium binding moiety e. g. C02H etc.

Formula 1 The present invention relates to compounds having general Formula 1. The following R Group substitution patterns

for R1, R2 and R3 are within the scope of this invention. Thus; Case A: When R1 is selected from Group I one of R2 or R3 must be selected from Group II, and one of R2 or R3 must be selected from Group III.

Case B: When R3 is selected from Group I one of R1 or R2 must be selected from Group II, and one of R1 or R2 must be selected from Group III.

Case C: When R2 is selected from Group I one of R1 or R3 must be selected from Group II, and one of R1 or R3 must be selected from Group III, where Groups I, II and III are defined below.

Definitions of Group I through Group III Group I is defined in Figure 1, Table 1, below: GO (OH or *N-linked amino acid) C02H Group I = X or ^ or fJ /

where R6 is selected from the following in Table 1: Figure 1 Table 1 R6 Atom or group Type Template X Y Z R7 RS R9 R8 I i R9 R7 C N CH =0 H (CH2) nx0H R9 ruz la P8 1 R9-zX' 0 CH2 00--*Aminoacid if R 2a (N-linked) R$ 1 F29-ZYX O CH2 C=O--NCH2CHS03H 111 tu 2b R Table 1 (cont.) w f R6 Atom or group Type Template x Z R7 R3 R9 R$ i iv R9 z, X ; O CH2 C=O N (Mc) CH2 2c R CO2H R8 R9-Z 0 CH2 C=O NHCH2- 7 2d R P (O) (OH) OEt , R8 i R9 0 CH2 C=O N (C (CH20H) 3 2e R7 2eR (CH2)2SO3H R8 1 Vii HO2C IX<} CH (CH2) n (cH2) ntco2H 3a R pu R8 viii H02CY\X N C H =0 3b R7 , R8 R8 ix HO CYX4g CH CH-OH OH 3c R 3c R R8 HOsCX N (CH2) n'--H 3d R$ 1 Xi HO2C lXt3Rg ° (CH2) n' Ruz 3e R Table 1 (cont.) R6 Atom or group Type Template x z R7 R8 R9 H02C, x". I 4a R C--=O-- all HOC,, xiv 4C R7 CH-OH xiii CH---OH xi 4c H02C, x"I 4d R CH--NH2 xv OH or *amino acid /\ - Sa Any a or (3, 0- _ _ linked _ _ Any a or p, 0- linked xvii 0-hnked Mon- 5b sacharide xviii HO2C x 6a In the above table n", and/or n'and/or n can be 0,1,2,3,4,5

or 6; *D or L natural or unnatural single aminoacid or dipeptide where the amino acid is selected from, 4-

hydroxyproline, cystein, serine, threonine, glycine, glutamine, asparagine, glutamic acid, aspartic acid, valine, alanine, iminodiacetic acid, 4-amino-2-hydroxy-butanoic acid and 4- amino-3-hydroxy-butanoic acid.

Group II is defined as one of the following: Group II : (i) Unsubstituted, mono-, di-, or tri-substituted aryl-Co ll alkyl wherein aryl is selected from the group consisting of phenyl, pyridino, wherein the substituents are selected from the group consisting of: (a) Halo, hydroxy; or (b)C0-6CO2R10, C0-6CONHR10, C0-6NHSO2R10, trans- CH=CHCO2R1°, or trans-CH=CHCONHRl°, wherein Rio is C1-16alkyl, C1-16 alkyloxyalkyl, C5-8 cycloalkyl, C1-11 alkylaryl, or Cl-5 alkylaryl C1 s alkyl in which the said alkyl group or said aryl group such as phenyl, thienyl, imidazoyl, indolyl, furyl or pyridyl, are unsubstituted, mono-or disubstituted with a member selected from the group consisting of hydroxy, carboxy, halo, Cl-6 alkyl and C16 alkyloxy, C16 cycloalkyloxy, C 1-C4 alkyl aryl or C1-C4 alkoxy aryl such as phenyl, or trans-2- phenylethenyl, 2-phenylethynyl, or 2-phenylethyl, in which said aryl group is either unsubstituted, mono- or disubstituted with a member selected from the group consisting of hydroxy, halo, C1-4 alkyl, or C1-4 alkyloxy; or Rio can be N-Boc-piperidino, or N- carboethoxypiperidino;

Group III is defined as either: Group III: (i) Hydrogen; or (ii) Unsubstituted, mono or disubstituted C1-16 alkyl, C0-16 alkylamino, Co-i6 alkyloxyalkyl or C2-16 alkenyl wherein the substituents are independently selected from the group consisting of hydroxy, C1-s alkyl, C1-s alkyloxyalkyl, C1-8 alkylthioalkyl, phenyl-C1-8 alkylamino, CI-8 alkoxycarbonyl; or C0-6 carboxyl, triazole, 2,3- (methylenedioxy) benzyl; or (iii) substituted or unsubstituted N or C-linked pyrrolidino, piperidino, piperidonyl, morpholino, piperazino, N-Boc- piperazino, N-C1-10 alkylpiperazino, N-C3-6 alkenylpiperazino, N-(C1-6 alkoxy C1-6 alkyl) piperazino, N- (C1-6 alkoxy C3-6 alkenyl) piperazino, N-(C16 alkylamino C 6 alkyl) piperazino, N-(C1-6 alkylamino C3-6 alkenyl) piperazino, wherein the substituents are N or C- linked as will be apparent to one skilled in the art, and are independently selected from: (a) substituted Cl-i6 alkyloxy, C3-16 alkenyloxy, substituted C3-16 alkynyloxy; or (b) substituted C16 alkyl-amino, di (substituted C1-6 alkyl) amino; or

(c) C3-6 alkenyl-amino, di (Cs-6 alkenyl) amino, substituted C3-6 alkenyl-amino, di (substituted C3-6 alkenyl) amino; or (d) CONHC1-Cl6 alkyl, COOCl-Cl6 alkyl, Co ll alkylCO2H, C0-11NHC(O)NHR11, C0-11NHSO2R11, trans- CH=CHCO2R11, or trans- CH=CHCONHR11 wherein R11 is C1-16 alkyl, or C1-16 alkyl aryl, in which the said aryl group such as phenyl, or pyridyl, is mono-or disubstituted with a member selected from the group consisting of hydroxy, halo, C1-6 alkyl and C1-6 alkyloxy, C16 cycloalkyloxy, or C1-C4 alkyl aryl or C1-C4 alkoxy aryl in which said aryl group is either unsubstituted, mono-or disubstituted with a member selected from the group consisting of hydroxy, halo, C1-4 alkyl, Ci- 4 alkyloxy, and aryl; or (e) pyrrolidino, piperidino, morpholino, imidazolyl, substituted, uracil or other purine or pyrimidine heterocycles, piperazino, N-C1-6 alkylpiperazino, N-C3-6 alkenylpiperazino, N- (C1-6 alkoxy C1-6 alkyl) piperazino, N-(C1-6 alkoxy C3-6 alkenyl) piperazino, N-(C1-6 alkylamino Ci-6

alkyl) piperazino, or N-(C1-6 alkylamino C3-6 alkenyl) piperazino, where the substituents are chosen from hydroxy, Cl-12 alkylalkoxy, Ci-12 alkylamino, Cs-12 alkenyloxy, or ¬3-12 alkenylamino; or (iv) either mono-, di, or tri-substituted aryl, or Co-Cl2 aryl such as phenyl, N-trityl-imidazolyl, furanoyl, pyrimidino, pyridino, or N or C-linked pyrrole or imidazolyl, wherein the substituents are independently selected from those listed above in Group III section (iii) (a) to (e), or a C-linked, N-substituted pyrrole substituted with either- CHaCONHCsHs-O-Fucose or-CH2CONHC2H5-O-Mannose.

And the corresponding pharmaceutically acceptable salts and esters thereof.

Detailed Description The present invention related to compounds of the general Formula 1.

More particularly, the present invention relates to compounds of formulas (46-67), as shown in Figure 2, or pharmaceutically acceptable salts or alternative esters thereof : Figure 2.

Example 1 Example 2 Example 3 Example 4 CO2Et 48 49

Figure 2 (cont.) H03S OH Example 7 0 HOC. ' Example 0 NH HN O C6H13 NH HO HO 52 HO Hp N 53 N S NH OH HO2C v NH H02C\ J H02C,, Example 9 o) Example 10 NH HN o O 0 C6H13 L tNH C6H N-S 02 f C6H13 S 54 N NtS 5 . r 55 nu 0 H O J w HO2CA Example 11 otH Example 12 O Hz O 0 HN-f \ N S 56 Nu NH \ NH Figure 2 (cont.) S03H cOzhbH o C02FbH Example 13 Example 14 N OEt HN O O HN 0 N S Nq, S O- \ \ Nez 58 59 NH NH HOZC, Example 16 OOH Examole 15 O H02C1',, \yH H NHC6H3 NNO p nu S r N S N 61 tNH OH H02C\ Example 17 HO2Co"I Example 18 HN, 0 0..,', HN O O O NHC6H3 HN\/p 'Y N H C6H13 N tS 62 N eS 63 I NH N-\ 0- Figure 2 (cont.) HO2C ° Example 19 HO C NH2 Example 20 , HNo4O O HN, o ° ro-NHC6Hl3 0 NHC6H13 64 -064 N S N S 65 NH NH OH \=/SH \=/ H02C) Example 21 H02C) \l Example 22 HN 0 NH HN 0 0 NHC6H T-ro-13 O O NtzS 66 N8, S 67 FNH ANH

The compounds depicted in Figure 2 are named as follows: Example 1 5- [4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-4- (4- ethoxycarbonyl methoxy-phenyl)-thiazole-2-carboxylic acid ethyl ester 46 Example 2 <BR> <BR> <BR> <BR> <BR> <BR> <BR> (4-f 5- [4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-thiazol-4-yl}- phenoxy)-acetic acid 47 Example 3 4- (4-Ethoxycarbonylmethoxy-phenyl)-5- (4- { (E)-2- [1- (4- pentyl-phenyl)-ethylcarbamoyl]-vinyl}-phenyl)-thiazole-2- carboxylic acid ethyl ester 48 Example 4 {4-[5-(4-{(E)-2-[1-(4-Pentyl-phenyl)-ethylcarbamoyl]-vinyl}- phenyl)-thiazol-4-yl]-phenoxy}-acetic acid 49 Example 5 <BR> <BR> <BR> <BR> <BR> <BR> <BR> (4- {5- [4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2-pyridin-3- yl-thiazol-4-yl}-phenoxy)-acetic acid ethyl ester 50 Example 6 <BR> <BR> <BR> <BR> <BR> <BR> (4- 5- [4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2-pyridin-3- yl-thiazol-4-yl}-phenoxy)-acetic acid 51

Example 7 <BR> <BR> 2- [ ( {4- [5- [4- (2-Hexylcarbamoylvinyl) phenyl]-2- (lH-pyrrol-2- yl) thiazol-4-yl] phenoxy} acetyl)- (2-hydroxy-1, 1- bishydroxymethylethyl) amino] ethanesulfonic acid 52 Example 8 (S)-3-Hydroxy-2- [2- [4- [5- [4- (2-octylcarbamoylvinyl) phenyl]-2- (lH-pyrrol-2-yl) thiazol-4-yl] phenoxy] acetylamino] propionic acid 53 Example 9 (R)-3-Hydroxy-2- [2- [4- [5- [4- (2-hexylcarbamoylvinyl) phenyl]- 2- (lH-pyrrol-2-yl) thiazol-4- yl] phenoxy] acetylamino] propionic acid 54 Example 10 (2S)-3-Hydroxy-2- [2- [4- [5- [4- (2-hexylcarbamoylvinyl) phenyl]- 2- (morpholin-4-yl) thiazol-4- yl] phenoxy] acetylamino] propionic acid 55 Example 11 (2- [4- [5- [4- (2-Octylcarbamoylvinyl) phenyl]-2- (lH-pyrrol-2- yl) thiazol-4-yl] phenoxy] acetylamino) acetic acid 56 Example 12 <BR> <BR> 3-Hydroxy-2- [2- (2- [4- [5- [4- (2-octylcarbamoylvinyl)-phenyl]-2- (lH-pyrrol-2-yl) thiazol-4- yl] phenoxy] acetylamino) acetylamino] propionic acid 57

Example 13 (2S)-4- [3- [4- [4- [4- [ (l-Carboxy-2-hydroxy- ethylcarbamoyl) methoxy-phenyl]-2- (lH-pyrrol-2-yl)-thiazol- 5-yl]-phenyl]-acryloylamino]-piperidine-1-carboxylic acid ethyl ester 58 Example 14 <BR> <BR> 2- (2- {4- [5- [4- (2-Hexylcarbamoyl-vinyl)-phenyl]-2- (lH-pyrrol-<BR> <BR> 2-yl)-thiazol-4-yl]-phenoxy}-acetylamino)-ethanesulfonic acid 59 Example 15 <BR> <BR> 2- (2- {4- [5- [4- (2-Hexylcarbamoylvinyl) phenyl]-2- (lH-pyrrol-2- yl) thiazol-4-yl] phenoxy} acetylamino)-3-hydroxybutyric acid 60 Example 16 (S)-3-Hydroxy-2- [2- [4- [5- [4- (2-Phenylbutylcarbamoyl) phenyl]- 2-(1H-pyrrol-2-yl] thiazol-4- yl] phenoxy] acetylamino] propionic acid 61 Example 17 (2- {4- [5- [4- (2-Hexylcarbamoylvinyl) phenyl]-2- (lH-pyrrol-2- yl) thiazol-4-yl] phenoxy} acetylamino) acetic acid 62 Example 18

2- (S)- (2- {4- [5- [4- (2-Hexylcarbamoyl-vinyl)-phenyl]-2- (l-<BR> <BR> methoxymethyl-lH-pyrrol-2-yl)-thiazol-4-yl]-phenoxy}- acetylamino)-3-hydroxy-propionic acid 63 Example 19 <BR> <BR> 4- (2- {4- [5- [4- (2-Hexylcarbamoyl-vinyl)-phenyl]-2- (lH-pyrrol-<BR> <BR> 2-yl)-thiazol-4-yl]-phenoxy}-acetylamino)-3-hydroxy-butyric acid 64 Example 20 4-Carbamoyl-2- (2- {4- [5- [4- (2-hexylcarbamoylvinyl) phenyl-2- (lH-pyrrol-2-yl) thiazol-4-yl] phenoxy} acetylamino) butyric acid 65 Example 21 3-Hydroxy-2- (2- {4- [5- [4- (2-phenethylcarbamoylvinyl) phenyl]- 2- (lH-pyrrol-2-yl) thiazol-4- yl] phenoxy} acetylamino) propionic acid 66 Example 22 <BR> <BR> 2- (2- {4- [5- [4- (2-hexylcarbamoylvinyl) phenyl]-2- (lH-pyrrol-2- yl) thiazol-4-yl] phenoxy} acetylamino)-3-mercaptopropionic acid 67

The compounds of the current invention may have asymmetric centers and can occur as racemates, racemic mixtures, and as individual enantiomers or diastereomers, with all isomeric forms being included in the present invention as well as mixtures thereof.

Pharmaceutically acceptable salts of the compounds above, where a basic or acidic group is present in the structure, are also included within the scope of this invention. When an acidic substituent is present, such as-CO2H, there can be formed the ammonium, sodium, potassium, calcium salt, and the like, for use as the dosage form. Basic groups, such as amino or basic heteroaryl radicals, or pyridyl and acidic salts, such as hydrochloride, hydrobromide, acetate, maleate, palmoate, methanesulfonate, p-toluenesulfonate, and the like, can be used as the dosage form.

Also, in the case of the-CO2H being present, pharmaceutically acceptable esters can be employed, e. g., methyl, ethyl, tert-butyl, pivaloyloxymethyl, acetoxymethyl, and the like, and those esters known in the art for modifying solubility or hydrolysis characteristics for use as sustained release or prodrug formulations.

In addition, some of the compounds of the instant invention may form solvates with water or common organic solvents. Such solvates are encompassed within the scope of the invention.

The term"therapeutically effective amount"shall mean that amount of drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor or other clinician.

Synthetic Procedures General references to methodologies for the synthesis of the compounds of the present invention are described in the following references 1) Drayton, C. J., Comprehensive Heterocyclic Chemistry, 1st ed; Pergamon: Oxford, 1984 and 2) Joule, J. A.; Mills, K.; and Smith, G. F., Heterocylic Chemistry, 3rd ed; Chapman and Hall, 1995.

More specific examples of and references to methodologies for the preparation of thiazoles can be found in Gauthier et al, Bioorg. & Med. Chem., 6,87-92, (1996); Maduskuie et al, J. Med. Chem., 38, 1067-1083 (1995).

Scheme 1 Thiazoles 9 can be synthesized through a common a- halocarbonyl intermediate as illustrated in Scheme 1 (the Hantzsch synthesis). The required intermediates for Scheme 1 are accessible from readily available starting materials, and can be synthesized as shown below (Scheme 2): carboxylic acids amino acids mandelic acids For ketones (Scheme 1) :

Scheme 2 A more specific example is shown in Scheme 3. Acid 16 is converted to the acyl chloride via treatment with thionyl chloride. Ketone 17 is then synthesized via a Friedel-Crafts alkylation reaction. Derivatization of ketone 18 can take place at this at this stage to give the phenoxyactetic acid derivative 19 in excellent yield. Ketobromide 20 can be synthesized via treatment of 19 with bromine. Reaction of 20 with a thioamide 21 gives the desired thiazole 22. This route is very high yielding and has been used routinely on a multigram scale. Br MeO Br HO Br 4 1. SOC12 t t L > 2. AIC13, Anisole Collidine 3 16 90% ° 17 81% Ó 18 BrCH2C02Et or BrCH2CO2fBu Cs2CO3, 82% brio Br R 02C Br2 R 02C quant. quant. 20 r R1 =tBu orEt o 19 NHsR r R R 02C > 4 00/-\ 22 N9, S 3 R

Scheme 3 Examples of how thiazole intermediates such as 22 or 23 can be derivatized, are shown in Scheme 4. General thiazole 23 can undergo a Heck reaction with an acrylate 24 to give the corresponding acid (or protected acid ester) 25 which can then be derivatized to thiazole 27. Alternatively thiazole 23 can undergo a Heck reaction with an acrylamide 26 to give thiazole 27 directly. 0, gO Br SoR2 Et02C / Heck Reaction Etp2C etc V5 8 I I 23 NYS p 24 25 N S R2'= H or ester R't R 3 Y, Heck Reaction 1. Deprotection (if necessary) <NHR3 2. EDCI, DMAP 26 NH2R2 0 SEHR2 /=/ Etc N S 27 R3 Ester Hydrolysis 0 NHR2 O HCC - ( H02C Nß, S N, S T,

Scheme 4 Lawesson's HO 0 Oxalyl Chloride Cl O Acetone H2N O Rea HN S _ R Drop DMF R3 CH3CO2NH4 R3 THF, Reflux R3 29 30 31 21 Scheme 5 The required thioamide 21 can be synthesized from the acid 29 via conversion of the acid to the corresponding amide 31 followed by treatment with Lawesson's reagent to give the thioamide 21 (Scheme 5).

Scheme 6 Thiazoles 39,40, and 41 can be synthesized through a coupling reaction with several amines such as amino acids, taurine, and N-tris (hydroxymethyl) methyl-2- aminoethanesulfonic acid in the presence of WSC and HOBt as shown in Scheme 6. The amine derivatives shown here are commercially available.

Scheme for Thioamide and Thiourea Synthesis Me00 R3 RUZ R 42 45 OEt EtO-) =S NH3 aq.-MeOH SH 4N HCI-EtOAc Lawesson's reagent Oxalyl Chloride Acetone HOO CIO H2N0 THF, Reflux H2NS Drop DMF R CH3C02NH4 R3 or R3 P205 29 30 31 29 30 31 Diglyme 21 NH4SCN n-BuOH or, NH3 aq . NHgaq 1)EtOCOSCN/THF 2) conc. HCI Ruz R N H HSS R3 43 43 44 Scheme 7

The required thioamide 21 can be synthesized from the nitrile 42, or dithiocarboxylic acid 44. In addition, the thiourea (21a, R3= NH-R) can be synthesized from the coupling of ammonium thiocyanate and amines 43 as shown in Scheme 7.

The methodologies for further derivatization of thiazoles for the synthesis of the compounds of the current invention will be apparent to those skilled in the art.

Experimental Synthetic Description To further illustrate the practice of this invention, the following examples are included along with the general methods employed to synthesize the compounds described.

General Experimental Information Nuclear magnetic resonance spectra (1H-NMR) were measured on either a Varian (300 MHz) or a Varian (400 MHz).

Chemical shifts (8) are reported in parts per million (ppm) downfield from tetramethylsilane (TMS). Data are reported as follows: chemical shift, multiplicity (br=broad, s=singlet, d=doublet, t=triplet, q=quadruplet, m=multiplet), coupling constant (Hz), integration and peak assignment.

Mass spectra were measured using Atmospheric Pressure Chemical Formation (APcI) looking at positive and negative modes on a Micromass LCZ (3 KeV with a probe temperature of 400 oc and a source block at 120 oc).

LC chromatograms for LC/MS were measured using an eluant of CH3CN (0.1% CFsCO2H)/H20 (0.1% CF3CO2H) (V: V)

on a Hewlett Packard HP1100 HPLC, in the range 200-300 nm with a Diode Array Detector (DAD); 5 jj. l per injection (Gilson 215 Autosampler) at an average concentration of 1 mg/mL; gradient: 10-100% CH3CN in 5 minutes, 100% CH3CN for 1 minute, 100-10% CHsCN in 0.2 minutes, 10% CHsCN for 1.4 minutes; LC element split 1: 4 directly into ion source (500 RL/min).

The chromatography columns used for LC in LC/MS and HPLC were 50 x 4.6 mm C-8 with 5 Rm particle sizes and Zorbax 150 x 4.6 mm C-8 with 5 Rm particle sizes, respectively.

The same gradient was used in HPLC as in LC for LC/MS.

Reactions in solution phase were monitored by thin layer chromatography (TLC) using Merck silica gel 60F-254-coated plates (0.25 mm thickness). Flash chromatography was performed using E. Merck silica gel 60 (230-400 mesh ASTM).

Synthetic Methods General Methods General Method 1 : Synthesis of keto-bromide intermediate 20 (where Ri = Me) (Scheme 3) 4-Bromobenzyl-4-methoxyphenylketone 20a To a mixture of p-bromo-phenylacetic acid 16 (51g, 237 mmol, 1 equiv.), and SOC12 (35 mL, 480 mmol, 2 equiv.), was added 1 drop of DMF. The mixture was stirred at 60°C for 30 min. then concentrated under reduced pressure. The residue was dissolved in CHC13 (140 mL), and Aids (35 g, 262 mmol, 1.1 equiv.) was added to the solution portionwise at 0° C. To this mixture was added anisole (30 g, 277 mmol, 1.2 equiv.) dropwise at 0° C, and the mixture stirred at 0° C for 15 min and r. t. for 1 h. The reaction mixture was poured onto ice- water, and extracted with CHC13 (150 x 3 mL). The combined extracts were washed with sat. NaHCOs (aq.) (2 x 200 mL), and water (3 x 200 mL), dried (MgS04), and concentrated under reduced pressure. The residue was suspended in hexane, and the insoluble material collected by filtration to give 4- Bromobenzyl-4-methoxyphenylketone 17 65g (90%).

Data for Compound 17 : 1H-NMR (300 MHz, CDC1s) : 7.98 (d, 2H, J= 9.0), 7.45 (d, 2H, J= 8.4), 7.15 (d, 2H, J= 8.4), 6.94 (d, 2H, J= 9.0), 4.20 (s, 2H), 3.88 (s, 3H).

4-Bromobenzyl-4-hydoxyphenylketone 18 A mixture of 4-Bromobenzyl-4-methyloxyphenylketone 17 (65 g, 213 mmol), LiI (50 g, 374 mmol) and collidine (100 mL) was stirred at 180° C for 3 h. The reaction mixture was diluted with ethylene glycol (100 mL) and stirred at 180 ° C for 30 min. The mixture was cooled, acidified to pH 1 with dilute (1N) HCl, and extracted with EtOAc (3 x 150 mL). The combined extracts were washed with water (3 x 200 mL), Sat.

NaHCOs (200 mL), and brine (3 x 200 mL), successively, dried (MgS04), and concentrated under reduced pressure. The residue was recrystallized using EtOAc to give 4-Bromobenzyl-4- hydoxyphenylketone 18 50 g (81%).

Data for compound 18 : 1H-NMR (300 MHz, CDC13) : 7.93 (d, 2H, J= 8.7), 7.50 (d, 2H, J= 8.4), 7.41 (d, 2H, J= 8.7), 6.89 (d, 2H, J= 9. 0), 6.29 (s, 2H).

4- [4-Bromophenylacetyl] phenoxyacetic ethyl ester (Rl = Et) 19 A mixture of 4-Bromobenzyl-4-hydoxyphenylketone 18 (50 g, 172 mmol, 1.0 equiv.), ethyl bromoacetate (30 g, 180 mmol, 1.05 equiv.), Cs2CO3 (60 g, 184 mmol, 1.07 equiv.) and DMF (300 mL) was stirred at r. t. for 1 hr. The reaction mixture was diluted with water (200 mL), and the resulting solid was collected by filtration. The solid was recrystallized from EtOH

to give 4-[4-Bromophenylacetyl]phenoxyacetic ethyl ester (R1 = Et) 19 53 g (82%).

Data for Compound 19 : 1H-NMR (300 MHz, CDCIs) : 7.98 (d, 2H, J= 9.3), 7.45 (d, 2H, J= 8.1), 7.13 (d, 2H, J= 8.4), 6.95 (d, 2H, J= 9.0), 4.69 (s, 2H), 4.29 (q, 2H, J= 7.2), 4.19 (s, 2H), 1.31 (t, 3H, J = 7. 2).

{4 [Bromo- (4-bromophenyl) acetyl] phenoxy} acetic acid ethyl ester 20 To a mixture of 4- [ (4-bromophenyl) acetyl] phenoxyacetic acid ethyl ester 19 (52 g, 136 mmol) and CHCIs (400 mL) was added Br2 (7.5 mL) dropwise at 40° C, and the mixture was stirred at r. t. for 1 h. The reaction mixture was washed with Sat. NaHCOs (aq) (2x 200 mL) and water (3 x 200 mL), dried (MgS04), and concentrated under reduced pressure. The desire product was recrystallized using ethyl acetate and hexane to give {4[Bromo-(4-bromophenyl) acetyl] phenoxy} acetic acid ethyl ester 20 56 g (90%).

Data for compound 20 ; lH-NMR (300 MHz, CDCl3) : 7.98 (d, 2H, J= 9.0), 7.50 (d, 2H, J= 8.4), 7.41 (d, 2H, J= 8.4), 6.94 (d, 2H, J= 8.7), 6.26 (s, 2H), 4.69 (s, 2H), 4.28 (q, 2H, J= 7.2), 1.30 (t, 3H, J= 7. 2).

General Method 2 : Synthesis of Thiazole 22 (Scheme 3), 23 (Rl=Et) (Scheme 4), using {4 [Bromo- (4-bromophenyl) acetyl] phenoxy} acetic acid ethyl ester 20 (Rl'=Et) as alpha keto- bromide source (Scheme 3): A mixture of alpha keto-bromide 20 (1 equiv.), thioamide 21 (1.3 equiv.), NaHCOs (1.3 equiv.) and CHsCN (0.3 M of alpha keto-bromide 20) was stirred at 80 °C for 5 h. The insoluble materials were filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Hexane-EtOAc, 4: 1) to give the desired intermediate thiazole 22,23 (Rl'=Et).

General Method 3: Heck Reaction with thiazole aryl bromide 23 and acrylamide 26 to give thiazole 27 (Scheme 4): A mixture of thiazole aryl bromide 23 (1 equiv.), acrylamide 26 (1.2 equiv.), Et3N (3 equiv.), Pd (OAc) 2 (0.06 equiv.

), P (o-tolyl) 3 (0.27 equiv.) and DMF (to make 0.5 M of the thiazole) was stirred at 100° C about 5 h. The reaction mixture was diluted with water and EtOAc, and the insoluble materials were filtered off. The filtrate was extracted with EtOAc, washed with water, dried (MgS04) and concentrated to dryness. The

thiazole derivative 27 was purified by silica gel column chromatography (Hexane-EtOAc, 4: 1).

General Method 4: Hydrolysis of thiazole ethyl ester 27 to give thiazole acid 28 (Scheme 4): A mixture of thiazole ethyl ester 27 (1 equiv.), IN LiOH (15 equiv.), and 1,4-Dioxane (0.3 M of thiazole ethyl ester) was stirred at r. t. overnight. The reaction mixture was acidified with IN HCl and extracted with ethyl acetate. The ethyl acetate solution was washed with water and brine, dried (MgS04) and concentrated to dryness. The thiazole acid 28 was recrystallized using isopropyl alcohol and ethyl acetate.

General Method 5: Preparation of Thioamide 21 from carboxylic acid 29 (Scheme 5): The carboxylic acid (1 equiv., dry) was placed into a flask fitted with a reflux condenser. Oxalyl Chloride (6 equiv.) was added followed by one drop of DMF. The reaction was stirred at 60 oC for 30 min. The excess oxalyl chloride was removed under reduced pressure. Ammonium acetate (4 equiv.) was added to the acid chloride in acetone (0.4 M) and stirred overnight at r. t.. The mixture was then filtered, the solvent

evaporated under reduced pressure and the residual amide 31 recrystallized using water and ethanol.

To a solution of amide 31 (1 equiv.) in THF was added Lawesson's reagent (2 equiv.) and the mixture refluxed for 4 h.

The reaction mixture was concentrated under reduced pressure and the thioamide 21 purified by silica gel chromatography (DCM: MeOH, 95: 5).

General Method 6: Synthesis of Acrylamide 26 (Scheme 4).

Acrylamides were prepared by adding acroyl chloride (1 equiv.) to a cooled solution (0 °C) of the desired amine in dichloromethane (0.5M) with triethylamine (1.5 equiv.) as base.

These acrylamides were used directly, without purification in the Heck reaction.

General Method 7 : Preparation of Thioamides using Lawesson's Reagent To a solution of amide in THF or toluene was added Lawesson's reagent and the mixture was refluxed for 2 h. The reaction mixture was concentrated in vacuo and the residue was purified by MPLC (acetone: n-hexane) to give the required thioamide.

CONH2 CSNH2 CONH2 CSNH2 N Lawesson Reagent !. L Lawesson Reagent toluene 0 toluene 0 '- '0 toene o quant. O-J O-J y. 43.6% CONH Lawesson Reagent CSNH2 CONH2 CSNH2 Lawesson Reagent toluène y. 92. 2% N toluene y. 88. 2% CONH2 CSNH2 CONH2 CSNH2 Lawesson Reagent Lawesson Reagent THF N Ngo THF N'Boc Boc Boc y 50% y. 95% General Method 8: Preparation of Thioamide using O, O'- Diethyldithiophosphate To a solution of nitrile (2.50ml, 23.6mmol) in 4N HCl- EtOAc (30ml) was added O, O'-Diethyldithiophosphate (4.21ml, 26.6mmol) and the mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated in vacuo and the residue was purified by MPLC (ethyl acetate: n-hexane) to give thioamide (2.59g, y. 74.2%) as pale yellow syrup.

Imidazole-4-carbothioamide A solution of imidazole-4-dithiocarboxylic acid (5.0 g, 34.7 mmol) in 28% ammonia water (25 ml) was stirred at 80 °C for 6 h. The reaction mixture was concentrated, and the residual oil 52

was dissolved in 1N HCl (100 ml). The brown precipitates were filtered off, and the filtrate was made basic with Na2CO3. The solution was decolorized by activated charcoal powder (ca. 2 g), and the solution was concentrated. The residue was dissolved in MeOH, and the insoluble salts were filtered off. The solvent was evaporated. Recrystallization from water afforded thioamide (1.1 g, y. 25%) as brown solids.

Procedure using Ammonium thiocyanate The mixture of 4-aminobutyric acid methyl ester hydrochloride (3.07g 20.0mM) and ammonium thiocyanate (1.52g 20.0mM) in H20 (lOmL) was refluxed for 5hr. The solvent was removed. To the residue in MeOH was added 2.0M TMSSCH2N2 with stirring at room temperature. After stirring for 10 min. at room temperature, the precipitates were filtered off, and the filtrate was concentrated to afford 4-Thioureido-butyric acid methyl ester (4.1 g with MeOH). react solvent product MeOOC'--N" NH H quant MeOOC------NUEH H20, (w! th ref) uxforshr n S MeOH Hooc----g- TMSC , gJL- H H20 1 N NAOH (I eq.) 1N HCI (1eq.) reflux overnight n-BuOH '-gCN HN O reflux overnight HN N<O 1 3 n-BuOH "°'"\/"'reftux overnight Boc-N N Nn 18%

*The product including 4-Thioureido-butyric acid which was de-esterified was re-esterified with TMSCH2N2.

**At first, the solution was neutralized with 1N NaOH, but re-acidified with 1NHC1.

Thioformamide A mixture of posphorous pentasulfide (1.02g, 2.2mmol), formamide (0.99ml, 22mmol) and diglyme (4ml) was stirred at room temperature for 5h. The mixture was filtered to remove off insoluble marterials. The filtrate, solution of thioformamide, was used without further purification.

Example 1 5- [4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-4- (4- ethoxycarbonyl methoxy-phenyl)-thiazole-2-carboxylic acid ethyl ester 46 The ethyl ester, 5-4- ( (E)-2-Dodecylcarbamoyl-vinyl)- phenyl]-4- (4-ethoxycarbonyl methoxy-phenyl)-thiazole-2- carboxylic acid ethyl ester 46, was synthesized according to General Method 2, from a mixture of alpha keto-bromide 20 (R1'= Et) (5.0 g, 11.0 mmol, lequiv.), ethyl thiooxamate (1.53, ll. Ommol, 1.0 equiv.), NaHCOs (2.8 g, 33.0 mmol, 3 equiv.) and CHsCN (30 mL) which, after purification by silica gel column chromatography (Hexane-EtOAc, 4: 1), gave the intermediate aryl bromide 22A 4.1 g (76%) (where R3 =ethylcarboxylate).

Data for 22A: 1H-NMR (300 MHz, Cd13) : 7.48 (d, 2H, J= 9.0), 7.45 (d, 2H, J= 9.0), 7.23 (d, 2H, J= 8.4), 6.85 (d, 2H, J= 8.7), 4.63 (s, 2H), 4.50 (q, 2H, J= 7.2), 4.28 (q, 2H, J=7. 2), 1.45 (t, 3H, J= 7.2), 1.30 (t, 3H, J= 7.2).

This intermediate aryl bromide 22A (where R3 =ethylcarboxylate) was then derivatized via a Heck reaction

according to General Method 3. A mixture of thiazole aryl bromide 23 (where R3 = ethylcarboxylate) (2.7g, 5.5. mmol, 1 equiv.), acrylamide 26A (where R2 =Dodecane) (1.48g, 6.1 mmol, 1.2 equiv.) prepared using General Method 6 from dodecylamine, EtsN (2.3 mL, 16.5 mmol, 3 equiv.), Pd (OAc) 2 (150 mg, 0.66 mmol, 0.12 equiv.), P (o-tolyl) 3 (900 mg, 2.97 mmol, 0.54 equiv.). The thiazole derivative was purified by silica gel column chromatography (Hexane-EtOAc, 4: 1) to give <BR> <BR> 5-4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-4- (4-ethoxycarbonyl methoxy-phenyl)-thiazole-2-carboxylic acid ethyl ester 46 3.3 g (92 %) (below).

Data for 5- (4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-4- (4- <BR> <BR> ethoxycarbon-yl methoxy-phenyl)-thiazole-2-carboxylic acid ethyl ester 46 : 1H-NMR (300 MHz, (CDCl3) : 7.62 (d, 1H, J = 15.6), 7.47 (d, 2H, J= 8.7), 7.46 (d, 2H, J= 8.4), 6.85 (d, 2H, J= 9.0), 6.41 (d, 1H, J= 15.6), 5.68 (t, 1H, J= 6.0), 4.68 (s, 2H), 4.51 (q, 2H, J = 7. 2), 4.28 (q, 2H, J= 7.2), 3.39 (dt, 2H, J= 6.75,6.50), 1.58 (m, 2H), 1.40-1.20 (br m, 21H), 0.88 (t, 3H, J= 6.3).

Example 2 (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-thiazol-4- yl}-phenoxy)-acetic acid 47 General Method 4 was used to hydrolyze the ethyl ester of 5- [4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-4- (4- ethoxycarbonylmethoxy-phenyl)-thiazole-2-carboxylic acid ethyl ester 47 (1.5 g, 2.3. mmol, 1 equiv.), with IN LiOH (11.5 equiv.), in 1,4-Dioxane (12 mL) which, after recrystallization using isopropyl alcohol and ethyl acetate spontaneously decarboxylated to give (4-5- 4- ( (E)-2-Dodecylcarbamoyl-vinyl)- phenyl]-thiazol-4-yl-phenoxy)-acetic acid 47 0.77 g (61 %). Data for (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-thiazol-4- yl}-phenoxy)-acetic acid 47 : 1H-NMR (300 MHz, DMSO-d6): 9.18 (s, 1H), 8.11 (t, 1H, J= 5.3), 7.57 (d, 2H, J= 8.1), 7.43-7.37 (m, 5H), 6.88 (d, 2H, J= 8. 7), 6.64 (d, 1H, J= 15.9), 4.68 (s, 2H), 3.15 (dt, 2H, J= 6.60,6.45), 1.44 (m, 2H), 1.24 (br s, 18H), 0.84 (t, 3H, J= 6.3). MS (APcI) : 549.2 (100, [M+H]); calcd C32H41N204S ( [M+H]) 549.3.

Example 3 4- (4-Ethoxycarbonylmethoxy-phenyl)-5- (4- (E)-2- [1- (4- pentyl-phenyl)-ethylcarbamoyl]-vinyl}-phenyl)-thiazole-2- carboxylic acid ethyl ester 48 The ethyl ester 4- (4-Ethoxycarbonylmethoxy-phenyl)-5- (4- {(E)-2-[1-(4-pentyl-phenyl)-ethylcarbamoyl]-vinyl}-phenyl)- thiazole-2-carboxylic acid ethyl ester 48 was synthesized according to General Method 2, from a mixture of alpha keto- bromide 22 (5. 0g, 11 mmol, 1 equiv.), ethyl thiooxamate (1.53 g, 11.0 mmol, 1.3 equiv.), NaHCOs (2.8 g, 33 mmol, 3 equiv.) and CHsCN (30 mL) which, after purification by silica gel column chromatography (Hexane-EtOAc, 4: 1), gave the the intermediate aryl bromide 22A (where R3 =ethylcarboxylate), 4.1 g (76 %).

Data for 22A-see Example 1.

This intermediate aryl bromide 22A (where R3 =ethylcarboxylate) was then derivatized via a Heck reaction according to General Method 3. A mixture of thiazole aryl bromide 23 (where R3 = ethylcarboxylate) (0.44 g, 0.9 mmol, 1 equiv.), acrylamide 26B (where R2 = 1- (4-pentyl-phenyl)-ethyl) (0.25 g, 1 mmol, 1.2 equiv.) prepared using General Method 6 from 1- (4-pentlyphenyl) ethylamine, EtsN (0.37 mL, 2.69 mmol, 3 equiv.), Pd (OAc) 2 (12 mg, 0.02 mmol, 0.02 equiv.), P (o-tolyl) 3

(20 mg, 0.08 mmol, 0.09 equiv.). The thiazole derivative was purified by silica gel column chromatography (Hexane-EtOAc, 4: 1) to give 4-(4-Ethoxycarbonylmethoxy-phenyl)-5-(4-{(E)-2-[1-(4- pentyl-phenyl)-ethylcarbamoyl]-vinyl}-phenyl)-thiazole-2- carboxylic acid ethyl ester 48 0.1 g (17 %).

Data for 4-(4-Ethoxycarbonylmethoxy-phenyl)-5-(4-{(E)-2-[1-(4- pentyl-phenyl)-ethylcarbamoyl]-vinyl}-phenyl)-thiazole-2- carboxylic acid ethyl ester 48 : 1H-NMR (400 MHz, CDC13) : 7.59 (d, 1H, J= 15.6), 7.44 (d, 2H, J= 8.8), 7.40 (d, 2H, J = 8.4), 7.31 (d, 2H, J = 8.0), 7.25 (d, 2H, J = 8.0), 7.13 (d, 2H, J = 7.6), 6.81 (d, 2H, J = 8.8), 6.42 (d, 1 H, J = 15.6), 6.15 (d, 1 H, J =8. 4), 5.27-5.20 (m, 1H), 4.59 (s, 2H), 4.48 (q, 2H, J= 7.2), 4.24 (q, 2H, J= 7.2), 2.55 (t, 2H, J= 7.6), 1.62-1.50 (m, 2H), 1.58 (m, 2H), 1.53 (d, 3H, J = 6.8), 1.43 (t, 3H, J =7. 2), 1.34-1.20 (m, 7H), 0.87 (t, 3H, J = 6. 8).

Example 4 {4- [5- (4- { (E)-2- [l- (4-Pentyl-phenyl)-ethylcarbamoyl]-vinyl}- phenyl)-thiazol-4-yl]-phenoxy}-acetic acid 49

General Method 4 was used to hydrolyze the ethyl ester of 4- (4- <BR> <BR> Ethoxycarbonylmethoxy-phenyl)-5- (4- f (E)-2- [1- (4-pentyl-<BR> <BR> phenyl)-ethylcarbamoyl]-vinyl}-phenyl)-thiazole-2-carboxylic acid ethyl ester 48 (0.1 g, 0.153 mmol, 1 equiv.), with 1N LiOH (1 mL, 0.918 mmol, 6 equiv.), in 1,4-Dioxane (2 mL), which after recrystallization using isopropyl alcohol and ethyl acetate spontaneously decarboxylated to give {4-[5-(4-{(E)-2-[1-(4-Pentyl- phenyl)ethylcarbamoyl]-vinyl}-phenyl)-thiazol-4-yl]-phenoxy} - acetic acid 49,12 mg (14 %).

Data for {4-[5-(4-{(E)-2-[1-(4-Pentyl-phenyl)-ethylcarbamoyl]- vinyl}-phenyl)-thiazol-4-yl]-phenoxy}-acetic acid 49: lH-NMR (300 MHz, DMSO-d6) : 9. 18 (s, 1H), 8.55 (d, 1H, J= 5.3), 7.57 (d, 2H, J = 8.4), 7.43-7.37 (m, 5H), 7.23 (d, 2H, J = 7.5), 7.14 (d, 2H, J = 8.1), 6.88 (d, 2H, J = 8.4), 6.71 (d, 1H, J= 15.9), 5.01 (m, 2H), 4.68 (s, 2H), 2.53 (t, 2H, J =7. 2), 1.59-1.49 (m, 2H), 1.38 (d, 3H, J =6.9), 1.34-1.20 (m, 4H), 0.85 (t, 3H, J

=6.6). MS (APcI): 555.5 (100, [M+H]) ; calcd C33H3sN204S ([M+H]) 555.2.

Example 5 <BR> <BR> (4-15- [4- ( (E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2-pyridin-3- yl-thiazol-4-yl}-phenoxy)-acetic acid ethyl ester 50 The ethyl ester (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)- phenyl]-2-pyridin-3-yl-thiazol-4-yl}-phenoxy)-acetic acid ethyl ester 50 was synthesized from a mixture of alpha keto-bromide 20A (Rl'= Et) (1 g, 2.2 mmol, 1 equiv.), thionicotinamide (390 mg, 2.9 mmol, 1.3 equiv.), NaHCOs (240 mg, 2.9 mmol, 1.3 equiv.) and CH3CN (8 mL) which, after purification by silica gel column chromatography (Hexane-EtOAc, 4: 1), gave the intermediate aryl bromide 22B, 600 mg (55%) (where R3 = pyridin-3-yl).

Data for 22B (where R3 = pyridin-3-yl) : 1H-NMR (300 MHz, CDCIs) : 9.19 (s, 1H), 8.66 (d, 1H, J= 3.9), 8.28 (d, 1H, J= 8.1), 7.51 (d, 2H, J= 8.7), 7.47 (d, 2H, J= 8.4), 7.41-7.37 (m, 1H),

7.26 (d, 2H, J= 8.4), 6.87 (d, 2H, J= 9.0), 4.63 (s, 2H), 4.27 (t, 2H, J= 7.2), 1.30 (t, 3H, J= 7.2).

This intermediate aryl bromide 22B (where R3 = pyridin- 3-yl) was then derivatized via a Heck reaction according to General Method 3. A mixture of thiazole aryl bromide 23 (where R3= pyridin-3-yl) (590 mg, 1.2 mmol, 1 equiv.), acrylamide 26A (where R2 =Dodecane) (340 mg, 1.43 mmol, 1.2 equiv.) prepared using General Method 6 from dodecylamine, EtsN (0.5 mL, 3.6 mmol, 3 equiv.), Pd (OAc) 2 (0.03 g, 0.12 mmol, 0.06 equiv.), P (o- tolyl) 3 (36 mg, 0.12 mmol, 0.27 equiv.). The thiazole derivative was purified by silica gel column chromatography (Hexane: EtOAc, 4: 1) to give (4-{5-[4-((E)-2-Dodecylcarbamoyl- vinylJ-phenyl]-2-pyridin-3-yl-thiazol-4-yl}-phenoxyJ-acetic acid ethyl ester 50 0.33 g (42 %) below.

Data for (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2- pyridin-3-yl-thiazol-4-yl-phenoxy)-acetic acid ethyl ester 50 : 1H- NMR (300 MHz, CDC1s) : 9.21 (d, 1H, J= 1.8), 8.68 (dd, 1H, J= 4.8,1.5), 8.31 (td, 1H, J= 8.7, J= 1.5), 7.63 (d, 1H, J= 15.6), 7.55 (d, 2H, J= 9.3), 7.48 (d, 2H, J= 8.1), 7.43-7.38 (m, 3H), 6.89 (d, 2H, J=9. 0), 6.40 (d, 1H, J=15. 6), 5.61 (t, 1H, J= 7.70), 4.65 (s, 2H), 4.29 (q, 2H, J= 7.2), 3.43-3.37 (m, 2H), 1.70-1.50 (br m, 2H), 1.40-1.20 (br m, 21H), 0.89 (t, 3H, J= 6.7).

Example 6 (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2-pyridin-3- yl-thiazol-4-yl}-phenoxy)-acetic acid 51 General Method 4 was used to hydrolyze the ethyl ester of (4-{5- [4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2-pyridin-3-yl-thi azol-4- yl-phenoxy)-acetic acid ethyl ester 50 (0.33 g, 0.5 mmol, 1 equiv.), with IN LiOH (2.5 mL), in 1,4-Dioxane (5 mL), to give, after recrystallization using isopropyl alcohol and ethyl acetate, {4-[5-(4-{(E)-2-[1-(4-Pentyl-phenyl)-ethylcarbamoyl]-vinyl}- phenyl)- thiazol-4-yl]-phenoxy-acetic acid 51 0.26 g (83 %).

Data for (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2- pyridin-3-yl-thiazol-4-yl-phenoxy)-acetic acid 51 : 1H-NMR (300 MHz, DMSO-d6) : 9.19 (d, 1H, J= 2.1), 8.71 (d, 1H, J= 4. 8), 8.36 (br d, 1H, J= 7.8), 8.12 (t, 1H, J= 6.0), 7.62-7.56 (m, 3H), 7.49-7.39 (m, 5H), 6.92 (d, 2H, J=8. 4), 6.66 (d, 1H, J = 15. 6), 4.69 (s, 2H), 3.19-3.13 (m, 2H), 1.50-1.40 (br m, 2H), 1.24 (br s, 18H), 0.85 (t, 3H, J = 6.5). MS (APcI): 626.4 (100, [M+H]); calcd for C37H44N204S [M+H] 626.3 Example 7 2-[ (f4- [5- [4- (2-Hexylcarbamoylvinyllphenyfl-2- (lH-pyrrol-2- yl)thiazol-4-yl]phenoxy}acetyl)-(2-hydroxy-1, 1- bishydroxymethylethyl)amino]ethanesulfonic acid 52 General method: A mixture of 28a (300 mg, 0.57 mmol), N- tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid (143

mg, 0.62 mmol), WSC HC1 (120 mg, 0.62 mmol), HOBt (96 mg, 0.62 mmol) and EtsN (63 mg, 0.62 mmol) in DMF (5 mL) was stirred at r. t. overnight. The reaction mixture was acidified with 1N HC1, and extracted with AcOEt-THF. The extracts were washed with water and brine, and dried over Na2SO4. The solvent was evaporated and the residue was purified by HPLC (ODS, CH3CN/0. 1% TFA 55: 45) to give 2-4-4-2- Hexylcarbamoylvingyl)phenyl]-2-(1H-pyrrol-2-yl)thiazol-4- yl]phenoxy}acetyl)-(2-hydroxy-1,1- bishydroxymethylethyl)amino]ethanesulfonic acid 52 as a yellow solid (85 mg, 20%).

Data for 2-[({4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2-(1H-pyrrol- 2-yl)thiazol-4-yl] phenoxygacetyl)-(2-hydroxy-1, 1- bishydroxymethylethyl) amino] ethanesulfonic acid 52 : lH-NMR (250 MHz, DMSO-d6) : 0.8-0.9 (m, 3H), 1.2-1.4 (m, 6H), 1.4-1.5 (m, 2H), 2.85 (t, J= 5.8,2H), 3.1-3.2 (m, 2H), 3.3-3.4 (m, 2H), 3.60 (s, 4H), 4.31 (s, 2H), 4.89 (s, 2H), 6.1-6.2 (m, 1H), 6.62 (d,

J= 15.8,1H), 6.6-6.7 (m, 1H), 6.9-7.0 (m, 1H), 6.95 (d, J= 8.9, 2H), 7.36 (d, J= 8.4,2H), 7.37 (d, J= 15.8,1H), 7.44 (d, J= 8.9,2H), 7.54 (d, J= 8.4,2H), 8.07 (t, J= 5.6,1H), 8.8-8.9 (br, 2H), 11.79 (d, J= 2. 1,1H).

Examples 8-23 were synthesized using a similar methodology to that used to synthesize to Example 7.

Example 8 (S)-3-Hydroxy-2-[2-[4-[5-[4-(2-octylcarbamoylvinyl)phenyl]-2 - (ihrthiazol-4- yl] phenoxy] acetylamino] propionic acid 53

Data for (S)-3-Hydroxy-2-[2-[4-[5-[4-(2- octylcarbamoylvinyl)phenyl]-2-(1H-pyrrol-2-yl)thiazol-4- yl]phenoxy]acetylamino]propionic acid 53:

1H-NMR (250 MHz, DMSO-d6) : 12.74 (br, 1H), 11.79 (br, 1H), 8.07-8.02 (m, 1H), 7.54 (d, 2H, J= 9.7), 7.47-7.34 (m, 5H), 6.98-6.94 (m, 3H), 6.71-6.69 (m, 1H), 6.61 (d, 1H, J=15.8), 6.19-6.16 (m, 1H), 4.37-4.34 (m, 1H), 3.75-3.64 (m, 2H), 3.29- 3.11 (m, 2H), 1.46-1.40 (m, 2H), 1.35-1.20 (m, 10H), 0.84 (t, 3H, J= 6.3).

Example 9 (R)-3-Hydroxy-2-[2-[4-[5-[4-(2-hexylcarbamoylvinyl)phenyl]- 2-(lH-pyrrol-2-vl) thiazol-4- yl] phenoxy] acetylamino] propionic acid 54 Data for (R)-3-Hydroxy-2-[2-[4-[5-[4-(2- hexylcarbamoylvinyl) phenyl]-2- (1 H-pyrrol-2-yl) thiazol-4- yl] phenoxy] acetylamino] propionic acid54: 1H-NMR (250 MHz, DMSO-d6) : 12.70 (br, 1H), 11.79 (br, 1H), 8.07-8.02 (m, 2H), 7.55-7.34 (m, 7H), 6.98-6.95 (m, 3H), 6.71- 6.68 (m, 1H), 6.61 (d, 1H, J= 15.8), 6.18-6.15 (m, 1H), 4.37-

4.33 (m, 1H), 3.77-3.60 (m, 2H), 3.16-3.14 (m, 2H), 1.46-1.40 (m, 2H), 1.35-1.20 (m, 6H), 0.87-0.83 (m, 3H).

Example 10 (2S)-3Hydroxy-2-[2-[4-[5-[4-(2-hexylcarbamoylvinyl)phenyl]- 2-thiazol-4- yllphenoxy] acetylamino] propionic acid 55 Data for (2S)-3Hydroxy-2-[2-[4-[5-[4-(2- hexylcarbamoylvinyl) phenyl]-2- (morpholin-4-yl) thiazol-4- yl] phenoxy] acetylamino] propionic acid55: 1H-NMR (250MHz, DMSO-d6): 0.80-0.95 (m, 3H), 1.20-1.40 (m, 6H), 1.40-1.55 (m, 2H), 3.15 (q, 2H, J= 6.0), 3.30-3.50 (m, 4H), 3.56-3.90 (m, 6H), 4.25-4.44 (m, 1H), 4.56 (s, 2H), 6.59 (d, 2H, J= 15.8), 6.92 (d, 2H, J= 8.9), 7.26 (d, 2H, J= 8.3), 7.28-7.44 (m, 3H), 7.48 (d, 2H, J= 8.4), 7.97-8.10 (m, 2H).

Example 11 <BR> <BR> (2- [4-fS- [4- (2-Octylcarbamoylvinyl) phenyll-2- (lH-pyrrol-2- yl)thiazol-4-yl]phenoxy]acetylamino)acetic acid 56 Data for (2-[4-[5-[4-(2-Octylcarbamoylvinyl)phenyl]-2-(1H-pyrrol-2- yl) thiazol-4-yl] phenoxy] acetylamino) acetic acid 56: 1H-NMR (250 MHz, DMSO-d6) : 12.65 (br, 1H), 11.80 (br, 1H), 8. 38 (t, 1H, J= 5.9), 8.05 (t, 1H, J= 6.0), 7.53 (d, 2H, J= 8.3), 7.46 (d, 2H, J=8. 9), 7.41-7.34 (m, 3H), 6.97 (d, 2H, J= 8.9), 6.95-6.93 (m, 1H), 6.71-6.69 (m, 1H), 6.61 (d, 1H, J= 15.8), 6.19-6.17 (m, 1H), 4.54 (s, 2H), 3.82 (d, 2H, J= 5.9), 3.14 (q, 2H, J= 6.0), 1.46-1.40 (m, 2H), 1.35-1.20 (m, 10H), 0.84 (t, 3H, J= 6. 4).

Example 12 3-Hydroxy-2-[2-(2-[4-[5-[4-(2-octylcarbamoylvinyl)-phenyl]-2 - (IH-pyrrol-2-yl)thiazol-4-yl] phenoxy] acetylamino) acetylamino] propionic acid 57

Data for 3-Hydroxy-2-[2-(2-[4-[5-[4-(2-octylcarbamoylvinyl)- phenyl]-2-(1H-pyrrol-2-yl)thiazol-4- yl] phenoxy] acetylamino) acetylaminopropionic acid 57: 1H-NMR (250 MHz, DMSO-d6) : 11.81 (br, 1H), 8.25 (t, 1H, J= 6.0), 8.10 (t, 1H, J= 6.0), 8. 03 (d, 2H, J= 7.6), 7.53 (d, 2H, J= 8.4), 7.45 (d, 2H, J= 8.8), 7.41-7.34 (m, 3H), 6.97 (d, 2H, J= 8.8), 6.95- 6.94 (m, 1H), 6.70-6.69 (m, 1H), 6.63 (d, 1H, J= 15.8), 6.19- 6.16 (m, 1H), 4.54 (s, 2H), 4.23-4.18 (m, 2H), 3.84 (d, 2H, J= 6.0), 3.67-3.52 (m, 2H), 3.14 (q, 2H, J= 6.0), 1.46-1.40 (m, 2H), 1.35-1.20 (m, 10H), 0.84 (t, 3H, J= 6.1).

Example 13 (2S)-4-[3-[4-[4-[4-[(1-Carboxy-2-hydroxy-ethylcarbamoyl) <BR> <BR> methoxy-phenyl]-2-{lH-pyrrol-2-yl)-thiazol-5-yll-phenyll- acryloylaminol-piperidine-l-carboxylic acid ethyl ester 58

Data for (2S)-4-[3-[4-[4-[4-[(1-Carboxy-2-hydroxy- ethylcarbamoyl)methoxy-phenyl]-2-(1H-pyrrol-2-yl)-thiazol-5- yl]- phenyl]-acryloylamino]-piperidine-1-carboxylic acid ethyl ester 58 : 1H NMR (DMSO-d6) : 1.18 (t, 3H, J=7. 1Hz), 1.20-1.40 (m, 2H), 1.72-1.88 (m, 2H), 2.83-3.10 (m, 2H), 3.60-3.86 (m, 7H), 4.03 (q, 2H, J= 7.0), 4.36 (qn, 1H, J= 4.1), 4.60 (s, 2H), 6.19 (dd, 1H, J= 2.3,5.9), 6.61 (d, 1H, J= 15. 6), 6.70-6.78 (m, 1H), 6.98 (d, 2H, J= 8.9), 7.38 (d, 2H, J= 8.3), 7.46 (d, 2H, J= 8.8), 7.56 (d, 2H, 8.3), 8.11 (t, 2H, J= 8.4), 11.78-11.88 (m, 1H).

Example 14 2-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(1H-pyrrol- <BR> <BR> 2-yl)-thiazol-4-yl]-phenoxy}-acetylamino)-ethanesulfonic acid 59

Data for 2-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl)-2-(1H- pyrrol-2-yl)-thiazol-4-yl]-phenoxy-acetylamino)-ethanesulfon ic acid 59: 1H-NMR (250 MHz, DMSO-d6) 0.85 (t, J= 7.0 Hz, 3H), 1.2-1.3 (m, 6H), 1.4-1.5 (m, 2H), 2.55 (t, J= 6.8 Hz, 2H), 3.13 (t, J= 7.0 Hz, 2H), 3.39 (t, J= 6.8 Hz, 2H), 4.67 (s, 2H), 6.1-6.2 (m, 1H), 6.65 (d, J= 15.8 Hz, 1H), 6. 6-6.7 (m, 1H), 6.93 (d, J= 8.8 Hz, 2H), 6.9-7.0 (m, 1H), 7.34 (d, J= 8.4 Hz, 2H), 7.37 (d, J= 15.8 Hz, 1H), 7.43 (d, J= 8.8 Hz, 2H), 7.54 (d, J= 8.4 Hz, 2H), 11.85 (d, J= 2. 5 Hz, 1H).

Example 15 2-(2-{4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2-(1H-pyrrol-2- yl)thiazol-4-yl]phenoxy}acetylamino)-3-hydroxybutyric acid 60

Data for 2-(2-{4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2- (l H-pyrrol-2-ylJthiazol-4-yl] phenoxy} acetylamino)-3- hydroxybutyric acid60: 1H NMR (DMSO-d6) : 0.96 (3H, t, J= 6.7), 1.12 (3H, d, J= 6.0), 1.30-1.40 (6H, m), 1.50-1.60 (2H, m), 3.20-3.30 (2H, m), 4.20-4.40 (2H, m), 4.74 (2H, s), 6.27-6.30 (1H, m), 6.71 (1H, d, J=15. 8), 6.79-6.83 (1H, m), 7.03-7.10 (3H, m), 7.46 (2H, d, J = 8.1), 7.48 (1H, d, J= 15.8), 7.55 (2H, d, J= 8.6), 7.64 (2H, d, J=8. 6), 7.89 (1H, d, J= 8.6), 8. 20 (1H, t, J= 5.8), 11.93 (1H, s).

Example 16 (S)-3-Hydroxy-2-[2-[4-[5-[4-(2-Phenylbutylcarbamoyl)phenyl]- 2- H-pyrrol-2-yl) thiazol-4-yll-phenoxy] acetylamino propionic acid 61 Data for (S)-3-Hydroxy-2-[2-[4-[5-[4-(2- Phenylbutylcarbamoyl) phenylj-2- (lH-pyrrol-2-yl) thiazol-4-ylj- phenoxyJacetylamino] propionic acid 61: 1H NMR (DMSO-d6) ; 1.30-1.70 (4H, m), 2.50-2.60 (2H, m), 3.10- 3.30 (2H, m), 3.60-3.80 (2H, m), 4.25-4.38 (1H, m), 4.59 (2H, s), 6.15-6.21 (1H, m), 6.55-6.72 (2H, m), 6.90-7.00 (3H, m), 7.10- 7.60 (12H, m), 8.00-8.20 (2H, m), 11.85 (lH, s).

Example 17 2-(2-{4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2-(1H-pyrrol-2- yl)thiazol-4-yl]phenoxy}acetyl- amino) acetic acid 62

Data for 2-(2-{4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2-(1H- pyrrol-2-yl) thiazol-4-yl] phenoxyacetyl-amino) acetic acid 62 : lH NMR (DMSO-d6) : 0.80-0.90 (3H, m), 1.20-1.37 (6H, m), 1.37- 1.50 (2H, m), 3.08-3.20 (2H, m), 3.80 (2H, d, J= 5.5), 4.54 (2H, s), 6.16-6.20 (1H, m), 6.61 (1H, d, J=15. 6), 6.68-6.72 (1H, m), 6.92-7.00 (3H, m), 7.30-7.50 (5H, m), 7.54 (2H, d, J= 7.5), 8.09 (1H, t, J= 5.5), 8.36-8.45 (1H, m), 11.82 (1H, s).

Example 18 2-(S)-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(1- methoxymethyl-1H-pyrrol-2-yl)-thiazol-4-yl]-phenoxy}- acetylamino)-3-hydroxy-propionic acid 63

Data for. 2-(S)-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(1- methoxymethyl-1H-pyrrol-2-yl)-thiazol-4-yl]-phenoxy}- acetylamino)-3-hydroxy-propionic acid 63: lH-NMR (250 MHz, DMSO-d6) : 0.8-0.9 (m, 3H), 1.2-1.4 (m, 6H), 1.4-1.5 (m, 2H), 3.1-3.2 (m, 5H), 3.6-3.7 (m, 2H), 4.3-4.4 (m, 1H), 4.58 (s, 2H), 5.80 (s, 2H), 6.21 (dd, J= 2.7 and 3.8,1H), 6.62 (d, J= 15.8, 1H), 6.80 (dd, J= 1.7 and 3.8,1H), 6.96 (d, J= 8.9,2H), 7.23 (dd, J= 1.7 and 2.7,1H), 7.38 (d, J= 8.4,2H), 7.39 (d, J= 15.8,1H), 7.44 (d, J= 8.9,2H), 7.56 (d, J= 8.4,2H), 8.0-8.1 (m, 2H).

Example 19 4-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(1H-pyrrol- <BR> <BR> 2-yl)-thiazol-4-yl]-phenoxf-acetolamino)-3-hydroxy-butYric acid 64

Data for 4-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(1H- <BR> <BR> pyrrol-2-yl)-thiazol-4-yl]-phenoxy-acetylamino)-3-hydroxy-bu tyric acid 64: 1H-NMR (250 MHz, DMSO-d6) : 0.8-0.9 (m, 3H), 1.2-1.4 (m, 6H), 1.4-1.5 (m, 2H), 2.16 (dd, J= 8.5 and 15.3 Hz, 1H), 2.35 (dd, J= 4.3 and 15.3 Hz, 1H), 3.0-3.2 (m, 4H), 3.8-4.0 (m, 1H), 4.50 (s, 2H), 5.01 (d, J= 4.8 Hz, 1H), 6.1-6.2 (m, 1H), 6.60 (d, J= 15.9 Hz, 1H), 6.6-6.7 (m, 1H), 6.94 (d, J= 8.8 Hz, 2H), 7.35 (d, J= 8.3 Hz, 2H), 7.38 (d, J= 15.9 Hz, 1H), 7.44 (d, J= 8.8 Hz, 2H), 7.53 (d, J= 8.3 Hz, 2H), 8.0- 8.1. (m, 2H), 11.82 (s, 1H), 12.07 (br s, 1H).

Example 20 4-Carbamoyl-2-(2-{4-[5-[4-(2-hexylcarbamoylvinyl)phenyl-2- (1H-pyrrol-2-yl)thiazol-4-yl]- phenoxy3acetolamino) butyric acid 65

Data for 4-carbamoyl-2-(2-{4-[5-[4-(2- hexylcarbamoylvinyl) phenyl-2- (1H-pyrrol-2-yl) thiazol-4-yl]- phenoxy} acetylamino) butyric acid 65: IH NMR (DMSO-d6) : 0.95 (3H, t, J= 6.5), 1.30-1.46 (6H, m), 1.47-1.60 (2H, m), 1.80-2.40 (4H, m), 3.20-3.30 (2H, m), 4.25-4.40 (1H, m), 4.64 (2H, s), 6.26-6.30 (1H, m), 6.71 (1H, d, J= 15.7), 6.78-6.82 (1H, m), 6.88-6.93 (1H, m), 7.00-7.10 (4H, m), 7.40-7.60 (5H, m), 7.64 (2H, d, J= 8.3), 8.17-8.23 (1H, m), 8.51 (1H, d, J= 8.4), 11. 9 (1H, s).

Example 21 3-Hydroxy-2-(2-{4-[5-[4-(2-phenethylcarbamoylvinyl)phenyl]- 2-[1H-pyrrol-2-yl)thiazol-4-yl]- phenoxy} acetylamino) propionic acid 66

Data for 3-Hydroxy-2-(2-{4-[5-[4-(2- phenethylcarbamoylvinyl)phenyl]-2-(1H-pyrrol-2-yl)thiazol-4- yl]- phenoxyacetylamino) propionic acid 66 :. 1H NMR (DMSO-d6) ; 2.70-2.80 (2H, m), 3.30-3.50 (2H, m), 3.60-3.90 (2H, m), 4.30- 4.50 (1H, m), 4.59 (2H, s), 5.00-5.20 (1H, m), 6.17-6.22 (1H, m), 6.61 (1H, d, J=15. 8), 6.70-6.74 (1H, m), 6.93-7.03 (3H, m), 7.10-7.40 (8H, m), 7.45 (2H, d, J=8. 8), 7.55 (2H, d, J=8. 3), 8.08 (1H, d, J=8. 3), 8.17-8.26 (1H, m), 11.8 (1H, s), 12.8 (lH, s).

Example 22 2-(2-{4-[5-[4-(2-hexylcarbamoylvinyl)phenyl]-2-(1H-pyrrol-2- yl)thiazol-4-yllphenoxylacetyl- amino)-3-mercaptopropionic acid 67

Data for 2-(2-{4-[5-[4-(2-hexylcarbamoylvinyl)phenyl]-2-(1H- pyrrol-2-yl) thiazol-4-yl] phenoxy} acetyl- amino)-3-mercaptopropionic acid 67: 1H NMR (DMSO-d6) : 0.80- 0.90 (3H, t, J= 6.9), 1.20-1.35 (6H, m), 1.35-1.50 (2H, m), 2.70- 3.00 (2H, m), 3.10-3.20 (2H, m), 4.40-4.50 (1H, m), 4.59 (2H, s), 6.15-6.20 (1H, m), 6.61 (1H, d, J= 15.8), 6.68-6.72 (1H, m), 6.90-7.00 (3H, m), 7.32-7.50 (5H, m), 7.57 (2H, d, J= 7.9), 8.05 (1H, t, J= 5.4), 8.29 (1H, d, J= 8. 1), 11.8 (lH, s).

Biological Assay The biological activity of Formula 1 is determined by the following procedures outlined in the materials and methods section. The results which show inhibitory activity as ICso in micromolar against P-selectin are tabulated below in Table 2.

Table 2 Compound Human ELISA - IC50 µM Cell Adhesion Number Assay-P-Selectin % Inhibition E-Selectin P-Selectin L-Selectin @ 30 HM IC50 µM 47 - 16 - - 91 49 23 40 51-2. 5--- 52 >500 5.1 1.6 86 - 53 3.0 0.55 0.71 80 - 54 >500 0.66 3.7 88 - 55 >500 16 20 78- 56 5. 4 0. 29 0. 25 84- 57 >500 0. 18 0. 16 95- 58 57 4. 2 4. 4 96- 59 7. 8 0. 46 0. 32 83 60 330 1. 2 1. 1 94 61 >500 6.3 9.0 92 - 62 >500 2. 9 3. 0 82 63 >500 5.5 10 85 - 64 69 3. 0 3. 1 98 65 >500 4.8 6.4 93 - 66 >500 6. 0 5. 0 88 67 >500 5. 8 5. 7 87

Materials and Methods (For compounds 47,49 and 51) P-selectin ELISA Assays ELISA-type assays were used to screen for inhibitors of selectin-ligand interactions. P-selectin-IgG chimera, constructed as described by Foxall and colleagues (Foxall et al., FASEB 117 : 895 (1992)), and sialyl-Lewisx pentaceramide were obtained from Kanebo, Ltd. (Osaka) well (Kiyoi et al., Bioorg. Med. Chem.

6: 587 (1998)). Assays were performed essentially as described (Ohmoto et al., J. Med. Chem. 39: 1339 (1996)). Polystyrene microtiter plates (Falcon Pro-Bind) were coated with the sialyl- LewisX analog at 40-100 pmol/. Coated wells were blocked with

5% bovine serum albumin (BSA) in 50 mM imidazole buffer, pH 7.2, for 1 hour at room temperature.

Compounds were diluted from DMSO stock solutions in assay buffer (50 mM imidazole buffer, pH 7.2, containing 1% BSA and 1 mM CaCl2). Compounds were always run in duplicate or triplicate. A complex consisting of P-selectin-IgG chimera, biotinylated goat F (ab) 2 anti-human IgG, and streptavidin- alkaline phosphatase conjugate was made in assay buffer.

Selectin chimera was omitted from the complex for negative control ("background") wells. The complex and the test compounds (or vehicle controls) were combined in wells of a polypropylene microtiter plate and incubated for 30 minutes at room temperature. The complex-compound mixture was then added to the blocked, sialyl-LewisX-ceramide coated plate and allowed to incubate for 45 minutes at 37°C. After washing 3-4 times with 50 mM imidazole, the bound complex was detected using the colorimetric phosphatase substrate, p- nitrophenylphosphate, at 1 mg/mL in 1 M diethanolamine pH 9.8 containing 0.01% MgCl2. After developing for 1-2 hours at room temperature, the absorbance at 405 nM was measured in a Molecular Devices microplate reader. Percent inhibition was calculated by comparing the test compound result with the vehicle control after subtracting the background from each. ICso

values were calculated by in-house data analysis software (OntoASSAY; Ontogen, Corp.) using standard algorithms.

Cell-Selectin Adhesion Assay The ability of compounds to inhibit the adhesion of HL60 cells to purified selectin protein was measured using a"cell- selectin"assay. Recombinant soluble P-selectin protein purchased from R&D Systems (Minneapolis, MN) was diluted to 2.5 J. g/mL in Dulbecco's PBS containing calcium and magnesium (PBS+). Falcon Pro-Bind microtiter plate wells were incubated with 50 pL of the P-selectin protein solution for 1 hr at 37°C or overnight at 4°C. The selectin protein was omitted from negative control ("background") wells. Coated wells were then washed three times with PBS+ and then blocked with 1% BSA in PBS+ for 1 hour at room temperature. After blocking, the plates were washed 3 times with PBS+. Compounds were diluted to 2x final test concentration in PBS+ and added to the blocked, selectin-coated wells in a volume of 50 ; je. Samples were always run in duplicate or triplicate. Compounds and vehicle controls were pre-incubated in the wells for ~20 minutes at room temperature.

HL60 cells obtained from the ATCC (Manassas, VA) were cultivated in RPMI medium containing 10% heat-inactivated fetal

bovine serum (FBS). For the assay, cells were harvested by centrifugation, washed once with PBS+, and resuspended in PBS at a concentration of 2 x 106 cells/mL. Cells were added directly to the compound-containing wells in a volume of 50 VLL per well, bringing the compound to its final test concentration in a total volume of 10011L. Cells and compound were incubated on the selectin-coated wells for, 45 minutes at 37°C. Unbound cells were removed using a vacuum manifold and a single wash with 200 J. L PBS+ (added slowly using a manual multichannel pipettor). Retained cells were labeled directly on the plate by adding 5 J. g/mL of the membrane-permeable fluorescent dye, calcein-AM, and incubating for 30 minutes at 37°C. Signal was quantified in a Wallac Victor fluorescent microplate reader using 485 nM excitation and 535 nM emission. Percent inhibition and ICso values were calculated as described above for the ELISA assay.

Materials RPMI medium, tissue culture grade BSA, and Dulbecco's PBS were purchased from Gibco-BRL (Rockville, MD). Fetal bovine serum was obtained from Gemini BioSystems (Calabasas, CA). Imidazole, BSA, calcium chloride, magnesium chloride, and diethanolamine were purchased from Wako Chemicals, Inc. (Richmond, VA). Calcein-AM and BCECF-AM

were from Molecular Probes, Inc. (Eugene, OR). Unless otherwise noted, all other reagents were purchased from Sigma Chemicals (St. Louis, MO).

For compounds 52-67 E-, P-and L-selectin ELISA Assays ELISA-type assays were used to screen for inhibitors of selectin-ligand interactions. E-, P-and L-selectin-IgG chimera, constructed as described by Foxwall and colleagues (Foxwall et al., FASEB 117: 895 (1992)), and sialyl-Lewisx pentasaccharide ceramide was synthesized as described (Kiyoi et al., Bioorg.

Med. Chem. 6: 587 (1998)). Assays were performed essentially as described (Ohmoto et al., J. Med. Chem. 39: 1339 (1996)).

Polystyrene microtiter plates (Falcon Pro-Bind) were coated with the sialyl-Lewisx analog at 40-100 pmole/well. Coated wells were blocked with 5% bovine serum albumin (BSA) in 50 mM imidazole buffer, pH 7.2, for 1 hour at room temperature.

Compounds were diluted from DMSO stock solutions in assay buffer (50 mM imidazole buffer, pH 7.2, containing 1% BSA and 1 mM CaCl2). Compounds were always run in duplicate or triplicate. A complex consisting of selectin-IgG chimera, biotinylated goat F (ab') 2 anti-human IgG (Biosource International, Camarillo, CA), and streptavidin-alkaline

phosphatase conjugate (Zymed Laboratories, South San Francisco, CA) was made in assay buffer. Selectin chimera was omitted from the complex for negative control ("background") wells. The complex and the test compounds (or vehicle controls) were combined in wells of a polypropylene microtiter plate and incubated for 30 minutes at room temperature. The complex- compound mixture was then added to the blocked, sialyl- Lewisx-ceramide coated plate and allowed to incubate for 45 minutes at 37°C. After washing 3-4 times with 50 mM imidazole, the bound complex was detected using the colorimetric phosphatase substrate, p-nitrophenylphosphate, at 1 mg/mL in 1 M diethanolamine pH 9.8 containing 0.01% MgCl2. After developing for 1-2 hours at room temperature, the absorbance at 405 nm was measured in a Molecular Devices microplate reader. Percent inhibition was calculated by comparing the test compound result with the vehicle control after. subtracting the background from each. ICso values were calculated probit method.

Cell-Selectin Chimera Adhesion Assay The ability of compounds to inhibit the adhesion of HL60 cells to P-selectin IgG chimera was measured using a"cell- selectin chimera"assay. Goat anti-human IgG Fc purchased

from Jackson Immuno Research Laboratories (West Grove, PA) was diluted to 10 ug/mL in Dulbecco's PBS (PBS-). Falcon Pro- Bind microtiter plate wells were coated with 100 uL of Goat anti-human IgG Fc solution overnight at 4°C. Coated wells were blocked with 2% BSA in PBS-for 1 hour at room temperature and then washed 3 times with PBS-containing 0.05% Tween20. P-selectin IgG chimera was diluted to 5 ug/mL in PBS-, and coated microplate well were incubated with 50 uL of the selectin chimera solution for 1 hour at room temperature. The selectin chimera protein was omitted from negative control ("background") wells. Selectin chimera coated wells were then washed 3 times with PBS-containing 0.05% Tween20. Compounds were diluted to 3x final test concentration in assay medium (RPMI-1640 containing 5.958 g/L of HEPES, 10% of heat-inactivated fetal calf serum (FCS) and 150 ug/ml of human IgG (ICN Pharmaceuticals, Costa Mesa, CA), pH 7.4), and added to the selectin chimera coated wells in a volume of 100 uL. Samples were always run in duplicate. Compounds and vehicle controls were preincubated in the wells for 30 minutes at 37°C.

HL60 cells obtained from ATCC (Manassas, VA) were cultivated in RPMI-1648 containing 10% FCS. For the assay, cells were harvested by centrifugation, washed once with RPMI-

1648 containing 10% FCS, and resuspended in assay medium at the concentration of 5 x 105 cells/mL Cells, prewarmed at 37°C, were added directly to the compound-containing wells in a volume of 200 pL per well, bringing the compound to its final test concentration in a total volume of 300 pL. Cells and compound were incubated on the selectin chimera coated wells for 30 minutes at 37°C. To remove unbound cells, assay plate was covered with PARAFILM (American National Can, Neenah, WI) and sealed with lid, and then the plate was inverted and incubated for 30 minutes at 37°C. After incubation, unbound cells were removed using a vacuum manifold. Retained cells were labeled directly on the plate by adding 2 MM of the membrane-permeable fluorescent dye, calcein-AM, and incubating for 30 minutes at 37°C. Signals were quantified in a PolarStar fluorescent microplate reader using 485 nm excitation and 520 nm emission. Percent inhibition was calculated as described above for the ELISA assay.

Materials Calcein-AM and HEPES were purchased from Dojindo Laboratories (Kumamoto). RPMI-1648, fetal calf serum and magnesium chloride were purchased from Nissui

Pharmaceutical (Tokyo), Gibco-BRL (Rockville, MD) and Kanto Chemical (Tokyo), respectively. Unless otherwise noted, all other reagents were purchased from Wako Pure Chemical Industries (Osaka).

Included within the scope of this invention are prodrugs of Formula 1. These include, but are not limited to, compounds such as Formula 2. n1 p2 R1 R2 Where at least one --< of R1, R2orR3 = EtO2CsG N-, S G R3 R where G is defined in Group I Formula 2 In the case of the-COOH being present, pharmaceutically acceptable esters can be employed, e. g., methyl, ethyl, tert-butyl, pivaloyloxymethyl, and the like, and those esters known in the art for modifying solubility or hydrolysis characteristics for use as sustained release or prodrug formulations.

Pharmaceutically acceptable salts of the compounds of Formula 1, where a basic or acidic group is present in the structure, are also included within the scope of this invention.

When an acidic substituent is present, such as-COOH, there

can be formed the ammonium, morpholinium, sodium, potassium, barium, calcium salt, and the like, for use as the dosage form. When a basic group is present, such as amino or a basic heteroaryl radical, such as pyridyl, an acidic salt, such as hydrochloride, hydrobromide, phosphate, sulfate, tri- fluoroacetate, trichloroacetate, acetate, oxalate, maleate, pyruvate, malonate, succinate, citrate, tartarate, fumarate, mandelate, benzoate, cinnamate, methanesulfonate, ethanesul- fonate, picrate and the like, and include acids related to the pharmaceutically acceptable salts listed in Journal of Pharmaceutical Science, 66,2 (1977) p. 1-19 and incorporated herein by reference, can be used as the dosage form.

In addition, some of the compounds of the present invention may form solvates with water or common organic solvents. Such solvates are encompassed within the scope of the invention.

The term"therapeutically effective amount"shall mean that amount of drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor or others.

The present invention provides a method of administering a compound selected from those defined in

Formula 1 above in cases where inhibition or modulating selectin activity in a body is needed. These conditions include but are not limited to the foregoing described diseases.

To administer Formula 1, the compounds may be administered orally as tablets, aqueous or oily suspensions, lozenges, troches, powders, granules, emulsions, capsules, syrups or elixirs. The composition for oral use may contain one or more agents selected from the group of sweetening agents, flavoring agents, coloring agents and preserving agents in order to produce pharmaceutically elegant and palatable preparations. The tablets contain the acting, ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets.

These excipients may be, for example, (1) inert diluents, such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; (2) granulating and disintegrating agents, such as corn starch or alginic acid; (3) binding agents, such as starch, gelatin or acacia; and (4) lubricating agents, such as magnesium stearate, stearic acid or talc. These tablets may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time

delay material such as glyceryl monostearate or glyceryl distearate may be employed. Coating may also be performed using techniques described in the U. S. Patent Nos. 4,256,108; 4,160,452; and 4,265,874 to form osmotic therapeutic tablets for control release.

Formulations for oral use may be in the form of hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin. They may also be in the form of soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.

Aqueous suspensions normally contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspension. Such excipients may be (1) suspending agent such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropylmethyl-cellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; (2) dispersing or wetting agents which may be (a) naturally occurring phosphatide such as lecithin; (b) a condensation product of ethylene oxide with a fatty acid, for example, polyoxyethylene stearate; (c) a condensation product of ethylene oxide with a long chain aliphatic alcohol, for example,

heptadecaethylen-oxycetanol ; (d) a condensation product of ethylene oxide with a partial ester derived from a fatty acid and hexitol such as polyoxyethylene sorbitol monooleate, or (e) a condensation product of ethylene oxide with a partial ester derived from fatty acids and hexitol anhydrides, for example polyoxyethylene sorbitan monooleate.

The pharmaceutical composition may be in the form of a sterile injectable aqueous or oleagenous suspension. This suspension may be formulated according to known methods using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1, 3-butanediol.

Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds of the invention may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperature but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

The compounds of the present invention may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidyl-cholines.

For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compounds of Formula 1 are employed.

The compounds of Formula 1 may also be administered directly into the lungs by inhalation or intranasal delivery when formulated in a solvent that is suitable for aerosol formation.

Such delivery would be useful for direct delivery to the site of action, as in asthma. However, because administration to the lungs may result in significant blood levels of the compound,

this route of administration can be also used in cases where systemic exposure is required.

The optium dosage level or regimen is to be determined or titrated by the clinician based on the patient's age, gender and the condition of the disease state, by methods known to the healing arts. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage will vary depending upon the host treated and the particular mode of administration. For example, a formulation intended for oral administration to humans may contain 5 mg to 1 g of an active compound with an appropriate and convenient amount of carrier material which may vary from about 5 to about 95 percent of the total composition. Dosage unit forms will generally contain between from about 5 mg to about 500 mg of active ingredient.

It will be understood however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, gender, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing therapy. The dosage needs to be individualized by the clinician.