WO2000034255A9 - Substituted thiazoles for treatment of human diseases involving modulators of p-, l- and e- selectin - Google Patents

Abstract

Compounds of Formula (1) (where at least one of R?1, R2 or R3¿ contains calcium binding moiety e.g. CO¿2?H etc.) are disclosed. Where R?1¿ is typically a moiety containing a terminal carboxylic acid group such as cinnamic acid or phenoxy acetic acid, R2 is typically a hydrophobic moiety such as functionalized alkyl chain or a functionalized aryl group, and R3 is typically a functionalized aryl group, are within the scope of this invention. These compounds exhibit inhibitory activity against P-, L- and E-selectin and are indicated in the treatment of human diseases involving P-, L- and E-selectin.

Description

Substituted Thiazoles for Treatment of Human Diseases Involving Modulators of P-, L- and E-selectin

This application claims the benefit of the filing date of provisional application serial no. 60/ 111,026, filed on December 4^th 1998, and provisional application serial no. 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 (sLe^x)) 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 Cellular Adhesion: 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. & Biology 2: 703 (1995)). The specific carbohydrate moieties necessary for selectin binding have been identified: the sialylated and fucosylated tetrasaccharide sialyl Lewis X (sLe^x), and a related structure sialyl Lewis a (sLe^a), 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, Cήt. 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. Physiol 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. Cordial 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. Endocήnol Diabetes 106: 183 (1998); Kopp et al, Exp.

Clin. Endocήnol Diabetes 106: 41 (1998); Albertini et al,

Diabetes Care 21: 1008 (1998); Bannan et al, 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. Cardiol 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. Vase. 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. Cancer Res. 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- Lewis analogs/ mimetics reported in the literature include: 'GSC-150' (Kanebo) which has been reported to have IC50 values of 280 μM, 100 μM, and 30 μM 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

IC50 values of 500 μM, 70 μM, and 560 μM 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 IC50 of 390 μM against

E-selectin (Kolb, Bioorg. Med. Chem. Lett. 7: 2629 (1997)); and C-mannose derivatives which have IC50 values of 100-

160 μM against E-selectin (Marron et al, Tet. Lett. 37: 9037

(1996)). Some of the most potent derivatives that have been reported are multivalent sialyl-Lewis^x analogs which have IC50 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 IC50 values of 160 μM and 2 μM, against P- and L-

selectin respectively, in competition ELISA assays (Cecconi et al, J. Biol Chem. 21: 15060 (1994)). Sulfatides have IC50 values in the 0.1-12 μM 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 IC50 values of 18 μM and 10 μM, 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 IC50 value of 60 μM (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 IC50 values of ~20 μM (Briggs et al, Glycobiology 5: 583

(1995)). Additional peptides, discovered by random

screening, have IC50 values of 5-10 μM 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 C12H25 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 "halo" include fluorine, chlorine, bromine, and iodine.

The term "alkyl" includes Ci-Ciβ straight chain saturated, C1-C16 branched saturated, C3-C8 cyclic saturated and Ci-Ciβ 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), teri-butyl (t-Bu), sec-butyl (s-Bu), isopentyl, neopentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropylmethyl, and the like. The term "alkenyl" includes C2-C16 straight chain unsaturated, C2-Cπ branched unsaturated, Cs-Cs unsaturated cyclic, and C2-C16 straight chain or branched unsaturated aliphatic hydrocarbon groups substituted with C3-C8 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 Ci-Ciδ alkyl, aryl Co-Ciβ alkyl, or Co-Ciβ alkylaiyl 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. me thoxy (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, Ci-iβ alkyl, arylCi-iβ alkyl, Co-iβ alkyloxy Co-ie alkyl, aryl Co-i6 alkyloxy Co-ie alkyl, Co-iβ alkylthio Co-i6 alkyl, aryl Co-iβ alkylthio Co-i6 alkyl, Co-iβ alkylamino Co-iβ alkyl, aryl Co-iβ alkylamino Co-iβ alkyl, di(aryl Ci-iβ alkyl)amino Co-i6 alkyl, Ci-iβ alkylcarbonyl Co-i6 alkyl, aryl Ci-iβ alkylcarbonyl Co- 16 alkyl, Ci-iβ alkylcarboxy Co-iβ alkyl, aryl Ci-iβ alkylcarboxy Co- 16 alkyl, Ci-iβ alkylcarbonylamino Co-iβ alkyl, aryl Ci-iβ alkylcarbonylamino Co-iβ alkyl, -C0-16 alkyl COORi, -Co-ie alkyl CONR2R3 wherein Ri, R2 and R3 are independently selected from hydrogen, Ci-Cnalkyl, arylCo-Cnalkyl, 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 Ci-Ciβ alkyl, aryl C0-C16 alkyl, or Co-Ciβ 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- aminonaphthyljhexyl, 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.

R¹ R²

M

^Nγ _R33^s

Where at least one of R¹, 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:

CO(OH or *N-linked amino acid)

Group I = or

where R⁶ is selected from the following in Table 1 :

Figure 1

Table 1

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-ii 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, Co-eCONHRⁱo, Co-₆NHSO₂R¹⁰, trans-

CH=CHCO₂R¹⁰, or trans-CH=CHCONHR¹⁰, wherein Rⁱo is d-iβ alkyl, Ci-ie alkyloxyalkyl, C5-8 cycloalkyl, Ci-n alkylaryl, or C1-5 alkylaryl Ci-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, Ci-β alkyl and C1-6 alkyloxy, C1-6 cycloalkyloxy, C1-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, d-4 alkyl, or C1-4 alkyloxy; or R¹⁰ can be N-Boc-piperidino, or N- carboethoxypiperidino; Group III is defined as either: Group III:

(i) Hydrogen; or

(ii) Unsubstituted, mono or disubstituted Ci-iδ alkyl, C0-16 alkylamino, Co-iβ alkyloxyalkyl or C2-16 alkenyl wherein the substituents are independently selected from the group consisting of hydroxy, Cι-8 alkyl, Ci-s alkyloxyalkyl, Ci-s alkylthioalkyl, phenyl- Ci-s alkylamino, Ci-s alkoxycarbonyl; or Co-6 carboxyl, triazole, 2,3- (methylenedioxy)ben2yl; or

(iii) substituted or unsubstituted N or C-linked pyrrolidino, piperidino, piperidonyl, morpholino, piperazino, N-Boc-

piperazino, N-Cι-10 alkylpiperazino, N-C3-6 alkenylpiperazino, N-(Cι-6 alkoxy C1-6 alkyl) piperazino, N- (C1-6 alkoxy C3-6 alkenyl) piperazino, N-(Cι-β alkylamino Ci-

6 alkyl) piperazino, N-(Cι-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 Ci-iθ alkyloxy, C3-16 alkenyloxy,

substituted C3-i6 alkynyloxy; or (b) substituted C1-6 alkyl-amino, di(substituted Ci-β alkyl) amino; or (c) C3-6 alkenyl- amino, di(C3-6 alkenyl) amino, substituted C3-6 alkenyl-amino, di(substituted

C3-6 alkenyl) amino; or

(d) CONHC1-C16 alkyl, COOCι-C₁₆ alkyl, Co-11

5 alkylCO₂H, Co-nNHC^NHR¹¹, C0-11NHSO2R¹¹,

trans- CH=CHCO₂Rⁿ, or trans- CH=CHCONHRⁿ wherein R¹¹ is Ci-iβ alkyl, or C1-16 alkyl aryl, in which the said aryl group

such as phenyl, or pyridyl, is mono- or

10 disubstituted with a member selected from the

group consisting of hydroxy, halo, Ci-β alkyl and

C1-6 alkyloxy, Cι-6 cycloalkyloxy, or Ci-C4 alkyl aryl or C1-C4 alkoxy aryl in which said aryl

group is either unsubstituted, mono- or

15 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

20 heterocycles, piperazino, N-Cι-6 alkylpiperazino,

N-C3-6 alkenylpiperazino, N-(Cι-β alkoxy Ci-β alkyl) piperazino, N-(Cι-β alkoxy C3-6 alkenyl) piperazino, N-(Cι-β alkylamino Ci-β alkyl) piperazino, or N-(Cι-6 alkylamino C3-6 alkenyl) piperazino, where the substituents are

chosen from hydroxy, C1-12 alkylalkoxy, C1-12 alkylamino,

C3-12 alkenyloxy, or C3-12 alkenylamino; or

(iv) either mono-, di, or tri-substituted aryl, or C0-C12 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 - CH₂CONHC₂H5-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

Figure 2 (cont.)

Figure 2 (cont.)

Figure 2 (cont.)

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 (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-thiazol-4-yl}- phenoxy)-acetic acid 47

Example 3 4-(4-Ethoxycarbonylmethoxy-ρhenyl)-5-(4-{(E)-2-[l-(4- pentyl-phenyl)-ethylcarbamoyl]~vinyl}-phenyl)-thiazole-2- carboxylic acid ethyl ester 48

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

Example 5 (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2-ρyridin-3- yl-thiazol-4-yl}-phenoxy)-acetic acid ethyl ester 50

Example 6 (4-{5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-2-pyridin-3- yl-thiazol-4-yl}-phenoxy)-acetic acid 51 Example 7

2-[({4-[5-[4-(2-Hexylcarbamoylvinyl)ρhenyl]-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)ρhenyl]-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)ρhenyl]- 2-( lH-ρyrrol-2-yl)thiazol-4- yl]phenoxy]acetylamino]propionic acid 54

Example 10 (2S)-3-Hydroxy-2-[2-[4-[5-[4-(2-hexylcarbamoylvinyl)ρhenyl]- 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

3-Hydroxy-2-[2-(2-[4-[5-[4-(2-octylcarbamoylvinyl)-phenyl]-2- ( lH-ρyrrol-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 2-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(lH-ρyrrol- 2-yl)-thiazol-4-yl]-phenoxy}-acetylamino)-ethanesulfonic

acid 59 Example 15

2-(2-{4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2-(lH-ρyrrol-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-( lH-pyrrol-2-yl)thiazol-4- yl]phenoxy]acetylamino]propionic acid 61

Example 17 (2-{4-[5-[4-(2-Hexylcarbamoylvinyl)ρhenyl]-2-(lH-ρyrrol-2- yl)thiazol-4-yl]phenoxy}acetylamino)acetic acid 62

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

Example 19 4-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-( lH-pyrrol- 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 2-(2-{4-[5-[4-(2-hexylcarbamoylvinyl)ρhenyl]-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, methane sulfonate, 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, 1^st ed; Pergamon: Oxford, 1984 and 2) Joule, J. A.; Mills, K.; and Smith, G.F., Heterocylic Chemistry, 3^rd 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. δs Med. Chem., 6, 87-92, (1996); Maduskuie et al, J. Med. Chem., 38, 1067-1083 (1995).

S 8 "

R R' Br₂ R R' „_RΛ R R

R NH,

O Br O Heat, H₂0 T

7 _R„ 9

Scheme 1

Thiazoles 9 can be synthesized through a common α-

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):

^•R ^A1Me3 'R NMe(OMe) '-Li '^R, .R"

^C0₂H ^ ^ ^

13 HNMe(OMe) O 14 15 O

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.

BrCH₂C02Et or BrCH₂C0₂ ^fBu Cs₂C0₃, 82%

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.

Scheme 4

Lawesson's

HO O ^0xa|y' Chloride C| O Acetone H₂N O Reagent

Y ^► ► ^H°^N

R³ Drop DMF s CH₃C0₂NH₄ R3 THF, Reflux R° 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(hydroxyme thy 1) 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

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 (Η-NMR) were measured on either a Varian (300 MHz) or a Varian (400 MHz).

Chemical shifts (δ) 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 °C and a source block at 120 °C).

LC chromatograms for LC/MS were measured using an eluant of CH₃CN (0.1% CF3CO₂H)/H₂O (0.1% CF3CO2H) (V:V) on a Hewlett Packard HP1100 HPLC, in the range 200-300 nm

with a Diode Array Detector (DAD); 5 μl 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% CH₃CN in 0.2 minutes, 10% CH CN for 1.4 minutes; LC element split 1:4 directly into ion source (500

μL/min).

The chromatography columns used for LC in LC/MS and

HPLC were 50 x 4.6 mm C-8 with 5 μm particle sizes and

Zorbax 150 x 4.6 mm C-8 with 5 μm 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 R¹ = Me) (Scheme 3) 4-Bromobenzyl-4-methoxyphenylketone 20a

To a mixture of p-bromo-phenylacetic acid 16 (51g, 237 mmol, 1 equiv.), and SOCk> (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 AICI3 (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 CHCI3 (150 x 3 mL). The combined extracts were washed with sat. NaHCO3 (aq.) (2 x 200 mL), and water (3 x 200 mL), dried (MgSO₄), 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: Η-NMR (300 MHz, CDCI3): 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), Lil (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 (IN) HC1, and extracted with EtOAc (3 x 150 mL). The combined extracts were washed with water (3 x 200 mL), Sat. NaHCO3 (200 mL), and brine (3 x 200 mL), successively, dried (MgSO4), 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: ⁱH-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 (R¹ = 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]phenoxy acetic ethyl ester (R¹ = Et) 19 53 g (82%).

Data for Compound 19: *H-NMR (300 MHz, CDC1₃): 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 CHCI3 (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. NaHCO₃ (_aq) (2x 200 mL) and water (3 x 200 mL), dried

(MgSO4), 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; Η-NMR (300 MHz, CDCI3): 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 (R^r=Et) (Scheme 4), using {4[Bromo-(4-bromophenyl) acetyl] phenoxy} acetic acid ethyl ester 20 (R^r=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 (R^r=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(OA (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 (MgSO4) 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 HC1 and extracted with ethyl acetate. The ethyl acetate solution was washed with water and brine, dried (MgSO4) 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 °C 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.

General Method 8: Preparation of Thioamide using O,O'- Diethyldithiophosphate

To a solution of nitrile (2.50ml, 23.6mmol) in 4N HC1- EtOAc(30ml) was added O,O'-Diethyldithioρhosphate(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.

OEt EtO-P=S SH

EtOOC CN EtOOC CSNHc

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 was dissolved in IN HC1 (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 H2O (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

H2O _{u nrϊ}^ 1 NaOH (1eq.) ,

^MeoocT^^ _{EH 1 N} HCI (1eq.) reflux overnight

ight X

3

c-N NH n-BuOH reflux overnight c-N N λ NH2 18%

*The product including 4-Thioureido-butyric acid which was de-esterified was re-esterified with TMSCH₂N₂. **At first , the solution was neutralized with IN NaOH, but re-acidified with IN HC1.

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 (RI' = Et) (5.0 g, 11.0 mmol, lequiv.), ethyl thiooxamate (1.53, l l .Ommol, 1.0 equiv.), NaHCO₃ (2.8 g, 33.0 mmol, 3 equiv.) and CH3CN (30 mL) which, after purification by silica gel column chromatography (Hexane-EtOAc, 4: 1), gave the intermediate aryl bromide 22 A 4.1g (76%) (where R³ =ethylcarboxylate) .

Data for 22A: Η-NMR (300 MHz, CDCls): 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(OA (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 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- ethoxy carbon -yl methoxy-phenyl)-thiazole-2-carboxylic acid ethyl ester 46: ⁱH-NMR (300 MHz, CDCls): 7.62 (d, IH, 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, IH, J= 15.6), 5.68 (t, IH, 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: ⁱH-NMR (300 MHz, DMSO-de): 9.18 (s, IH), 8.11 (t, IH, 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, IH, 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 C32H41N2O4S ([M+H]) 549.3. Example 3 4-(4-Ethoxycarbonylmethoxy-phenyl)-5-(4-{(E)-2-[l-(4- pentyl-phenyl)-ethylcarbamoyl]-vinyl}-phenyl)~thiazole-2- carboxylic acid ethyl ester 48 The ethyl ester 4-(4-Ethoxycarbonylmethoxy-phenyl)-5-(4-

{(E)-2-[l-(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.), NaHCO3 (2.8 g, 33 mmol, 3 equiv.) and CH3CN (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 22 A - 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 = l-(4-pentyl-phenyl)-ethyl) (0.25 g, 1 mmol, 1.2 equiv.) prepared using General Method 6 from l-(4-pentlyphenyl)ethylamine, Et3N (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-[l-(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-[l-(4- pentyl-phenyl)-ethylcarbamoyl]-vinyl}-phenyl)-thiazole-2- carboxylic acid ethyl ester 48: *H-NMR (400 MHz, CDC1₃): 7.59 (d, IH, 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, IH, J = 15.6), 6.15 (d, IH, J =8.4), 5.27-5.20 (m, IH), 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- Ethoxycarbonylmethoxy-phenyl)-5-(4-{(E)-2-[l-(4-pentyl- phenyl)-ethylcarbamoyl]-vinyl}-phenyl)-thiazole-2-carboxylic acid ethyl ester 48 (0.1 g, 0.153 mmol, 1 equiv.), with IN 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-[l-(4-Pentyl- phenyl)-ethylcarbamoyl]-vinyl}-phenyl)-thiazol-4-ylJ-phenoxy}- acetic acid 49, 12 mg (14 %).

Data for {4-[5-(4-{(E)-2-[l-(4-Pentyl-phenyl)-ethylcarbamoyl]- vinyl}-phenyl)-thiazol-4-yl]-phenoxy}-acetic acid 49: ²H-NMR (300 MHz, DMSO-de): 9.18 (s, IH), 8.55 (d, IH, 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, IH, 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 C33H35N2O4S ([M+H]) 555.2.

Example 5 (4-{5-[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 (RI' = Et) (1 g, 2.2 mmol, 1 equiv.), thionicotinamide (390 mg, 2.9 mmol, 1.3 equiv.), NaHCO₃ (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): ⁱH-NMR (300 MHz, CDCI3): 9.19 (s, IH), 8.66 (d, IH, J= 3.9), 8.28 (d, IH, J= 8.1), 7.51 (d, 2H, J = 8.7), 7.47 (d, 2H, J= 8.4), 7.41-7.37 (m, IH), 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- vinyl)-phenyl]-2-pyridin-3-yl-thiazol-4-yl}-phenoxy)-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: ⁱH- NMR (300 MHz, CDCls): 9.21 (d, IH, J = 1.8), 8.68 (dd, IH, J = 4.8, 1.5), 8.31 (td, IH, J = 8.7, J = 1.5), 7.63 (d, IH, 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, IH, J = 15.6), 5.61 (t, IH, 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-thiazol-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-[l-(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: ⁱH-NMR (300 MHz, DMSO-de): 9.19 (d, IH, J = 2.1), 8.71 (d, IH, J = 4.8), 8.36 (br d, IH, J = 7.8), 8.12 (t, IH, 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, IH, 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 C37H44N2O4S [M+H] 626.3

Example 7 2-r({4-r5-y4-(2-Hexylcarbamoylvinyl)ρhenvn-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 Et₃N (63 mg, 0.62 mmol) in DMF (5 mL) was stirred at r.t. overnight. The reaction mixture was acidified with IN 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, CHsCN/0.1% TFA 55:45) to give 2-[({4-[5-[4-(2- Hexylcarbamoylvinyl)phenyl]-2-(lH-pyrrol-2-yl)thiazol-4- yl]phenoxy}acetyl)-(2-hydroxy-l, 1 - bishydroxymethylethyl)amino]ethanesulfonic acid 52 as a yellow solid (85 mg, 20%).

₃

Data for 2-[({4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2-(lH-pyrrol- 2-yl)thiazol-4-yl]phenoxy}acetyl)-(2-hydroxy-l, 1 - bishydroxymethylethyl)amino]ethanesulfonic acid 52: ⁱH-NMR (250 MHz, DMSO-de): 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, IH), 6.62 (d, J= 15.8, IH), 6.6-6.7 (m, IH), 6.9-7.0 (m, IH), 6.95 (d, J = 8.9, 2H), 7.36 (d, J = 8.4, 2H), 7.37 (d, J = 15.8, IH), 7.44 (d, J = 8.9, 2H), 7.54 (d, J= 8.4, 2H), 8.07 (t, J = 5.6, IH), 8.8-8.9 (br, 2H), 11.79 (d, J = 2.1, IH).

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

Example 8 (S)-3-Hydroxy-2-f2-r4-r5-r4-(2-octylcarbamoylvinvHphenvn-2- (lH-pyrrol-2-vUthiazol-4- yl|phenoxy|acetylamino]propionic acid 53

Data for (S)-3-Hydroxy-2-[2-[4-[5-[4-(2- octylcarbamoylvinyl)phenyl]-2-(lH-pyrrol-2-yl)thiazol-4- yl]phenoxy]acetylamino]propionic acid 53: ⁱH-NMR (250 MHz, DMSO-de): 12.74 (br, IH), 11.79 (br, IH), 8.07-8.02 (m, IH), 7.54 (d, 2H, J= 9.7), 7.47-7.34 (m, 5H), 6.98-6.94 (m, 3H), 6.71-6.69 (m, IH), 6.61 (d, IH, J = 15.8), 6.19-6.16 (m, IH), 4.37-4.34 (m, IH), 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 rR)-3-Hvdroxy-2-f2-r4-r5-r4-(2-hexylcarbamoylvinyl)phenyn- 2-( lH-pyrrol-2-yl)thiazol-4- yl]phenoxylacetylaminolpropionic acid 54

Data for (R)-3-Hydroxy-2-[2-[4-[5-[4-(2- hexylcarbamoylvinyl)phenyl]-2-(lH-pyrrol-2~yl)thiazol-4- yljphenoxyjacetylaminojpropionic acid 54: ⁱH-NMR (250 MHz, DMSO-de): 12.70 (br, IH), 11.79 (br, IH), 8.07-8.02 (m, 2H), 7.55 -7.34 (m, 7H), 6.98-6.95 (m, 3H), 6.71- 6.68 (m, IH), 6.61 (d, IH, J= 15.8), 6.18-6.15 (m, IH), 4.37- 4.33 (m, IH), 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)-3Hvdroxy-2-r2-r4-r5-r4-(2-hexylcarbamoylvinyl)phenyn- 2-(morpholin-4-yl)thiazol-4- yl]phenoxy]acetylaminolpropionic acid 55

Data for (2S)-3Hydroxy-2-[2-[4-[5-[4-(2- hexylcarbamoylvinyl)phenyl]-2-(morpholin-4-yl)thiazol-4- yl]phenoxy]acetylamino]propionic acid 55: ⁱH-NMR (250MHz, DMSO-de): 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, IH), 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 (2-r4-r5-r4-(2-Octylcarbamoylvinyl)phenvn-2-(lH-ρyrrol-2- yl)thiazol-4-yl]phenoxy|acetylamino)acetic acid 56

Data for (2-[4-[5-[4-(2-Octylcarbamoylvinyl)phenyl]-2-(lH-pyrrol-2- yl)thiazol-4-yl]phenoxy]acetylamino)acetic acid 56: ⁱH-NMR (250 MHz, DMSO-de): 12.65 (br, IH), 11.80 (br, IH), 8.38 (t, IH, J= 5.9), 8.05 (t, IH, 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, IH), 6.71-6.69 (m, IH), 6.61 (d, IH, J= 15.8), 6.19-6.17 (m, IH), 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-Hvdroxy-2-r2-(2-r4-r5-r4-(2-octylcarbamoylvinyl)-phenvn-2- (lH-pyrrol-2-yl)thiazol-4-yl1phenoxylacetylamino) acetylamino propionic acid 57

Data for 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: ⁱH-NMR (250 MHz, DMSO-de): 11.81 (br, IH), 8.25 (t, IH, J= 6.0), 8.10 (t, IH, 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, IH), 6.70-6.69 (m, IH), 6.63 (d, IH, J= 15.8), 6.19- 6.16 (m, IH), 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-r3-r4-r4-r4-r(l-Carboxy-2-hydroxy-ethylcarbamoyl) methoxy-phenyl]-2-(lH-pyrrol-2-yl)-thiazol-5-yl -phenyl]- acryloylaminol-piperidine-1 -carboxylic acid ethyl ester 58

Data for (2S)-4-[3-[4-[4-[4-[(l-Carboxy-2-hydroxy- ethylcarbamoyl)methoxy-phenyl]-2-(lH-pyrrol-2-yl)-thiazol-5-yl]- phenyl]-acryloylamino]-piperidine-l -carboxylic acid ethyl ester 58;ⁱH 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, IH, J= 4.1), 4.60 (s, 2H), 6.19 (dd, IH, J= 2.3, 5.9), 6.61(d, IH, J= 15.6), 6.70-6.78 (m, IH), 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, IH). Example 14 2-(2-{4-r5-r4-(2-Hexylcarbamoyl-vinyl)-phenvn-2-(l-H^r-pyrrol- 2-yl)-thiazol-4-yl]-phenoxy}-acetylamino)-ethanesulfonic acid 59

Data for 2-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(lH- pyrrol-2-yl)-thiazol-4-yl]-phenoxy}-acetylamino)-ethanesulfonic

acid 59: ⁱH-NMR (250 MHz, DMSO-de) 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, IH), 6.65 (d, J = 15.8 Hz, IH), 6.6-6.7 (m, IH), 6.93 (d, J = 8.8 Hz, 2H), 6.9-7.0 (m, IH), 7.34 (d, J= 8.4 Hz, 2H), 7.37 (d, J= 15.8 Hz, IH), 7.43 (d, J = 8.8 Hz, 2H), 7.54 (d, J = 8.4 Hz, 2H), 11.85 (d, J = 2.5 Hz, IH). Example 15 2-(2-{4-r5-f4-(2-Hexylcarbamoylvinyl)phenvn-2-(lH-pyrrol-2- yl)thiazol-4-yl|phenoxy}acetylamino)-3-hydroxybutyric acid 60

Data for 2-(2-{4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2- (lH-pyrrol-2-yl)thiazol-4-yl]phenoxy}acetylamino)-3-

hydroxy butyric acid 60: ⁱH NMR (DMSO-de): 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 (IH, m), 6.71 (IH, d, J=15.8), 6.79-6.83 (IH, m), 7.03-7.10 (3H, m), 7.46 (2H, d, J = 8.1), 7.48 (IH, d, J= 15.8), 7.55 (2H, d, J= 8.6), 7.64 (2H, d, J=8.6), 7.89 (IH, d, J= 8.6), 8.20 (IH, t, J= 5.8), 11.93 (IH, s). Example 16 (S)-3-Hydroxy-2-r2-r4-r5-r4-(2-Phenylbutylcarbamoyl)phenvn- 2-(lH-pyrrol-2-yl)thiazol-4-yl -phenoxy]acetylamino propionic acid 61

Data for (S)-3-Hydroxy-2-[2-[4-[5-[4-(2-

Phenylbutylcarbamoyl)phenyl]-2-(lH-pyrrol-2-yl)thiazol-4-yl]-

phenoxy]acetylamino]propionic acid 61: ⁱH NMR (DMSO-de); 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 (IH, m), 4.59 (2H,s), 6.15-6.21 (IH, 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-r5-r4-(2-Hexylcarbamoylvinyllphenvn-2-(lH-ρyrrol-2- yl)thiazol-4-yl1phenoxy}acetyl- amino) acetic acid 62

Data for 2-(2-{4-[5-[4-(2-Hexylcarbamoylvinyl)phenyl]-2-(lH- pyrrol-2-yl)thiazol-4-yl]phenoxy}acetyl-amino) acetic acid 62: ^XH NMR (DMSO-de): 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 (IH, m), 6.61 (IH, d, J=15.6), 6.68-6.72 (IH, m), 6.92-7.00 (3H, m), 7.30-7.50 (5H, m), 7.54 (2H, d, J= 7.5), 8.09 (IH, t, J= 5.5), 8.36-8.45 (IH, m), 11.82 (IH, s).

Example 18 2-^-(2-{4-r5-f4-(2-Hexylcarbamoyl-vinyl)-phenyl -2-(l- methoxymethyl-lff-pyrrol-2-yl)-thiazol-4-yπ-phenoxy}- acetylamino)-3-hydroxy-propionic acid 63

Data for .2-(S)-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(l - methoxymethyl-lH-pyrrol-2-yl)-thiazol-4-yl]-phenoxy}- acetylamino)-3-hydroxy-propionic acid 63: ⁱH-NMR (250 MHz, DMSO-de): 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, IH), 4.58 (s, 2H), 5.80 (s, 2H), 6.21 (dd, J= 2.7 and 3.8, IH), 6.62 (d, J = 15.8, IH), 6.80 (dd, J = 1.7 and 3.8, IH), 6.96 (d, J = 8.9, 2H), 7.23 (dd, J = 1.7 and 2.7, IH), 7.38 (d, J = 8.4, 2H), 7.39 (d, J = 15.8, IH), 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-r5-f4-(2-Hexylcarbamoyl-vinyl)-ρhenvn-2-(li-r-ρyrrol- 2-yl)-thiazol-4-yl]-phenoxy}-acetylamino)-3-hydroxy-butyric acid 64

Data for 4-(2-{4-[5-[4-(2-Hexylcarbamoyl-vinyl)-phenyl]-2-(lH- pyrrol-2-yl)-thiazol-4-yl]-phenoxy}-acetylamino)-3-hydroxy-butyric

acid 64: ⁱH-NMR (250 MHz, DMSO-de): 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, IH), 2.35 (dd, J= 4.3 and 15.3 Hz, IH), 3.0-3.2 (m, 4H), 3.8-4.0 (m, IH), 4.50 (s, 2H), 5.01 (d, J = 4.8 Hz, IH), 6.1-6.2 (m, IH), 6.60 (d, J = 15.9 Hz, IH), 6.6-6.7 (m, IH), 6.94 (d, J = 8.8 Hz, 2H), 7.35 (d, J= 8.3 Hz, 2H), 7.38 (d, J= 15.9 Hz, IH), 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, IH), 12.07 (br s, IH). Example 20 4-Carbamoyl-2-(2-{4-[5-f4-(2-hexylcarbamoylvinyl)phenyl-2- ( lH-pyrrol-2-yl)thiazol-4-vn- phenoxy}acetylamino)butyric acid 65

Data for 4-Carbamoyl-2-(2-{4-[5-[4-(2- hexylcarbamoylvinyl)phenyl-2-(lH-pyrrol-2-yl)thiazol-4-yl]- phenoxy}acetylamino)butyric acid 65: ^lH NMR (DMSO-dβ): 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 (IH, m), 4.64 (2H, s), 6.26-6.30 (IH, m), 6.71 (IH, d, J= 15.7), 6.78-6.82 (IH, m), 6.88-6.93 (IH, m), 7.00-7.10 (4H, m), 7.40-7.60 (5H, m), 7.64 (2H, d, J= 8.3), 8.17-8.23 (IH, m), 8.51 (IH, d, J= 8.4), 11.9 (IH, s). Example 21 3-Hydroxy-2-(2-{4-f5-f4-(2-phenethylcarbamoylvinyl)phenyl - 2-(lH-pyrrol-2-vHthiazol-4-vn- phenoxy}acetylamino)propionic acid 66

Data for 3-Hydroxy-2-(2-{4-[5-[4-(2- phenethylcarbamoylvinyl)phenyl]-2-(lH-pyrrol-2-yl)thiazol-4-yl]-

phenoxy}acetylamino)propionic acid 66: ^:H NMR (DMSO-dβ); 2.70-2.80 (2H, m), 3.30-3.50 (2H, m),3.60-3.90 (2H, m), 4.30- 4.50 (IH, m), 4.59 (2H, s), 5.00-5.20 (IH, m), 6.17-6.22 (IH, m), 6.61 (IH, d, J= 15.8), 6.70-6.74 (IH, 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 (IH, d, J=8.3), 8.17-8.26 (IH, m), 11.8 (IH, s), 12.8 (lH,s).

Example 22 2-(2-{4-r5-r4-(2-hexylcarbamoylvinvUρhenvn-2-(lH-pyrrol-2- yl)thiazol-4-yl]phenoxy}acetyl- amino)-3-mercaptopropionic acid 67

Data for 2-(2-{4-[5-[4-(2-hexylcarbamoylvinyl)phenyl]-2-(lH- pyrrol-2-yl)thiazol-4-yl]phenoxy}acetyl- amino)-3-mercaptopropionic acid 67: ⁱH NMR (DMSO-dβ): 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(lH, m), 4.59 (2H, s), 6.15-6.20 (IH, m), 6.61 (IH, d, J= 15.8), 6.68-6.72 (IH, m), 6.90-7.00 (3H, m), 7.32-7.50 (5H, m), 7.57 (2H, d, J= 7.9), 8.05 (IH, t, J= 5.4), 8.29 (IH, 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 IC50 in micromolar against P-selectin are tabulated below in Table 2. Table 2

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-Lewis^x 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- Lewis^x 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 CaCk>). 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-Lewis^x -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% MgCk>. 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. IC50 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 RδδD Systems (Minneapolis, MN) was diluted to

2.5 μg/mL in Dulbecco's PBS containing calcium and

magnesium (PBS⁺). Falcon Pro-Bind microtiter plate wells were

incubated with 50 μL 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 μL. 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 μL

per well, bringing the compound to its final test concentration in

a total volume of 100 μL. 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 μL PBS⁺ (added slowly using a manual multichannel

pipettor). Retained cells were labeled directly on the plate by

adding 5 μg/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 IC50 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-Lewis^x 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-Lewis^x 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 CaCk>). 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- Lewis^x-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% MgC . 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. IC50 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 μg/mL in Dulbecco's PBS (PBS-). Falcon Pro- Bind microtiter plate wells were coated with 100 μL 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 μg/mL in PBS-, and coated microplate well were incubated with 50 μL 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 μg/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 μL. 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 centrifαgation, 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 μL per well, bringing the compound to its final test concentration in a total volume of 300 μL. 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 μM 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.

R¹ R² w Where at least one of R¹ R² or R³ = Et0₂C

T _R3 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 pho sphatidyl-choline s .

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.

Claims

We claim:

1. A compound having the structural Formula 1 :

R¹ R² x ^Nr R₃ ^3s

Where at least one of R¹, R² or R³ contains calcium binding moiety e.g. CO_aH etc.

Formula 1

In which the said calcium binding moiety is selected from Group I as shown.

CO (OH or *N- linked amino acid)

Group I = or

where

following in Table 1 : Table 1

Table 1 (cont.)

In the above table n", and/or n¹ 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.

In Case A: When R¹ is selected from Group I, one of R² or R³ must be selected from Group II as shown, Group II:

(i) Unsubstituted, mono-, di-, or tri-substituted aryl-Co-ii 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, Co-eCONHRⁱo, C0-6NHSO2R¹⁰, trans- CH=CHCO₂R¹⁰, or trαns-CH=CHCONHR¹⁰, wherein R¹⁰ is d-iβ alkyl, Ci-iβ alkyloxyalkyl, C5-8 cycloalkyl, Ci-n alkylaryl, or C1-5 alkylaryl Cι-8 alkyl in which the said alkyl group or said aryl group is unsubstituted, mono- or disubstituted with a member selected from the group consisting of hydroxy, carboxy, halo, d-β alkyl and Ci-6 alkyloxy, C].^ cycloalkyloxy, 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, or C1-4 alkyloxy; or R¹⁰ can be N-Boc- piperidino, or N-carboethoxypiperidino;

and one of R² or R³ must be selected from Group III as shown. Group III:

(i) Hydrogen; or

(ii) Unsubstituted, mono or disubstituted Ci-iβ alkyl, Co-iβ alkylamino, Co-iβ alkyloxyalkyl or C2-16 alkenyl wherein the substituents are independently selected from the group consisting of hydroxy, Cι-8 alkyl, Ci-s alkyloxyalkyl,

Ci-s alkylthioalkyl, phenyl- C 1-8 alkylamino, Ci-s alkoxycarbonyl; or Co-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~Ci-io alkylpiperazino, N-C3-6 alkenylpiperazino, N-(Cι-6 alkoxy C 1-6 alkyl) piperazino, N- (Ci-6 alkoxy C3-6 alkenyl) piperazino, N-(Cι_6 alkylamino Ci- 6 alkyl) piperazino, N-(Cι-6 alkylamino C3-6 alkenyl) piperazino, wherein the substituents are N or C- linked and are independently selected from:

(a) substituted Ci-iβ alkyloxy, C3-16 alkenyloxy, substituted C3-16 alkynyloxy; or (b) substituted d-6 alkyl-amino, di(substituted Cι-6 alkyl) amino; or

(c) C3-6 alkenyl-amino, di(C3-6 alkenyl) amino, substituted C3-6 alkenyl-amino, di(substituted

5 C3-6 alkenyl) amino; or

(d) CONHC1-C16 alkyl, COOCι-C₁₆ alkyl, Co-11 alkylCO₂H, C₀-nNHC(O)NHR^H, C0-11NHSO2R¹¹, trans- CH=CHCO₂Rⁿ, or trans-

CH=CHCONHRⁿ wherein R¹¹ is C1-16 alkyl, or

10 Ci-iβ alkyl aryl, in which the said aryl group is mono- or disubstituted with a member selected

from the group consisting of hydroxy, halo, C1-6

alkyl and C1-6 alkyloxy, C1-6 cycloalkyloxy, or Ci-

C4 alkyl aryl or C1-C4 alkoxy aryl in which said

15 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,

20 substituted, uracil or other purine or pyrimidine

heterocycles, piperazino, N-Ci-β alkylpiperazino,

N-C3-6 alkenylpiperazino, N-(Cι-β alkoxy C1-6 alkyl) piperazino, N-(Cι-6 alkoxy C3-6 alkenyl) piperazino, N-(Cι-6 alkylamino Cι-6 alkyl) piperazino, or N-(Cι-β alkylamino C3-6 alkenyl) piperazino, where the substituents are chosen from hydroxy, C1-12 alkylalkoxy, C1-12 alkylamino,

C3-12 alkenyloxy, or C3-12 alkenylamino; or (iv) either mono-, di, or tri-substituted aryl, or C0-C12 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 - CH₂CONHC₂H₅-O-Fucose or -CH2CONHC₂H₅-O-Mannose.

In 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 as defined above.

In 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 as defined above.

And the corresponding pharmaceutically acceptable salts and esters thereof.

2. A compound according to claim 1, by the name of 5- [4-((E)-2-Dodecylcarbamoyl-vinyl)-phenvf|-4-(4- ethoxycarbonyl methoxy-phenyl)-thiazole-2-carboxylic acid ethyl ester having the following structural formula:

And the corresponding pharmaceutically acceptable

salts thereof.

3. A compound according to claim 1, by the name of (4- {5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl]-thiazol-4-yl)- phenoxy] -acetic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

4. A compound according to claim 1, by the name of 4- (4-Ethoxycarbonylmethoxy-phenyl)-5-(4-((E)-2-[l-(4-pentyl- phenyl)-ethylcarbamoyll-vinyl}-phenyl)-thiazole-2 -carboxylic acid ethyl ester having the following structural formula:

1

And the corresponding pharmaceutically acceptable salts thereof.

5. A compound according to claim 1, by the name of (4- r5-(4-{(E)-2-[l-(4-Pentyl-phenyll-ethylcarbamoyll-vinyl}- phenyl) -thiazol-4-yl|-phenoxy}-acetic acid having the following structural formula: 1

And the corresponding pharmaceutically acceptable salts and esters thereof.

6. A compound according to claim 1, by the name of (4- {5-[4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl1-2-pyridin-3-yl- thiazol-4-yl|-phenoxy) -acetic acid ethyl ester having the following structural formula:

And the corresponding pharmaceutically acceptable salts thereof.

7. A compound according to claim 1, by the name of (4- {5-r4-((E)-2-Dodecylcarbamoyl-vinyl)-phenyl1-2-pyridin-3-yl- thiazol-4-yll-phenoxy) -acetic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

8. A compound according to claim 1, by the name of c f({4- (5- [4- (2-Hexylcarbamoylvinyl)phenyl|-2- ( 1 H-pyrrol-2- yl)thiazol-4-yl]phenoxy|acetyl)-(2-hydroxy- 1, 1- bishydroxymethylethyl) ammo] ethanesulfonic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

9. A compound according to claim 1 , by the name of (S)- 3-Hydroxy-2-[2-[4-[5-[4-(2-octylcarbamoylvinyl)phenyl1-2^ ( 1 H-pyrrol-2-yl)thiazol-4-yllphenoxy1acetylamino1propionic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

10. A compound according to claim 1, by the name of (R)- 3-Hydroxy-2-[2-[4-[5-[4-(2-hexylcarbamoylvinyl)phenyl]-2-(lH- pyrrol-2-yl)thiazol-4-yl]phenoxy]acetylamino]propionic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

11. A compound according to claim 1 , by the name of (2S)-3Hydroxy-2-[2-r4-[5-r4-(2-hexylcarbamoylvinyllρhenyll- 2-(morpholin-4-yl)thiazol-4- yllphenoxylacetylaminolpropionic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

12. A compound according to claim 1, by the name of (2- [4-[5- 4-(2-Octylcarbamoylvinyl)phenyl]-2-(lH-pyrrol-2- yl)thiazol-4-yl]phenoxy]acetylamino)acetic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

13. A compound according to claim 1, by the name of 3^ Hydroxy-2- 2-(2-[4-[5-[4-(2-octylcarbamoylvinyl)-phenyl1-2- ( 1 H-pyrrol-2 -yl) thiazol-4- yl] phenoxy] acetylamino) acetylamino]propionic acid having the following structural formula:

no

And the corresponding pharmaceutically acceptable salts and esters thereof.

14. A compound according to claim 1, by the name of (2S)-4-[3-r4-r4-[4-r(l-Carboxy-2-hvdroxy- ethylcarbamoyl)methoxy-phenyl]-2-(lH-pyrrol-2-yl)-thiazol- 5-yl]-phenyl]-acryloylamino]-piperidine- 1-carboxylic acid ethyl ester having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

il l

15. A compound according to claim 1, by the name of 2^ (2-{4-r5-r4-(2-Hexylcarbamoyl-vinyl)-phenyl1-2-(lH-pyrrol-2- yl)-thiazol-4-yl]-phenoxy}-acetylamino) -ethanesulfonic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

16. A compound according to claim 1, by the name of 2- (2-{4-[5- 4-(2-Hexylcarbamoylvinyl phenyl]-2-(lH-pyrrol-2- yl)thiazol-4-yl]phenoxy|acetylamino)-3-hydroxybutyric acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

17. A compound according to claim 1, by the name of (S)- 3-Hvdroxy-2-r2-r4-r5-r4-(2-Phenylbutylcarbamoyl)phenyl1-2- (lH-pyrrol-2-yl)thiazol-4-yl]-phenoxy]acetylamino]propionic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

18. A compound according to claim 1, by the name of 2- (2-{4-[5-[4-f2-Hexylcarbamoylvinyl)phenyl1-2-(lH-pyrrol-2- yl)thiazol-4-yl1phenoxy|acetyl-amino) acetic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

19. A compound according to claim 1, by the name of 2

(S)-(2-(4-[5-r4-(2-Hexylcarbamoyl-vinyl)-phenyll-2-f l- methoxymethyl- 1 H-pyrrol-2-yl) -thiazol-4-yll -phenoxy}- acetylamino)-3-hvdroxy-propionic acid having the following structural formula:

₃

And the corresponding pharmaceutically acceptable salts and esters thereof.

20. A compound according to claim 1, by the name of 4^ (2-{4-r5-r4-(2-Hexylcarbamoyl-vinyl)-phenyll-2-f lH-ρyrrol-2- yl)-thiazol-4-yl]-phenoxy}-acetylamino)-3-hydroxy-butyric acid having the following structural formula:

₃

And the corresponding pharmaceutically acceptable salts and esters thereof.

21. A compound according to claim 1 , by the name of 4; Carbamoyl-2-(2-{4-[5-[4-(2-hexylcarbamoylvinyl)phenyl-2- ( 1 H-pyrrol-2-yl)thiazol-4-yl]-phenoxy|acetylamino)butyric acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

22. A compound according to claim 1, by the name of 3;

Hydroxy-2-(2-(4- 5-[4-(2-phenethylcarbamoylvinyl)phenyl1-

2-(lH-pyrrol-2-yl)thiazol-4-yll- phenoxy|acetylamino)propionic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

23. A compound according to claim 1, by the name of 2- (2-{4-[5-f4-(2-hexylcarbamoylvinyl)phenyn-2-(lH-pyrrol-2- yl)thiazol-4-yl]phenoxy}acetyl-amino)-3-mercaptopropionic acid having the following structural formula:

And the corresponding pharmaceutically acceptable salts and esters thereof.

24. A method for treating human diseases involving P-, L- and E-selectin in a subject, which comprises the administration of an effective therapeutic amount of a compound selected from those defined in Claim 1 or the pharmaceutically acceptable salts and esters thereof.