WO1997049399A1

WO1997049399A1 - Il-8 receptor antagonists

Info

Publication number: WO1997049399A1
Application number: PCT/US1997/010904
Authority: WO
Inventors: Katherine L. Widdowson
Original assignee: Smithkline Beecham Corporation
Priority date: 1996-06-27
Filing date: 1997-06-24
Publication date: 1997-12-31
Also published as: EP0907362A4; JP2000513359A; EP0907362A1

Abstract

This invention relates to the use of phenyl ureas of formula (I) in the treatment of disease states mediated by the chemokine, Interleukin-8 (IL-8). The variables of formula (I) are defined herein.

Description

LL-8 RECEPTOR ANTAGONISTS

FIELD OF THE INVENTION

This invention relates to a novel group of phenyl urea compounds, processes for the preparation thereof, the use thereof in treating IL-8, GROα, GROβ, GROγ NAP-2 and ENA-78 mediated diseases and pharmaceutical compositions for use in such therapy.

BACKGROUND OF THE INVENTION

Many different names have been applied to Interleukin-8 (IL-8), such as neutrophil attractant/activation protein- 1 (NAP-1), monocyte derived neutrophil chemotactic factor (MDNCF), neutrophil activating factor (NAF), and T-cell lymphocyte chemotactic factor. Interleukin-8 is a chemoattractant for neutrophils, basophils, and a subset of T-cells. It is produced by a majority of nucleated cells including macrophages, fibroblasts, endothelial and epithelial cells exposed to TNF, IL-lα, IL-lβ or LPS, and by neutrophils themselves when exposed to LPS or chemotactic factors such as FMLP. M. Baggiolini et al, J. Clin. Invest. 84, 1045 (1989); J. Schroder et al, J. Immunol. 139, 3474 (1987) and J. Immunol. 144. 2223 (1990) ; Strieter, et al, Science 243. 1467 (1989) and J. Biol. Chem. 264. 10621 (1989); Cassatella et al, J. Immunol. 148. 3216 (1992).

Groα, GROβ, GROγ and NAP-2 also belong to the chemokine α family. Like IL-8 these chemokines have also been referred to by different names. For instance GROα, β, γ have been referred to as MGSAα, β and γ respectively (Melanoma Growth Stimulating Activity), see Richmond et al, J. Cell Physiology 129, 375 (1986) and Chang et al, J. Immunol 148, 451 (1992). All of the chemokines of the α-family which possess the ELR motif directly preceding the CXC motif bind to the IL-8 B receptor.

IL-8, GROα, GROβ, GROγ, NAP-2 and ENA-78 stimulate a number of functions in vitro. They have all been shown to have chemoattractant properties for neutrophils, while LL-8 and GROα have demonstrated T-lymphocytes, and basophiles chemotactic activity. In addition LL-8 can induce histamine release from basophils from both normal and atopic individuals GRO-α and LL-8 can in addition, induce lysozomal enzyme release and respiratory burst from neutrophils. LL-8 has also been shown to increase the surface expression of Mac- 1 (CD1 lb/CD 18) on neutrophils without de novo protein synthesis. This may contribute to increased adhesion of the neutrophils to vascular endothelial cells. Many known diseases are characterized by massive neutrophil infiltration. As IL-8, Groα, GROβ, GROγ and NAP-2 promote the accumulation and activation of neutrophils, these chemokines have been implicated in a wide range of acute and chronic inflammatory disorders including psoriasis and rheumatoid arthritis, Baggiolini et al, FEBS Lett. 307, 97 (1992); Miller et al, Crit. Rev. Immunol. 12. 17 (1992); Oppenheim et al, Annu. Rev. Immunol. 9. 617 (1991 ): Seitz et al., J. Clin. Invest. 87. 463 (1991): Miller et al, Am. Rev. Respir. Pis. 146. 427 (1992); Donnely et al., Lancet 341. 643 (1993). In addition the ELR chemokines (those containing the amino acids ELR motif just prior to the CXC motif) have also been implicated in angiostasis. Strieter et al, Science 258, 1798 (1992).

In vitro, IL-8, Groα, GROβ, GROγ and NAP-2 induce neutrophil shape change, chemotaxis, granule release, and respiratory burst, by binding to and activating receptors of the seven-transmembrane, G-protein-linked family, in particular by binding to IL-8 receptors, most notably the B-receptor. Thomas et al., J. Biol. Chem. 266. 14839 (1991 ); and Holmes et al., Science 253. 1278 (1991). The development of non-peptide small molecule antagonists for members of this receptor family has precedent. For a review see R. Freidinger in: Progress in Drug Research. Vol. 40, pp. 33-98, Birkhauser Verlag, Basel 1993. Hence, the IL-8 receptor represents a promising target for the development of novel anti- inflammatory agents.

Two high affinity human LL-8 receptors (77% homology) have been characterized: LL-8Rα, which binds only IL-8 with high affinity, and IL-8Rβ, which has high affinity for IL-8 as well as for GRO-α, GROβ, GROγ and NAP-2. See Holmes et al., supra; Murphy et al., Science 253. 1280 (1991 ); Lee et al., J. Biol. Chem. 267, 16283 (1992); LaRosa et al., J. Biol. Chem. 267. 25402 (1992); and Gayte et al., J. Biol. Chem. 268. 7283 (1993). There remains a need for treatment, in this field, for compounds which are capable of binding to the LL-8 α or β receptor. Therefore, conditions associated with an increase in IL-8 production (which is responsible for chemotaxis of neutrophil and T-cells subsets into the inflammatory site) would benefit by compounds which are inhibitors of LL-8 receptor binding.

SUMMARY OF THE INVENTION This invention provides for a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an LL-8 α or β receptor and which method comprises administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In particular the chemokine is LL-8.

This invention also relates to a method of inhibiting the binding of LL-8 to its receptors in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula (I).

Compounds of Formula (I) useful in the present invention are represented by the structure:

wherein

X is oxygen or sulfur; R is any functional moiety having an ionizable hydrogen and a pKa of 10 or less; Rl is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted

Ci- io alkyl; Ci- io alkyl; C2-10 alkenyl; Ci- io alkoxy; halosubstituted C i-10 alkoxy; azide; (CRgRg)q S(O)_tR4; hydroxy; hydroxy C i-4alkyl; aryl; aryl C i-4 alkyl; aryloxy; aryl Ci-4 alkyloxy; heteroaryl; heteroarylalkyl; heterocyciic, heterocyciic C l-4alkyl; heteroaryl Ci-4 alkyloxy; aryl C2-10 alkenyl; heteroaryl

C2- 10 alkenyl; heterocyciic C2-10 alkenyl; (CR₈R₈)qNR4R5; C2- 10 alkenyl C(O)NR4R5; (CR₈R8)q C(O)NR4R5; (CR₈R₈)q C(O)NR₄Rl0; S(O)3H; S(O)3R8; (CRgR8)q C(O)Rι i ; C2- 10 alkenyl C(O)R] 1 ; C2- 10 alkenyl C(O)ORn(CR₈R₈)q C(O)ORι₂; (CR₈R₈)q OC(O) Rn ; (CR₈R₈)qNR4C(O)Ri 1 , (CR₈R₈)q NHS(O)₂Rι₇, (CR₈R₈)q S(O)₂NR₄R₅; or two Rl moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; q is 0, or an integer having a value of 1 to 10; m is an integer having a value of 1 to 3; t is 0, or an integer having a value of 1 or 2; s is an integer having a value of 1 to 3; v is 0, or an integer having a value of 1 to 4; R4 and R5 are independently hydrogen, optionally substituted Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C] -4alkyl, heterocyciic, heterocyciic C1-.4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from oxygen, nitrogen or sulfur;

HET is an optionally substituted heteroaryl moiety;

R₈ is hydrogen or Ci-4 alkyl;

RlO is C ι_ιo alkyl C(O)2R8; Rl 1 is hydrogen, Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl Cι_4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC 1 - 4alkyl, optionally substituted heterocyciic, or optionally substituted heterocyclicC 1 _4alky 1 ;

Rl2 is hydrogen, Ci-io alkyl, optionally substituted aryl or optionally substituted arylalkyl;

Rl3 and R 14 are independently hydrogen or C 1.4 alkyl;

Rl7 is Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC ι_4alkyl, heterocyciic, or heterocyclicC i_4alkyl, wherein the aryl, heteroaryl and heterocyciic rings may all be optionally substituted; E is optionally selected from

Rt ; the asterix * denoting point of attachment of the ring; or a pharmaceutically acceptably salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of Formula (I) may also be used in association with the veterinary treatment of mammals, other than humans, in need of inhibition of IL-8 or other chemokines which bind to the IL-8 α and β receptors. Chemokine mediated diseases for treatment, therapeutically or prophylactically, in animals include disease states such as those noted herein in the Methods of Treatment section.

In compounds of Formula (I), R is suitably any functional moiety which provides an ionizable hydrogen having a pKa of 10 or less, preferably from about 3 to 9, more preferably from about 3 to 7. Such functional groups include, but are not limited to, hydroxy, carboxylic acid, thiol, -SR2 -OR2, -NH-C(O)R_a, -C(O)NR6R7, a substituted sulfonamides of the formula -NHS(O)2Rb, -S(O)2NHR_c, NHC(X2)NHRb, or a tetrazolyl; wherein X2 is oxygen or sulfur, preferably oxygen.

Preferably, the functional group is other than a sulfonic acid, either directly or as a substituent group on the aryl, heteroaryl, or heterocyciic moiety ring, such as in SR2 or OR2- More preferably R is OH, SH, or NHS(O)2Rb- Suitably, R2 is a substituted aryl, heteroaryl, or heterocyciic ring which ring contains the functional moiety providing the ionizable hydrogen having a pKa of 10 or less.

Suitably, R6 and R7 are independently hydrogen or a Ci-4 alkyl group, or R6 and R7 together with the nitrogen to which they are attached form a 5 to 7 member ring which ring may optionally contain an additional heteroatom which heteroatom is selected from oxygen, nitrogen or sulfur. This heteroring may be optionally substituted as defined herein.

Suitably R_a is an alkyl, aryl, arylCi-4alkyl, heteroaryl, heteroarylC i-4alkyl, heterocyciic, or a heterocyciic C]-4alkyl moiety, all of which may be optionally substituted, as defined herein below.

Suitably, R_D is a NR6R7, alkyl, aryl, arylCi-4alkyl, arylC2-4alkenyl, heteroaryl, heteroarylC ] -4alkyl, heteroarylC2-4 alkenyl, heterocyciic, or heterocyciic C 1 _4alkyl, or a heterocyciic C2-4alkenyl moiety, camphor, all of which may be optionally substituted one to three times independently by halogen; nitro; halosubstituted Ci-4 alkyl, such as CF3; Q-4 alkyl, such as methyl; Ci-4 alkoxy, such as methoxy; NR9C(O)R_a; C(O)NR6R7, S(O)3H, or C(O)OCi-4 alkyl.

Preferably, R_D is an optionally substituted phenyl, benzyl, or styryl. When Rb is a heteroaryl ring, it is preferably an optionally substituted thiazole, an optionally substituted thienyl, or an optionally substituted quinolinyl ring. Suitably, R9 is hydrogen or a Ci-4 alkyl. Preferably R9 is hydrogen. When Rb is the substituent group NR9C(O)R_a, then R_a is preferably an alkyl group, such as methyl.

Suitably Rς is hydrogen, alkyl, aryl, arylCi-4alkyl, arylCi-4alkenyl, heteroaryl, heteroarylC i-4alkyl, heteroarylC 1 -4alkenyl, heterocyciic, or heterocyciic Ci-4alkyl, or a heterocyciic Ci-4alkenyl moiety, all of which may be optionally substituted one to three times independently by halogen, nitro, halosubstituted Ci-4 alkyl, C1.4 alkyl, C1-4 alkoxy, NRαC(O)R_a, C(O)NR6R7, S(O)3H, or C(O)OCi-4 alkyl, wherein R9 is hydrogen or a Ci-4 alkyl. Preferably, R_c is an optionally substituted phenyl.

When R is an OR2 or SR2 moiety it is recognized by one of skill in the art that the aryl ring must, therefore, contain the required ionizable hydrogen. The aryl ring may also be additionally substituted, independently, by one to three groups, which groups may also contain an additional ionizable group, and which include but are not limited to, halogen, nitro, halosubstituted Ci-4 alkyl, Ci-4 alkyl, Cj-4 alkoxy, hydroxy, SH, -C(O)NR6R7, -NH-C(O)R_a, -NHS(O)2Rb, S(O)2NRβR7- C(O)OR_8> or a tetrazolyl ring.

In compounds of Formula (I), suitably Rl is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Cj-io alkyl, such as CF3; Ci-io alkyl, such as methyl, ethyl, isopropyl, or n-propyl; C2-10 alkenyl; Ci-io alkoxy, such as methoxy, or ethoxy; halosubstituted Cι_ιo alkoxy, such as trifluoromethoxy; azide; (CRgRg)q S(O)_tR4, wherein l is 0, 1 or 2; hydroxy; hydroxy Ci-4alkyl, such as methanol or ethanol; aryl, such as phenyl or naphthyl; aryl C]-4 alkyl, such as benzyl; aryloxy, such as phenoxy; aryl Ci-4 alkyloxy, such as benzyloxy; heteroaryl; heteroarylalkyl; heteroaryl Ci-4 alkyloxy; aryl C2-10 alkenyl ; heteroaryl C2-10 alkenyl; heterocyciic C2-10 alkenyl; (CRgRg)qNR4Rs; C2-10 alkenyl C(O)NR4R5; (CR₈R₈)q C(O)NR4R5; (CR₈R₈)q C(O)NR4Rlθ; S(O)₃H; S(O)₃R8; (CR₈R₈)q C(O)Rn ; C₂-10 alkenyl C(O)Rι 1; C₂-10 alkenyl C(O)ORn; C(O)Rn; (CR₈R₈)q C(O)ORl2; (CR₈R₈)qOC(O)Rn; (CR₈R₈)q NR4C(O)Rι 1, (CR₈R₈)q NHS(O)₂Rπ, (CR₈R₈)qS(O)₂NR₄R₅; or two Rl moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; and s is an integer having a value of 1 to 3. The aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl, heterocyciic, heterocyclicalkyl, and heterocyclicalkenyl moieties may all be optionally substituted as defined herein below.

Suitably, q is 0, or an integer having a value of 1 to 10.

When R] forms a dioxybridge, s is preferably 1. When Ri forms an additional unsaturated ring, it is preferably 6 membered resulting in a naphthylene ring system. This naphthylene ring may be substituted independently, 1 to 3 times by the other Rl moieties as defined above.

10

Suitably, R4 and R5 are independently hydrogen, optionally substituted Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl Ci-4alkyl, heterocyciic, heterocyclicC 1-4 alkyl, or R4 and R5 together with the nitrogen to which they are 15 attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from O/N/S.

R₈ is suitably independently selected from hydrogen or Cj-4 alkyl.

~> .0 R 10 is suitably C 1.10 alkyl C(O)2R₈, such as CH2C(O)2H or

CH₂C(O)₂CH₃.

Rl 1 is suitably hydrogen, Ci-4 alkyl, aryl, aryl C}-4 alkyl, heteroaryl, heteroaryl Ci-4alkyl, heterocyciic, or heterocyciic Ci-4alkyl. i 5

Rl2 is suitably hydrogen, Cl-10 alkyl, optionally substituted aryl or optionally substituted arylalkyl.

Rl7 is suitably Cj-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC 1 -4alkyl, 30 heterocyciic, or heterocyclicC i-4alkyl, wherein the aryl, heteroaryl and heterocyciic rings may all be optionally substituted.

Preferably Rl is halogen, cyano, nitro, CF3, C(O)NR4R5, alkenyl C(O)NR4R5, C(O) R4R1O, alkenyl C(O)ORi2, heteroaryl, heteroarylalkyl, 35 heteroaryl alkenyl, or S(O)NR4R5, and preferably R4 and R5 are both hydrogen or one is phenyl. A preferred ring substitution for Rj is in the 4-position of the phenyl ring.

When R is OH, SH or NHS(O)2Rb than Rl is preferably substituted in the 3-position, the 4- position or di substituted in the 3,4- position. The substituent group is suitably an electron withdrawing moiety. Preferably when R is OH, SH or NSO2Rb_> ^tnan Rl is nitro, halogen, cyano, trifluoromethyl group, or C(O)NR4R5.

When R is carboxylic acid, than Rl is preferably hydrogen, or Rl is preferably substituted in the 4-position, more preferably substituted by trifluoromethyl or chloro.

In compounds of Formula (I), suitably R13 and R14 are independently hydrogen or C1-4 alkyl which may be straight or branched as defined herein; v is 0, or an integer having a value of 1 to 4, preferably v = 0.

The E ring denoted by its point of attachment through the asterix (*) may optionally be present. If if it is not present the ring is a phenyl moiety which is substituted by the R and Rl terms as shown. The Rl moiety may be substituted in any ring, saturated or unsaturated, including any of the E rings, and is shown for purposes herein substituted only in the unsaturated phenyl ring containing the R moiety.

In compounds of Formula (I), HET is suitably a heteroaryl ring or ring system. If the HET moiety is a multi ring system, the ring containing the heteroatom does not need to be directly attached to the urea moiety. All the rings in this ring system may be optionally substituted as defined herein. Preferably the HET moiety is a pyridyl, which may be 2-, 3- or 4-pyridyl, more preferably a 3- or 4-pyridyl. If the ring is a multi system ring it is preferably a benzimidazole, a dibenzothiophene, or an indole ring. Other heterocyciic rings of interest for use herein include, but are not limited to thiophene, furan, or pyrimidine rings.

In compounds of Formula (I), the HET ring may be optionally substituted independently one to three times by Y, i.e. (Y(_n) ). wherein n is an integer having a value of 1 to 3. Y is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Cj-io alkyl; Ci-io alkyl; C2-10 alkenyl; Cj- io alkoxy; halosubstituted Ci-io alkoxy; azide; (CR₈R₈)q S(O)tR4; hydroxy; hydroxyCi-4alkyl; aryl; aryl C]-4 alkyl; aryloxy; arylCi-4 alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl Ci-4 alkyloxy; heterocyciic, heterocyciic Ci-4alkyl; aryl

C2-10 alkenyl; heteroaryl C2-10 alkenyl; heterocyciic C2-10 alkenyl; (CR₈R₈)q NR4R5; C2-10 alkenyl C(O)NR4R5; (CR₈R₈)q C(O)NR4R5; (CR₈R₈)q

C(O)NR4Rl0; S(O)₃H; S(O)3R8; (CR₈R₈)q C(O)Rι l ; C2-10 alkenyl C(O)Rl l ;

C2-10 alkenyl C(O)ORι 1 ; (CR₈R₈)q C(O)ORi2; (CR₈R₈)q OC(O) Rl 1;

(CR₈R₈)q NR4C(O)Ri 1, (CR₈R₈)q NHS(O)₂R_d, (CR₈R₈)q S(O)₂NR₄R₅or two

Y moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring When Y forms a dioxybridge, s is preferably 1. When Y forms an additional unsaturated ring, it is preferably a 5 or 6 membered ring. The aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl, heterocyciic, heterocyclicalkyl, and heterocyclicalkenyl moieties noted above may all be optionally substituted as defined herein.

Suitably, R<j is a NR6R7, alkyl, aryl C1.4 alklyl, arylC 2-4 alkenyl, heteroaryl, hetroaryl-C]_4alkyl, heteroarylC2-4 alkenyl, heterocyciic, heterocyclicC j .4 alkyl, or heterocyciic C 2-4 alkenyl moiety, wherein the aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl, heterocyciic, and heterocyclicalkyl, and heterocyclicalkenyl moieties noted above may all be optionally substituted as defined herein.

In compounds of Formula (I), X is suitably oxygen or sulfur, preferably oxygen.

Exemplified compounds of Formula (I) include: N-(2-hydroxy-4-nitro phenyl)-N'-(2-pyridyl) urea N-(2-hydroxy-4-nitro phenyl)-N'-(3-pyridyl) urea N-(2-hydroxy-4-nitro phenyl)-N'-(4-pyridyl) urea N-[2-hydroxy-3-cyanophenyl]-N'-[2-chloro-3-pyridyl] urea

Other compounds within the scope of Formula (I) include: N-(2-hydroxy-4-nitro phenyl)-N'-(4-dibenzothiophene) urea N-(2-hydroxy-4-nitro phenyl)-N'-(2-benzimidazole) urea N-(2-hydroxy-4-nitro phenyl)-N'-(4-indole) urea As used herein, "optionally substituted" unless specifically defined shall mean such groups as halogen, such as fluorine, chlorine, bromine or iodine; hydroxy; hydroxy substituted Ci-ioalkyl; Ci- io alkoxy, such as methoxy or ethoxy; S(O)_m' Ci-io alkyl, wherein m' is 0, 1 or 2, such as methyl thio, methyl sulfinyl or methyl sulfonyl; amino, mono & di-substituted amino, such as in the NR4R5 group; NHC(O)R4; C(O)NR4R5; C(O)OH; S(O)2NR4R5; NHS(O)2Rl5, Ci-io alkyl, such as methyl, ethyl, propyl, isopropyl, or t-butyl; halosubstituted Ci-io alkyl, such CF3; an optionally substituted aryl, such as phenyl, or an optionally substituted arylalkyl, such as benzyl or phenethyl, optionally substituted heterocylic, optionally substituted heterocylicalkyl, optionally substituted heteroaryl, optionally substituted heteroaryl alkyl, wherein these aryl , hetroaryl, or heterocyciic moieties may be substituted one to two times by halogen; hydroxy; hydroxy substituted alkyl; Ci-io alkoxy; S(O)_m'Ci-io alkyl; amino, mono & di- substituted amino, such as in the NR4R5 group; Cj-io alkyl, or halosubstituted Cj-io alkyl, such as CF3.

Rl5 is suitably Ci-4 alkyl, aryl, aryl Ci-4alkyl, heteroaryl, heteroarylC j-4alkyl, heterocyciic, or heterocyclicC i-4alkyl.

Suitable pharmaceutically acceptable salts are well known to those skilled in the art and include basic salts of inorganic and organic acids, such as hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, methane sulphonic acid, ethane sulphonic acid, acetic acid, malic acid, tartaric acid, citric acid, lactic acid, oxalic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid, phenylacetic acid and mandelic acid. In addition, pharmaceutically acceptable salts of compounds of Formula (I) may also be formed with a pharmaceutically acceptable cation, for instance, if a substituent group comprises a carboxy moiety. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkaline, alkaline earth, ammonium and quaternary ammonium cations.

The following terms, as used herein, refer to:

• "halo" - all halogens, that is chloro, fluoro, bromo and iodo.

• "Cι_ιøalkyl" or "alkyl" - both straight and branched chain radicals of 1 to

10 carbon atoms, unless the chain length is otherwise limited, including, but not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, j^'.vo-butyl, tert- butyl, rc-pentyl and the like. • The term "cycloalkyl" is used herein to mean cyclic radicals, preferably of 3 to 8 carbons, including but not limited to cyclopropyl, cyclopentyl, cyclohexyl, and the like.

• The term "alkenyl" is used herein at all occurrences to mean straight or branched chain radical of 2- 10 carbon atoms, unless the chain length is limited thereto, including, but not limited to ethenyl, 1-propenyl, 2-propenyl, 2-methyl-l- propenyl, 1-butenyl, 2-butenyl and the like.

• "aryl" - phenyl and naphthyl;

• "heteroaryl" (on its own or in any combination, such as "heteroaryloxy", or "heteroaryl alkyl") - a 5-10 membered aromatic ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, O or S, such as, but not limited, to pyrrole, pyrazole, furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine, pyrimidine, oxazole, thiazole, thiadiazole, triazole, imidazole, or benzimidazole. • "heterocyciic" (on its own or in any combination, such as

"heterocyclicalkyl") - a saturated or partially unsaturated 4-10 membered ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, O, or S; such as, but not limited to, pyrrolidine, piperidine, piperazine, morpholine, tetrahydropyran, or imidazolidine. • The term "arylalkyl" or "heteroarylalkyl" or "heterocyclicalkyl" is used herein to mean Cj-io alkyl, as defined above, attached to an aryl, heteroaryl or heterocyciic moiety, as also defined herein, unless otherwise indicated.

• "sulfinyl" - the oxide S (O) of the corresponding sulfide, the term "thio" refers to the sulfide, and the term "sulfonyl" refers to the fully oxidized S(O)2 moiety.

• The term "wherein two Rl moieties (or two Y moieties) may together form a 5 or 6 membered unsaturated ring" is used herein to mean the formation of a napthylene ring system or a phenyl moiety having attached a 6 membered partially unsaturated ring such as a C6 cycloalkenyl, i.e hexene, or a C5 cyloalkenyl moiety, cyclopentene.

The compounds of Formula (I) may be obtained by applying synthetic procedures, some of which are illustrated in the Schemes below. The synthesis provided for in these Schemes is applicable for the producing compounds of Formula (I) having a variety of different R, Rj , and Aryl groups which are reacted, employing optional substituents which are suitably protected, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, in those cases, then affords compounds of the nature generally disclosed. Once the urea nucleus has been established, further compounds of these formulas may be prepared by applying standard techniques for functional group interconversion, well known in the art. While the schemes are shown with compounds only of Formula (I), or specific moieties, this is merely for illustration purposes only.

Scheme 1

R=NH₂, OH, C0₂H, SH a)HET-NCO

NHSQ₂Rb

I () Ortho substituted phenyl ureas shown in 2-scheme 1 may be prepared by standard conditions involving the condensation of commercially available ortho substituted aniline (Aldrich Chemical Co., Milwaukee, Wi) with the commercially available optionally substituted heteroaryl isocyanate (Aldrich Chemical Co., Milwaukee, Wi) in an aprotic solvent (DMF, toluene). When the l-(RSO2NH)2-

15 (NH2)Ph is not commercially available it can be made by treating the commercially available RSO2CI with the cooresponding 2-phenylene diamine in the presence of an base like triethyl amine or NaH in an aprotic solvent (like methylene chloride or DMF).

Scheme 2

a)TBSCI, imid b)triphosgene, NaHCOg c)RNH₂ d)Et₃NΗF

R-NH2 as used in Scheme 2 refers to R as a HET- moeity as defined in Formula (I). Alternatively, as shown above in Scheme 2 the 2-hydroxy aniline can be protected by reagents known in the art such as tert(butyl)dimethylsilyl chloride and imidazole in an aprotic solvent like DMF (Scheme 2). The aniline can then be reacted with a phosgene equivalent like triphosgene or carbonyl diimidazole in the presence of a base such as sodium bicarbonate to form the isocyanate 4. This isocyanate can then be condensed with the desired heterocyciic amine which can be purchased commercially. The protected phenol can then be deprotected by standard conditions such as triethylamine hydrofluoride to form the urea 5_.

Scheme 3

R"=OH, NH₂, NHS0₂R_b a)HNQ₃ b)SnCI₂ If the desired 2-substituted aniline 7-scheme 3. is not commercially available the corresponding nitro compound can be prepared from 6-scheme 3. under standard nitration conditions (using HNO3 or BF4NO3) at 23 °C. The nitro compound is then reduced to the corresponding aniline using SnCl2 in EtOH(or alternately with H2/Pd or LiAlH4).

Scheme 4

l ^{o r} a)NH₄SCN, Br b)NaOH

If the desired 2-amino benzenethiol 8-scheme 4 is not commercially available it can be synthesized by reaction of the phenyl aniline with the thiocyanate anion in the presence of an oxidant(like bromine) to produce the 2- amino benzthiazole. This thiazole can then be hydrolyzed to the desired 2-amino benzenethiol 9-scheme 4 with a strong base like NaOH in a protic solvent (i.e., EtOH).

Scheme 5

a)DPPA, Et₃N b)l wherein R is HET.

Alternatively the heterocyciic isocyanate can be synthesized from the corresponding carboxylic acid using the Curtius rearrangement (dppa and triethyl amine, oxalyl chloride followed by sodium azide, scheme 5). This isocyanate can then be condensed with the commercially available hydroxy aniline to form urea JJ. (scheme 5). S Pharmaceutically acceptable salts of compounds of Formula (I) may be obtained in known manner, for example by treatment thereof with an appropriate amount of acid or base in the presence of a suitable solvent.

Numerous conversions of aryl halides to aryl cyano derivatives with copper

(I) cyanide have been published. However, no examples of an aryl ring with a hydroxy group present were mentioned. Several attempts to obtain a cyano phenol moiety with published results failed. Using known conditions of elevated temperatures, greater than 170 °C, such as from 180 to 210° did not yield displacement of the halogen to a cyano moiety. Standard bases, such as DMF and pyridine further provided no desired product. Intermediates such as 2-amino-5- fluorophenol, 2-nitro-5-fluorophenol, 2-nitro-5-methyl-6-bromophenol were tried with a change of halogens, from fluorine to chlorine to bromine, and with use of copper (1) cyanide. The use of a bromine derivative, such as 2-nitro-5-methyl-6- bromophenol, with dimethylformamide and using triethylamine with a catalytic amount of dimethylamino pyridine and copper (I) cyanide at reduced temperatures,, i.e. <100°C, preferably 60 to about 80°C for reduced times from strandarized procedures, i.e., < 18 hours, preferably about 4 to 6 hours yielded the desired products. In the Examples, all temperatures are in degrees Centigrade (°C). Mass spectra were performed upon a VG Zab mass spectrometer using fast atom bombardment, unless otherwise indicated. 1H-NMR (hereinafter "NMR") spectra were recorded at 250 MHz or 400MHz using a Bruker AM 250 or Am 400 spectrometer, respectively. Multiplicities indicated are: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet and br indicates a broad signal. Sat. indicates a saturated solution, equiv. indicates the proportion of a molar equivalent of reagent relative to the principal reactant.

Flash chromatography is run over Merck Silica gel 60 (230 - 400 mesh).

SYNTHETIC EXAMPLES

The invention will now be described by reference to the following examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention. All temperatures are given in degrees centigrade, all solvents used herein are of the highest available purity and all reactions are run under anhydrous conditions in an argon atmosphere unless otherwise indicated. Example 1 Preparation of N-(2-hydroxy-4-nitro phenyl)-N'-(2-pyridyl) urea

The urea was synthesized by the treatment of 2-pyridyl carboxylic acid(2 mmol) with diphenyl phosphoryl azide(.475 mL) and triethyl amine(.14 mL) in DMF at 80 °C. After 24 h 5-nitro 2-amino phenol (1 equiv.) was added. The reaction was heated for 24 h at 80°C. The reaction product was oiled out with hexane. The residue was dissolved in methanol and the solid was precipitated out with water.(180 mg, 32%) EI-MS m/z 273(M-H) ^'

Example 2 Preparation of N-f2-hydroxy-4-nitro phenyP-N'-G-pyridyl) urea

a)Preparation of 2-(tert-butyldimethylsiloxy)-4-nitroaniline 2-(tert-Butyldimethylsiloxy)-4-nitroaniline was prepared by treating a solution of 2- amino 5-nitro phenol(17 g, .11 mol) in DMF with tert-butyldimethylsilyl chloride and imidazole for 64 hours at 23 °C. The reaction mixture was then partitioned between 0.1 N HCL and 1: 1 EtO Ac/ether. The aqueous phase was separated and the organic phase was washed once with 0.1N HCI, twice with sat NaHCO3 and twice more with distilled water. The organic phase was dried over magnesium sulfate and concentrated in vacuo to a volume of 30 mL. The resulting solid was collected by filtration and washed with ether. The residual starting material was removed by titruating with 3: 1 methanohwater and filtering. The title compound was collected as a pale orange solid(15.3 grams, 50%). ^lH NMR (CDC1₃): δ 7.82 (d,lH), 7.67(s, 1H), 6.67 (d, 1H), 4.52(s, 2H), 1.16(s, 9H), 0.38(s, 6H).

b)Preparation of 2-tert-Butyldimethysilyloxy-4-nitrophenyl isocyanate 2-(tert-Butyldimethylsiloxy)-4-nitroaniline(2.0 g, 7.5 mmol) was added to a solution of phosgene(2 eq) and triethyl amine(2 eq) in toluene. The reaction mixture was stirred at 23°C for 12 hours. The solid was filtered off. The filterate was concentrated in vacuo to afford desired which was used immediately without further purification. H NMR (CDCl₃): δ 7.83 (d,lH), 7.75(s, 1H), 7.1 1 (d, 1H) 1.03(s, 9H), 0.43(s, 6H).

c) Preparation of N-[(2-terr-butyldimethylsiloxy)-4-nitrophenyl]-N'-[(3-pyridyl] urea A solution of 3-amino pyridine(180 mg, 1.9 mmol) in DMF was treated with 2-tert- butyldimethysilyloxy-4-nitrophenyl isocyanate at 80 °C overnight. The reaction mixture was partitioned between methylene chloride and water. The organic layer was separated and washed 2-times with water, then it was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by flash chromatography

(30% EtOAc/hexanes) to afford desired as a pale yellow solid(52 mg, 7%). H NMR (DMSO): δ 9.9(s, 1H), 8.65 (s,lH), 8.28(m, 3H), 7.95 (m, 2H) 7.19(s, 1H), 7.35(m, 1H), 1.03(s, 9H), 0.43(s, 6H).

d) Preparation of N-(2-hydroxy-4-nitro phenyl)-N'-(3-pyridyl) urea

N-(2-Hydroxy-4-nitro phenyl)-N'-(3-pyridyl) urea was prepared by treating a solution of N-[(2-tc?r/-butyldimethylsiloxy)-4-nitrophenyl]-N'-[(3-pyridyl] urea(45 mg, .1 12 mmol) in methanol with acetic acid. After 30 minutes the reaction mixture was partitioned between water and methylene chloride. The organic layer was separated, dried over sodium sulfate and concentrated in vacuo to afford desired(23 mg, 74%). H NMR (CD₃OD): δ 8.6(s, 1H, br), 8.34(d,lH), 8.20(s, 1H, br), 8.05(d, 1H), 7.75( d, 1H),

7.67(s, lH), 7.38(m, br)

Example 3

Preparation of N-Q-hvdroxy^-nitro phenyl)-N'-(4-pyridyl) urea a) Preparation of N-[(2-tø^butyldimethylsiloxy)-4-nitrophenyl]-N'-[(4-pyridyl] urea A solution of 4-amino pyridine in DMF was treated with 2-tert-butyldimethysilyloxy-4- nitrophenyl isocyanate(example 2b) at 80 °C overnight. The reaction mixture was partitioned between methylene chloride and water. The organic layer was separated and washed 2 times with water. It was then dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by flash chromatography (30% EtOAc/hexanes) to afford desired as a pale yellow solid.

b) Preparation of N-(2-hydroxy-4-nitro phenyl)-N'-(4-pyridyl) urea

N-(2-Hydroxy-4-nitro phenyl)-N'-(4-pyridyl) urea was prepared by treating a solution of N-[(2-/e?r/-butyldimethylsiloxy)-4-nitrophenyl]-N'-f(3-pyridyl] urea(25 mg, .065 mmol) in methanol with acetic acid. After 30 minutes the reaction mixture was partitioned between water and methylene chloride. The organic layer was separated, dried over sodium sulfate and concencetrated in vacuo to afford desired(16 mg, 90%). ^]H NMR (DMSO): δ 10.38(s, 1H, br), 9.14(s,lH), 8.08(s, 1H), 8.05(d, 1H), 7.59( d, 2H), 7.55(d, 1H), 6.95(d, 1H), 6.86(s, 1H), 6.64(d, 2H).

Example 4 Preparation of N-[2-hydroxy-3-cyanophenyl]-N'-[2-chloro-3-pyridyl] urea

A mixture of 2-chloronicotinic acid (310mg, 2mmol), diphenylphosphory azide (550mg, 2mmol) and trieyhyl amine(200mg, 2mmol) was heated to 80° C in

50ml of toluene for 2 hrs. Addition of 2-amino-6-cyanophenol was then made and the reaction stirred overnight. After 12 hrs the toluene was removed by rotoevaporation and chromatography of the resulting solid on silica gel (4%MeOH/ CH₂C1₂) gave the desired product(260 mg, 45 %). Η NMR (CD₃SO₂CD₃): δ 10.89

(s, 1H), 9.37 (s,lH), 9.1 1 (s,lH), 8.49 (d, 1H), 8.19 (d, 1H), 8.05 (dd, 1H), 7.40 (dd, 1H), 7.29 (d, 1H), 6.99 (t, 1H).

METHOD OF TREATMENT

The compounds of Formula (I), or a pharmaceutically acceptable salt thereof can be used in the manufacture of a medicament for the prophylactic or therapeutic treatment of any disease state in a human, or other mammal, which is exacerbated or caused by excessive or unregulated LL-8 cytokine production by such mammal's cell, such as but not limited to monocytes and/or macrophages, or other chemokines which bind to the IL-8 a or b receptor, also referred to as the type I or type II receptor.

Accordingly, the present invention provides a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an IL-8 a or b receptor and which method comprises administering an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In particular, the chemokines are LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78. The compounds of Formula (I) are administered in an amount sufficient to inhibit cytokine function, in particular LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78, such that they are biologically regulated down to normal levels of physiological function, or in some case to subnormal levels, so as to ameliorate the disease state. Abnormal levels of LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78 for instance in the context of the present invention, constitute: (i) levels of free IL-8 greater than or equal to 1 picogram per mL; (ii) any cell LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78 above normal physiological levels; or (iii) the presence of LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78 above basal levels in cells or tissues in which LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78 respectively, is produced.

There are many disease states in which excessive or unregulated LL-8 production is implicated in exacerbating and/or causing the disease. Chemokine mediated diseases include psoriasis, atopic dermatitis, arthritis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, inflammatory bowel disease, Crohn's disease, ulcerative colitis, stroke, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, graft vs. host reaction, alzheimers disease, allograft rejections, malaria, restinosis, angiogenesis or undesired hematopoietic stem cells release.

These diseases are primarily characterized by massive neutrophil infiltration, T-cell infiltration, or neovascular growth, and are associated with LL-8, GROα, GROβ, GROγ , or NAP-2 production which is responsible for the chemotaxis of neutrophils into the inflammatory site or the directional growth of endothelial cells. In contrast to other inflammatory cytokines (IL-1, TNF, and IL-6), LL-8, GROα, GROβ, GROγ or NAP-2 has the unique property of promoting neutrophil chemotaxis, enzyme release including but not limited to elastase release as well as superoxide production and activation. The α-chemokines but particularly, GROα, GROβ, GROγ or NAP-2,^' working through the LL-8 type I or II receptor can promote the neovascularization of tumors by promoting the directional growth of endothelial cells. Therefore, the inhibition of LL-8 induced chemotaxis or activation would lead to a direct reduction in the neutrophil infiltration.

Recent evidence also implicates the role of chemokines in the treatment of HIV infections, Littleman et al., Nature 381, pp661 (1996) and Koup et al., Nature 381, pp 667 (1996).

The present invention also provides for a means of treating, in an acute setting, as well as preventing, in those individuals deemed susceptible to, CNS injuries by the chemokine receptor antagonist compounds of Formula (I). CNS injuries as defined herein include both open or penetrating head trauma, such as by surgery, or a closed head trauma injury, such as by an injury to the head region. Also included within this definition is ischemic stroke, particularly to the brain area.

Ischemic stroke may be defined as a focal neurologic disorder that results from insufficient blood supply to a particular brain area, usually as a consequence of an embolus, thrombi, or local atheromatous closure of the blood vessel. The role of inflammatory cytokines in this are has been emerging and the present invention provides a mean for the potential treatment of these injuries. Relatively little treatment, for an acute injury such as these has been available.

TNF-α is a cytokine with proinflammatory actions, including endothelial leukocyte adhesion molecule expression. Leukocytes infiltrate into ischemic brain lesions and hence compounds which inhibit or decrease levels of TNF would be useful for treatment of ischemic brain injury. See Liu et al.. Stoke, Vol. 25., No. 7, pp 1481-88 (1994) whose disclosure is incorporated herein by reference.

Models of closed head injuries and treatment with mixed 5-LO/CO agents is discussed in Shohami et al., J. of Vaisc & Clinical Physiology and Pharmacology, Vol. 3, No. 2, pp. 99-107 (1992) whose disclosure is incorporated herein by reference. Treatment which reduced edema formation was found to improve functional outcome in those animals treated.

The compounds of Formula (I) are administered in an amount sufficient to inhibit IL-8, binding to the IL-8 alpha or beta receptors, from binding to these receptors, such as evidenced by a reduction in neutrophil chemotaxis and activation. The discovery that the compounds of Formula (I) are inhibitors of IL-8 binding is based upon the effects of the compounds of Formulas (I) in the in vitro receptor binding assays which are described herein. The compounds of Formula (I) have been shown, in some instances, to be dual inhibitors of both recombinant type I and type II IL-8 receptors. Preferably the compounds are inhibitors of only one receptor, more preferably Type II.

As used herein, the term "LL-8 mediated disease or disease state" refers to any and all disease states in which LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78 plays a role, either by production of LL-8, GROα, GROβ, GROγ , NAP-2 or ENA- 78 themselves, or by LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78 causing another monokine to be released, such as but not limited to IL-1, LL-6 or TNF. A disease state in which, for instance, IL- 1 is a major component, and whose production or action, is exacerbated or secreted in response to LL-8, would therefore be considered a disease stated mediated by LL-8. As used herein, the term "chemokine mediated disease or disease state" refers to any and all disease states in which a chemokine which binds to an LL-8 a or b receptor plays a role, such as but not limited LL-8, GROα, GROβ, GROγ , NAP-2 or ENA-78. This would include a disease state in which, LL-8 plays a role, either by production of IL-8 itself, or by IL-8 causing another monokine to be released, such as but not limited to IL-1, LL-6 or TNF. A disease state in which, for instance, IL-1 is a major component, and whose production or action, is exacerbated or secreted in response to IL-8, would therefore be considered a disease stated mediated by LL-8. As used herein, the term "cytokine" refers to any secreted polypeptide that affects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response. A cytokine includes, but is not limited to, monokines and lymphokines, regardless of which cells produce them. For instance, a monokine is generally referred to as being produced and secreted by a mononuclear cell, such as a macrophage and/or monocyte. Many other cells however also produce monokines, such as natural killer cells, fibroblasts, basophils, neutrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epideral keratinocytes and B-lymphocytes. Lymphokines are generally referred to as being produced by lymphocyte cells. Examples of cytokines include, but are not limited to, Interleukin-1 (IL- 1), Interleukin-6 (LL-6), Interleukin-8 (LL-8), Tumor Necrosis Factor-alpha (TNF-α) and Tumor Necrosis Factor beta (TNF-β).

As used herein, the term "chemokine" refers to any secreted polypeptide that affects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response, similar to the term "cytokine" above. A chemokine is primarily secreted through cell transmembranes and causes chemotaxis and activation of specific white blood cells and leukocytes, neutrophils, monocytes, macrophages, T-cells, B-cells, endothelial cells and smooth muscle cells. Examples of chemokines include, but are not limited to, LL-8, GRO-α, GRO-β, GRO-γ, NAP-2, ENA-78, IP- 10, MlP-lα, MlP-β, PF4, and MCP 1 , 2, and 3.

In order to use a compound of Formula (I) or a pharmaceutically acceptable salt thereof in therapy, it will normally be formulated into a pharmaceutical composition in accordance with standard pharmaceutical practice. This invention, therefore, also relates to a pharmaceutical composition comprising an effective, non- toxic amount of a compound of Formula (I) and a pharmaceutically acceptable carrier or diluent.

Compounds of Formula (I), pharmaceutically acceptable salts thereof and pharmaceutical compositions incorporating such may conveniently be administered by any of the routes conventionally used for drug administration, for instance, orally, topically, parenterally or by inhalation. The compounds of Formula (I) may be administered in conventional dosage forms prepared by combining a compound of Formula (I) with standard pharmaceutical carriers according to conventional procedures. The compounds of Formula (I) may also be administered in conventional dosages in combination with a known, second therapeutically active compound. These procedures may involve mixing, granulating and compressing or dissolving the ingredients as appropriate to the desired preparation. It will be appreciated that the form and character of the pharmaceutically acceptable character or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables. The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The pharmaceutical carrier employed may be, for example, either a solid or liquid. Exemplary of solid carriers are lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Exemplary of liquid carriers are syrup, peanut oil, olive oil, water and the like. Similarly, the carrier or diluent may include time delay material well known to the art, such as glyceryl mono-stearate or glyceryl distearate alone or with a wax.

A wide variety of pharmaceutical forms can be employed. Thus, if a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form or in the form of a troche or lozenge. The amount of solid carrier will vary widely but preferably will be from about 25mg. to about lg. When a liquid carrier is used, the preparation will be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampule or nonaqueous liquid suspension.

Compounds of Formula (I) may be administered topically, that is by non- systemic administration. This includes the application of a compound of Formula (I) externally to the epidermis or the buccal cavity and the instillation of such a compound into the ear, eye and nose, such that the compound does not significantly enter the blood stream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.

Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site of inflammation such as liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose. The active ingredient may comprise, for topical administration, from 0.001 % to 10% w/w, for instance from 1% to 2% by weight of the Formulation. It may however comprise as much as 10% w/w but preferably will comprise less than 5% w/w, more preferably from 0.1% to 1% w/w of the Formulation.

Lotions according to the present invention include those suitable for application to the skin or eye. An eye lotion may comprise a sterile aqueous solution optionally containing a bactericide and may be prepared by methods similar to those for the preparation of drops. Lotions or liniments for application to the skin may also include an agent to hasten drying and to cool the skin, such as an alcohol or acetone, and/or a moisturizer such as glycerol or an oil such as castor oil or arachis oil.

Creams, ointments or pastes according to the present invention are semi- solid formulations of the active ingredient for external application. They may be made by mixing the active ingredient in finely-divided or powdered form, alone or in solution or suspension in an aqueous or non-aqueous fluid, with the aid of suitable machinery, with a greasy or non-greasy base. The base may comprise hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, a metallic soap; a mucilage; an oil of natural origin such as almond, corn, arachis, castor or olive oil; wool fat or its derivatives or a fatty acid such as steric or oleic acid together with an alcohol such as propylene glycol or a macrogel. The formulation may incorporate any suitable surface active agent such as an anionic, cationic or non-ionic surfactant such as a sorbitan ester or a polyoxyethylene derivative thereof. Suspending agents such as natural gums, cellulose derivatives or inorganic materials such as silicaceous silicas, and other ingredients such as lanolin, may also be included.

Drops according to the present invention may comprise sterile aqueous or oily solutions or suspensions and may be prepared by dissolving the active ingredient in a suitable aqueous solution of a bactericidal and/or fungicidal agent and/or any other suitable preservative, and preferably including a surface active agent. The resulting solution may then be clarified by filtration, transferred to a suitable container which is then sealed and sterilized by autoclaving or maintaining at 98-100 C. for half an hour. Alternatively, the solution may be sterilized by filtration and transferred to the container by an aseptic technique. Examples of bactericidal and fungicidal agents suitable for inclusion in the drops are phenyl mercuric nitrate or acetate (0.002%), benzalkonium chloride (0.01 %) and chlorhexidine acetate (0.01 %). Suitable solvents for the preparation of an oily solution include glycerol, diluted alcohol and propylene glycol.

Compounds of formula (I) may be administered parenterally, that is by intravenous, intramuscular, subcutaneous intranasal, intrarectal, intravaginal or intraperitoneal administration. The subcutaneous and intramuscular forms of parenteral administration are generally preferred. Appropriate dosage forms for such administration may be prepared by conventional techniques. Compounds of Formula (I) may also be administered by inhalation, that is by intranasal and oral inhalation administration. Appropriate dosage forms for such administration, such as an aerosol formulation or a metered dose inhaler, may be prepared by conventional techniques.

For all methods of use disclosed herein for the compounds of Formula (I), the daily oral dosage regimen will preferably be from about 0.01 to about 80 mg/kg of total body weight. The daily parenteral dosage regimen about 0.001 to about 80 mg/kg of total body weight. The daily topical dosage regimen will preferably be from 0.1 mg to 150 mg, administered one to four, preferably two or three times daily. The daily inhalation dosage regimen will preferably be from about 0.01 mg/kg to about 1 mg/kg per day. It will also be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a compound of

Formula (1) or a pharmaceutically acceptable salt thereof will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the particular patient being treated, and that such optimums can be determined by conventional techniques. It will also be appreciated by one of skill in the art that the optimal course of treatment, i.e., the number of doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof given per day for a defined number of days, can be ascertained by those skilled in the art using conventional course of treatment determination tests. The invention will now be described by reference to the following biological examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention.

BIOLOGICAL EXAMPLES

The LL-8, and Gro-α chemokine inhibitiory effects of compounds of the present invention were determined by the following in vitro assay: Receptor Binding Assays:

[*25i] IL_₈ (human recombinant) was obtained from Amersham Corp., Arlington Heights, LL, with specific activity 2000 Ci/mmol. Gro-α was obtained from NEN- New England Nuclear. All other chemicals were of analytical grade. High levels of recombinant human LL-8 type α and β receptors were individually expressed in Chinese hamster ovary cells as described previously (Holmes, et al. Science, 1991, 253, 1278). The Chinese hamster ovary membranes were homogenized according to a previously described protocol (Haour, et al., J Biol Chem., 249 pp 2195-2205 (1974)). Except that the homogenization buffer was changed to lOmM Tris-HCL, lmM MgS04, 0.5mM EDTA (ethylene-diaminetetra- acetic acid), ImMPMSF (α-toluenesulphonyl fluoride), 0.5 mg/L Leupeptin, pH 7.5. Membrane protein concentration was determined using Pierce Co. micro-assay kit using bovine serum albumin as a standard. All assays were performed in a 96- well micro plate format. Each reaction mixture contained 125 j JL_₈ (0.25 nM) or ¹²⁵I Gro-α and 0.5 μg/mL of IL-8Rα or 1.0 μg/mL of IL-8Rβ membranes in 20 mM Bis-Trispropane and 0.4 mM Tris HCI buffers, pH 8.0, containing 1.2 mM MgSO4, 0.1 mM EDTA, 25 mM NaCI and 0.03% CHAPS. In addition, drug or compound of interest was added which had been pre-dissolved in DMSO so as to reach a final concentration of between O.OlnM and 100 uM. The assay was initiated by addition of ^^I-IL-S. After 1 hour at room temperature the plate was harvested using a Tomtec 96- well harvester onto a glass fiber filtermat blocked with 1 % polyethylenimine/0.5% BSA and washed 3 times with 25 mM NaCI, 10 mM TrisHCl, 1 mM MgSO4, 0.5 mM EDTA, 0.03 % CHAPS, pH 7.4. The filter was then dried and counted on the Betaplate liquid scintillation counter. The recombinant IL-8 Rα, or Type I, receptor is also referred to herein as the non- permissive receptor and the recombinant LL-8 Rβ, or Type II, receptor is referred to as the permissive receptor. Exemplified compounds of Formulas (I) noted herein, as Examples 1 to 4 in the Synthetic Chemistry Section demonstrated an IC50 from about 30 to about <1 μg/mL in the permissive models for LL-8 receptor inhibition. The compound N-(6- Methyl-2-pyridine)-N'-(2-hydroxy-4-nitrophenyl)urea was found inactive in this assay.

Chemotaxis Assay :

The in vitro inhibitory properties of these compounds are determined in the neutrophil chemotaxis assay as described in Current Protocols in Immunology, vol I, Suppl 1 , Unit 6.12.3., whose disclosure is incorporated herein by reference in its entirety. Neutrophils where isolated from human blood as described in Current Protocols in Immunology Vol I, Suppl 1 Unit 7.23.1, whose disclosure is incorporated herein by reference in its entirety. The chemoattractants IL-8, GRO-α, GRO-β, GRO-γ and NAP-2 are placed in the bottom chamber of a 48 multiwell chamber (Neuro Probe, Cabin John, MD) at a concentration between 0.1 and 100 nM. The two chambers are separated by a 5um polycarbonate filter. When compounds of this invention are tested, they are mixed with the cells (0.001 - 1000 nM) just prior to the addition of the cells to the upper chamber. Incubation is allowed to proceed for between about 45 and 90 min at about 37°C in a humidified incubator with 5% CO2- At the end of the incubation period, the polycarbonate membrane is removed and the top side washed, the membrane then stained using the Diff Quick staining protocol (Baxter Products, McGaw Park, IL, USA). Cells which have chemotaxed to the chemokine are visually counted using a microscope. Generally, four fields are counted for each sample, these numbers are averaged to give the average number of cells which had migrated. Each sample is tested in triplicate and each compound repeated at least four times. To certain cells (positive control cells) no compound is added, these cells represent the maximum chemotactic response of the cells. In the case where a negative control (unstimulated) is desired, no chemokine is added to the bottom chamber. The difference between the positive control and the negative control represents the chemotactic activity of the cells.

Elastase Release Assay:

The compounds of this invention are tested for their ability to prevent Elastase release from human neutrophils. Neutrophils are isolated from human blood as described in Current Protocols in Immunology Vol I, Suppl 1 Unit 7.23.1. PMNs 0.88 x 10⁶ cells suspended in Ringer's Solution (NaCI 1 18, KC1 4.56, NaHCO3 25, KH2PO4 1.03, Glucose 11.1, HEPES 5 mM, pH 7.4) are placed in each well of a 96 well plate in a volume of 50 ul. To this plate is added the test compound (0.001 - 1000 nM) in a volume of 50 ul, Cytochalasin B in a volume of 50 ul (20ug/ml) and Ringers buffer in a volume of 50 ul. These cells are allowed to 5 warm (37 °C, 5% CO2, 95% RH) for 5 min before LL-8, GROα, GROβ, GROγ or NAP-2 at a final concentration of 0.01 - 1000 nM was added. The reaction is allowed to proceed for 45 min before the 96 well plate is centrifuged (800 xg 5 min) and 100 ul of the supernatant removed. This suppernatant is added to a second 96 well plate followed by an artificial elastase substrate (MeOSuc-Ala-Ala-Pro- Val¬ id AMC, Nova Biochem, La Jolla, CA) to a final concentration of 6 ug/ml dissolved in phosphate buffered saline. Immediately, the plate is placed in a fluorescent 96 well plate reader (Cytofluor 2350, Millipore, Bedford, MA) and data collected at 3 min intervals according to the method of Nakajima et al J. Biol Chem 254 4027 (1979). The amount of Elastase released from the PMNs is calculated by measuring 15 the rate of MeOSuc-Ala-Ala-Pro-Val-AMC degradation.

TNF-α in Traumatic Brain Injury Assay

The present assay provides for examination of the expression of tumor necrosis factor mRNA in specfic brain regions which follow experimentally induced lateral fluid- 0 percussion traumatic brain injury (TBI) in rats. Adult Sprague-Dawley rats (n=42) were anesthetized with sodium pentobarbital (60 mg/kg, i.p.) and subjected to lateral fluid- percussion brain injury of moderate severity (2.4 atm.) centered over the left temporaparietal cortex (n=18), or "sham" treatment (anesthesia and surgery without injury, n=18). Animals are sacrificed by decapitation at 1, 6 and 24 hr. post injury, brains 5 removed, and tissue samples of left (injured) parietal cortex (LC), corresponding area in the contralateral right cortex (RC), cortex adjacent to injured parietal cortex (LA), corresponding adjacent area in the right cortex (RA), left hippocampus (LH) and right hippocampus (RH) are prepared. Total RNA was isolated and Northern blot hybridization is performed and quantitated relative to an TNF-α positive control RNA (macrophage =

30 100%). A marked increase of TNF- α mRNA expression is observed in LH (104±17% of positive control, p < 0.05 compared with sham), LC (105±21%, p< 0.05) and LA (69±8%, p < 0.01) in the traumatized hemisphere 1 hr. following injury. An increased TNF- α mRNA expression is also observed in LH (46±8%, p < 0.05), LC (30±3%, p < 0.01) and LA (32±3%, p < 0.01) at 6 hr. which resolves by 24 hr. following injury. In the

35 contralateral hemisphere, expression of TNF- α mRNA is increased in RH (46±2%, p < 0.01), RC (4±3%) and RA (22±8%) at 1 hr. and in RH (28±11%), RC (7±5%) and RA (26±6%, p < 0.05) at 6 hr. but not at 24 hr. following injury. In sham (surgery without injury) or naive animals, no consistent changes in expression of TNF- α mRNA are observed in any of the 6 brain areas in either hemisphere at any times. These results indicate that following parasagittal fluid-percussion brain injury, the temporal expression of TNF-α mRNA is altered in specific brain regions, including those of the non- traumatized hemisphere. Since TNF- α is able to induce nerve growth factor (NGF) and stimulate the release of other cytokines from activated astrocytes, this post-traumatic alteration in gene expression of TNF-α plays an important role in both the acute and regenerative response to CNS trauma.

CNS Injury model for IL-b mRNA

This assay characterizes the regional expression of interleukin- 1 β (IL-lβ) mRNA in specific brain regions following experimental lateral fluid-percussion traumatic brain injury (TBI) in rats. Adult Sprague-Dawley rats (n=42) are anesthetized with sodium pentobarbital (60 mg/kg, i.p.) and subjected to lateral fluid-percussion brain injury of moderate severity (2.4 atm.) centered over the left temporaparietal cortex (n=18), or "sham" treatment (anesthesia and surgery without injury). Animals are sacrificed at 1 , 6 and 24 hr. post injury, brains removed, and tissue samples of left (injured) parietal cortex (LC), corresponding area in the contralateral right cortex (RC), cortex adjacent to injured parietal cortex (LA), corresponding adjacent area in the right cortex (RA), left hippocampus (LH) and right hippocampus (RH) are prepared. Total RNA is isolated and Northern blot hybridization was performed and the quantity of brain tissue IL- lβ mRNA is presented as percent relative radioactivity of IL-lβ positive macrophage RNA which was loaded on same gel. At 1 hr. following brain injury, a marked and significant increase in expression of IL-lβ mRNA is observed in LC (20.0±0.7% of positive control, n=6, p < 0.05 compared with sham animal), LH (24.5±0.9%, p < 0.05) and LA (21.5±3.1 %, p < 0.05) in the injured hemisphere, which remained elevated up to 6 hr. post injury in the LC (4.0±0.4%, n=6, p < 0.05) and LH (5.0+1.3%, p < 0.05). In sham or naive animals, no expression of IL- lβ mRNA is observed in any of the respective brain areas. These results indicate that following TBI, the temporal expression of IL-lβ mRNA is regionally stimulated in specific brain regions. These regional changes in cytokines, such as IL-lβ play a role in the post-traumatic.

All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as though fully set forth.

The above description fully discloses the invention including preferred embodiments thereof. Modifications and improvements of the embodiments specifically disclosed herein are within the scope of the following claims. Without further elaboration, it is believed that one skilled in the are can, using the preceding description, utilize the present invention to its fullest extent. Therefore the Examples herein are to be construed as merely illustrative and not a limitation of the scope of the present invention in any way. The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows.

Claims

What is claimed is:

1. A method of treating a chemokine mediated disease state, wherein the chemokine binds to an LL-8 α or β receptor in a mammal, which comprises administering to said mammal an effective amount of a compound of the formula:

wherein

X is oxygen or sulfur; R is any functional moiety having an ionizable hydrogen and a pKa of 10 or less; Rl is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Ci-io alkyl; C2-10 alkenyl; Ci- io alkoxy; halosubstituted Ci-io alkoxy; azide; (CR₈R₈)q S(O)_tR4; hydroxy; hydroxy Cι_4alkyl; aryl; aryl Cj-4 alkyl; aryloxy; aryl Cι_4 alkyloxy; heteroaryl; heteroarylalkyl; heterocyciic, heterocyciic Ci-4alkyl; heteroaryl Ci-4 alkyloxy; aryl C2-10 alkenyl; heteroaryl

C2-10 alkenyl; heterocyciic C2-10 alkenyl; (CR₈R₈)qNR4Rs; C2-10 alkenyl C(O)NR4R5; (CR₈R₈)q C(O)NR₄R5; (CR₈R₈)q C(O)NR₄Rlθ; S(O)₃H; S(O)3R₈; (CR₈R₈)q C(O)Rι i ; C2-10 alkenyl C(O)Rι l ; C2-10 alkenyl C(O)ORi ι (CR₈R₈)q C(O)ORi2; (CR₈R₈)q OC(O) Rn ; (CR₈R₈)qNR4C(O)Ri 1, (CR₈R₈)q NHS(O)₂Rι₇, (CR₈R₈)q S(O)₂NR₄R₅; or two Rl moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; q is 0, or an integer having a value of 1 to 10; m is an integer having a value of 1 to 3; t is 0, or an integer having a value of 1 or 2; s is an integer having a value of 1 to 3; v is 0, or an integer having a value of 1 to 4; v is 0, or an integer having a value of 1 to 4;

R4 and R5 are independently hydrogen, optionally substituted Cj-4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl Ci-4alkyl, heterocyciic, heterocyciic Ci-4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from oxygen, nitrogen or sulfur;

HET is an optionally substituted heteroaryl moiety;

R₈ is hydrogen or C 1.4 alkyl;

RlO is Ci-io alkyl C(O)2R_8;

Rl 1 is hydrogen, Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC ι_4alkyl, optionally substituted heterocyciic, or optionally substituted heterocyclicC 1 -4alkyl;

Rl2 is hydrogen, Ci-io alkyl, optionally substituted aryl or optionally substituted arylalkyl; Rl3 and R 14 are independently hydrogen or Ci-4 alkyl; Rl7 is Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC i-4alkyl, heterocyciic, or heterocyclicC i-4alkyl, wherein the aryl, heteroaryl and heterocyciic rings may all be optionally substituted; E is optionally selected from

"1 ; the asterix * denoting point of attachment of the ring; or a pharmaceutically acceptably salt thereof.

2. The method according to Claim 1 wherein the ionizable hydrogen has a pKa of 3 to 10.

3. The method according to Claim 2 wherein R is hydroxy, carboxylic acid, thiol, -SR2 -OR2, -NH-C(O)R_a, -C(O)NR6R7, -NHS(O)2Rb, -S(O)2NHR_c, NHC(X₂)NHRb, or tetrazolyl; wherein R2 is a substituted aryl, heteroaryl, or heterocyciic ring which ring contains the functional moiety providing the ionizable hydrogen having a pKa of 10 or less;

R_a is an alkyl, aryl, aryl C]-4alkyl, heteroaryl, heteroaryl Ci-4alkyl, heterocyciic, or a heterocyciic Ci-4alkyl moiety, all of which may be optionally substituted;

Rb is a NR6R7, alkyl, aryl, arylCi-4alkyl, arylC2-4alkenyl, heteroaryl, heteroarylC i-4alkyl, heteroarylC2-4 alkenyl, heterocyciic, heterocyciic Cι_4alkyl, heterocyciic C2-4alkenyl moiety, camphor, all of which may be optionally substituted one to three times independently by halogen; nitro; halosubstituted Ci-4 alkyl; C1-4 alkyl; C1.4 alkoxy; NR9C(O)R_a; C(O)NR6R7, S(O)3H, or C(O)OC] -4 alkyl;

R6 and R7 are independently hydrogen or a Ci-4 alkyl group, or R6 and R7 together with the nitrogen to which they are attached form a 5 to 7 member ring which ring may optionally contain an additional heteroatom which heteroatom is selected from oxygen, nitrogen or sulfur;

R9 is hydrogen or a C1-4 alkyl;

R_c is alkyl, aryl, arylCi-4alkyl, arylC2-4alkenyl, heteroaryl, heteroarylC i-4alkyl, heteroarylC2-4alkenyl, heterocyciic, heterocyciic Cι_4alkyl, or a heterocyciic C2-4alkenyl moiety, all of which may be optionally substituted one to three times independently by halogen, nitro, halosubstituted Ci-4 alkyl, Ci -4 alkyl, C1-4 alkoxy, NR9C(O)R_a, C(O)NR6R7, S(O)3H, or C(O)OCi_4 alkyl; and

X2 is oxygen or sulfur.

4. The method according to Claim 3 wherein the R2 is optionally substituted one to three times by halogen, nitro, halosubstituted Ci-]0 alkyl, Ci-io alkyl, Ci-10 alkoxy, hydroxy, SH, -C(O)NR6R7, -NH-C(O)R_a, -NHS(O)₂Rb, S(O)NR6R7, C(O)OR₈, or a tetrazolyl ring.

5. The method according to Claim 3 wherein R is OH, -NHS(O)2Rb or C(O)OH.

6. The method according to Claim 1 wherein R\ is halogen, cyano, nitro, CF3, C(O)NR4R5, alkenyl C(O)NR4R5, C(O) R4RIO, alkenyl C(O)ORi2, heteroaryl, heteroarylalkyl , heteroaryl alkenyl, or S(O)NR4R5.

7. The method according to Claim 1 wherein the HET ring is substituted by (Y)n; wherein n is an integer having a value of 1 to 3;

Y is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Ci-io alkyl; C2-10 alkenyl; Ci-io alkoxy; halosubstituted Ci-io alkoxy; azide; (CR₈R₈)q S(O)tR4; hydroxy; hydroxyCi-4alkyl; aryl; aryl Q-4 alkyl; aryloxy; arylCi-4 alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl Ci-4 alkyloxy; heterocyciic, heterocyciic Ci-4alkyl; aryl C2-10 alkenyl; heteroaryl C2-10 alkenyl; heterocyciic C2-10 alkenyl; (CR₈R₈)q NR4R5; C2-IO alkenyl C(O)NR4R5; (CR₈R₈)q C(O)NR4R5; (CR₈R₈)q

C(O)NR4Rlθ; S(O)₃H; S(O)₃R_8; (CR₈R₈)q C(O)Rn; C₂-10 alkenyl C(O)Ri 1; C2-10 alkenyl C(O)ORn ; C(O)Rn_; (CR₈R₈)q C(O)ORι₂; (CR₈R₈)q OC(O) Rn; (CR₈R₈)q NR₄C(O)Ri 1, (CR₈R₈)q NHS(O)₂R_d, (CR₈R₈)q S(O)₂NR₄R5; or two Y moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; and

R_fj is NRgR7, alkyl, arylCl -4alklyl, arylC 2-4 alkenyl, heteroaryl, hetroaryl-

C^alkyl, heteroarylC2_4 alkenyl, heterocyciic, heterocyclicC 1.4 alkyl, wherein the aryl, heteoaryl and heterocyciic rings may all be optionally substituted.

8. The method according to Claim 7 wherein Y is halogen, C 1.4 alkoxy, optionally substituted aryl, optionally substituted arylalkoxy, methylene dioxy, NR4R5, thioCi -4alkyl, thioaryl, halosubstituted alkoxy, optionally substituted Ci-4alkyl, or hydroxy alkyl.

9. The method according to Claim 1 wherein R is OH, SH, or NHS(O)2Rb and

Rj is substituted in the 3-position, the 4- position or di substituted in the 3,4- position by an electron withdrawing moiety.

10. The method according to any of Claims 1 to 9 wherein the mammal is afflicted with a chemokine mediated disease selected from psoriasis, or atopic dermatitis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, stroke, cardiac and renal reperfusion injury, glomerulo-nephritis, or thrombosis, alzheimers disease, graft vs. host reaction, or allograft rejections.

11. A compound of the formula:

wherein X is oxygen or sulfur;

R is any functional moiety having an ionizable hydrogen and a pKa of 10 or less;

Rl is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Ci-io alkyl; C2-10 alkenyl; Ci-io alkoxy; halosubstituted Ci-io alkoxy; azide; (CR₈R₈)q S(O)_tR4; hydroxy; hydroxy Ci-4alkyl; aryl; aryl Cl-4 alkyl; aryloxy; aryl Cl-4 alkyloxy; heteroaryl; heteroarylalkyl; heterocyciic, heterocyciic Ci-4alkyl; heteroaryl Cj-4 alkyloxy; aryl C2- 10 alkenyl; heteroaryl C2-10 alkenyl; heterocyciic C2-10 alkenyl; (CR₈R₈)qNR4R5; C2-10 alkenyl C(O)NR4R5; (CR₈R₈)q C(O)NR4R5; (CR₈R₈)q C(O)NR4Rlθ; S(O)₃H; S(O)3R8; (CR₈R₈)q C(O)Ri j ; C2-10 alkenyl C(O)Rι i ; C2-10 alkenyl C(O)ORn(CR₈R₈)q C(O)ORl₂; (CR₈R₈)q OC(O) Rn;

(CR₈R₈)qNR4C(O)Rn, (CR₈R₈)q NHS(O)₂Rι₇, (CR₈R₈)q S(O)₂NR₄R₅; or two R 1 moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; q is 0, or an integer having a value of 1 to 10; m is an integer having a value of 1 to 3; t is 0, or an integer having a value of 1 or 2; s is an integer having a value of 1 to 3; v is 0, or an integer having a value of 1 to 4; v is 0, or an integer having a value of 1 to 4; R4 and R5 are independently hydrogen, optionally substituted C 1.4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl Cι_4alkyl, heterocyciic, heterocyciic Cl-4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from oxygen, nitrogen or sulfur;

HET is an optionally substituted heteroaryl moiety; R₈ is hydrogen or Cl-4 alkyl;

RlO is Ci-10 alkyl C(O)2R8- Rl 1 is hydrogen, Cι_4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC i -4alkyl, optionally substituted heterocyciic, or optionally substituted heterocyclicC 1 -4alkyl;

Rl2 is hydrogen, Ci-io alkyl, optionally substituted aryl or optionally substituted arylalkyl; Rl3 and R 14 are independently hydrogen or Cl-4 alkyl; Rl7 is Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC i-4alkyl, heterocyciic, or heterocyclicC i-4alkyl, wherein the aryl, heteroaryl and heterocyciic rings may all be optionally substituted; E is optionally selected from

wherein the asterix * denotes the point of attachment of the ring; or a pharmaceutically acceptably salt thereof.

12. The compound according to Claim 1 1 wherein the ionizable hydrogen has a pKa of 3 to 10.

13. The compound according to Claim 1 1 wherein R is hydroxy, carboxylic acid, thiol, -SR2 -OR2, -NH-C(O)R_a, -C(O)NRόR7, -NHS(O)2Rb, -S(O)2NHR_c, NHC(X₂)NHRb, or tetrazolyl; wherein R2 is a substituted aryl, heteroaryl, or heterocyciic ring which ring contains a functional moiety providing an ionizable hydrogen having a pKa of 10 or less;

R_a is an alkyl, aryl, aryl Cι_4alkyl, heteroaryl, heteroaryl Ci-4alkyl, heterocyciic, or a heterocyciic Ci-4alkyl moiety, all of which may be optionally substituted; Rb is a NR6R7, alkyl, aryl, arylCi-4alkyl, arylC2-4alkenyl, heteroaryl, heteroarylC i-4alkyl, heteroarylC2-4 alkenyl, heterocyciic, heterocyciic Ci-4alkyl, heterocyciic C2-4alkenyl moiety, camphor, all of which may be optionally substituted one to three times independently by halogen; nitro; halosubstituted Cl-4 alkyl; Cl-4 alkyl; C M alkoxy; NR9C(O)R_a; C(O)NR6R7, S(O)3H, or C(O)OCi-4 alkyl;

R6 and R7 are independently hydrogen or a C M alkyl group, or R6 and R7 together with the nitrogen to which they are attached form a 5 to 7 member ring which ring may optionally contain an additional heteroatom which heteroatom is selected from oxygen, nitrogen or sulfur;

R9 is hydrogen or a C 1-4 alkyl;

R_c is alkyl, aryl, arylCi-4alkyl, arylC2-4alkenyl, heteroaryl, heteroarylC l-4alkyl, heteroarylC2-4alkenyl, heterocyciic, heterocyciic CMalkyl, or a heterocyciic C2-4alkenyl moiety, all of which may be optionally substituted one to three times independently by halogen, nitro, halosubstituted C M alkyl, C]-4 alkyl, C1 -4 alkoxy, NR9C(O)R_a, C(O)NR6R7, S(O)3H, or C(O)OC M alkyl; and

X2 is oxygen or sulfur.

14. The compound according to Claim 13 wherein the R2 is optionally substituted one to three times by halogen, nitro, halosubstituted Ci-io alkyl, Ci-io alkyl, Q-io alkoxy, hydroxy, SH, -C(O)NR6R7, -NH-C(O)R_a, -NHS(O)₂Rb, S(O)NR6R7, C(O)OR₈, or a tetrazolyl ring.

15. The compound according to Claim 13 wherein R is OH, -NHS(O)2Rb or C(O)OH.

16. The compound according to Claim 11 wherein R] is halogen, cyano, nitro, CF3, C(O)NR4R5, alkenyl C(O)NR4R5, C(O) R4R10, alkenyl C(O)ORi2, heteroaryl, heteroarylalkyl , heteroaryl alkenyl, or S(O)NR4R5.

17. The compound according to Claim 1 1 wherein the HET ring is substituted by (Y)n; and wherein n is an integer having a value of 1 to 3;

Y is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Cj-io alkyl; C2-10 alkenyl; Ci-io alkoxy; halosubstituted Ci-io alkoxy; azide; (CR₈R₈)q S(O)tR4; hydroxy; hydroxyCi-4alkyl; aryl; aryl Cl-4 alkyl; aryloxy; arylCi-4 alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl Cl-4 alkyloxy; heterocyciic, heterocyciic CMalkyl; aryl

C2-10 alkenyl; heteroaryl C2-10 alkenyl; heterocyciic C2-10 alkenyl; (CR₈R₈)q

NR4R5; C2-10 alkenyl C(O)NR4R5; (CR₈R₈)q C(O)NR4R5; (CR₈R₈)q C(O)NR4Rlθ; S(O)3H; S(O)3R8; (CR₈R₈)q C(O)Rπ; C₂-10 alkenyl C(O)Rn ;

C2-10 alkenyl C(O)ORn; C(O)Rn _; (CR₈R₈)q C(O)ORι₂; (CR₈R₈)q OC(O) Rn ;

(CR₈R₈)q NR4C(O)Ri 1, (CR₈R₈)q NHS(O)₂Rd, (CR₈R₈)q S(O)₂NR₄R₅; or two

Y moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; and Rd is NR6R7, alkyl, arylCl-4alklyl, arylC 2.4 alkenyl, heteroaryl, hetroaryl-

Cj_4alkyl, heteroarylC2-4 alkenyl, heterocyciic, heterocyclicC _j .4 alkyl, wherein the aryl, heteoaryl and heterocyciic rings may all be optionally substituted.

18. The compound according to Claim 17 wherein Y is halogen, C M alkoxy, optionally substituted aryl, optionally substituted arylalkoxy, methylene dioxy, NR4R5, thioCi-4alkyl, thioaryl, halosubstituted alkoxy, optionally substituted Ci-4alkyl, hydroxy alkyl.

19. The compound according to Claim 1 1 wherein R is OH, SH, or NHS(O)2Rb and Rl is substituted in the 3-position, the 4- position or di substituted in the 3,4- position by an electron withdrawing moiety.

20. A pharmaceutical composition comprising an effective amount of a compound according to any of Claims 1 1 to 19, and a pharmaceutically acceptable carrier or diluent.