WO1999065310A1 - Il-8 receptor antagonists - Google Patents

Abstract

This invention relates to novel compounds of Formula (I), and compositions thereof, useful in the treatment of disease states mediated by the chemokine, Interleukin-8 (IL-8).

Description

IL-8 RECEPTOR ANTAGONISTS

FIELD OF THE INVENTION

This invention relates to novel benzoisothiazole substituted compounds, pharmaceutical compositions, processes for their preparation, and use thereof in treating IL-8, GROα, GROβ, GROγ and NAP-2 mediated diseases.

BACKGROUND OF THE INVENTION

Many different names have been applied to Interleukin-8 (LL-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 LL-8, these chemokines have also been referred to by different names. For instance GROα, β, γ have been referred to as MGS Aα, 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. LL-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 IL-8 and GROα have demonstrated T-lymphocytes, and basophilic 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. IL-8 has also been shown to increase the surface expression of Mac- 1 (CD 1 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, see Strieter et al., Science 258, 1798 (1992).

In vitro, LL-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 IL-8 receptors (77% homology) have been characterized: LL-8Rα, which binds only IL-8 with high affinity, and LL-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 Gayle 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 IL-8 α or β receptor. Therefore, conditions associated with an increase in LL-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 (II) 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 IL-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) or (II).

The present invention also provides for the novel compounds of Formula (I), and (II) and pharmaceutical compositions comprising a compound of Formula (I), and (II) and a pharmaceutical carrier or diluent. Compounds of Formula (I) useful in the present invention are represented by the structure:

wherein

R is -NH -C(X₂)-NH- (CR₁₃R₁₄)_V - Z;

(CR₁₅R₁₆)P Z is W, HET, , optionally substituted C i .i Q alkyl, optionally substituted C2-10 alkenyl, or optionally substituted C2-I0 alkynyl; X is C(X ₂, O, N-Rig, C=0, or S(O)_mn Xi is independently hydrogen, halogen, Ci-io alkyl, NR4R5, C(O)NR4R5, optionally substituted Ci-io alkyl, Ci-io alkoxy, halosubstituted Ci-io alkoxy, hydroxy, aryl, aryl C 1-4 alkyl, aryloxy, aryl Ci-4 alkyloxy, heteroaryl, heteroarylalkyl, heterocyclic, heterocyclic Cι_4alkyl, or heteroaryl Ci-4 alkyloxy; provided that both of X\ are not hydrogen; X2 is =O, or =S; A is CH₂, C(O) or C(S); R\ is independently selected from hydrogen, halogen, nitro, cyano, halosubstituted Ci-io alkyl, Ci-io alkyl, C2-10 alkenyl, Ci-io alkoxy, halosubstituted -io alkoxy, azide, (CRgRg)q S(O)_tR4, hydroxy, hydroxyCi-ioalkyl, aryl, arylCi-4 alkyl, aryloxy, aryl Ci-4 alkyloxy, heteroaryl, heteroarylalkyl, heterocyclic, heterocyclic Ci-4alkyl, heteroaryl C1-4 alkyloxy, aryl C2-10 alkenyl, heteroaryl C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CRgRg)qNR4R5, C2-10 alkenyl

C(O)NR4R5, (CR₈Rg)q C(O)NR4R5, (CR₈Rg)q C(O)NR4Rl0, S(O)₃H, S(O)₃R8, (CR₈R₈)q C(O)Rι i, C2-10 alkenyl C(O)Ri 1, C2-10 alkenyl C(O)ORl 1, C(O)Ri 1, (CR₈R₈)q C(O)ORl₂, (CR₈R₈)q OC(O) Ri 1, (CR₈R₈)q NR4C(O)Ri 1, (CR₈R₈)qC(NR₄)NR4R₅, (CR₈R₈)q NR₄C(NR₅)Rι 1 , (CR₈R₈)q NHS(O)2Rπ, ^or (CR₈R₈)q S(O)2NR₄R5, or two Ri moieties together may form O-(CH2)sO- or a 5 to 6 membered saturated or unsaturated ring; n is an integer having a value of 1 to 3; m is an integer having a value of 1 to 3; m' is an integer having a value of 1 or 2; p is an integer having a value of 1 to 3; q is 0, or an integer having a value of 1 to 10; s is an integer having a value of 1 to 3; t is 0, or an integer having a value of 1 or 2; v is 0, or an integer having a value of 1 to 4;

HET is an optionally substituted heteroaryl; R4 and R5 are independently hydrogen, optionally substituted Ci .4 alkyl, optionally substituted aryl, optionally substituted aryl Cι_4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl Cj-4alkyl, heterocyclic, or heterocyclic C i-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 O/N/S;

Y is independently selected from hydrogen, halogen, nitro, cyano, halosubstituted Ci-io alkyl, Ci-io alkyl, C2-10 alkenyl, Ci-io alkoxy, halosubstituted -io alkoxy, azide, (CR₈R )q S(O)tR4, hydroxy, hydroxyCi-ioalkyl, aryl, aryl Ci-4 alkyl, aryloxy, arylC ι.4 alkyloxy, heteroaryl, heteroarylalkyl, heteroaryl Ci-4 alkyloxy, heterocyclic, heterocyclic Cι_4alkyl, aryl C2-10 alkenyl, heteroaryl

C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CR R₈)q NR4R5, C2-10 alkenyl C(O)NR4R5, (CR₈R₈)q C(O)NR4R5, (CRgR₈)q C(O)NR4Rlθ, S(O)₃R₈, (CR₈R₈)q C(O)Rι 1, C2-10 alkenyl C(O)Rι 1, C2-10 alkenyl C(O)ORi 1 , (CR₈R₈)q C(O)ORi2, (CR₈R₈)q OC(O) Ri 1, (CR₈R₈)q NR4C(O)Rι 1, (CR₈R₈)qC(NR₄)NR₄R₅, (CR₈R₈)q NR₄C(NR₅)Rι _h (CR₈R₈)q NHS(O)₂R_a, or (CR₈R₈)q S(O)2NR4R5, or two Y moieties together may form O-(CH2)s -O or a 5 to 6 membered saturated or unsaturated ring; R and R7 are independently hydrogen or a Cι_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; R₈ is independently hydrogen or Cj-4 alkyl;

RlO is C i-io alkyl C(O)₂R₈;

Rl 1 is hydrogen, Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl

Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylCi-4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC l -4alkyl; Rl2 is hydrogen, Ci-io alkyl, optionally substituted aryl or optionally substituted arylalkyl; Rl₃ and R 14 are independently hydrogen, optionally substituted Cj_4 alkyl, or one of Rι₃ and R14 may be an optionally substituted aryl;

Rj5 and Ri g are independently hydrogen, or an optionally substituted Ci-4 alkyl; Rl7 is Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylCι_4alkyl, heterocyclic, or heterocyclicC i-4alkyl, wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted; Rj₈ is hydrogen, Ci-4 alkyl, aryl, arylCι.4 alkyl, heteroaryl, heteroarylCι .4 alkyl, heterocyclic, or heterocyclicC \ .4 alkyl, all of which may be optionally substituted; R_a is NRgR7, alkyl, arylCι .4 alkyl, arylC2_4 alkenyl, heteroaryl, heteroaryl-

C^alkyl, heteroarylC2-4 alkenyl, heterocyclic, or heterocyclicC .A. alkyl, wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted;

the E containing ring is optionally selected from

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

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

wherein

R is -NH -C(X )-NH- (CR₁₃R₁₄)_V - Z;

Z is W, HET, (^γ'ⁿ , optionally substituted Cι _ 0 alkyl, optionally substituted C2-10 alkenyl, or optionally substituted C2-10 alkynyl; X is C(Xι);

X1 is hydrogen, halogen, Ci-io alkyl, NR4R5, C(O)NR4R5, optionally substituted Ci-io alkyl, Ci-io alkoxy, halosubstituted Ci-io alkoxy, hydroxy, aryl, aryl Ci-4 alkyl, aryloxy, aryl Ci-4 alkyloxy, heteroaryl, heteroaryl Ci-4 alkyl, heterocyclic, heterocyclic Cι_4alkyl, or heteroaryl Ci-4 alkyloxy; X2 is =O, or =S; A is CR₁₈;

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-ioalkyl, aryl, aryl Cι_4 alkyl, aryloxy, aryl C]-4 alkyloxy, heteroaryl, heteroaryl C i-4 alkyl, heterocyclic, heterocyclic Cj-4alkyl, heteroaryl Ci-4 alkyloxy, aryl C2-10 alkenyl, heteroaryl C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CR₈R )qNR4R5, C2-10 alkenyl C(O)NR4R5, (CR₈R₈)q C(O)NR4R5, (CR₈R₈)q C(O)NR4Rl0, S(O)₃H, S(O)₃Rg, (CR₈R₈)q C(O)Rι 1, C -10 alkenyl C(O)Rι 1, C₂-10 alkenyl C(O)ORi 1, C(O)Rl l, (CR₈R₈)q C(O)ORi2, (CR₈R₈)q OC(O) Rl 1, (CR₈R₈)q

NR4C(O)Ri 1, (CR₈R₈)qC(NR₄)NR₄R₅, (CR₈R₈)q NR₄C(NR₅)Rι _h (CR₈R₈)q NHS(O)2Ri7, or (CR₈R₈)q S(O)2NR4R5, or two Ri moieties together may form O-(CH2)sO or a 5 to 6 membered saturated or unsaturated ring; m is an integer having a value of 1 to 3; n is an integer having a value of 1 to 3; p is an integer having a value of 1 to 3; q is 0, or an integer having a value of 1 to 10; s is an integer having a value of 1 to 3; t is 0, or an integer having a value of 1 or 2; v is 0, or an integer having a value of 1 to 4;

HET is an optionally substituted heteroaryl;

R4 and R5 are independently hydrogen, optionally substituted Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl Cι_4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl Ci-4alkyl, heterocyclic, or heterocyclic 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 O/N/S;

Y is independently selected from hydrogen, halogen, nitro, cyano, halosubstituted Ci-io alkyl, Cj-io alkyl, C2-10 alkenyl, -io alkoxy, halosubstituted Ci-io alkoxy, azide, (CR R )q S(O)tR4, hydroxy, hydroxyCi-ioalkyl, aryl, aryl Ci-4 alkyl, aryloxy, arylCi-4 alkyloxy, heteroaryl, heteroaryl C1.4 alkyl, heteroaryl Ci-4 alkyloxy, heterocyclic, heterocyclic Ci-4alkyl, aryl C2-10 alkenyl, heteroaryl C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CR₈R₈)q NR4R5, C2-10 alkenyl C(O)NR4R5, (CR₈R₈)q C(O)NR4R5, (CR₈R₈)q C(O)NR4Rι₀, S(O)₃R₈, (CR₈R₈)q C(O)Rι l, C2-10 alkenyl C(O)Rn, C2-10 alkenyl

C(O)ORl l, (CR₈R₈)q C(O)ORi2, (CR₈R₈)q OC(O) Rn, (CR₈R₈)q NR4C(O)Rι 1, (CR₈R₈)qC(NR₄)NR₄R5, (CR₈R₈)q NR₄C(NR₅)R_{1 l f} (CR₈R₈)q NHS(O)2R_a, or (CR₈R₈)q S(O)₂NR R₅, or two Y moieties together may form O-(CH2)sO or a 5 to 6 membered saturated or unsaturated ring; R6 and R7 are independently hydrogen or a -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;

R₈ is independently hydrogen or C 1-4 alkyl; Rio is Cι_ιo alkyl C(O)2R₈;

Rl 1 is hydrogen, Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl

Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylCι_4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC 1 -4alkyl; R 2 is hydrogen, Ci-io alkyl, optionally substituted aryl or optionally substituted arylalkyl; Rl₃ and R14 are independently hydrogen, optionally substituted Ci-4 alkyl, or one of Rι₃ and R14 may be an optionally substituted aryl; Rl5 and Rig are independently hydrogen, or an optionally substituted Ci-4 alkyl; Rl7 is Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC i-4alkyl, heterocyclic, or heterocyclicC _4alkyl, wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted; Rl₈ is hydrogen, Ci-4 alkyl, aryl, arylCι.4 alkyl, heteroaryl, heteroarylC 1.4 alkyl, heterocyclic, or heterocyclicC 1.4 alkyl, all of which may be optionally substituted; R_a is NRgR7, alkyl, arylCι.4 alkyl, arylC2-4 alkenyl, heteroaryl, heteroaryl-

Cι _4alkyl, heteroarylC2-4 alkenyl, heterocyclic,or heterocyclicC 1.4 alkyl, wherein the aryl, heteroaryl and heterocyclic rings may all be optionally substituted;

the E containing ring is optionally selected from

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

DETAILED DESCRIPTION OF THE INVENTION

The compounds of Formula (I) and (II) 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 LL-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. As readily seen, the difference between compounds of Formula (I) and (II) lies in the unsaturation of the A containing ring, and hence the substitutions on the X and A moieties. The remaining terms, defined below, are the same for both compounds of Formula (I) and (II) unless otherwise indicated. Suitably, Rl is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl, such as CF₃; C i-io alkyl, such as methyl, ethyl, isopropyl, or n-propyl; C2-10 alkenyl; Ci-io alkoxy, such as methoxy, or ethoxy; halosubstituted Ci-io alkoxy, such as trifluoromethoxy; azide; (CR₈R )q S(O)_tR4, wherein t is 0, 1 or 2; hydroxy; hydroxy C i-ioalkyl, such as methanol or ethanol; aryl, such as phenyl or naphthyl; aryl Ci-4 alkyl, such as benzyl; aryloxy, such as phenoxy; aryl Ci-4 alkyloxy, such as benzyloxy; heteroaryl; heteroaryl C 1.4 alkyl; heteroaryl Ci-4 alkyloxy; aryl C2-10 alkenyl; heteroaryl C2-10 alkenyl; heterocyclic C2-10 alkenyl; (CR₈R₈)qNR4R5; C2-10 alkenyl C(O)NR4R5; (CR₈R₈)q C(O)NR4R5; (CRgR₈)q C(O)NR4Rlθ; S(O)₃H; S(0)₃R₈; (CR₈R₈)q C(O)Rn ; C2-10 alkenyl C(O)Rn; C2-10 alkenyl C(O)ORn ; C(O)Rn ; (CR₈R₈)q C(O)ORi2; (CR₈R₈)q OC(O) Rl 1 ; (CR₈R₈)q NR4C(O)Ri 1 ; (CR₈R₈)qC(NR4)NR₄R5; (CR₈R₈)q NR₄C(NR₅)Rι 1 ; (CR₈R₈)q NHS(O)₂Ri₇; or (CR₈R₈)q S(O)₂NR₄R5; or two R 1 moieties together may form O-(CH2)s-O or a 5 to 6 membered saturated or unsaturated ring. All of the aryl, heteroaryl, and heterocyclic containing moieties above may be optionally substituted as defined herein below. Preferably, Rj is other than azido for methods of use herein.

It is recognized that the Ri moiety may be substituted on the benzene ring or the A containing ring, if possible. Suitably, s is an integer having a value of 1 to 3, and m is an integer having a value of 1 to 3. When Ri forms a dioxybridge, s is preferably 1. When Ri forms an additional saturated or unsaturated ring, it is preferably 6 membered ring, preferably resulting in a naphthylene ring system. These additional rings may be substituted independently, 1 to 3 times, by the other Ri moieties as defined above.

Preferably, Ri is hydrogen, halogen, cyano, nitro, CF₃, (CR₈R )q C(O)NR4R5 , C2- 10 alkenyl C(O)NR4R5 , (CR₈R₈)q C(O)R4R 10, C2- 10 alkenyl

C(O)OR 2, heteroaryl, heteroaryl Cι _4 alkyl, heteroaryl C2-10 alkenyl, or

S(O)₂NR4R5.

Suitably, R4 and R5 are independently hydrogen, optionally substituted Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl C ι_4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl Ci-4alkyl, heterocyclic, or heterocyclicC 1-4 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 O/N/S.

Suitably, R6 and R7 are independently hydrogen or a Ci-4 alkyl group, or R 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.

Suitably, R₈ is independently hydrogen or Cι_4 alkyl.

Suitably, q is 0 or an integer having a value of 1 to 10. Suitably, Rio is Ci-io alkyl C(O)2R₈, such as CH2C(O)2H or CH2C(O)2CH₃.

Suitably, Rl 1 is hydrogen, Ci-4 alkyl, aryl, aryl Ci-4 alkyl, heteroaryl, heteroaryl Ci-4alkyl, heterocyclic, or heterocyclic Ci-4alkyl.

Suitably, R12 is hydrogen, C J.IQ alkyl, optionally substituted aryl or optionally substituted aryl Ci-4 alkyl. Suitably, Ri₃ and R 14 are independently hydrogen, an optionally substituted

Ci-4 alkyl which may be straight or branched as defined herein, or one of Rj₃ and R 14 are an optionally substituted aryl; v is 0, or an integer having a value of 1 to 4. When Rj or R14 are an optionally substituted alkyl, the alkyl moiety may be substituted one to three times independently by halogen; halosubstituted Ci .4 alkyl such as trifluoromethyl; hydroxy; hydroxy Ci-4alkyl, Ci-4 alkoxy; such as methoxy, or ethoxy, halosubstituted Cι_ 10 alkoxy, S(O)tR4; aryl; NR4R5; NHC(O)R4; C(O)NR4Rs; or C(O)OR₈.

Suitably, R17 is Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC i-4alkyl, heterocyclic, or heterocyclicC i-4alkyl, wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted.

Suitably, 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-ioalkyl, aryl, aryl Ci-4 alkyl, aryloxy, arylCi-4 alkyloxy, heteroaryl, heteroaryl Ci-4 alkyl, heteroaryl Ci-4 alkyloxy, heterocyclic, heterocyclic Ci-4alkyl, aryl C2-10 alkenyl, heteroaryl C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CR₈R₈)q NR4R5, C₂-10 alkenyl C(O)NR4R5, (CR₈R₈)q C(O)NR4R5, (CR₈R₈)q C(O)NR4Rl0, S(O)₃R₈, (CR₈R₈)q C(O)Rι 1, C₂-10 alkenyl C(O)Rι 1, C2-10 alkenyl C(O)ORi 1, (CR₈R₈)q C(O)ORι₂, (CR₈R₈)q OC(O) Rl 1, (CR₈R₈)q NR4C(O)Rι ι, (CR₈R₈)qC(NR4)NR4R5, (CR₈R₈)q NR₄C(NR₅)Rι ι, (CR₈R₈)q NHS(O)2R_a, or (CR₈R )q S(O)2NR4R5, or two Y moieties together may form O-(CH2)s -O or a 5 to 6 membered saturated or unsaturated ring. Preferably, Y is other than azido for methods of use herein.

Suitably, n is an integer having a value of 1 to 3.

When Y forms a dioxybridge, s is preferably 1. The aryl, heteroaryl and heterocyclic containing moieties noted above for Y may all be optionally substituted as defined herein. When Y forms an additional saturated or unsaturated ring, it is preferably 6 membered ring, more preferably resulting in a naphthylene ring system.

These additional rings may be optionally substituted 1 to 3 times by other Y moieties as defined above. Suitably, R_a is NRgR7, alkyl, aryl C1.4 alkyl, arylC 2-4 alkenyl, heteroaryl, heteroaryl-Ci-4alkyl, heteroarylC2-4 alkenyl, heterocyclic, or heterocyclicC 1.4 alkyl, wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted.

Y is preferably a halogen, C i-4 alkoxy, optionally substituted aryl, optionally substituted aryloxy or aryl Ci-4 alkoxy, methylenedioxy, NR4R5, thio Ci-4alkyl, thioaryl, halosubstituted Cj-io alkoxy, Ci-io alkyl, or hydroxy Ci-io alkyl. Y is more preferably mono-substituted halogen, disubstituted halogen, mono-substituted alkoxy, disubstituted alkoxy, methylenedioxy, aryl, or alkyl. More preferably these groups are mono or di-substituted in the 2'- position or 2'-, 3 '-position when Z is W and W is a phenyl ring (such as when no E group is present).

While Y may be substituted in any of the 5 ring positions when W is a phenyl moiety, Y is preferably mono-substituted in the 2'-position or 3'- position, with the 4'- preferably being unsubstituted. If the phenyl ring is disubstituted, the substituents are preferably in the 2 ' or 3' position of a monocyclic ring. While both Ri and Y can both be hydrogen, it is preferred that at least one of the rings be substituted, preferably both rings are substituted.

In compounds of Formula (I), A is suitably CH2, C(S) or C(S). It is noted that in Formula (I) the A containing ring is saturated. In compounds of Formula (II), A is suitably CRι₈. It is also noted that in Formula (II) the A containing ring contains unsaturation.

Suitably, Rl₈ is hydrogen, -4 alkyl, aryl, arylCι.4 alkyl, heteroaryl, heteroarylC i-4 alkyl, heterocyclic, or heterocyclicC 1.4 alkyl, all of which may be optionally substituted.

R is suitably -NH -C(X₂)-NH- (CRj ₃Ri4)_v - Z.

Suitably, Z is W, HET, (^γ)n , an optionally substituted C i . IQ alkyl, an optionally substituted C2-10 alkenyl, or an optionally substituted C2-10 alkynyl. Suitably p is an integer having a value of 1 to 3.

Suitably, W is

, or

Suitably, the E containing ring is optionally selected from

the asterix * denoting point of attachment of the ring.

Preferably, Z is

The E ring, denoted by its point of attachment through the asterix (*) may optionally be present. If it is not present, the ring is a phenyl moiety which is substituted by the Y terms as shown above. The E ring may be substituted by the Y moiety in any ring, saturated or unsaturated, and is shown for purposes herein substituted only in the unsaturated ring(s). Suitably X2 is =O, or =S.

In compounds of Formula (I), X is suitably C(Xι)2, N-R ₈, O, C=O, or S(O)_m', and m' is an integer having a value of 1 or 2. Preferably X is O, N-Rι₈, C=O, or

is S(O)_m', and m' is 2, when A is CH2. When X is C(Xι)2, both of Xi can not be hydrogen.

In compounds of Formula (II), X is suitably C(Xι). Suitably, Xi is independently hydrogen, halogen, NR4R5, C(O)NR4R5_; optionally substituted i-io alkyl, Ci-io alkoxy, halosubstituted Ci-io alkoxy, aryl, aryl C i-4 alkyl, aryloxy, aryl Ci-4 alkyloxy, heteroaryl, heteroaryl Ci-4 alkyl, heterocyclic, heterocyclic Ci-4alkyl; or heteroaryl Ci-4 alkyloxy. The alkyl group may be optionally substituted one or more times by hydroxy, NR4R5, or halogen. Preferably, for compounds of Formula (I), when X is C(Xι)2, at least one of X\ is hydrogen.

HET is an optionally substituted heteroaryl moiety, as defined below both for optional substituents and for particular heteroaryl moieties. Suitably R 5 and R are independently hydrogen, or an optionally substituted Ci-4 alkyl as defined above for Rι₃ and R14.

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; Cj-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)2Rl9; C i-io alkyl, such as methyl, ethyl, propyl, isopropyl, or t-butyl; halosubstituted C i-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 heterocyclicalkyl; optionally substituted heteroaryl; optionally substituted heteroaryl alkyl; and wherein these aryl , heteroaryl, or heterocyclic containing 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 C i-io alkyl, such as CF₃.

Rl9 is suitably C1-4 alkyl, aryl, aryl Ci-4alkyl, heteroaryl, heteroarylC i-4alkyl, heterocyclic, 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_ιoalkyl" 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, π-propyl, wσ-propyl, π-butyl, sec-butyl, wø-butyl, tert- butyl, rc-pentyl and the like.

• "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.

• "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. • "heterocyclic" (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. • "arylalkyl" or "heteroarylalkyl" or "heterocyclicalkyl" is used herein to mean Cι_ιo alkyl, as defined above, attached to an aryl, heteroaryl or heterocyclic 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. • "wherein two Ri moieties (or two Y moieties) may together form a 5 or 6 membered saturated or unsaturated ring" is used herein to mean the formation of a bicyclo ring system or a phenyl moiety having attached a 5 or 6 membered saturated or unsaturated ring such as a C cycloalkenyl, i.e. hexene, or a C5 cycloalkenyl moiety, cyclopentene, or a fully unsaturated ring 5 or 6 membered ring such as benzene, i.e. resulting in a naphtylene ring.

Exemplified compounds of Formula (I) include:

N-(2-Bromophenyl) -N' -(1, 3-Dihydro-2, 2-dioxo-2,l-benzisothiazol-7-yl-)urea N-[(l,3-dihydro-2,2-dioxo-4-floro-2,l-benzisothiazo)-7-yl]-N'-(2- bromophenyl)urea N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-(2,3- dichlorophenyl)urea N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-(2- chlorophenyl)urea

N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-(2- methoxyphenyl)urea N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-isopropylurea N-[(l,3-Dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'- (2-bromophenyl)urea

N-[( 1 ,3-Dihydro-2,2-dioxo-4-cyano-2, 1 -benzisothiazo)-7-yl]-N'-

(2-bromophenyl)urea N-[(l,3-Dihydro-2,2-dioxo-4-bromo-2,l-benzisothiazo)-7-yl]-N'- (2-bromophenyl)urea

Additional exemplified compounds of Formula (I) include: N-(4-Bromophenyl) -N' -(1, 3-Dihydro-2, 2-dioxo-2,l-benzisothiazol-7-yl-)urea

Exemplified compounds of Formula (II) include: N-(2-indazole)-N'-(2-bromophenyl)urea

Methods of Preparation

The compounds of Formula (I) and (II) 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 of Formula (I) and (II) having a variety of different Z, R\, and E 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 various compounds of Formula (I) and (II) this is merely for illustration purposes only and not a limitation on the extent of synthesis available using these methods.

Scheme 1

a)SnC , EtOAc b)Et₃N, CH₂CI₂

If the desired cyclic sulfonamide 2-scheme 1 is not commercially available the corresponding heterocyclic sulfamide can be prepared from the commercially available compound 1 -scheme 1 with reduction of the nitro group using suitable reducing conditions such as SnCl2, hydrogen and Pd C or zinc metal in a suitable organic solvent such as, ethyl acetate and cyclization using triethylamine in methylene chloride. Alternative reaction conditions for the cyclic sulfonamide β scheme 1 can be achieved by cyclization of 2-chlorobenzylsulfonamide with potassium carbonate and copper-bronze powder in under pressure using a polar solvent or using a high boiling solvent such as 2, 3-dimethylaniline at 100-180°C, preferably about 180°C, or by reacting the 2-aminobenzylsulfonic acid sodium salt with phosphorus oxychloride at 30-170°c, preferably about 170°C, for between 1-24 hours, preferably about 3 hours. Scheme 2

a) Ph₃P oxide, EtgN, TFAA, 1 ,2 dichloroethane

If the desired heterocyclic compound 2-scheme 2 is not commercially available then it can be prepared from the commercially available compound A scheme 2 with triphenyl phosphine oxide, triethylamine, trifluoroacetic anhydride in an aprotic solvent such as 1, 2 - dichloroethane.

Scheme 3

a) Zn, NH₄CI, THF/H₂0

If the desired heterocyclic compound 2-scheme 3 is not commercially available then it can be prepared from the commercially available compound _ scheme 3 with zinc and ammonium chloride in THF/H2O.

Scheme 4

a) Aluminum amalgam, Et₂0

If the desired heterocyclic compound 2-scheme 4 is not commercially available then it can be prepared from the commercially available compound h scheme 4 with aluminum amalgam in diethyl ether or THF.

Scheme 5

a) HONO b) Na₂NO_s c)H₂S0₄

If the desired heterocyclic compound 3-scheme 5 is not commercially available then it can be prepared from the commercially available compound A scheme 5 with nitrous acid to the diazonium salt, followed by treating with sodium sulfite to the hydrazine 2-scheme 5. Cyclization can be achieved with sulfuric acid to give 3-scheme 5. An alternative method to obtain 3-scheme 5 would be reduction of indazolinone with lithium aluminum hydride.

Scheme 6

a) NaNO,, 3M H₂S0₄, CH₂CI₂, 23°C b) Pd/C, MeOH

If the desired aniline 3-scheme 6 is not commercially available the corresponding nitro compound can be prepared from 1 -scheme 6, under standard nitration conditions (using HNO₃ or NaNO₃) at 0-100°C, preferably about 23°C, under acid conditions such as acetic acid, acetic anhydride, or under biphasic condition such as aqueous sulphuric acid and a chlorinated solvent such as methylene chloride. The nitro compound is then reduced to the corresponding aniline using suitable reducing agents, such as H2 P in an organic solvent, such as

MeOH, DMF or ethylacetate (alternately SnCl2 in EtOH, or LiAlH4 or zinc metal in acetic acid ) at 0-100°C. Other commercially available heterocylic compounds such as Indoline, Indole, Oxindole, Isatin, Indazole and Indazolinone can be converted to the desired anilines through this procedure.

Scheme 7

a) PhNCO, DMF, 80°C

Ortho substituted heterocyclic phenyl ureas in 2-scheme 7 may be prepared by standard conditions involving the condensation of the commercially available optionally substituted aryl isocyanate(Aldrich Chemical Co., Milwaukee, Wi) with the corresponding aniline 1 -scheme 7 in an aprotic solvent such as DMSO, DMF, toluene or methylene chloride at room temperature or elevated temperature from 1 h to 24 h.

Alternatively the desired isocyanates can be made by condensing the amine with triphosgene in the presence of base (such as potassium carbonate) or by reacting the carboxylic acid with diphenyl phosphoazide in the presence of a base (such as triethyl amine).

Another aspect of the invention is the novel process for making compounds of Formula (I), in particularly when A is CH2 and X is S(O)m which process comprises a process for producing a compound of Formula (I), as defined above, wherein A is CH2 and X is S(O)_m, which process comprises a) reacting a compound of the formula

with a compound of the formula:

C(X₂)-N- (CR₁₃Ri4)_v - Z;

to yield a compound of Formula (I).

Another aspect of the present invention is the novel process to make a compound of Formula (A) which process comprises reacting the corresponding nitro compound under reducing conditions to yield the corresponding aniline derivative.

Another aspect of the present invention is the novel process of producing novel compounds of the formula:

which process comprises reacting a compound of the formula

under nitrating conditions to yield a compound of Formula (B).

Yet another aspect of the present invention is a process for producing a compound of the Formula (C), which process comprises reacting a compound of the formula

under reducing conditions and cyclization of the corresponding reduced amine to yield a compound of Formula (D).

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 are highest available purity and all reactions run under anhydrous conditions in an argon atmosphere unless otherwise indicated.

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. ^H-NMR (hereinafter "NMR") spectra were recorded at 250 MHz using a Bruker AM 250 or Am 400 spectrometer. Multiplicities indicated are: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet and br indicates a broad signal. Sat. indicates a saturated solution, eq indicates the proportion of a molar equivalent of reagent relative to the principal reactant. General Method: Synthesis of N, N'- phenyl urea To a solution of phenyl isocyanate(1.0 equiv.) in dimethyl formamide (1ml) the corresponding aniline (1.0 equiv.) was added. The reaction mixture was stirred at 80°C until complete (3-16 hrs.), then removed solvent under vacuum. The purification, yields and spectral characteristics for each individual compound are listed below. Additional synthetic methods are provided for in PCT US96/02260 filed 16 February 1996 whose disclosure is incorporated herein by reference. Example 1 Preparation of N-(2-Bromophenyl) -N' -(l, 3-Dihydro-2, 2-dioxo-2.1-benzisothiazol- 7-yl-)urea a)Preparation of 1, 3-Dihydro-l, 2-benzisothiazole-2, 2-dioxide To a solution of 2-nitro-alpha-toluenesulfonyl chloride(5.0g, 21.3mmol) in

250ml of ethyl acetate was added tin II chloride(19.2g, 85mmol). The reaction was stirred at 70°C overnight then poured into ice and neutralized with sodium bicarbonate. The solution was then extracted with ethyl acetate and the solvents were evaporated. The crude reaction mixture was then diluted with methylene chloride and excess triethyl amine was added. The solution was stirred at 25°C overnight and the solvent was evaporated and the product was obtained by chromatography of the resulting solid on silica gel(EtOAc/ hexane(lequiv./lequiv.)). (500mg, 14%). Η NMR (CDC1₃): δ 7.25 (d, 1H), 7.24

(t, 1H), 7.07 (t, 1H), 6.91 (d, 1H), 6.62 (s, 1H), 4.40 (s, 2H). b)Preparation of 4-nitro-l, 3-Dihydro-l. 2-benzisothiazole-2, 2-dioxide

1, 3-Dihydro-l, 2-benzisothiazole-2, 2-dioxide (400mg, 2.40mmol) was dissolved in methylene chloride(40ml) followed by the addition of sodium nitrate (0.22g, 2.60mmol). The addition of sulfuric acid (5.0ml/ 3M) is then made, followed by addition of a catalytic amount of sodium nitrite. The mixture is allowed to stir. After 24 hr., the reaction mixture is diluted with methylene chloride and extracted with water. The organic layer is dried over MgSO₄ and filtered. The solvent was evaporated and chromatography of the resulting solid on silica gel (4%MeOH/ CH,C1₂) gave the desired product(150 mg, 29 %). Η NMR (CD₃OD): δ

8.11 (d, 1H), 7.57 d, 1H), 7.09 (t, 1H), 4.50 (s, 2H). c)Preparation of 4-amino- 1 , 3-Dihydro- 1 , 2-benzisothiazole-2, 2-dioxide

To a solution of 4-nitro-l, 3-Dihydro-l, 2-benzisothiazole-2, 2-dioxide (lOOmg, 5.0mmol) in methanol (50ml) and was added 10% Pd/C (50mg). The mixture was flushed with argon, then hydrogen was bubbled through the solution for 10 min. and a hydrogen atmosphere was maintained at balloon pressure overnight. The mixture was filtered through celite and the celite was washed with methanol. The solvent was evaporated and chromatography of the resulting solid on silica gel (10%MeOH/ CH₂C1₂) gave the desired product (64mg, 74 %). Η NMR (CD₃OD): δ 6.86 (t, 1H), 6.62 (d, 1H), 6.55 (d, 1H), 4.35 (s, 2H). d)Preparation of N-[l, 3-Dihydro-l, 2-benzisothiazole-3, 3 -dioxide] -N'- [2- bromophenyl] urea N-[l, 3-Dihydro-l, 2-benzisothiazole-3, 3-dioxide]-N'-[2-bromophenyl] urea was prepared from 4-amino-l, 3-Dihydro-l, 2-benzisothiazole-2, 2-dioxide (64mg, 0.35 mmol) according to the procedure in General Method B. The product was purified by chromatography of the resulting solid on silica gel(EtOAc/ hexane (lequivJlequiv.)). (45mg, 34%). Η NMR (CD₃SO₂CD₃): δ 9.70 (s, IH), 9.04 (s,

IH), 8.51 (s, IH), 8.08 (d,lH), 7.67 (t, IH), 7.60 (d, IH), 7.34 (t, IH), 7.04 (d, 2H), 6.96 (t, IH), 4.58 (s, 2H).

Using analogous methods to those described above or in the schematics, the following compounds may be synthesized:

Example 2: N-(l, 3-Dihydro-4-bromo-l, 2-benzisothiazole-3, 3-dioxide) N - phenylurea Example 3: N-[(l,3-dihydro-2,2-dioxo-4-floro-2,l-benzisothiazo)-7-yl]-N'-(2- bromophenyl)urea; (M^": 398.1, 400.1) Example 4: N-[( l,3-dihydro-2,2-dioxo-4-chloro-2, l-benzisothiazo)-7-yl]-N'-(2,3- dichlorophenyl)urea; (M^" : 403.9, 406.2, 408.0) Example 5: N-[(l,3-dihydro-2,2-dioxo-4-chloro-2, l-benzisothiazo)-7-yl]-N'-(2- chlorophenyl)urea; (M^" : 370.1, 372.1) Example 6: N-[( l,3-dihydro-2,2-dioxo-4-chloro-2, l-benzisothiazo)-7-yl]-N'-(2- methoxyphenyl)urea; (M^" : 366.2, 358.1)

Example 7: N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N - isopropylurea; (M^" : 304.1, 306.2) Example 8: N-[( l,3-Dihydro-2,2-dioxo-4-chloro-2.1-benzisothiazo)-7-yl]-N'- (2-bromophenyl)urea; (M- : 414, 416) Example 9: N-[( l,3-Dihydro-2,2-dioxo-4-cyano-2, l-benzisothiazo)-7-yl]-N'- (2-bromophenyl)urea; (M^" : 404.9, 407.1) Example 10: N-[(l,3-Dihydro-2,2-dioxo-4-bromo-2,l-benzisothiazo)-7-yl]-N -

(2-bromophenyl)urea; (M^" : 457.9, 459.9, 461.9) Example 1 1: N-(2-indazole)-N'-(2-bromophenyl)urea; MS (M +H = 331.1 ; M-H = 329.2)

METHOD OF TREATMENT

The compounds of Formula (I), and (II) 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 LL-8 α or β receptor, also referred to as the type I or type II receptor.

For purposes herein, the term Formula (I) will also refer to compounds of Formula (II) unless otherwise indicated.

Accordingly, the present invention provides a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an IL-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 chemokines are IL-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 IL-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 IL-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 associated IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 above normal physiological levels; or (iii) the presence IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 above basal levels in cells or tissues in which IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 respectively, is produced.

There are many disease states in which excessive or unregulated IL-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, Alzheimer's disease, atherosclerosis, gingivitis, allograft rejections, malaria, restinosis, angiogenesis or undesired hematopoietic stem cells release, rhinovirus infections, and various bone resorptive indications, such as osteoporosis or osteoarthritis, and diseases caused by respiratory viruses, including but not limited to rhinovirus and influenza virus, herpesvirus, including but not limited to herpes simplex I and II, and hepatitis viruses, including but not limited to Hepatitis B and C virus. The association of interleukin-8 and rhinovirus may be found in articles such as: Turner,et al., Clin. Infect. Dis. (1998), 26(4), 840-846; Sanders, et al., J. Virol. (1998), 72(2), 934-942; Sethi, et al., Clin. Exp. Immunol. (1997), 110(3), 362-369; Zhu, et al., Am. J. Physiol. (1997), 273(4, Pt. 1), L814-L824; Terajima, et al., Am. J. Physiol. (1997), 273(4, Pt. 1), L749-L759; Grunberg, et al., Clin. Exp. Allergy (1997), 27(1), 36-45; and Johnston, et al., J. Infect. Dis. (1997), 175(2), 323-329. The association of interleukin-8 and osteoporosis may be found in articles such as: Streckfus et al., J. Gerontol., Ser. A (1997), 52A(6), M343-M351; Hermann, T. WO 95/31722; and Chaudhary, et al., Endocrinology (Baltimore) (1992), 130(5), 2528-34.

These diseases are primarily characterized by massive neutrophil infiltration, T-cell infiltration, or neovascular growth, and are associated with IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 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 (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 IL-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, pp. 661 (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.

Present evidence also indicates the use of IL-8 inhibitors in the treatment of atherosclerosis. The first reference, Boisvert et al., J Clin Invest, 1998, 101:353- 363 shows, through bone marrow transplantation, that the absence of IL-8 receptors on stem cells (and, therefore, on monocytes/macrophages) leads to a reduction in the development of atherosclerotic plaques in LDL receptor deficient mice. Additinal supporting references are: Apostolopoulos, et al., Arterioscler Thromb Vase Biol. 1996, 16: 1007-1012; Liu, et al., Arterioscler Thromb Vase Biol, 1997, 17:317-323; Rus, et al., Atherosclerosis. 1996, 127:263-271.; Wang et al., J Biol Chem. 1996, 271:8837-8842; Yue, et al., Eur J Pharmacol. 1993, 240:81-84; Koch, et al., Am J Pathol, 1993, 142: 1423-1431.; Lee, et al., Immunol Lett, 1996, 53, 109-113.; and Terkeltaub et al., Arterioscler Thromb, 1994, 14:47-53.

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 LL-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 to be inhibitors of type II LL-8 receptors.

As used herein, the term "LL-8 mediated disease or disease state" refers to any and all disease states in which IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 plays a role, either by production of IL-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, LL-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 "chemokine mediated disease or disease state" refers to any and all disease states in which a chemokine which binds to an IL-8 α or β receptor plays a role, such as but not limited IL-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 LL-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, LL-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 IL-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 (IL-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, IL-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 phenylmercuric 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 (I) 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 IL-8, and GRO-α chemokine inhibitory effects of compounds of the present invention are determined by the following in vitro assay: Receptor Binding Assays:

[125j IL_₈ (human recombinant) is obtained from Amersham Corp., Arlington Heights, LL, with specific activity 2000 Ci/mmol. Gro-α is obtained from NEN- New England Nuclear. All other chemicals are of analytical grade. High levels of recombinant human IL-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 is changed to lOmM Tris-HCL, ImM MgS04, 0.5mM EDTA (ethylene-diaminetetra- acetic acid), lm MPMSF (α-toluenesulphonyl fluoride), 0.5 mg/L Leupeptin, pH 7.5. Membrane protein concentration is determined using Pierce Co. micro-assay kit using bovine serum albumin as a standard. All assays are performed in a 96-well micro plate format. Each reaction mixture contains ^5j IL_₈ (0.25 nM) or 125j_ GRO-α and 0.5 μg/mL of LL-8Rα or 1.0 μg/mL of IL-8Rβ membranes in 20 mM Bis-Trispropane and 0.4 mM Tris HC1 buffers, pH 8.0, containing 1.2 mM MgSO4,

0.1 mM EDTA, 25 mM NaCl and 0.03% CHAPS. In addition, drug or compound of interest is added which has been pre-dissolved in DMSO so as to reach a final concentration of between O.OlnM and 100 uM. The assay is initiated by addition of 125τ-iL_g After 1 hour at room temperature the plate is 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 NaCl, 10 mM TrisHCl, 1 mM MgSO4,

0.5 mM EDTA, 0.03 % CHAPS, pH 7.4. The filter is 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.

Representative compounds of Formula (I), Example 1, 3 to 10, and a representative compound of Formula (II), Example 11. have been found to have positive inhibitory activity of < 30 μmg 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 LL-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, LL, 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 (NaCl 118, KC1 4.56,

NaHCO3 25, KH2PO4 1.03, Glucose 1 1.1, 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 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 supernatant is added to a second 96 well plate followed by an artificial elastase substrate (MeOSuc- Ala- Ala-Pro- Val- 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 the rate of MeOSuc- Ala-Ala-Pro- Val-AMC degradation. TNF-α in Traumatic Brain Injury Assay

This assay provides for examination of the expression of tumor necrosis factor mRNA in specific brain regions which follow experimentally induced lateral fluid- percussion traumatic brain injury (TBI) in rats. 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. A suitable assay may be found in WO 97/35856 or WO 97/49286 whose disclosures are incorporated herein by reference.

CNS Injury model for IL-β mRNA

This assay characterizes the regional expression of interleukin-lβ (IL-lβ) mRNA in specific brain regions following experimental lateral fluid-percussion traumatic brain injury (TBI) in rats. Results from these assays 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 pathologic or regenerative sequelae of brain injury. A suitable assay may be found in WO 97/35856 or WO 97/49286 whose disclosures are incorporated herein by reference.

In vivo - athereoschlerosis assay:

In vivo models for measuring atherosclerosis in mice is based on the assay of Paigen et al with small modifications as described below. See Paigen B, Morrow A, Holmes PA, Mitchell D, Williams RA. Quantitative assessment of atherosclerotic lesions in mice. Atherosclerosis 68: 231-240 (1987); and Groot PHE, van Vlijmen BJM, Benson GM, Hofker MH, Schiffelers R, Vidgeon-Hart M, Havekes LM. Quantitative assessment of aortic atherosclerosis in APOE*3 Leiden transgenic mice and its relationship to serum cholesterol exposure. Arterioscler Thromb Vase Biol. 16: 926-933 (1996). Sectioning and staining of the aortic sinus

Cross-sections of the aortic root are taken as has been described previously (1,2). Briefly, the hearts are bisected just below the level of the atria and the base of the heart plus aortic root are taken for analysis. After equilibrating the tissue in OCT compound overnight the hearts are immersed in OCT compound on a cryostat chuck (Bright Instrument Company Ltd., UK) with the aorta facing the chuck. The tissue is frozen by surrounding the chuck with dry ice. The hearts are then sectioned peφendicular to the axis of the aorta, starting within the heart and working in the direction of the aorta. Once the aortic root has been identified by the appearance of the three valve leaflets, alternate 10 mm sections are taken and mounted on gelatinised slides. Sections are air dried for 1 hour and subsequently rinsed briefly in 60% isopropyl alcohol. The sections are stained with Oil Red O, counterstained with Mayer's haematoxylin, cover slipped using glycerol gelatine and sealed with nail varnish.

Quantification of atherosclerosis in the aortic root

Ten alternate sections of the aortic root are imaged using an Olympus BH-2 microscope equipped with an 4x objective and a video camera (Hitachi, HV-C10). Twenty-four bit colour images are acquired and analysed using a PC (Datacell Pentium P5-133, Datacell, Berks, U.K.) fitted with a framegrabbing board (Snapper, Active Imaging Ltd, Berks, U.K.) and running Optimas software (version 5.1, Optimas Coφ., WA, U.S.A.). The images are captured under identical lighting, microscope, camera and PC conditions. Quantification of the atherosclerotic lesion areas is performed by drawing around the lesions by hand using the Optimas software. Colour thresholds are set that quantify the areas that are stained red within the lesions. Absolute values for the cross-sectional areas of the lesions and the areas stained red are obtained by calibrating the software using an image of the grid on a haemocytometer slide.

All publications, including but not limited to patents and patent applications, cited in this specification are herein incoφorated by reference as if each individual publication were specifically and individually indicated to be incoφorated 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 disease selected from the group consisting of gingitivitis, diseases caused by respiratory viruses, heφesviruses, and hepatitis viruses, and atheroschlerosis in a mammal in need thereof, which method comprises administering to said mammal an effective amount of a compound according to the formula:

wherein

R is -NH -C(X )-NH- (CR ₃R╬╣₄)_v - Z;

Z is W, HET, ^{( )n} , optionally substituted C i-io alkyl, optionally substituted C2-10 alkenyl, or optionally substituted C2-10 alkynyl;

X is O, N-R╬╣₈, C=O, or S(O)_m';

X2 is =O, or =S;

A is CH₂, C(O) or C(S);

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-ioalkyl, aryl, aryl C1-4 alkyl, aryloxy, aryl Cj-4 alkyloxy, heteroaryl, heteroaryl Ci-4 alkyl, heterocyclic, heterocyclic Ci-4alkyl, heteroaryl Ci-4 alkyloxy, aryl C2-10 alkenyl, heteroaryl C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CR₈Rg)qNR4R5, C2-10 alkenyl C(O)NR4R5, (CR₈R₈)q C(O)NR4R5, (CR₈R₈)q C(O)NR4RlO, S(O)₃R₈, (CR₈R₈)q C(O)R╬╣ 1, C2-10 alkenyl C(O)R╬╣ 1, C2-10 alkenyl C(O)ORi 1, C(O)R╬╣ 1, (CR₈R₈)q C(O)OR╬╣₂, (CR₈R₈)q OC(O)R╬╣ 1, (CR₈R₈)q NR4C(O)Rn, (CR₈R₈)qC(NR4)NR4R₅; (CR₈R₈)q NR4C(NR₅)Rn; (CR₈R₈)q NHS(O)₂Rn, or (CR₈R₈)q S(O)2NR4R₅, or two Rl moieties together may form O-(CH2)sO- or a 5 to 6 membered saturated or unsaturated ring; and wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted; n is an integer having a value of 1 to 3; m is an integer having a value of 1 to 3; m' is an integer having a value of 1 or 2; q is 0, or an integer having a value of 1 to 10; s is an integer having a value of 1 to 3; t is 0, or an integer having a value of 1 or 2; v is 0, or an integer having a value of 1 to 4; p is an integer having a value of 1 to 3;

HET is an optionally substituted heteroaryl;

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, heterocyclic, or heterocyclic 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 O/N/S; Y is independently selected from hydrogen, halogen, nitro, cyano, halosubstituted Ci-io alkyl, -io alkyl, C2-10 alkenyl, Ci-io alkoxy, halosubstituted Ci-io alkoxy, azide, (CR₈R )q S(O)_tR4, hydroxy, hydroxyCi-ioalkyl, aryl, aryl -4 alkyl, aryloxy, arylCi-4 alkyloxy, heteroaryl, heteroaryl Ci-4 alkyl, heteroaryl Ci-4 alkyloxy, heterocyclic, heterocyclic Ci-4alkyl, aryl C2-10 alkenyl, heteroaryl C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CR₈R₈)q NR4R5, C2-10 alkenyl C(O)NR4R5, (CR₈R₈)q C(O)NR4R5, (CR₈R₈)q C(O)NR4Rl0, S(O)₃R₈, (CR₈R₈)q C(O)Rn, C₂-10 alkenyl C(O)Rn, C -10 alkenyl C(O)ORn, (CR₈R₈)q C(O)ORl2, (CR₈R₈)q OC(O)Rn, (CR₈R₈)q NR4C(O)R╬╣ 1, (CR₈R₈)qC(NR₄)NR₄R₅, (CR₈R₈)q NR₄C(NR5)R! _h (CR₈R₈)q NHS(O)2R_a, or (CR R₈)q S(O)2NR4R5, or two Y moieties together may form O-(CH2)s -O or a 5 to 6 membered saturated or unsaturated ring; and wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted;

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; R₈ is independently hydrogen or Ci-4 alkyl;

RlO is Ci-10 alkyl C(O)2R₈; Rl 1 is hydrogen, Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC i-4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC i-4alkyl; Rl2 is hydrogen, Ci-io alkyl, optionally substituted aryl, or optionally substituted arylalkyl; Rl₃ and R14 are independently hydrogen, optionally substituted Ci .4 alkyl, or one of R╬╣₃ and R14 may be an optionally substituted aryl; Rl5 and R g are independently hydrogen, or an optionally substituted Ci-4 alkyl; Rl7 is C╬╣_4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC i-4alkyl, heterocyclic, or heterocyclicC i-4alkyl, and wherein the aryl, heteroaryl and heterocyclic rings may all be optionally substituted; Rl₈ is hydrogen, Ci-4 alkyl, aryl, arylC╬╣.4 alkyl, heteroaryl, heteroarylC 1.4 alkyl, heterocyclic, or heterocyclicC 1.4 alkyl, all of which may be optionally substituted; R_a is NR6R7, alkyl, aryl C1.4 alkyl, arylC2-4 alkenyl, heteroaryl, heteroaryl- Cj.4 alkyl, heteroarylC 2-4 alkenyl, heterocyclic, heterocyclicC 1.4 alkyl, and wherein the aryl, heteroaryl and heterocyclic rings may all be optionally substituted;

the E containing ring is optionally selected from

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

2. The method according to Claim 1 wherein Ri is halogen, cyano, nitro, CF₃, (CR₈R₈)q C(O)NR4R5, C₂-io alkenyl C(O)NR4R₅, (CR₈R₈)q

C(O)R4R╬╣o, C2-IO alkenyl C(O)ORi2, heteroaryl, heteroaryl C1.4 alkyl, heteroaryl C2-10 alkenyl, or S(O)₂NR4R5.

3. The method according to Claim 1 wherein X is S(O)m'.

4. The method according to Claim 1 wherein X is N-R╬╣ .

5. The method according to Claim 1 wherein X is O.

6. The method according to Claim 1 wherein X is C=O.

7. The method according to any of Claims 1 to 6 wherein A is CH2-

8. The method according to Claim 1 wherein Z is W.

9. The method according to Claim 8 wherein Y is halogen, Ci-ioalkoxy, optionally substituted aryl, optionally substituted aryl Cj_4 alkoxy, methylenedioxy, NR4R5, thio Ci-4 alkyl, thioaryl, halosubstituted Ci-io alkoxy, C╬╣_╬╣oalkyl, or hydroxy Ci-io alkyl.

10. The method according to Claim 1 which is: N-( 1 ,3-Dihydro- 1 ,2-benzisothizole-3,3-dioxide) N - (2-bromophenyl)urea; N-[( 1 ,3-dihydro-2,2-dioxo-4-floro-2, 1 -benzisothiazo)-7-yl]-N'-(2- bromopheny 1 )urea N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-(2,3- dichlorophenyl)urea N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-(2- chlorophenyl)urea N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-(2- methoxyphenyl)urea N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-isopropylurea N-[(l,3-Dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N - (2-bromophenyl)urea N-[( 1 ,3-Dihydro-2,2-dioxo-4-cyano-2, 1 -benzisothiazo)-7-yl]-N'-

(2-bromophenyl)urea N-[(l,3-Dihydro-2,2-dioxo-4-bromo-2,l-benzisothiazo)-7-yl]-N'- (2-bromophenyl)urea; or a pharmaceutically acceptable salt thereof.

1 1. A method of treating osteoporosis in a mammal in need thereof, which method comprises administering to said mammal an effective amount of a compound according to the formula:

wherein

R is -NH -C(X₂)-NH- (CR╬╣₃R╬╣₄)_v - Z;

Z is W, HET, ^(╬│'ⁿ , optionally substituted C╬╣_ 10 alkyl, optionally substituted C2-10 alkenyl, or optionally substituted C2-10 alkynyl;

X is O, N-R ₈, C=O, or S(O)_m'; X2 is =O, or =S; A is CH₂, C(O) or C(S);

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, (CR₈R )q S(O)_tR4, hydroxy, hydroxy Cj-ioalkyl, aryl, aryl Cj-4 alkyl, aryloxy, aryl Ci-4 alkyloxy, heteroaryl, heteroaryl C╬╣_4 alkyl, heterocyclic, heterocyclic C╬╣_4alkyl, heteroaryl Ci-4 alkyloxy, aryl C2-10 alkenyl, heteroaryl C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CR₈R )qNR4R5, C2-10 alkenyl C(O)NR4R5, (CR₈R₈)q C(O)NR4R₅, (CR₈R₈)q C(O)NR4Rl0,

S(O)₃R₈, (CR₈R₈)q C(O)R╬╣ 1, C2-10 alkenyl C(O)R╬╣ 1, C -10 alkenyl C(O)ORl l, C(O)Rl 1, (CR₈R₈)q C(O)ORl₂, (CR₈R₈)q OC(O)Rl l, (CR₈R₈)q NR4C(O)Rn, (CR₈R₈)qC(NR4)NR₄R5; (CR₈R₈)q NR₄C(NR₅)R╬╣ _{i ;} (CR₈R₈)q NHS(O)2Ri7, or (CR₈R )q S(O)₂NR R₅, or two R\ moieties together may form O-(CH2)sO- or a 5 to 6 membered saturated or unsaturated ring; and wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted; n is an integer having a value of 1 to 3; m is an integer having a value of 1 to 3; m' is an integer having a value of 1 or 2; q is 0, or an integer having a value of 1 to 10; s is an integer having a value of 1 to 3; t is 0, or an integer having a value of 1 or 2; v is 0, or an integer having a value of 1 to 4; p is an integer having a value of 1 to 3; HET is an optionally substituted heteroaryl;

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 C╬╣_4alkyl, heterocyclic, or heterocyclic 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 O/N/S;

Y is independently selected from hydrogen, halogen, nitro, cyano, halosubstituted Ci-io alkyl, Cj-io alkyl, C2-10 alkenyl, C i-io alkoxy, halosubstituted Ci-io alkoxy, azide, (CR₈R₈)q S(O)tR4, hydroxy, hydroxyCj-ioalkyl, aryl, aryl Ci-4 alkyl, aryloxy, arylCi-4 alkyloxy, heteroaryl, heteroaryl Ci-4 alkyl, heteroaryl

Ci-4 alkyloxy, heterocyclic, heterocyclic Ci-4alkyl, aryl C2-10 alkenyl, heteroaryl C2-10 alkenyl, heterocyclic C2-10 alkenyl, (CR₈R )q NR4R5, C2-10 alkenyl C(O)NR4R₅, (CR₈R₈)q C(O)NR4R5, (CR₈R₈)q C(O)NR4R╬╣₀, S(O)₃R₈, (CR₈R₈)q C(O)Rn , C₂-10 alkenyl C(O)Rn, C2-10 alkenyl C(O)ORi 1, (CR₈R₈)q C(O)ORi₂, (CR₈R₈)q OC(O)Ri 1, (CR₈R₈)q

NR4C(O)R╬╣ 1, (CR₈R₈)qC(NR₄)NR₄R5, (CR₈R₈)q NR₄C(NR₅)R╬╣ (CR₈R₈)q NHS(O)2R_a, or (CR₈R₈)q S(O)2NR4R₅, or two Y moieties together may form O-(CH2)s -O or a 5 to 6 membered saturated or unsaturated ring; and wherein the aryl, heteroaryl and heterocyclic containing rings may all be optionally substituted;

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; R₈ is independently hydrogen or C1-.4 alkyl;

RlO is Ci-10 alkyl C(O)2R₈;

Rl 1 is hydrogen, Ci_4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC ╬╣_4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC 1 _4alky 1 ; Rl2 is hydrogen, Ci-io alkyl, optionally substituted aryl, or optionally substituted arylalkyl; Rl₃ and R14 are independently hydrogen, optionally substituted Ci-4 alkyl, or one of R╬╣₃ and R14 may be an optionally substituted aryl; Rl5 and R15 are independently hydrogen, or an optionally substituted Ci-4 alkyl; Rl7 is Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC i-4alkyl, heterocyclic, or heterocyclicC i-4alkyl, and wherein the aryl, heteroaryl and heterocyclic rings may all be optionally substituted; Rl₈ is hydrogen, Ci-4 alkyl, aryl, arylC╬╣_4 alkyl, heteroaryl, heteroarylC 1.4 alkyl, heterocyclic, or heterocyclicC .4 alkyl, all of which may be optionally substituted; R_a is NRgR7, alkyl, aryl C .4 alkyl, arylC2-4 alkenyl, heteroaryl, heteroaryl- C╬╣_4alkyl, heteroarylC2-4 alkenyl, heterocyclic, heterocyclicC 1.4 alkyl, and wherein the aryl, heteroaryl and heterocyclic rings may all be optionally substituted;

the E containing ring is optionally selected from

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

12. The method according to Claim 11 wherein Ri is halogen, cyano, nitro, CF₃, (CR₈R₈)q C(O)NR4R5, C₂-io alkenyl C(O)NR4R5, (CR₈R₈)q C(O)R4R╬╣o, C2-10 alkenyl C(O)ORi2, heteroaryl, heteroaryl C╬╣_4 alkyl, heteroaryl C₂-io alkenyl, or S(O)2NR4R5.

13. The method according to Claim 11 wherein X is S(O)m'.

14. The method according to Claim 11 wherein X is N-R╬╣₈.

15. The method according to Claim 1 lwherein X is O.

16. The method according to Claim 1 wherein X is C=O.

17. The method according to any of Claims 11 to 16 wherein A is CH2-

18. The method according to Claim 11 wherein Z is W.

19. The method according to Claim 18 wherein Y is halogen, Ci-ioalkoxy, optionally substituted aryl, optionally substituted aryl Ci-4 alkoxy, methylenedioxy, NR4R5, thio Ci-4 alkyl, thioaryl, halosubstituted Ci-io alkoxy, Ci-ioalkyl, or hydroxy Ci-io alkyl.

20. The method according to Claim 11 which is:

N-( 1 ,3-Dihydro- 1 ,2-benzisothizole-3,3-dioxide) N- (2-bromophenyl)urea; N-[( 1 ,3-dihydro-2,2-dioxo-4-floro-2, l-benzisothiazo)-7-yl]-N'-(2- bromophenyl)urea

N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-(2,3- dichlorophenyl)urea N-[(l,3-dihydro-2,2-dioxo-4-chloro-2,l-benzisothiazo)-7-yl]-N'-(2- chlorophenyl)urea N-[( 1 ,3-dihydro-2,2-dioxo-4-chloro-2, 1 -benzisothiazo)-7-yl]-N'-(2- methoxyphenyl)urea N-[( 1 ,3-dihydro-2,2-dioxo-4-chloro-2, 1 -benzisothiazo)-7-yl]-N'-isopropylurea N-[( 1 ,3-Dihydro-2,2-dioxo-4-chloro-2, l-benzisothiazo)-7-yl]-N'-

(2-bromophenyl)urea N-[( 1 ,3-Dihydro-2,2-dioxo-4-cyano-2, 1 -benzisothiazo)-7-yl]-N'-

(2-bromophenyl)urea N-[( 1 ,3-Dihydro-2,2-dioxo-4-bromo-2, 1 -benzisothiazo)-7-yl]-N'-

(2-bromophenyl)urea; or a pharmaceutically acceptable salt thereof.