MODULATORS

FIELD OF THE INVENTION

[01] This invention provides benzoxazine aryl sulfonamide compounds and related pharmaceutical compositions, methods for preventing and/or treating conditions associated with potassium channel activity, and methods of making the subject compounds.

BACKGROUND OF THE INVENTION

[02] Potassium channels represent a complex class of voltage-gated ion channels from both functional and structural standpoints. Their functions include regulating neurotransmitter release, heart rate, insulin secretion, neuronal excitability, epithelial electrolyte transport, smooth muscle contraction, and cell volume. In general, four sequence-related potassium channel genes - shaker, shaw, shab, and shal - have been identified in Drosophila, and each has been shown to have human homolog(s).

[03] KCNA3 encodes the voltage-gated K _v l .3 potassium channel, which is shaker-related and is expressed in lymphocytes (T and B lymphocytes), the central nervous system, fat and other tissues. The functional channel is composed of four identical K _v l .3 a-sub units. The K _v 1.3 potassium channel regulates membrane potential and thereby indirectly influences calcium signaling in human effector-memory T cells (Grissmer S. et al, Proc. Natl. Acad. Sci. U.S.A. 87(23): 9411-5; DeCoursey T.E. et al, Nature 307 (5950): 465-8; Chandy K.G. et al, Trends Pharmacol. Sci. 25(5): 280-9; Wulff H. et al, J. Clin. Invest. I l l (11): 1703-13). Effector memory T cells are important mediators of multiple sclerosis, Type I diabetes mellitus, psoriasis, and rheumatoid arthritis.

[04] The K _v l .3 channel is expressed in T and B lymphocytes in a distinct pattern that depends on the state of lymphocyte activation and differentiation. Upon activation, naive and central memory T cells increase expression of the Kc _a 3.1 channel per cell, while effector-memory T cells increase expression of the K _v l .3 channel. Amongst human B cells, naive and early memory B cells express small numbers of K _v l .3 and Kc _a 3.1 channels when they are quiescent, and augment Kc _a 3.1 expression after activation. In contrast, class-switched memory B cells express high numbers of K _v l .3 channels per cell (about 1500/cell) and this number increases after activation (Chandy K.G. et al, Trends Pharmacol. Sci. 25(5): 280-9; Wulff H. et al, J. Clin. Invest. 1 11 (11): 1703-13; Wulff H. et al, J. Immunol. 173(2): 776-86). The K _v 1.3 channel promotes the calcium homeostasis required for T-cell receptor-mediated cell activation, gene transcription, and proliferation (Panyi, G et al (2004) Trends Immunol 25:565-569).

[05] K _v l .3 is physically coupled through a series of adaptor proteins to the T- cell receptor signaling complex and it traffics to the immunological synapse during antigen presentation. However, blockade of the channel does not prevent immune synapse formation (Panyi G. et al, Proc. Natl. Acad. Sci. U.S.A., 101(5): 1285-90; Beeton C. et al, Proc. Natl. Acad. Sci. U.S.A., 103(46): 17414-9).

[06] K _v l .3 and Kc _a 3.1 regulate membrane potential and calcium signaling of T cells. Calcium entry through the CRAC channel is promoted by potassium efflux through the K _v l .3 and Kc _a 3.1 potassium channels. Blockade of K _v 1.3 channels in effector- memory T cells suppresses activities like calcium signaling, cytokine production

(interferon-gamma, interleukin 2) and cell proliferation. Effector-memory T cells (TEM) were originally defined by their expression of cell surface markers, and can enter sites of inflammation in non-lymphoid tissues, while not participating in the process of lymphoid recirculation carried out by most other lymphocytes. TEMs have been shown to uniquely express high numbers of the K _v l .3 potassium channel and depend on these channels for their function. In vivo, K _v l .3 blockers paralyze effector-memory T cells at the sites of inflammation and prevent their reactivation in inflamed tissues. In contrast, K _v l .3 blockers do not affect the homing to and motility within lymph nodes of naive and central memory T cells, most likely because these cells express the Kc _a 3.1 channel and are therefore protected from the effect ofK _v 1.3 blockade. Suppressing the function of these cells by selectively blocking the K _v l .3 channel offers the potential for highly effective therapy of autoimmune diseases with minimal effects on either beneficial immune responses or other organs (Chandy K.G. et al, Trends Pharmacol. Sci. 25(5): 280-9;

Wulff H. et al, J. Clin. Invest. 11 1 (11): 1703-13; Beeton C. et al, Proc. Natl. Acad. Sci. U.S.A., 103(46): 17414-9; Matheu M. P. et al, Immunity 29(4): 602-14).

[07] K _v l .3 has been reported to be expressed in the inner mitochondrial membrane in lymphocytes. The apoptotic protein Bax has been suggested to insert into the outer membrane of the mitochondria and occlude the pore of K _v l .3 via a lysine residue. Thus, K _v l .3 blockade may be one of many mechanisms that contribute to apoptosis (Szabo I. et al, J. Biol. Chem. 280(13): 12790-8; Szabo I. et al, Proc. Natl. Acad. Sci. U.S.A. 105(39): 14861-6)

[08] Autoimmune Disease is a family of disorders resulting from tissue damage caused by a malfunctioning immune system, affecting tens of millions of people worldwide. Such diseases may be restricted to a single organ, as e.g. in multiple sclerosis and Type I diabetes mellitus, or may involve multiple organs as in the case of rheumatoid arthritis and systemic lupus erythematosus. Treatment is generally palliative and typically includes anti-inflammatory and immunosuppressive drugs. The severe side effects of many of these therapies have fueled a continuing search for more effective and selective immunosuppressive drugs. Among these are those which can selectively inhibit the function of effector-memory T cells, known to be involved in the etiology of many of these autoimmune diseases and thereby ameliorate many autoimmune diseases without compromising the protective immune response.

[09] Multiple sclerosis is a disease caused by autoimmune damage to the central nervous system including the brain, which affects roughly two and a half million people worldwide. Symptoms include muscle weakness and paralysis, and the disease can progress rapidly and unpredictably and may eventually lead to death. Treatment usually includes the use of anti-inflammatory and immunosuppressive drugs which have potentially severe side effects. K _v l .3 has been shown to be highly expressed in autoreactive effector memory T cells from MS patients (Wulff, H et al (2003) J Clin Invest 11 1 : 1703-1713; Rus H et al (2005) PNAS 102: 11094-11099). Animal models of multiple sclerosis have been successfully treated using blockers of the K _v l .3 potassium channel. In patients with multiple sclerosis, disease-associated myelin-specific T cells from the blood are predominantly co-stimulation independent effector-memory T cells that express high numbers of K _v l .3 channels. T cells in MS lesions in postmortem brain lesions are also predominantly effector-memory T cells that express high levels of the K _v 1.3 channel (Wulff H. et al, J. Clin. Invest. 11 1(11): 1703-13; Beeton C. et al, Proc. Natl. Acad. Sci. U.S.A. 103(46): 17414-9).

[10] Type 1 diabetes mellitus is a disease caused by autoimmune destruction of insulin-producing cells in the pancreas, resulting in high blood sugar and other metabolic abnormalities. Type 1 diabetes mellitus affects close to four hundred thousand people in the US alone, and is usually diagnosed before age 20. Its long-term consequences may include blindness, nerve damage and kidney failure, and left untreated is rapidly fatal. Treatment involves life-long administration of insulin or pancreas transplantation, both of which may entail serious side effects (Beeton C. et al, Proc. Natl. Acad. Sci. U.S.A.

103(46): 17414-9).

[11] K _v l .3 is also considered a therapeutic target for the treatment of obesity, for enhancing peripheral insulin sensitivity in patients with type-2 diabetes mellitus, and for preventing bone resorption in periodontal disease (Tucker K. et al, Int. J. Obes.

(Lond) 32(8): 1222-32; Xu J. et al, Hum. Mol Genet. 12(5): 551-9; Xu J. et al, Proc. Natl. Acad. Sci. U.S.A. 101(9): 3112-7; Valverde P. et al, J. Dent. Res 84(6): 488-99;

Tschritter O. et al, J. Clin. Endocrinol. Metab. 91(2): 654-8).

[12] Compounds which are selective K _v l .3 blockers are thus potential therapeutic agents as immunosuppressants or immune system modulators including for the prevention of graft rejection, and the treatment of autoimmune and inflammatory disorders. K _v 1.3 modulators may be used alone or in conjunction with other immunosuppressants, such as selective K _ca 3.1 Blockers or cyclosporin, in order to achieve synergism and/or to reduce toxicity, especially of cyclosporin.

[13] ShK toxin has been shown to both prevent and treat experimental autoimmune encephalomyelitis in Lewis rats, an animal model for human multiple sclerosis (Beeton C. et al 2001, Proc. Natl. Acad, Sci. U.S.A. 98:13942-7), by selectively targeting T cells chronically activated by the myelin antigen, MBP (myelin basic protein). The same study also indicated that chronically activated encephalitogenic rat T cells express a unique channel phenotype characterised by high expression of K _v 1.3 channels (approximately 1500 per cell) and low numbers of Kc _a 3.1 channels (approximately 120 per cell). This channel phenotype is distinct from that seen in quiescent and acutely activated cells and may be a functionally relevant marker for chronically activated rat T- lymphocytes.

[14] There is accordingly a need for improved and specific therapies for immune diseases, including autoimmune diseases, and for immunosuppressive agents which lack problematic side effects. The compounds, compositions and methods disclosed herein help to satisfy this need.

SUMMARY OF THE INVENTION

[15] The invention provides compounds of the formula I:

or pharmaceutically acceptable salts thereof,

wherein:

E is C, CH or N;

Z is: a bond; or -C(O)-;

m is from 0 to 2;

R ¹ is: optionally substituted aryl; optionally substituted C ₃ _ ₆ cycloalkyl; or optionally substituted heteroaryl;

each R is independently; Ci-6-alkyl; Ci-6-alkoxy; halo-Ci-6-alkyl; or halo;

R ³ is: hydrogen; or Ci-6-alkyl;

R ⁴ is: Ci_ ₆ alkyl; Ci- ₆ alkoxy-Ci_ ₆ alkyl; Ci_ ₆ alkylsulfonyl-Ci_ ₆ alkyl;

hydroxy-Ci_ ₆ alkyl; amino-Ci_ ₆ alkyl; optionally substituted aryl; aryl-Ci_ ₆ alkyl wherein the aryl portion is optionally substituted; optionally substituted C ₃ _ ₆ cycloalkyl; optionally substituted heteroaryl; heteroaryl-Ci_ ₆ alkyl wherein the heteroaryl portion is optionally substituted; aryl-sulfonamido wherein the aryl portion may be optionally substituted; optionally susbstituted heterocycyl; or heterocyclyl-Ci_ ₆ alkyl wherein the heterocyclyl portion may be optionally substituted; and

R ⁵ is: Ci-6-alkyl; optionally substituted aryl; optionally substituted C ₃ _ ₆ cycloalkyl; or optionally substituted heteroaryl.

[16] The invention also provides and pharmaceutical compositions comprising the compounds, methods of using the compounds, and methods of preparing the compounds. DETAILED DESCRIPTION OF THE INVENTION

Definitions

[17] Unless otherwise stated, the following terms used in this Application, including the specification and claims, have the definitions given below. It must be noted that, as used in the specification and the appended claims, the singular forms "a", "an," and "the" include plural referents unless the context clearly dictates otherwise.

[18] "Alkyl" means the monovalent linear or branched saturated hydrocarbon moiety, consisting solely of carbon and hydrogen atoms, having from one to twelve carbon atoms. "Lower alkyl" refers to an alkyl group of one to six carbon atoms, i.e. d- Cealkyl. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, sec-butyl, tert-butyl, pentyl, n-hexyl, octyl, dodecyl, and the like.

[19] "Alkenyl" means a linear monovalent hydrocarbon radical of two to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms, containing at least one double bond, e.g., ethenyl, propenyl, and the like.

[20] "Alkynyl" means a linear monovalent hydrocarbon radical of two to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms, containing at least one triple bond, e.g., ethynyl, propynyl, and the like.

[21] "Alkylene" means a linear saturated divalent hydrocarbon radical of one to six carbon atoms or a branched saturated divalent hydrocarbon radical of three to six carbon atoms, e.g., methylene, ethylene, 2,2-dimethylethylene, propylene, 2- methylpropylene, butylene, pentylene, and the like.

[22] "Alkoxy" and "alkyloxy", which may be used interchangeably, mean a moiety of the formula -OR, wherein R is an alkyl moiety as defined herein. Examples of alkoxy moieties include, but are not limited to, methoxy, ethoxy, isopropoxy, and the like.

[23] "Alkoxyalkyl" means a moiety of the formula R ^a -0-R ^b -, where R ^a is alkyl and R ^b is alkylene as defined herein. Exemplary alkoxyalkyl groups include, by way of example, 2 -methoxy ethyl, 3-methoxypropyl, l-methyl-2-methoxyethyl, l -(2- methoxyethyl)-3-methoxypropyl, and 1 -(2-methoxyethyl)-3-methoxypropyl.

[24] "Alkoxyalkoxy' means a group of the formula -O-R-R' wherein R is alkylene and R' is alkoxy as defined herein. [25] "Alkylcarbonyl" means a moiety of the formula -C(0)-R, wherein R is alkyl as defined herein.

[26] "Alkoxycarbonyl" means a group of the formula -C(0)-R wherein R is alkoxy as defined herein.

[27] "Alkylcarbonylalkyl" means a group of the formula -R-C(0)-R' wherein R is alkylene and R' is alkyl as defined herein.

"Alkoxycarbonylalkyl" means a group of the formula -R-C(0)-R' wherein R is alkylene and R' is alkoxy as defined herein.

[28] "Alkoxycarbonylalkoxy"means a group of the formula -0-R-C(0)-R' wherein R is alkylene and R' is alkoxy as defined herein.

[29] "Hydroxycarbonylalkoxy" means a group of the formula -0-R-C(0)-OH wherein R is alkylene as defined herein.

[30] "Alkylaminocarbonylalkoxy" means a group of the formula -O-R-C(O)-

NHR' wherein R is alkylene and R' is alkyl as defined herein.

[31] "Dialkylaminocarbonylalkoxy" means a group of the formula -O-R-C(O)-

NR'R" wherein R is alkylene and R' and R" are alkyl as defined herein.

[32] "Alkylaminoalkoxy" means a group of the formula -O-R-NHR' wherein R is alkylene and R' is alkyl as defined herein.

[33] "Dialkylaminoalkoxy" means a group of the formula -O-R-NR'R' wherein R is alkylene and R' and R" are alkyl as defined herein.

[34] "Alkylsulfonyl" means a moiety of the formula - S0 ₂ -R, wherein R is alkyl as defined herein.

[35] "Alkylsulfonylalkyl means a moiety of the formula -R'-S0 ₂ -R" where where R' is alkylene and R" is alkyl as defined herein.

[36] "Alkylsulfonylalkoxy" means a group of the formula -0-R-S0 ₂ -R' wherein R is alkylene and R' is alkyl as defined herein.

[37] "Amino" means a moiety of the formula -NRR wherein R and R' each independently is hydogen or alkyl as defined herein, or one of R and R' may be tert- butoxycarbonyl or other protecting group. "Amino thus includes "alkylamino (where one of R and R' is alkyl and the other is hydrogen) and "dialkylamino (where R and R' are both alkyl. [38] "Aminocarbonyl" means a group of the formula -C(0)-R wherein R is amino as defined herein.

[39] "Alkoxyamino" means a moiety of the formula -NR-OR' wherein R is hydrogen or alkyl and R' is alkyl as defined herein.

[40] "Alkylsulfanyl" means a moiety of the formula -SR wherein R is alkyl as defined herein.

[41] "Aminoalkyl" means a group -R-R' wherein R is amino and R is alkylene as defined herein. "Aminoalkyl" includes aminomethyl, aminoethyl, 1 -aminopropyl, 2- aminopropyl, and the like. The amino moiety of "aminoalkyl" may be substituted once or twice with alkyl to provide "alkylaminoalkyl" and "dialkylaminoalkyl" respectively.

"Alkylaminoalkyl" includes methylaminomethyl, methylaminoethyl, methylaminopropyl, ethylaminoethyl and the like. "Dialkylaminoalkyl" includes dimethylaminomethyl, dimethylaminoethyl, dimethylaminopropyl, N-methyl-N-ethylaminoethyl, and the like.

[42] "Aminoalkoxy" means a group -OR-R' wherein R is amino and R is alkylene as defined herein.

[43] "Alkylsulfonylamido" means a moiety of the formula -NRSO ₂ -R wherein

R is alkyl and R' is hydrogen or alkyl.

[44] "Aminocarbonyloxyalkyl" or "carbamylalkyl" means a group of the formula -R-0-C(0)-NR'R" wherein R is alkylene and R, R" each independently is hydrogen or alkyl as defined herein.

[45] "Alkynylalkoxy" means a group of the formula -O-R-R' wherein R is alkylene and R is alkynyl as defined herein.

[46] "Aryl" means a monovalent cyclic aromatic hydrocarbon moiety consisting of a mono-, bi- or tricyclic aromatic ring. The aryl group can be optionally substituted as defined herein. Examples of aryl moieties include, but are not limited to, phenyl, naphthyl, phenanthryl, fluorenyl, indenyl, pentalenyl, azulenyl, oxydiphenyl, biphenyl, methylenediphenyl, aminodiphenyl, diphenylsulfidyl, diphenylsulfonyl, diphenylisopropylidenyl, benzodioxanyl, benzofuranyl, benzodioxylyl, benzopyranyl, benzoxazinyl, benzoxazinonyl, benzopiperadinyl, benzopiperazinyl, benzopyrrolidinyl, benzomorpholinyl, methylenedioxyphenyl, ethylenedioxyphenyl, and the like, including partially hydrogenated derivatives thereof, each being optionally substituted. [47] "Arylalkyl" and "Aralkyl", which may be used interchangeably, mean a radical-R ^a R ^b where R ^a is an alkylene group and R ^b is an aryl group as defined herein; e.g., phenylalkyls such as benzyl, phenylethyl, 3-(3-chlorophenyl)-2-methylpentyl, and the like are examples of arylalkyl.

[48] "Arylsulfonyl means a group of the formula -SO2-R wherein R is aryl as defined herein.

[49] "Aryloxy" means a group of the formula -O-R wherein R is aryl as defined herein.

[50] "Aralkyloxy" means a group of the formula -O-R-R" wherein R is alkylene and R is aryl as defined herein.

[51] "Carboxy" means a group of the formula -C(0)-OR wherein R is hydrogen or alkyl as defined herein.

[52] "Cyanoalkyl" " means a moiety of the formula -R'-R", where R' is alkylene as defined herein and R" is cyano or nitrile.

[53] "Cycloalkyl" means a monovalent saturated carbocyclic moiety consisting of mono- or bicyclic rings. Preferred cycloalkyl are unsubstituted or substituted with alkyl. Cycloalkyl can optionally be substituted with one or more substituents, wherein each substituent is independently hydroxy, alkyl, alkoxy, halo, haloalkyl, amino, monoalkylamino, or dialkylamino, unless otherwise specifically indicated. Examples of cycloalkyl moieties include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like, including partially unsaturated (cycloalkenyl) derivatives thereof.

[54] "Cycloalkylalkyl" means a moiety of the formula -R'-R", where R' is alkylene and R" is cycloalkyl as defined herein.

[55] "Cycloalkylalkoxy" means a group of the formula -O-R-R' wherein R is alkylene and R' is cycloalkyl as defined herein.

[56] "Heteroaryl" means a monocyclic or bicyclic radical of 5 to 12 ring atoms having at least one aromatic ring containing one, two, or three ring heteroatoms selected from N, O, or S, the remaining ring atoms being C, with the understanding that the attachment point of the heteroaryl radical will be on an aromatic ring. The heteroaryl ring may be optionally substituted as defined herein. Examples of heteroaryl moieties include, but are not limited to, optionally substituted imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, pyrazinyl, thienyl, benzothienyl, thiophenyl, furanyl, pyranyl, pyridyl, pyrrolyl, pyrazolyl, pyrimidyl, quinolinyl, isoquinolinyl, benzofuryl, benzothiophenyl, benzothiopyranyl, benzimidazolyl, benzooxazolyl, benzooxadiazolyl, benzothiazolyl, benzothiadiazolyl, benzopyranyl, indolyl, isoindolyl, triazolyl, triazinyl, quinoxalinyl, purinyl, quinazolinyl, quinolizinyl, naphthyridinyl, pteridinyl, carbazolyl, azepinyl, diazepinyl, acridinyl and the like, including partially hydrogenated derivatives thereof, each optionally substituted.

[57] Heteroarylalkyl" or "heteroaralkyl" means a group of the formula -R-R' wherein R is alkylene and R' is heteroaryl as defined herein.

[58] "Heteroarylsulfonyl means a group of the formula -S0 ₂ -R wherein R is heteroaryl as defined herein.

[59] "Heteroaryloxy" means a group of the formula -O-R wherein R is heteroaryl as defined herein.

[60] "Heteroaralkyloxy" means a group of the formula -O-R-R' wherein R is alkylene and R' is heteroaryl as defined herein.

[61] The terms "halo", "halogen" and "halide", which may be used

interchangeably, refer to a substituent fiuoro, chloro, bromo, or iodo.

[62] "Haloalkyl" means alkyl as defined herein in which one or more hydrogen has been replaced with same or different halogen. Exemplary haloalkyls include -CH ₂ C1, -CH ₂ CF ₃ , -CH ₂ CCI ₃ , perfiuoroalkyl (e.g., -CF ₃ ), and the like.

[63] "Haloalkoxy" means a moiety of the formula -OR, wherein R is a haloalkyl moiety as defined herein. An exemplary haloalkoxy is difiuoromethoxy.

[64] "Heterocycloamino" means a saturated ring wherein at least one ring atom is N, NH or N-alkyl and the remaining ring atoms form an alkylene group.

[65] "Heterocyclyl" means a monovalent saturated moiety, consisting of one to three rings, incorporating one, two, or three or four heteroatoms (chosen from nitrogen, oxygen or sulfur). The heterocyclyl ring may be optionally substituted as defined herein. Examples of heterocyclyl moieties include, but are not limited to, optionally substituted piperidinyl, piperazinyl, homopiperazinyl, azepinyl, pyrrolidinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyridinyl, pyridazinyl, pyrimidinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinuclidinyl, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazolylidinyl, benzothiazolidinyl, benzoazolylidinyl, dihydrofuryl, tetrahydrofuryl, dihydropyranyl, tetrahydropyranyl, thiamorpholinyl, thiamorpholinylsulfoxide, thiamorpholinylsulfone, dihydroquinolinyl,

dihydrisoquinolinyl, tetrahydroquinolinyl, tetrahydrisoquinolinyl, and the like.

[66] "Heterocyclylalkyl" means a moiety of the formula -R-R' wherein R is alkylene and R is heterocyclyl as defined herein.

[67] "Heterocyclyloxy" means a moiety of the formula -OR wherein R is heterocyclyl as defined herein.

[68] "Heterocyclylalkoxy" means a moiety of the formula -OR-R wherein R is alkylene and R' is heterocyclyl as defined herein.

[69] "Hydroxyalkoxy" means a moiety of the formula -OR wherein R is hydroxyalkyl as defined herein.

[70] "Hydroxyalkylamino" means a moiety of the formula -NR-R' wherein R is hydrogen or alkyl and R' is hydroxyalkyl as defined herein.

[71] "Hydroxyalkylaminoalkyl" means a moiety of the formula -R-NR'-R" wherein R is alkylene, R is hydrogen or alkyl, and R" is hydroxyalkyl as defined herein.

[72] "Hydroxycarbonylalkyl" or "carboxyalkyl" means a group of the formula -

R-(CO)-OH where R is alkylene as defined herein.

[73] "Hydroxycarbonylalkoxy" means a group of the formula -0-R-C(0)-OH wherein R is alkylene as defined herein.

[74] "Hydroxyalkyloxycarbonylalkyl" or "hydroxyalkoxycarbonylalkyl" means a group of the formula -R-C(0)-0-R-OH wherein each R is alkylene and may be the same or different.

[75] "Hydroxyalkyl" means an alkyl moiety as defined herein, substituted with one or more, preferably one, two or three hydroxy groups, provided that the same carbon atom does not carry more than one hydroxy group. Representative examples include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, l-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2,3-dihydroxypropyl, 2 -hydroxy- 1 -hydroxymethylethyl, 2,3-dihydroxybutyl,

3,4-dihydroxybutyl and 2-(hydroxymethyl)-3-hydroxypropyl [76] "Hydroxycycloalkyl" means a cycloalkyl moiety as defined herein wherein one, two or three hydrogen atoms in the cycloalkyl radical have been replaced with a hydroxy substituent. Representative examples include, but are not limited to, 2-, 3-, or 4-hydroxycyclohexyl, and the like.

[77] "Alkoxy hydroxyalkyl" and "hydroxy alkoxyalkyl", which may be used interchangeably, means an alkyl as defined herein that is substituted at least once with hydroxy and at least once with alkoxy. "Alkoxy hydroxyalkyl" and "hydroxy

alkoxyalkyl" thus encompass, for example, 2-hydroxy-3-methoxy-propan-l -yl and the like.

[78] "Urea"or "ureido" means a group of the formula -NR'-C(0)-NR"R ^m wherein R, R" and R'" each independently is hydrogen or alkyl.

[79] "Carbamate" means a group of the formula -0-C(0)-NR'R" wherein R and R" each independently is hydrogen or alkyl.

[80] "Carboxy" means a group of the formula -0-C(0)-OH.

[81] "Sulfonamido" means a group of the formula -S0 ₂ -NR'R" or -NR'S0 ₂ - wherein R and R" each independently is hydrogen or alkyl.

[82] "Optionally substituted", when used in association with "aryl", phenyl",

"heteroaryl" "cycloalkyl" or "heterocyclyl", means an aryl, phenyl, heteroaryl, cycloalkyl or heterocyclyl which is optionally substituted independently with one to four substituents, preferably one or two substituents selected from alkyl, cycloalkyl, aryl, cycloalkylalkyl, heteroalkyl, hydroxyalkyl, halo, nitro, cyano, hydroxy, alkoxy, amino, acylamino, mono-alkylamino, di-alkylamino, haloalkyl, haloalkoxy, -COR, -S0 ₂ R (where R is hydrogen, alkyl, phenyl or phenylalkyl), -(CR'R") _n -COOR (where n is an integer from 0 to 5, R' and R" are independently hydrogen or alkyl, and R is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl), or -(CR'R") _n -CONR ^a R ^b

(where n is an integer from 0 to 5, R' and R" are independently hydrogen or alkyl, and R ^a and R ^b are, independently of each other, hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl). Certain preferred optional substituents for "aryl", phenyl", "heteroaryl" "cycloalkyl" or 'Tieterocyclyl" include alkyl, halo, haloalkyl, alkoxy, cyano, amino and alkylsulfonyl. More preferred substituents are methyl, fluoro, chloro, trifiuoromethyl, methoxy, amino and methanesulfonyl.

[83] "Leaving group" means the group with the meaning conventionally associated with it in synthetic organic chemistry, i.e., an atom or group displaceable under substitution reaction conditions. Examples of leaving groups include, but are not limited to, halogen, alkane- or arylenesulfonyloxy, such as methanesulfonyloxy, ethanesulfonyloxy, thiomethyl, benzenesulfonyloxy, tosyloxy, and thienyloxy, dihalophosphinoyloxy, optionally substituted benzyloxy, isopropyloxy, acyloxy, and the like.

[84] "Modulator" means a molecule that interacts with a target. The interactions include, but are not limited to, agonist, antagonist, and the like, as defined herein.

[85] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not.

[86] "Disease" and "Disease state" means any disease, condition, symptom, disorder or indication.

[87] "Inert organic solvent" or "inert solvent" means the solvent is inert under the conditions of the reaction being described in conjunction therewith, including for example, benzene, toluene, acetonitrile, tetrahydrofuran, N,N-dimethylformamide, chloroform, methylene chloride or dichloromethane, dichloro ethane, diethyl ether, ethyl acetate, acetone, methyl ethyl ketone, methanol, ethanol, propanol, isopropanol, tert- butanol, dioxane, pyridine, and the like. Unless specified to the contrary, the solvents used in the reactions of the present invention are inert solvents.

[88] "Pharmaceutically acceptable" means that which is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable and includes that which is acceptable for veterinary as well as human pharmaceutical use.

[89] "Pharmaceutically acceptable salts" of a compound means salts that are pharmaceutically acceptable, as defined herein, and that possess the desired

pharmacological activity of the parent compound. Such salts include: acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or formed with organic acids such as acetic acid, benzenesulfonic acid, benzoic, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydroxynaphtoic acid, 2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, 2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinic acid, tartaric acid, p- toluenesulfonic acid, trimethylacetic acid, and the like; or

salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic or inorganic base. Acceptable organic bases include diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, tromethamine, and the like. Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and sodium hydroxide.

[90] The preferred pharmaceutically acceptable salts are the salts formed from acetic acid, hydrochloric acid, sulphuric acid, methanesulfonic acid, maleic acid, phosphoric acid, tartaric acid, citric acid, sodium, potassium, calcium, zinc, and magnesium.

[91] It should be understood that all references to pharmaceutically acceptable salts include solvent addition forms (solvates) or crystal forms (polymorphs) as defined herein, of the same acid addition salt.

[92] "Protective group" or "protecting group" means the group which selectively blocks one reactive site in a multifunctional compound such that a chemical reaction can be carried out selectively at another unprotected reactive site in the meaning conventionally associated with it in synthetic chemistry. Certain processes of this invention rely upon the protective groups to block reactive nitrogen and/or oxygen atoms present in the reactants. For example, the terms "amino-protecting group" and "nitrogen protecting group" are used interchangeably herein and refer to those organic groups intended to protect the nitrogen atom against undesirable reactions during synthetic procedures. Exemplary nitrogen protecting groups include, but are not limited to, trifluoro acetyl, acetamido, benzyl (Bn), benzyloxycarbonyl (carbobenzyloxy, CBZ), p- methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, iert-butoxycarbonyl (BOC), and the like. The artisan in the art will know how to chose a group for the ease of removal and for the ability to withstand the following reactions.

[93] "Solvates" means solvent additions forms that contain either

stoichiometric or non stoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate, when the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one of the substances in which the water retains its molecular state as H ₂ 0, such combination being able to form one or more hydrate.

[94] "Subject" means mammals and non-mammals. Mammals means any member of the mammalia class including, but not limited to, humans; non-human primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, and swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice, and guinea pigs; and the like. Examples of non-mammals include, but are not limited to, birds, and the like. The term "subject" does not denote a particular age or sex.

[95] "Arthritis" means diseases or conditions damage to joints of the body and pain associated with such joint damage. Arithritis includes rheumatoid arthritis, osteoarthritis, psoriatic arthritis, septic arthritis and gouty arthritis.

[96] "Pain" includes, without limitation, inflammatory pain; surgical pain; visceral pain; dental pain; premenstrual pain; central pain; pain due to burns; migraine or cluster headaches; nerve injury; neuritis; neuralgias; poisoning; ischemic injury;

interstitial cystitis; cancer pain; viral, parasitic or bacterial infection; post-traumatic injury; or pain associated with irritable bowel syndrome.

[97] "Therapeutically effective amount" means an amount of a compound that, when administered to a subject for treating a disease state, is sufficient to effect such treatment for the disease state. The "therapeutically effective amount" will vary depending on the compound, disease state being treated, the severity or the disease treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending medical or veterinary practitioner, and other factors. [98] The terms "those defined above" and "those defined herein" when referring to a variable incorporates by reference the broad definition of the variable as well as preferred, more preferred and most preferred definitions, if any.

[99] "Treating" or "treatment" of a disease state includes:

(i) preventing the disease state, i.e. causing the clinical symptoms of the disease state not to develop in a subject that may be exposed to or predisposed to the disease state, but does not yet experience or display symptoms of the disease state.

(ii) inhibiting the disease state, i.e., arresting the development of the disease state or its clinical symptoms, or

(iii) relieving the disease state , i.e., causing temporary or permanent regression of the disease state or its clinical symptoms.

[100] The terms "treating", "contacting" and "reacting" when referring to a chemical reaction means adding or mixing two or more reagents under appropriate conditions to produce the indicated and/or the desired product. It should be appreciated that the reaction which produces the indicated and/or the desired product may not necessarily result directly from the combination of two reagents which were initially added, i.e., there may be one or more intermediates which are produced in the mixture which ultimately leads to the formation of the indicated and/or the desired product. Nomenclature and Structures

[101] In general, the nomenclature used in this Application is based on

AUTONOM™ v.4.0, a Beilstein Institute computerized system for the generation of IUPAC systematic nomenclature. Chemical structures shown herein were prepared using ISIS ^® version 2.2. Any open valency appearing on a carbon, oxygen sulfur or nitrogen atom in the structures herein indicates the presence of a hydrogen atom unless indicated otherwise. Where a chiral center exists in a structure but no specific stereochemistry is shown for the chiral center, both enantiomers associated with the chiral center are encompassed by the structure. Where a structure shown herein may exist in multiple tautomeric forms, all such tautomers are encompassed by the structure. The atoms represented in the structures herein are intended to encompass all naturally occurring isotopes of such atoms. Thus, for example, the hydrogen atoms represented herein are meant to include deuterium and tritium, and the carbon atoms are meant to include C and C ¹⁴ isotopes.

[102] All patents and publications identified herein are incorporated herein by reference in their entirety.

Compounds of the Invention

[103] Preferably, the invention provides compounds of the formula I:

or pharmaceutically acceptable salts thereof,

wherein:

E is C or ;

Z is: a bond; or -C(O)-;

m is from 0 to 2;

R ¹ is: optionally substituted aryl; optionally substituted C ₃ - ₆ cycloalkyl; or optionally substituted heteroaryl;

each R ² is independently; Ci_ ₆ -alkyl; Ci- ₆ -alkoxy; halo-Ci- ₆ -alkyl; or halo;

R is: hydrogen; or Ci- ₆ -alkyl;

R ⁴ is: Ci_ ₆ alkyl; Ci_ ₆ alkoxy-Ci_ ₆ alkyl; Ci_ ₆ alkylsulfonyl-Ci_ ₆ alkyl; hydroxy-Ci-

₆ alkyl; amino-Ci_ ₆ alkyl; optionally substituted aryl; aryl-Ci_ ₆ alkyl wherein the aryl portion is optionally substituted; optionally substituted C ₃ _ ₆ cycloalkyl; optionally substituted heteroaryl; heteroaryl-Ci_ ₆ alkyl wherein the heteroaryl portion is optionally substituted; aryl-sulfonamido wherein the aryl portion may be optionally substituted; optionally susbstituted heterocycyl; or heterocyclyl-Ci_ ₆ alkyl wherein the heterocyclyl portion may be optionally substituted; and

R ⁵ is: Ci- ₆ -alkyl; optionally substituted aryl; optionally substituted C ₃ _ ₆ cycloalkyl; or optionally substituted heteroaryl.

[104] In certain embodiments of formula I, Z is a bond.

[105] In certain embodiments of formula I, Z is -C(O)-. [106] In certain embodiments of formula I, E is C.

[107] In certain embodiments of formula I, E is CH.

[108] In certain embodiments of formula I, E is N.

[109] In certain embodiments of formula I, m is 0 or 1.

[110] In certain embodiments of formula I, m is 0.

[111] In certain embodiments of formula I, m is 1.

[112] In certain embodiments of formula I, R ¹ is optionally substituted aryl.

[113] In certain embodiments of formula I, R ¹ is optionally substituted phenyl.

[114] In certain embodiments of formula I, R ¹ is phenyl optionally substituted one, two or three times with a group or groups independently selected from: Ci_ ₆ alkyl; Ci_ ₆ alkoxy; halo-Ci_ ₆ alkyl; halo-Ci_ ₆ alkoxy; halo; carboxy; and cyano.

[115] In certain embodiments of formula I, R ¹ is phenyl optionally substituted once or twice with a group or groups independently selected from: Ci_ ₆ alkyl; Ci_ ₆ alkoxy; halo-Ci- ₆ alkyl; halo; carboxy; and cyano.

[116] In certain embodiments of formula I, R ¹ is phenyl optionally substituted once with Ci_ ₆ alkyl; Ci_ ₆ alkoxy; halo; or halo-Ci_ ₆ alkyl.

[117] In certain embodiments of formula I, R ¹ is phenyl optionally substituted once with methyl, methoxy, fluoro, chloro or trifluoromethyl.

[118] In certain embodiments of formula I, R ¹ is phenyl optionally substituted once with Ci_ ₆ alkyl or halo.

[119] In certain embodiments of formula I, R ¹ is phenyl optionally substituted with fluoro.

[120] In certain embodiments of formula I, R ¹ is unsubstituted phenyl.

[121] In certain embodiments of formula I, R ¹ is optionally substituted heteroaryl.

[122] In certain embodiments of formula I, R ¹ is heteroaryl selected from:

pyridinyl, pyrimidinyl, pyrazinyl, furanyl; pyridonyl, benzimidazolyl, thiazolyl;

isothiazolyl; tetrahydroisoquinolinyl; benzothiazolyl, pyrazolyl; imidazo[l ,2- ]pyridinyl, oxazolyl; isoxazolyl; or imidazolyl, each of which may be optionally substituted.

[123] In certain embodiments of formula I, R ¹ is pyridinyl. [124] In certain embodiments of formula I, R ¹ is C ₃ _ ₆ cycloalkyl optionally substituted with Ci- ₆ alkyl.

[125] In certain embodiments of formula I, R ¹ is cyclohexyl.

[126] In certain embodiments of formula I, R ² is hydrogen.

[127] In certain embodiments of formula I, each R is independently methyl, methoxy, trifiuoromethyl, fiuoro or chloro.

[128] In certain embodiments of formula I, R ³ is hydrogen.

[129] In certain embodiments of formula I, R ³ is Ci_ ₆ alkyl.

[130] In certain embodiments of formula I, R ³ is hydrogen or methyl.

[131] In certain embodiments of formula I, R ³ is methyl.

[132] In certain embodiments of formula I, R ⁴ is:

Ci- ₆ alkyl;

C i_ ₆ alkoxy-C i - ₆ alkyl;

Ci- ₆ alkylsulfonyl-Ci- ₆ alkyl;

hydro xy-Ci_ ₆ alkyl; amino-Ci_ ₆ alkyl;

optionally substituted phenyl;

phenyl-Ci_ ₆ alkyl wherein the phenyl portion is optionally substituted;

C ₃ _ ₆ cycloalkyl optionally substituted with Ci_ ₆ alkyl or phenyl;

heteroaryl selected from pyridinyl; pyrimidinyl; pyrazinyl; furanyl; pyridonyl; benzimidazolyl; thiazolyl; isothiazolyl; tetrahydroisoquinolinyl; benzothiazolyl, pyrazolyl; imidazo[ l ,2- ]pyridinyl; oxazolyl; isoxazolyl; thiophenyl; imidazo[2,l- 0]thiazolyl; or imidazolyl, each of which may be optionally substituted;

heteroaryl-Ci- ₆ alkyl wherein the heteroaryl portion is selected from: pyridinyl; pyrimidinyl; pyrazinyl; furanyl; pyridonyl; benzimidazolyl; thiazolyl; isothiazolyl;

tetrahydroisoquinolinyl; benzothiazolyl, pyrazolyl; imidazo[l ,2- ]pyridinyl; oxazolyl; isoxazolyl; thiophenyl; imidazo[2,l -0]thiazolyl; or imidazolyl, each of which may be optionally substituted;

phenyl-sulfonamido wherein the phenyl portion may be optionally substituted; heterocycyl selected from: tetrahydropyranyl; morpholinyl; oxetanyl; piperidinyl; piperazinyl; pyrrolidinyl; thiazolidinyl; or azetidinyl, each optionally substituted with Ci_ ₆ alkyl or carboxy; or heterocyclyl-Ci-6alkyl wherein the heterocyclyl portion is selected from:

tetrahydropyranyl; morpholinyl; oxetanyl; piperidinyl; piperazinyl; pyrrolidinyl;

thiazolidinyl; or azetidinyl, each optionally substituted with Ci_6alkyl or carboxy.

[133] In certain embodiments of formula I, R ⁴ is:

Ci-6alkyl;

C i_6alkoxy-C i -6alkyl;

Ci-6alkylsulfonyl-Ci-6alkyl;

hydro xy-Ci_6alkyl; amino-Ci_6alkyl;

phenyl optionally substituted one, two or three times with a group or groups independently selected from: Ci_6alkyl; Ci_6alkoxy; halo-Ci_6alkyl; halo; amino; carboxy; sulfonamido; and cyano.

phenyl-Ci-6alkyl wherein the phenyl portion is optionally substituted one, two or three times with a group or groups independently selected from: Ci_6alkyl; Ci_6alkoxy; halo-Ci-6alkyl; halo; carboxy; amino; and cyano.

C3_6cycloalkyl optionally substituted with Ci_6alkyl or phenyl;

heteroaryl selected from pyridinyl; pyrimidinyl; pyrazinyl; furanyl; pyridonyl; benzimidazolyl; thiazolyl; isothiazolyl; tetrahydroisoquinolinyl; benzothiazolyl, pyrazolyl; imidazo[ l ,2- ]pyridinyl; oxazolyl; isoxazolyl; thiophenyl; or imidazolyl, each of which may be optionally substituted once or twice with a group or groups

independently selected from: Ci_6alkyl; Ci_6alkoxy; halo-Ci_6alkyl; halo; oxo; and cyano. heteroaryl-Ci-6alkyl wherein the heteroaryl portion is selected from: pyridinyl; pyrimidinyl; pyrazinyl; furanyl; pyridonyl; benzimidazolyl; thiazolyl; isothiazolyl;

tetrahydroisoquinolinyl; benzothiazolyl, pyrazolyl; imidazo[ l ,2- ]pyridinyl; oxazolyl; isoxazolyl; thiophenyl; or imidazolyl, each of which may be optionally substituted once or twice with a group or groups independently selected from: Ci-6alkyl; Ci-6alkoxy; halo- Ci_6alkyl; halo; oxo; and cyano;

phenyl-sulfonamido wherein the phenyl portion may be optionally substituted one, two or three times with a group or groups independently selected from: Ci_6alkyl; Ci_ 6alkoxy; halo-Ci_6alkyl; halo; and cyano; heterocycyl selected from: tetrahydropyranyl; morpholinyl; oxetanyl; piperidinyl; pyrrollidinyl; thiazolidinyl; or azetidinyl, each optionally substituted with Ci-6alkyl or carboxy; or

heterocyclyl-Ci_6alkyl wherein the heterocyclyl portion is selected from:

tetrahydropyranyl; morpholinyl; oxetanyl; piperidinyl; pyrrolidinyl; thiazolidinyl; or azetidinyl, each optionally substituted with Ci_6alkyl or carboxy.

[134] In certain embodiments of formula I, R ⁴ is:

Ci_ ₆ alkyl;

C i_6alkoxy-C i -6alkyl;

Ci-6alkylsulfonyl-Ci-6alkyl;

hydro xy-Ci_6alkyl; amino-Ci_6alkyl;

phenyl optionally substituted once or with a group or groups independently selected from: Ci_6alkyl; Ci_6alkoxy; halo-Ci_6alkyl; halo; carboxy; amino; and sulfonamido.

phenyl-Ci_6alkyl wherein the phenyl portion is optionally substituted one, two or three times with a group or groups independently selected from: Ci_6alkyl; Ci_6alkoxy; halo-Ci-6alkyl; halo; amino; and carboxy;

C3_6cycloalkyl optionally substituted with Ci_6alkyl or phenyl;

heteroaryl-Ci-6alkyl wherein the heteroaryl portion is selected from: pyridinyl; pyrimidinyl; pyrazinyl; furanyl; pyridonyl; benzimidazolyl; thiazolyl; isothiazolyl;

tetrahydroisoquinolinyl; benzothiazolyl, pyrazolyl; imidazo[l ,2- ]pyridinyl; oxazolyl; isoxazolyl; thiophenyl; or imidazolyl, each of which may be optionally substituted once or twice with a group or groups independently selected from: Ci_6alkyl; Ci_6alkoxy; halo- Ci_6alkyl; oxo; and halo;

phenyl-sulfonamido wherein the phenyl portion may be optionally substituted one, two or three times with a group or groups independently selected from: Ci_6alkyl; Ci_ 6alkoxy; halo-Ci_6alkyl; halo; and carboxy;

heterocycyl selected from: tetrahydropyranyl; oxetanyl; pyrrolidinyl; and azetidinyl, each optionally substituted with Ci_6alkyl or carboxy; or heterocyclyl-Ci- ₆ alkyl wherein the heterocyclyl portion is selected from:

piperidinyl; pyrrolidinyl; morpholinyl; thiazolidinyl; or azetidinyl, each optionally substituted with Ci_6alkyl or carboxy.

[135] In certain embodiments of formula I, R ⁴ is Ci_ ₆ alkyl.

[136] In ermbodiments of formula I wherein R ⁴ is R ⁴ is Ci-6alkyl, such Ci-6alkyl may be propyl, isopropyl, butyl, isobutyl or tert-butyl.

[137] In certain embodiments of formula I, R ⁴ is Ci- ₆ alkoxy-Ci_ ₆ alkyl.

[138] In embodiments of formula I wherein R ⁴ is Ci_ ₆ alkoxy-Ci_ ₆ alkyl, such Ci_

₆ alkoxy-Ci_ ₆ alkyl may be 2-methoxy-ethyl, 2 -methoxy-1 -methyl-ethyl, 3-methoxy-propyl, 2-ethoxy-ethyl, or 2-methoxy-propyl.

[139] In certain embodiments of formula I, R ⁴ is Ci_ ₆ alkylsulfonyl-Ci_ ₆ alkyl;

[140] In embodiments of formula I wherein R ⁴ is Ci-6alkylsulfonyl-Ci-6alkyl, such Ci- ₆ alkylsulfonyl-Ci- ₆ alkyl may be 2-(methanesulfonyl)-ethyl, 2-(methanesulfonyl)-

1 -methyl-ethyl or 3-(methanesulfonyl)-propyl.

[141] In certain embodiments of formula I, R ⁴ is hydroxy-Ci- ₆ alkyl.

[142] In embodiments of formula I wherein R ⁴ is hydroxy-Ci_ ₆ alkyl, such hydroxy-Ci- ₆ alkyl may be 2-hydroxy-ethyl, 2-hydroxy-propyl, 2 -hydroxy- 1 -methyl-ethyl or 2-hydroxy-2-methyl-propyl.

[143] In certain embodiments of formula I, R ⁴ is optionally substituted phenyl.

[144] In embodiments of formula I wherein R ⁴ is optionally substituted phenyl, such optionally substituted phenyl may be unsubstituted phenyl, 2-chloro-phenyl, 3- chloro-phenyl, 4-chloro-phenyl, 2-methyl-phenyl, 3-trifluoromethyl-phenyl, 4- trifluoromethyl-phenyl, 2-fluoro-phenyl, 3 -fluoro -phenyl, 4-fluoro-phenyl, 2,4-difluoro- phenyl, 2-methoxy-phenyl, 3-methoxy-phenyl, 4-methoxy-phenyl, 3-carboxy-phenyl, or 4-amino-phenyl.

[145] In certain embodiments of formula I, R ⁴ is phenyl-Ci_ ₆ alkyl wherein the phenyl portion is optionally substituted.

[146] In embodiments of formula I wherein R ⁴ is phenyl-Ci_ ₆ alkyl, the phenyl portion may be unsubstituted phenyl, 2-chloro-phenyl, 3-chloro-phenyl, 4-chloro-phenyl, 2-methyl-phenyl, 3-trifluoromethyl-phenyl, 4-trifluoromethyl-phenyl, 2-fluoro-phenyl, 3- fluoro -phenyl, 4-fluoro-phenyl, 2,4-difluoro-phenyl, 2-methoxy-phenyl, 3-methoxy- phenyl, 4-methoxy-phenyl, 3-carboxy-phenyl, or 4-amino-phenyl.

[147] In embodiments of formula I wherein R ⁴ is phenyl-Ci_6alkyl, the Ci_6alkyl portion may be methylene, ethylene, propylene or butylene.

[148] In embodiments of formula I wherein R ⁴ is phenyl-Ci-6alkyl, the Ci-6alkyl portion may be methylene or ethylene.

[149] In embodiments of formula I wherein R ⁴ is phenyl-Ci_6alkyl, the Ci_6alkyl portion may be methylene.

[150] In certain embodiments of formula I, R ⁴ is C3_6cycloalkyl optionally substituted with Ci_6alkyl or phenyl.

[151] In embodiments of formula I wherein R ⁴ is C3_ ₆ cycloalkyl, such C ₃ _

6cycloalkyl may be 2-phenyl-cyclopropyl or 3-phenyl-cyclobutyl.

[152] In certain embodiments of formula I, R ⁴ is optionally substituted heteroaryl.

[153] In embodiments of formula I wherein R ⁴ is optionally substituted heteroaryl, such optionally substituted heteroaryl may be pyrazin-2-yl, 5-methyl-pyrazin-

2- yl, pyrimidin-5-yl, pyridin-2-yl, 6-methyl-pyridin-2-yl, 6-chloro-pyridin-3-yl, pyridin-

3- yl, pyridin-4-yl, furan-2-yl, 6-oxo-pyridin-3-yl, benzimidazol-2-yl, thiazol-2-yl, benzothiazol-2-yl, pyrazol-4-yl, imidazo[l ,2-a]pyridin-3-yl, isoxazol-5-yl, imidazol-4-yl, imidazol-l -yl, pyrazol-3-yl, isoxazol-3-yl, thiophen-2-yl, 3-chloro-5-trifluoromethyl-pyridin-2-yl, or imidazo[2,l -0]thiazol-6-yl.

[154] In certain embodiments of formula I, R ⁴ is heteroaryl-Ci_6alkyl wherein the heteroaryl portion is optionally substituted.

[155] In embodiments of formula I wherein R ⁴ is heteroaryl-Ci_6alkyl wherein the heteroaryl portion is optionally substituted, the heteroaryl portions thereof may be pyrazin-2-yl, 5-methyl-pyrazin-2-yl, pyrimidin-5-yl, pyridin-2-yl, 6-methyl-pyridin-2-yl, 6-chloro-pyridin-3-yl, pyridin-3-yl, pyridin-4-yl, furan-2-yl, 6-oxo-pyridin-3-yl, benzimidazol-2-yl, thiazol-2-yl, benzothiazol-2-yl, pyrazol-4-yl, imidazo[l,2-a]pyridin-3-yl, isoxazol-5-yl, imidazol-4-yl, imidazol-l-yl, pyrazol-3-yl, isoxazol-3-yl, thiophen-2-yl, 3-chloro- 5-trifluoromethyl-pyridin-2-yl, l-tert-butoxycarbonyl-3,4-dihydroisoquinolin-2-yl, 3,4- dihydroisoquinolin-2-yl, or imidazo[2,l -0]thiazol-6-yl. [156] In embodiments of formula I wherein R ⁴ is heteroaryl-Ci_ ₆ alkyl, the Ci_

6alkyl portion may be methylene, ethylene, propylene or butylene.

[157] In embodiments of formula I wherein R ⁴ is heteroaryl-Ci_ ₆ alkyl, the Ci_

₆ alkyl portion may be methylene or ethylene.

[158] In embodiments of formula I wherein R ⁴ is heteroaryl-Ci- ₆ alkyl, the Ci-

₆ alkyl portion may be methylene.

[159] In certain embodiments of formula I, R ⁴ is phenyl-sulfonamido wherein the phenyl portion is optionally substituted.

[160] In embodiments of formula I wherein R ⁴ is phenyl-sulfonamido, the phenyl portion may be unsubstituted phenyl, 2-chloro-phenyl, 3-chloro-phenyl, 4-chloro- phenyl, 2-methyl-phenyl, 3-trifiuoromethyl-phenyl, 4-trifiuoromethyl-phenyl, 2-fluoro- phenyl, 3 -fiuoro -phenyl, 4-fluoro-phenyl, 2,4-difluoro-phenyl, 2-methoxy-phenyl, 3- methoxy-phenyl, 4-methoxy-phenyl, 3-carboxy-phenyl, or 4-amino-phenyl.

[161] In embodiments of formula I wherein R ⁴ is phenyl-sulfonamido, the sulfonamido portion may be of the formula -SO ₂ NH-.

[162] In embodiments of formula I wherein R ⁴ is phenyl-sulfonamido, the sulfonamido portion may be of the formula -NHSO ₂ -.

[163] In certain embodiments of formula I, R ⁴ is optionally substituted heterocycyl.

[164] In embodiments of formula I wherein R ⁴ is optionally substituted heterocycyl, such heterocycly may be tetrahydropyran-4-yl, oxetan-3-yl, 1-methyl- pyrrolidin-3-yl, l -tert-butoxycarbonyl-pyrrolidin-3-yl, pyrrolidin-3-yl, or 1-tert- butoxycarbonyl-azetidin-3-yl.

[165] In certain embodiments of formula I, R ⁴ is heterocyclyl-Ci_ ₆ alkyl wherein the heterocyclyl portion is optionally substituted with Ci-6alkyl or carboxy.

[166] In embodiments of formula I wherein R ⁴ is heterocyclyl-Ci_ ₆ alkyl wherein the heterocyclyl portion is optionally substituted, the heteroaryl portions thereof may be morpholin-4-yl, pyrrolidin-l -yl, l-tert-butoxycarbonyl-piperidin-4-yl, piperidin-4-yl, 1 - tert-butoxycarbonyl-pyrrolidin-3-yl, 3-tert-butoxycarbonyl-thiazolidin-5-yl, thiazolidine- 4-yl, l -tert-butoxycarbonyl-piperidin-3-yl, or piperidin-3-yl. [167] In embodiments of formula I wherein R ⁴ is heterocyclyl-Ci_ ₆ alkyl, the Ci_

6alkyl portion may be methylene, ethylene, propylene or butylene.

[168] In embodiments of formula I wherein R ⁴ is heterocycyl-Ci_ ₆ alkyl, the Ci_

₆ alkyl portion may be methylene or ethylene.

[169] In embodiments of formula I wherein R ⁴ is heterocycyl-Ci- ₆ alkyl, the Ci-

₆ alkyl portion may be methylene.

[170] In certain embodiments of formula I, R ⁴ is benzyl; phenyl; 4-phenyl-butyl;

2-chlorobenzyl; 3-chlorobenzyl; 4-chlorobenzyl; 2-phenyl-cyclopropyl; pyrazin-2- ylmethyl; pyrimidin-5-ylmethyl; phenylsulfonamido; 2-methyl-phenyl; n-butyl; isobutyl; tert-butyl; 3-trifluoromethy-benzyl; 4-trifluoromethy-benzyl; 2-fluoro-benzyl; 3-fluoro- benzyl; 4-fiuoro-benzyl; 4-fiuoro-phenyl; 2,4-difluoro-phenyl; 3-phenyl-cyclobutyl; 2- methoxy-benzyl; 3-methoxy-benzyl; 4-methoxy-benzyl; 2-methoxy-ethyl; l -(phenyl)- ethyl; 3-(methanesulfonyl)-propyl; 3-carboxy-benzyl; 2-(morpholin-4-yl)-ethyl; 2- (pyrrolidin-1 -yl)-ethyl; 1 -methyl-pyrrolidin-3-yl; 1 -(tert-butoxycarbonyl)-azetidin-3-yl; 2- hydroxy-propyl; 2-hydroxy-l -methyl-ethyl; 2-hydroxy-2-methyl-propyl; 2 -hydro xy- ethyl; tetrahydropyran-4-yl; 2-(ethoxy)-ethyl; 2-(methanesulfonyl)-l -methyl-ethyl;

oxetan-3-yl; 2 -(ethoxy)-l -methyl-ethyl; 2-(pyridin-2-yl)-l -methyl-ethyl; 2-(6-methyl- pyridin-2-yl)- 1 -methyl-ethyl; 2-( ,N-dimethylamino)- 1 -methyl-ethyl; 2-(furan-2-yl)-l - methyl-ethyl; pyridin-2-ylmethyl; pyridin-3-ylmethyl; pyridin-4-ylmethyl; 2-( pyridin-2- yl)-l -methyl-ethyl; 2-( pyridin-4-yl)-l -methyl-ethyl; l -(tert-butoxycarbonyl)-piperidin-4- ylmethyl; 1 -(tert-butoxycarbonyl)-pyrrolidin-3-ylmethyl; 1 -ethyl-pyrrolidin-2- ylmethyl;2-chloro-pyridin-5-ylmethyl; 6-oxo- 1 ,6,-dihydropyridin-3-ylmethyl; 3-(tert- butoxycarbonyl)-thiazolidin-5-ylmethyl; 5-methyl-pyrazin-2-ylmethyl; benzimidazol-2- ylmethyl; thiazol-2-ylmethyl; 4-sulfmamoyl-benzyl.1 -(tert-butoxycarbonyl)-3,4-dihydro- isoquinolin-2-ylmethyl; 4-(tert-butoxycarbonylamino)-benzyl; 3,4-dihydro-isoquinolin-2- ylmethyl; 4-amino-benzyl; thiazolidin-4-ylmethyl; benzothiazol-2-ylmethyl; 2-(pyrazol- 4-yl)-ethyl; imidazo[l,2-a]pyridm-3-ylmethyl; isoxazol-5-ylmethyl;imidazol-4-ylmethyl;

imidazol-l -ylmethyl; pyrazol-3-ylmethyl; isoxazol-3-ylmethyl; 2-(thiophen-2-yl)-l -methyl-ethyl; 3 -chloro-5 -trifluoromethyl-pyridin-2-ylmethyl; 2-(methanesulfonyl)-ethyl; 3 -methoxycarbonyl- benzyl; 6-trifluoromethyl-pyridin3-ylmethyl; imidazo[2,l -0]thiazol-6-ylmethyl; l -(tert- butoxycarbonyl)-piperidin-3-ylmethyl; l-(tert-butoxycarbonyl)-pyrrolidin-3-yl;

orpyrrolidin-3-ylmethyl.

[171] In certain embodiments of formula , R ⁴ is benzyl.

[172] In certain embodiments of formula ] , R ⁴ is phenyl.

[173] In certain embodiments of formula ] , R ⁴ is 4-phenyl-butyl.

[174] In certain embodiments of formula ] , R ⁴ is 2-chlorobenzyl.

[175] In certain embodiments of formula ] , R ⁴ is 3-chlorobenzyl.

[176] In certain embodiments of formula ] , R ⁴ is 4-chlorobenzyl.

[177] In certain embodiments of formula ] , R ⁴ is 2-phenyl-cyclopropyl.

[178] In certain embodiments of formula ] , R ⁴ is pyrazin-2-ylmethyl.

[179] In certain embodiments of formula ] , R ⁴ is pyrimidin-5-ylmethyl.

[180] In certain embodiments of formula ] , R ⁴ is phenylsulfonamido.

[181] In certain embodiments of formula ] , R ⁴ is 2-methyl-phenyl.

[182] In certain embodiments of formula ] , R ⁴ is n-butyl.

[183] In certain embodiments of formula ] , R ⁴ is isobutyl.

[184] In certain embodiments of formula ] , R ⁴ is tert-butyl.

[185] In certain embodiments of formula ] , R ⁴ is 3-trifiuoromethy-benzyl.

[186] In certain embodiments of formula ] , R ⁴ is 4-trifiuoromethy-benzyl.

[187] In certain embodiments of formula ] , R ⁴ is 2-fluoro-benzyl.

[188] In certain embodiments of formula 1 , R ⁴ is 3-fluoro-benzyl.

[189] In certain embodiments of formula 1 , R ⁴ is 4-fluoro-benzyl.

[190] In certain embodiments of formula 1 , R ⁴ is 4-fluoro-phenyl.

[191] In certain embodiments of formula 1 , R ⁴ is 2, 4-difluoro -phenyl.

[192] In certain embodiments of formula ] , R ⁴ is 3-phenyl-cyclobutyl.

[193] In certain embodiments of formula I , R ⁴ is 2-methoxy-benzyl.

[194] In certain embodiments of formula I , R ⁴ is 3-methoxy-benzyl.

[195] In certain embodiments of formula I , R ⁴ is 4-methoxy-benzyl.

[196] In certain embodiments of formula I , R ⁴ is 2-methoxy-ethyl.

[197] In certain embodiments of formula I , R ⁴ is l-(phenyl)-ethyl.

[198] In certain embodiments of formula I , R ⁴ is 3-(methanesulfonyl)-propyl.

[199] In certain embodiments of formula I , R ⁴ is 3-carboxy-benzyl. [200] In certain embodiments of formula I, R ⁴ is 2-(morpholin-4-yl)-ethyl.

[201] In certain embodiments of formula I, R ⁴ is 2-(pyrrolidin-l-yl)-ethyl.

[202] In certain embodiments of formula I, R ⁴ is l-methyl-pyrrolidin-3-yl.

[203] In certain embodiments of formula I, R ⁴ is 1 -(tert-butoxycarbonyl)- azetidin-3-yl.

[204] In certain embodiments of formula I, R ⁴ is 2-hydroxy-propyl.

[205] In certain embodiments of formula I, R ⁴ is 2-hydroxy-l -methyl-ethyl.

[206] In certain embodiments of formula I, R ⁴ is 2-hydroxy-2-methyl-propyl.

[207] In certain embodiments of formula I, R ⁴ is 2-hydroxy-ethyl.

[208] In certain embodiments of formula I, R ⁴ is tetrahydropyran-4-yl.

[209] In certain embodiments of formula I, R ⁴ is 2-(ethoxy)-ethyl.

[210] In certain embodiments of formula I, R ⁴ is 2-(methanesulfonyl)-l -methyl- ethyl.

[211] In certain embodiments of formula I, R ⁴ is oxetan-3-yl.

[212] In certain embodiments of formula I, R ⁴ is 2-(ethoxy)-l -methyl-ethyl.

[213] In certain embodiments of formula I, R ⁴ is 2-(pyridin-2-yl)-l -methyl-ethyl.

[214] In certain embodiments of formula I, R ⁴ is 2-(6-methyl-pyridin-2-yl)-l- methyl-ethyl.

[215] In certain embodiments of formula I, R ⁴ is 2-( ,N-dimethylamino)-l- methyl-ethyl.

[216] In certain embodiments of formula I, R ⁴ is 2-(furan-2-yl)-l -methyl-ethyl.

[217] In certain embodiments of formula I, R ⁴ is pyridin-2-ylmethyl.

[218] In certain embodiments of formula I, R ⁴ is pyridin-3-ylmethyl.

[219] In certain embodiments of formula I, R ⁴ is pyridin-4-ylmethyl.

[220] In certain embodiments of formula I, R ⁴ is 2-( pyridin-2-yl)-l -methyl- ethyl.

[221] In certain embodiments of formula I, R ⁴ is 2-( pyridin-4-yl)-l -methyl- ethyl.

[222] In certain embodiments of formula I, R ⁴ is 1 -(tert-butoxycarbonyl)- piperidin-4-ylmethyl. [223] In certain embodiments of formula I, R ⁴ is 1 -(tert-butoxycarbonyl)- pyrrolidin-3 -ylmethyl.

[224] In certain embodiments of formula I, R ⁴ is 1 -ethyl-pyrrolidin-2 -ylmethyl.

[225] In certain embodiments of formula I, R ⁴ is 2-chloro-pyridin-5-ylmethyl.

[226] In certain embodiments of formula I, R ⁴ is 6-oxo-l,6,-dihydropyridin-3- ylmethyl.

[227] In certain embodiments of formula I, R ⁴ is 3-(tert-butoxycarbonyl)- thiazo lidin-5 -ylmethyl.

[228] In certain embodiments of formula I, R ⁴ is 5 -methyl-pyrazin-2 -ylmethyl;

[229] In certain embodiments of formula I, R ⁴ is benzimidazol-2 -ylmethyl.

[230] In certain embodiments of formula I, R ⁴ is thiazol-2-ylmethyl.

[231] In certain embodiments of formula I, R ⁴ is 4-sulfmamoyl-benzyl.

[232] In certain embodiments of formula I, R ⁴ is l-(tert-butoxycarbonyl)-3,4- dihydro-isoquinolin-2-ylmethyl.

[233] In certain embodiments of formula I, R ⁴ is 4-(tert-butoxycarbonylamino)- benzyl.

[234] In certain embodiments of formula I, R ⁴ is 3,4-dihydro-isoquinolin-2- ylmethyl.

[235] In certain embodiments of formula I, R ⁴ is 4-amino-benzyl.

[236] In certain embodiments of formula I, R ⁴ is thiazolidin-4-ylmethyl.

[237] In certain embodiments of formula I, R ⁴ is benzothiazol-2-ylmethyl.

[238] In certain embodiments of formula I, R ⁴ is 2-(pyrazol-4-yl)-ethyl.

[239] In certain embodiments of formula I, R ⁴ is imidazo[l,2-a]pyridin-3 -ylmethyl.

[240] In certain embodiments of formula I, R ⁴ is isoxazol-5 -ylmethyl.

[241] In certain embodiments of formula I, R ⁴ is imidazol-4 -ylmethyl.

[242] In certain embodiments of formula I, R ⁴ is imidazol-1 -ylmethyl.

[243] In certain embodiments of formula I, R ⁴ is pyrazol-3 -ylmethyl.

[244] In certain embodiments of formula I, R ⁴ is isoxazol-3 -ylmethyl.

[245] In certain embodiments of formula I, R ⁴ is 2-(thiophen-2-yl)-l -methyl-ethyl.

[246] In certain embodiments of formula I, R ⁴ is 3-chloro-5-trifluoromethyl- pyridin-2 -ylmethyl. [247] In certain embodiments of formula I, R ⁴ is 2-(methanesulfonyl)-ethyl.

[248] In certain embodiments of formula I, R ⁴ is 3 -methoxycarbonyl-b enzyl.

[249] In certain embodiments of formula I, R ⁴ is 6-trifluoromethyl-pyridin3 - ylmethyl.

[250] In certain embodiments of formula I, R ⁴ is imidazo [2, 1 -0]thiazol-6-ylmethy

[251] In certain embodiments of formula I, R ⁴ is 1 -(tert-butoxycarbonyl)- piperidin ■3 -ylmethyl.

[252] In certain embodiments of formula I, R ⁴ is 1 -(tert-butoxycarbonyl)- pyrrolidin-3-yl.

[253] In certain embodiments of formula I, R ⁴ is pyrrolidin-3 -ylmethyl.

[254] In certain embodiments of formula I, R ⁵ is Ci_ ₆ alkyl.

[255] In certain embodiments of formula I, R ⁵ is methyl.

[256] In certain embodiments of formula I, R ⁵ is propyl.

[257] In certain embodiments of formula I, R ⁵ is optionally substituted aryl.

[258] In certain embodiments of formula I, R ⁵ is optionally substituted phenyl.

[259] In certain embodiments of formula I, R ⁵ is phenyl optionally substituted one, two or three times with a group or groups independently selected from: Ci_ ₆ alkyl; Ci_ ₆ alkoxy; halo-Ci_ ₆ alkyl; halo; carboxy; and cyano.

[260] In certain embodiments of formula I, R ⁵ is phenyl optionally substituted once or twice with a group or groups independently selected from: Ci_ ₆ alkyl; Ci_ ₆ alkoxy; halo-Ci- ₆ alkyl; halo; carboxy; and cyano.

[261] In certain embodiments of formula I, R ⁵ is phenyl optionally substituted once with Ci_ ₆ alkyl; Ci_ ₆ alkoxy; halo; or halo-Ci_ ₆ alkyl.

[262] In certain embodiments of formula I, R ⁵ is phenyl optionally substituted once with methyl, methoxy, fluoro, chloro or trifluoromethyl.

[263] In certain embodiments of formula I, R ⁵ is phenyl optionally substituted once with fluoro, chloro or methyl.

[264] In certain embodiments of formula I, R ⁵ is phenyl optionally substituted once with Ci- ₆ alkyl or halo.

[265] In certain embodiments of formula I, R ⁵ is unsubstituted phenyl. [266] In certain embodiments of formula I, R ⁵ is optionally substituted heteroaryl.

[267] In certain embodiments of formula I, R ⁵ is heteroaryl selected from:

pyridinyl, pyrimidinyl, pyrazinyl, furanyl; pyridonyl, benzimidazolyl, thiazolyl;

isothiazolyl; tetrahydroisoquinolinyl; benzothiazolyl, pyrazolyl; imidazo[l ,2- ]pyridinyl, oxazolyl; isoxazolyl; or imidazolyl, each of which may be optionally substituted.

[268] In certain embodiments of formula I, R ⁵ is pyridinyl.

[269] In certain embodiments of formula I, R ⁵ is pyridin-2-yl.

[270] In certain embodiments of formula I, R ⁵ is pyridin-3-yl.

[271] In certain embodiments of formula I, R ⁵ is C3_ ₆ cycloalkyl optionally substituted with Ci_ ₆ alkyl.

[272] In certain embodiments of formula I, R ⁵ is cyclohexyl.

[273] In certain embodiments of formula I, the compounds may be more specifically represented by formula II

or pharmaceutically acceptable salts thereof,

wherein, Z, m, R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined herein.

[274] In certain embodiments of formula I, the subject compounds may be more specifically represented by formula III

wherein:

n is from 0 to 3; p is from 0 to 3;

each of R ⁶ and R ⁷ is independently: Ci-6alkyl; Ci-6alkoxy; halo-Ci-6alkyl; halo; carboxy; or cyano; and

R ³ and R ⁴ are as described above.

[275] In certain embodiments of formula III, n is from 0 to 2.

[276] In certain embodiments of formula III, n is 0 or 1.

[277] In certain embodiments of formula III, n is 0.

[278] In certain embodiments of formula III, n is 1.

[279] In certain embodiments of formula III, p is from 0 to 2.

[280] In certain embodiments of formula III, p is 0 or 1.

[281] In certain embodiments of formula III, p is 0.

[282] In certain embodiments of formula III, p is 1.

[283] In certain embodiments of formula III, R ⁶ is Ci_ ₆ alkyl; C

halo-Ci- ₆ alkyl.

[284] In certain embodiments of formula III, R ⁶ is methyl, methoxy, fluoro, chloro or trifiuoromethyl.

[285] In certain embodiments of formula III, R ⁶ is fluoro, chloro or methyl.

[286] In certain embodiments of formula III, R ⁷ is Ci_6alkyl; Ci_6alkoxy; halo; or halo-Ci_ ₆ alkyl.

[287] In certain embodiments of formula III, R ⁷ is methyl, methoxy, fluoro, chloro or trifiuoromethyl.

[288] In certain embodiments of formula III, R ⁷ is fluoro, chloro or methyl.

[289] In certain embodiments of formula III, R ⁷ is fluoro.

[290] In certain embodiments of formula I, the subject compounds may be more specifically represented by formula IV

wherein n, p, R ³ , R ⁴ , R ⁶ and R ⁷ are as described above.

[291] Where any of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ or R ⁷ is alkyl or contains an alky- moiety, such alkyl is preferably lower alkyl, i.e. Ci-Cealkyl, and in many embodiments is Ci-C ₄ alkyl.

Methods

[292] The invention also provides methods for treating a disease or condition mediated by or otherwise associated with potassium channel activity, the method comprising administering to a subject in need thereof an effective amount of a compound of the invention.

[293] The disease or condition may be selected from autoimmune disorders, inflammatory disorders, immune-mediated disorders, or other disorders, including multiple sclerosis, type-1 diabetes mellitus, type-2 diabetes mellitus, rheumatoid arthritis, psoriasis, contact dermatitis, obesity, systemic lupus erythematosus, graft-versus host disease, transplant rejection, and delayed type hypersensitivity.

[294] The invention also provides methods for treating an inflammatory, respiratory or diabetes condition, the method comprising administering to a subject in need thereof an effective amount of a compound of the invention.

[295] The disease may be an inflammatory disease such as arthritis, and more particularly rheumatoid arthritis, osteoarthritis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airways hyper-responsiveness, septic shock, glomerulonephritis, irritable bowel disease, and Crohn's disease.

[296] The disease may be muscular sclerosis.

[297] The disease may be a respiratory disorder, such as chronic obstructive pulmonary disorder (COPD), asthma, or bronchospasm, or a gastrointestinal (GI) disorder such as Irritable Bowel Syndrome (IBS), Inflammatory Bowel Disease (IBD), biliary colic and other biliary disorders, renal colic, diarrhea-dominant IBS, pain associated with GI distension.

[298] Representative compounds in accordance with the methods of the invention are shown in Table 1 , together with selected IC50 values for Kvl .3.

tructure Name IC50

4-(4-Fluoro-benzenesulfonyl)- 3 -phenyl-3 ,4-dihydro-2H-

0.359 b enzo [1,4] oxazine-7 -carb oxylic acid (2-methoxy-ethyl)-amide

3 -(4-Chloro-phenyl)-4-(4- fluoro-benzenesulfonyl)-3 ,4- dihydro-2H-benzo[l,4]oxazine- 0.242 7-carboxylic acid (2-methoxy- ethyl)-amide

3 -(4-Chloro-phenyl)-4- (pyridine-2-sulfonyl)-3 ,4- dihydro-2H-benzo[l,4]oxazine- 0.645 7-carboxylic acid (2-methoxy- ethyl)-amide

3 -(4-Chloro-phenyl)-4- (pyridine-2-sulfonyl)-3 ,4-

0.597 dihydro-2H-benzo[l,4]oxazine- 7-carboxylic acid benzylamide

3 -(4-Chloro-phenyl)-4- (pyridine-3 -sulfonyl)-3 ,4- dihydro-2H-benzo[l,4]oxazine- 0.648 7-carboxylic acid (2-methoxy- ethyl)-amide

3 -(4-Chloro-phenyl)-4- (propane- 1 -sulfonyl)-3 ,4- dihydro-2H-benzo[l,4]oxazine- 4.916 7-carboxylic acid (2-methoxy- ethyl)-amide

Cl ^/N/ o ' ^^CH ₃

# Structure Name IC50

O

1 -(4-Fluoro-benzenesulfonyl)- 2-phenyl-2,3 -dihydro- 1 H-

23

pyrido [2,3 -b] [1,4] oxazine-6- carboxylic acid benzylamide

0

1 -(4-Fluoro-benzenesulfonyl)- 2-phenyl-2,3 -dihydro- 1 H-

24 pyrido [2,3 -b] [1,4] oxazine-6- carboxylic acid (2-methoxy- ethyl)-amide

TABLE 1

Synthesis

[299] Compounds of the present invention can be made by a variety of methods depicted in the illustrative synthetic reaction schemes shown and described below.

[300] The starting materials and reagents used in preparing these compounds generally are either available from commercial suppliers, such as Aldrich Chemical Co., or are prepared by methods known to those skilled in the art following procedures set forth in references such as Fieser and Fieser's Reagents for Organic Synthesis; Wiley & Sons: New York, 1991, Volumes 1-15; Rodd's Chemistry of Carbon Compounds, Elsevier Science Publishers, 1989, Volumes 1-5 and Supplemental; and Organic Reactions, Wiley & Sons: New York, 1991, Volumes 1-40. The following synthetic reaction schemes are merely illustrative of some methods by which the compounds of the present invention can be synthesized, and various modifications to these synthetic reaction schemes can be made and will be suggested to one skilled in the art having referred to the disclosure contained in this Application.

[301] The starting materials and the intermediates of the synthetic reaction schemes can be isolated and purified if desired using conventional techniques, including but not limited to, filtration, distillation, crystallization, chromatography, and the like. Such materials can be characterized using conventional means, including physical constants and spectral data. [302] Unless specified to the contrary, the reactions described herein preferably are conducted under an inert atmosphere at atmospheric pressure at a reaction temperature range of from about -78 °C to about 150 °C, more preferably from about 0 °C to about 125 °C, and most preferably and conveniently at about room (or ambient) temperature, e.g., about 20 °C.

[303] Scheme A below illustrates one synthetic procedure usable to prepare specific compounds of formula I, wherein X is a leaving group such as halo and may be the same or different for each occurrence, R is lower alkyl, and m, R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined herein.

SCHEME A

[304] In step 1 of Scheme A, nitro phenyl ester compound a undergoes an O- alkylation by reaction with ketone compound b to ether compound c . Ketone b may comprise, for example, an acetophenone or other aryl or heteroaryl ketone in accordance with the invention.

[305] In step 2 the carbonyl group of ether compound c undergoes reduction to afford the corresponding hydroxy compound d. This reduction may be carried out using sodium borohydride or like reducing agent. [306] In step 3, the nitro group of compound d is reduced to provide the corresponding amino compound e. The nitro reduction of step 3 may be achieved by reduction in the presence of metallic iron and a chlorine salt such as ammonium chloride.

[307] The amino compound e is then reacted with sulfonyl halide compound f in step 4 to yield sulfonamido compound g. Sulfonyl halide compound f may comprise, for example, a phenyl sulfonyl chloride or like aryl or heteroaryl sulfonyl chloride.

[308] A cyclization reaction occurs in step 5 wherein ring closure provides benzoxazine compound h. This cyclization may be carried out, for example, using diethyl azodicarboxylate reagent in the presence of triphenyl phosphene.

[309] In step 6, the ester group of compound h undergoes hydrolysis to afford the corresponding carboxylic acid compound i. The hydrolysis of step 6 may be achieved under aqueous basic conditions using an alkali metal hydroxide or like base.

[310] In step 7, an amide formation occurs by reaction of carboxylic acid compound i with amine , to afford benzoxazine amide compound k. Benzoxazine k is a compound of formula I in accordance with the invention. Various amide coupling reagents, such as EDCI, HATU, BOP, PyBOP or carbodiimides, or an acid chloride intermediate, may be used in the reaction of step 7.

[311] Scheme B below illustrates another synthetic procedure usable to prepare specific compounds of formula I, wherein X is a leaving group such as halo and may be the same or different for each occurrence, and m, R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined herein.

SCHEME B

[312] Steps 1 through 5 of Scheme B proceed in a manner similar to that described above for Scheme A. In this regard, nitro phenyl ester compound a undergoes an O-alkylation by reaction with ketone compound b to provide ether compound c in step 1. The carbonyl group of compound c is reduced in step 2 to afford the hydroxy compound d. In step 3, the nitro group of compound d is reduced to provide amino compound e. Amino compound e is then reacted with sulfonyl halide compound f in step 4 to yield sulfonamido compound g. Compound g is cyclized in step 5 give benzoxazine compound h.

[313] In step 6, benzoxazine compound h is reacted with amine in the presence of palladium catalyst i to afford benzoxazine amine s, which is a compound of formula I in accordance with the invention.

[314] Many variations on the procedure of Scheme A and Scheme B are possible and will suggest themselves to those skilled in the art. Specific details for producing compounds of the invention are described in the Examples section below.

Utility

[315] The compounds of the invention are usable for the treatment of a wide range of inflammatory diseases and conditions such as arthritis, including but not limited to, rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, systemic lupus erythematosus and juvenile arthritis, osteoarthritis, gouty arthritis and other arthritic conditions. The subject compounds would be useful for the treatment of pulmonary disorders or lung inflammation, including adult respiratory distress syndrome, pulmonary sarcoidosis, asthma, silicosis, and chronic pulmonary inflammatory disease.

[316] The disease or condition may be selected from autoimmune disorders, inflammatory disorders, immune-mediated disorders, or other disorders, including multiple sclerosis, type-1 diabetes mellitus, type-2 diabetes mellitus, rheumatoid arthritis, psoriasis, contact dermatitis, obesity, systemic lupus erythematosus, graft-versus host disease, transplant rejection, and delayed type hypersensitivity.

[317] Further, compounds of the invention are useful for treating respiratory disorders, including chronic obstructive pulmonary disorder (COPD), asthma, broncho spasm, and the like.

Administration and Pharmaceutical Composition

[318] The invention includes pharmaceutical compositions comprising at least one compound of the present invention, or an individual isomer, racemic or non-racemic mixture of isomers or a pharmaceutically acceptable salt or solvate thereof, together with at least one pharmaceutically acceptable carrier, and optionally other therapeutic and/or prophylactic ingredients.

[319] In general, the compounds of the invention will be administered in a therapeutically effective amount by any of the accepted modes of administration for agents that serve similar utilities. Suitable dosage ranges are typically 1-500 mg daily, preferably 1 -100 mg daily, and most preferably 1 -30 mg daily, depending upon numerous factors such as the severity of the disease to be treated, the age and relative health of the subject, the potency of the compound used, the route and form of administration, the indication towards which the administration is directed, and the preferences and experience of the medical practitioner involved. One of ordinary skill in the art of treating such diseases will be able, without undue experimentation and in reliance upon personal knowledge and the disclosure of this Application, to ascertain a therapeutically effective amount of the compounds of the present invention for a given disease.

[320] Compounds of the invention may be administered as pharmaceutical formulations including those suitable for oral (including buccal and sub-lingual), rectal, nasal, topical, pulmonary, vaginal, or parenteral (including intramuscular, intraarterial, intrathecal, subcutaneous and intravenous) administration or in a form suitable for administration by inhalation or insufflation. The preferred manner of administration is generally oral using a convenient daily dosage regimen which can be adjusted according to the degree of affliction.

[321] A compound or compounds of the invention, together with one or more conventional adjuvants, carriers, or diluents, may be placed into the form of

pharmaceutical compositions and unit dosages. The pharmaceutical compositions and unit dosage forms may be comprised of conventional ingredients in conventional proportions, with or without additional active compounds or principles, and the unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range to be employed. The pharmaceutical compositions may be employed as solids, such as tablets or filled capsules, semisolids, powders, sustained release formulations, or liquids such as solutions, suspensions, emulsions, elixirs, or filled capsules for oral use; or in the form of suppositories for rectal or vaginal administration; or in the form of sterile injectable solutions for parenteral use. Formulations containing about one (1) milligram of active ingredient or, more broadly, about 0.01 to about one hundred (100) milligrams, per tablet, are accordingly suitable representative unit dosage forms.

[322] The compounds of the invention may be formulated in a wide variety of oral administration dosage forms. The pharmaceutical compositions and dosage forms may comprise a compound or compounds of the present invention or pharmaceutically acceptable salts thereof as the active component. The pharmaceutically acceptable carriers may be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier may be one or more substances which may also act as diluents, flavouring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier generally is a finely divided solid which is a mixture with the finely divided active component. In tablets, the active component generally is mixed with the carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain from about one (1) to about seventy (70) percent of the active compound. Suitable carriers include but are not limited to magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatine, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term "preparation" is intended to include the formulation of the active compound with encapsulating material as carrier, providing a capsule in which the active component, with or without carriers, is surrounded by a carrier, which is in association with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges may be as solid forms suitable for oral administration.

[323] Other forms suitable for oral administration include liquid form

preparations including emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions, or solid form preparations which are intended to be converted shortly before use to liquid form preparations. Emulsions may be prepared in solutions, for example, in aqueous propylene glycol solutions or may contain emulsifying agents, for example, such as lecithin, sorbitan monooleate, or acacia. Aqueous solutions can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents. Aqueous suspensions can be prepared by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well known suspending agents. Solid form preparations include solutions, suspensions, and emulsions, and may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

[324] The compounds of the invention may be formulated for parenteral administration (e.g., by injection, for example bolus injection or continuous infusion) and may be presented in unit dose form in ampoules, pre-filled syringes, small volume infusion or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, for example solutions in aqueous polyethylene glycol. Examples of oily or nonaqueous carriers, diluents, solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate), and may contain formulatory agents such as preserving, wetting, emulsifying or suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution for constitution before use with a suitable vehicle, e.g., sterile, pyrogen-free water.

[325] The compounds of the invention may be formulated for topical

administration to the epidermis as ointments, creams or lotions, or as a transdermal patch. Ointments and creams may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents. Lotions may be formulated with an aqueous or oily base and will in general also containing one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents. Formulations suitable for topical administration in the mouth include lozenges comprising active agents in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatine and glycerine or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[326] The compounds of the invention may be formulated for administration as suppositories. A low melting wax, such as a mixture of fatty acid glycerides or cocoa butter is first melted and the active component is dispersed homogeneously, for example, by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool, and to solidify.

[327] The compounds of the invention may be formulated for vaginal administration. Pessaries, tampons, creams, gels, pastes, foams or sprays containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

[328] The subject compounds may be formulated for nasal administration. The solutions or suspensions are applied directly to the nasal cavity by conventional means, for example, with a dropper, pipette or spray. The formulations may be provided in a single or multidose form. In the latter case of a dropper or pipette, this may be achieved by the patient administering an appropriate, predetermined volume of the solution or suspension. In the case of a spray, this may be achieved for example by means of a metering atomizing spray pump. [329] The compounds of the invention may be formulated for aerosol administration, particularly to the respiratory tract and including intranasal administration. The compound will generally have a small particle size for example of the order of five (5) microns or less. Such a particle size may be obtained by means known in the art, for example by micronization. The active ingredient is provided in a pressurized pack with a suitable propellant such as a chlorofiuorocarbon (CFC), for example,

dichlorodifluoromethane, trichlorofluoromethane, or dichlorotetrafluoroethane, or carbon dioxide or other suitable gas. The aerosol may conveniently also contain a surfactant such as lecithin. The dose of drug may be controlled by a metered valve. Alternatively the active ingredients may be provided in a form of a dry powder, for example a powder mix of the compound in a suitable powder base such as lactose, starch, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidine (PVP). The powder carrier will form a gel in the nasal cavity. The powder composition may be presented in unit dose form for example in capsules or cartridges of e.g., gelatine or blister packs from which the powder may be administered by means of an inhaler.

[330] When desired, formulations can be prepared with enteric coatings adapted for sustained or controlled release administration of the active ingredient. For example, the compounds of the present invention can be formulated in transdermal or subcutaneous drug delivery devices. These delivery systems are advantageous when sustained release of the compound is necessary and when patient compliance with a treatment regimen is crucial. Compounds in transdermal delivery systems are frequently attached to an skin- adhesive solid support. The compound of interest can also be combined with a penetration enhancer, e.g., Azone (l-dodecylazacycloheptan-2-one). Sustained release delivery systems are inserted subcutaneously into the subdermal layer by surgery or injection. The subdermal implants encapsulate the compound in a lipid soluble membrane, e.g., silicone rubber, or a biodegradable polymer, e.g., polylactic acid.

[331] The pharmaceutical preparations are preferably in unit dosage forms. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

[332] Other suitable pharmaceutical carriers and their formulations are described in Remington: The Science and Practice of Pharmacy 1995, edited by E. W. Martin, Mack Publishing Company, 19th edition, Easton, Pennsylvania. Representative pharmaceutical formulations containing a compound of the present invention are described below.

EXAMPLES

[333] The following preparations and examples are given to enable those skilled in the art to more clearly understand and to practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof.

[334] Unless otherwise stated, all temperatures including melting points (i.e.,

MP) are in degrees Celsius (°C). It should be appreciated that the reaction which produces the indicated and/or the desired product may not necessarily result directly from the combination of two reagents which were initially added, i.e., there may be one or more intermediates which are produced in the mixture which ultimately leads to the formation of the indicated and/or the desired product. The following abbreviations may be used in the Preparations and Examples.

ABBREVIATIONS

BOP benzotriazol- 1 -yloxytris(dimethylamino)phosphonium hexafluoropho sphate

DEAD diethyl azodicarboxylate

DME 1 ,2-dimethoxy-ethane

dppf 1 , 1 '-Bis(diphenylphosphino)ferrocene

EDCI 1 -ethyl-3-(3 '-dimethylaminopropyl)carbodiimide

EtOAc ethyl acetate

EtOH ethanol

HOAc acetic acid

HOBt N-Hydroxybenzotriazole

hplc high performance liquid chromatography

IPA isopropanol

MeOH methanol

Pd ₂ (dba) ₃ Tris(dibenzylideneacetone)dipalladium(0)

TEA triethylamine

THF tetrahy dro furan

TLC thin layer chromatography Example 1

3-(4-Chloro-phenyl)-4-(4-fluoro-benzenesulfonyl)-3,4-dihydro -2H-benzo[l,4]oxazine-7- carboxylic acid (2-methoxy-ethyl)-amide

[335] The synthetic procedure used in this preparation is outlined in Scheme C.

Step 1 Methyl 3-(2-(4-chlorophenyl)-2-oxoethoxy)-4-nitrobenzoate

[336] To a solution of methyl 3-hydroxy-4-nitrobenzoate (4.0 g, 20.3 mmol, Eq:

1.00) and 2-bromo-4'-chloroacetophenone (5.21 g, 22.3 mmol, Eq: 1.1) was added cesium carbonate (7.27 g, 22.3 mmol, Eq: 1.1) to give a bright orange mixture. The reaction mixture was stirred for 3 h at room temperature. The reaction mixture was then poured into 100 mL sat. aqueous NaHC0 ₃ and extracted with EtOAc. The combined organic layers were washed with sat. aqueous NaCl and then concentraetd in vacuo. The crude material was purified by flash chromatography (silica gel, 40 g, 100% CH ₂ C1 ₂ ) and then treated with 10% EtOAc in hexanes to give methyl 3-(2-(4-chlorophenyl)-2- oxoethoxy)-4-nitrobenzoate (5.8 g, 16.6 mmol, 81.7 % yield) as a solid.

[337] Similarly prepared, using 4'-chloroacetophenone in place of 2-bromo-4'- chloroacetophenone, was methyl 3-(2-phenyl-2-oxoethoxy)-4-nitrobenzoate.

Step 2 Methyl 3-(2-(4-chlorophenyl)-2-hydroxyethoxy)-4-nitrobenzoate

[338] A suspension of methyl 3-(2-(4-chlorophenyl)-2-oxoethoxy)-4- nitrobenzoate (5.8 g, 16.6 mmol, Eq: 1.00) in methanol (150 mL) was cooled in an ice bath. Sodium borohydride (1.0 g, 26.4 mmol, Eq: 1.59) was added, the cooling bath removed, and the reaction mixture stirred at room temperature for 1 h. The reaction was quenched by adding sat. aqueous NH4CI (40 mL) and the product was extracted into EtOAc. The combined organic layers were washed with sat. aqueous NaCl and then concentrated in vacuo. The crude material was purified by flash chromatography (silica gel, 40 g, 0% to 40% EtOAc in hexanes) to give methyl 3-(2-(4-chlorophenyl)-2- hydroxyethoxy)-4-nitrobenzoate (5.76 g, 16.4 mmol, 98.7 % yield) as a yellow solid. Step 3 Methyl 4-amino-3-(2-(4-chlorophenyl)-2-hydroxyethoxy)benzoate

[339] In a 500 mL round-bottomed flask equipped with a 2" magnetic stir bar, methyl 3-(2-(4-chlorophenyl)-2-hydroxyethoxy)-4-nitrobenzoate (1.1 g, 3.13 mmol, Eq: 1.00), iron (0.88 g, 15.8 mmol, Eq: 5.04) and ammonium chloride (0.83 g, 15.5 mmol, Eq: 4.96) were combined in a mixture of ethanol (25 mL) and water (10 mL). The mixture was then heated at 80°C with rapid stirring for 2 h. The hot reaction mixture was then filtered through celite and the filtrate was concentrated in vacuo. The resulting residue was treated with sat. aqueous Na ₂ C0 ₃ (50 mL), and the product was then extracted into EtOAc. The combined organic layers were washed with brine and concentrated in vacuo. The crude methyl 4-amino-3-(2-(4-chlorophenyl)-2- hydroxyethoxy)benzoate was used directly in the next step without further purification. Step 4 Methyl 3-(2-(4-chlorophenyl)-2-hydroxyethoxy)-4-(4- fluorophenylsulfonamido)benzoate

[340] Methyl 4-amino-3-(2-(4-chlorophenyl)-2-hydroxyethoxy)benzoate (997 mg, 3.1 mmol, Eq: 1.00), 4-fluorobenzene-l -sulfonyl chloride (782 mg, 0.8 mL, 4.02 mmol, Eq: 1.3) and pyridine (313 mg, 0.32 mL, 3.96 mmol, Eq: 1.28) were combined in CH ₂ CI ₂ (80 mL) and the resulting solution was stirred at rt for 72 h at room temperature. The reaction mixture was then concentrated to ~20 mL in vacuo, diluted with EtOAc, washed with sat. aqueous NH ₄ CI, 1 M aqueous HC1, and sat. aqueous NaCl, and then concentrated in vacuo. The resulting crude material was purified by flash

chromatography (silica gel, 40 g, 0% to 80% EtOAc in hexanes) to give methyl 3-(2-(4- chlorophenyl)-2-hydroxyethoxy)-4-(4-fluorophenylsulfonamido) benzoate (1.43 g, 2.98 mmol, 96.1 % yield).

[341] Similarly prepared, but replacing 4-fluorobenzene-l -sulfonyl chloride with the appropriate sulfonyl chloride, were the following:

methyl 3-(2-(4-chlorophenyl)-2-hydroxyethoxy)-4-

(phenylsulfonamido)benzoate;

methyl 3-(2-(4-chlorophenyl)-2-hydroxyethoxy)-4-(pyridin-2- sulfonamido)benzoate; and

methyl 3-(2-(4-chlorophenyl)-2-hydroxyethoxy)-4-(pyridin-3- sulfonamido)benzoate.

Step 5 Methyl 3-(4-chlorophenyl)-4-(4-fluorophenylsulfonyl)-3,4-dihydro-2H - benzo[b] [ 1 ,4]oxazine-7-carboxylate

[342] Diethyl azodicarboxylate (1.87 g, 1.7 mL, 10.7 mmol, Eq: 3.6) was added dropwise to a solution of methyl 3-(2-(4-chlorophenyl)-2-hydroxyethoxy)-4-(4- fluorophenylsulfonamido)benzoate (1.43 g, 2.98 mmol, Eq: 1.00) and triphenylphosphine (2.5 g, 9.54 mmol, Eq: 3.2) in THF (40 mL). The reaction mixture was stirred at room temperature for 1 h and then was concentrated in vacuo. The bulk of the

triphenylphosphine oxide was removed by a quick flash column (silica gel, 40 g, 100% CH ₂ CI ₂ ). A second flash column was then run (silica gel, 40 g, 0% to 30% EtOAc in hexanes) and the resulting product was treated with EtOAc/hexanes to give methyl 3-(4- chlorophenyl)-4-(4-fluorophenylsulfonyl)-3,4-dihydro-2H-benz o[b][l,4]oxazine-7- carboxylate (1.18 g, 2.55 mmol, 85.7 % yield) as a white solid.

Step 6 3-(4-Chlorophenyl)-4-(4-fluorophenylsulfonyl)-3,4-dihydro-2H - benzo[b][l ,4]oxazine-7-carboxylic acid

[343] A solution of lithium hydroxide monohydrate (0.93 g, 22.1 mmol, Eq:

10.2) in water (40 mL) was added to a solution of methyl 3-(4-chlorophenyl)-4-(4- fluorophenylsulfonyl)-3,4-dihydro-2H-benzo[b][l ,4]oxazine-7-carboxylate (1.0 g, 2.17 mmol, Eq: 1.00) in a mixture of THF (40 mL) and methanol (25 mL). The resulting mixture was stirred at room temperature for 4 h, then poured into 100 mL 1 M aqueous HCl and extracted with EtOAc. The combined organic layers were concentrated in vacuo and the resulting residue treated with EtOAc and hexanes to give 3-(4-chlorophenyl)-4- (4-fluorophenylsulfonyl)-3,4-dihydro-2H-benzo[b][l,4]oxazine -7-carboxylic acid (0.97 g, 2.17 mmol, 100 % yield) as a white solid.

Step 7 3-(4-Chloro-phenyl)-4-(4-fluoro-benzenesulfonyl)-3 ,4-dihydro-2H- benzo[ 1 ,4]oxazine-7-carbonyl chloride

[344] Thionyl chloride (1.63 g, 1 mL, 13.7 mmol, Eq: 30.7) was added to a solution of 3-(4-chlorophenyl)-4-(4-fiuorophenylsulfonyl)-3,4-dihydro-2H - benzo[b][l,4]oxazine-7-carboxylic acid (0.20 g, 0.45 mmol, Eq: 1.00) in DME (1 mL) and the resulting mixture was stirred at room temperature for 3 h. The mixture was then concentrated in vacuo and the residue azeotroped with toluene (5 mL). The resulting crude acid chloride (0.20 g) was used in the next step without further purification.

Step 8 (3-(4-chlorophenyl)-4-(4-fluorophenylsulfonyl)-N-(2-methoxye thyl)-3,4- dihydro-2H-benzo[b] [ 1 ,4]oxazine-7-carboxamide

[345] A solution of 3-(4-chlorophenyl)-4-(4-fluorophenylsulfonyl)-3 ,4-dihydro-

2H-benzo[b][l ,4]oxazine-7-carbonyl chloride (50 mg, 107 umol, Eq: 1.00) in CH ₂ CI ₂ (2 mL) was added to a solution of 2-methoxyethanamine (35.0 mg, 40 uL, 465 umol, Eq:

4.34) in CH ₂ CI ₂ (2 mL) and the resulting mixture stirred for 30 min at room temperature. The reaction mixture was poured into 20 mL EtOAc and washed with 1 M aqueous HCl, sat. aqueous NaHC0 ₃ , and brine. The organic layer was concentrated in vacuo, and the resulting crude material was purified by flash chromatography (silica gel, 4 g, 0% to 40% EtOAc in hexanes) to give (3-(4-chlorophenyl)-4-(4-fiuorophenylsulfonyl)-N-(2- methoxyethyl)-3,4-dihydro-2H-benzo[b][l ,4]oxazine-7-carboxamide (52 mg, 103 umol, 96.0 % yield), MS (M+H) = 506.

[346] Replacement of 2-methoxyethanamine with various other amines allowed preparation of various corresponding amides. Additional compounds made by the above procedure are shown in Table 1. Example 2

[3-(4-Chloro^henylV4-(2-fluoro-benzenesulfonylV3.4-dihydro-2 H-benzo[1.4]oxazin-7- yl] - (2 -methoxy-ethyl)- amine

[347] The synthetic procedure used in this preparation is outlined in Scheme D.

SCHEME D

Step 1 2-(5-Bromo-2-nitrophenoxy)-l-phenylethanone

[348] To a mixture of 5-bromo-2-nitrophenol (719 mg, 3.3 mmol, Eq: 1.00) and cesium carbonate (1.29 g, 3.96 mmol, Eq: 1.2) was added 2-bromoacetophenone (788 mg, 3.96 mmol, Eq: 1.2) to give a bright orange mixture. The reaction mixture was stirred for 3 h at room temperature. The reaction mixture was diluted with 200 mL EtOAc, washed with sat. aqueous NaHC0 ₃ and brine, and then concentrated in vacuo. The resulting residue was stirred with EtOAc for 30 min, then hexanes were added and the resulting precipitate was collected by filtration. Additional product was obtained from the mother liquor by concentration and purification by flash chromatography (silica gel, 40 g, 50% CH ₂ CI ₂ in hexanes), giving 2-(5-bromo-2-nitrophenoxy)-l-phenylethanone (1.1 g, 3.27 mmol, 99.2 % yield) as a pale yellow solid.

Step 2 2-(5-Bromo-2-nitrophenoxy)-l -phenylethanol

[349] A suspension of 2-(5-bromo-2-nitrophenoxy)-l -phenylethanone (1.01 g, 3 mmol, Eq: 1.00) in methanol (70 mL) was cooled in an ice bath. Sodium borohydride (0.17 g, 4.49 mmol, Eq: 1.5) was added, the cooling bath removed, and the reaction mixture stirred at room temperaturefor 2 h. The reaction mixture was concentrated to a final volume of ~ 20 mL, and then sat. aqueous NH ₄ CI (40 mL) was added. The product was extracted into EtOAc, and the combined organic layers were washed with sat.

aqueous NaCl and then concentrated in vacuo to give 2-(5-bromo-2-nitrophenoxy)- 1 - phenylethanol (1.0 g, 2.96 mmol, 98.6 % yield) as a yellow oil.

Step 3 2-(2-Amino-5-bromophenoxy)-l-phenylethanol

[350] In a 500 mL round-bottomed flask equiped with a 2" magnetic stir bar, 2-

(5-bromo-2-nitrophenoxy)-l -phenylethanol (1.0 g, 2.96 mmol, Eq: 1.00), iron (0.83 g, 14.9 mmol, Eq: 5.03) and ammonium chloride (0.80 g, 15.0 mmol, Eq: 5.06) were combined in ethanol (25.0 ml) and water (10.0 ml). The mixture was then heated at 80°C with rapid stirring for 2.5 h. The hot reaction mixture was filtered through celite and the filtrate was concentrated in vacuo. The resulting residue was treated with sat. aqueous Na ₂ C0 ₃ , and the product was then extracted into EtOAc. The combined organic layers were washed with brine and concentrated in vacuo. The crude material was purified by flash chromatography (silica gel, 40 g, 0% to 50% EtOAc in hexanes then (silica gel, 40 g, 0% to 30% acetone in hexanes) to give 2-(2-amino-5-bromophenoxy)-l -phenylethanol (0.73 g, 2.37 mmol, 80.1 % yield) as an oil.

Step 4 N-(4-bromo-2-(2-hydroxy-2-phenylethoxy)phenyl)-2- fluorobenzenesulfonamide

[351] 2-(2-amino-5-bromophenoxy)-l -phenylethanol (0.25 g, 81 1 μιηοΐ, Eq:

1.00), 2-fluorobenzene-l -sulfonyl chloride (0.21 g, 1.08 mmol, Eq: 1.33) and pyridine (88.0 mg, 90 μL, 1.11 mmol, Eq: 1.37) were combined in CH ₂ C1 ₂ (10 mL) and the resulting solution was stirred at room temperature for 3 h. The reaction mixture was diluted with EtOAc (100 mL), washed with 1 aqueous M HC1, sat. aqueous NaHC0 ₃ , and sat. aqueous NaCl, and then concentrated in vacuo. The crude material was sonicated with 10% EtOAc in hexanes for 10 min, stirred for 30 min at room temperature and then allowed to stand at room temperatur for 72 h. The mother liquor was decanted off and the product ( -(4-bromo-2-(2-hydroxy-2-phenylethoxy)phenyl)-2- fluorobenzenesulfonamide) was used directly in the next step without further purification. Step 5 7-Bromo-4-(2-fluorophenylsulfonylV3-phenyl-3.4-dihydro-2H- benzo [b] [ 1 ,4] oxazine

[352] Diethyl azodicarboxylate (308 mg, 0.28 mL, 1.77 mmol, Eq: 2.21) was added dropwise to a solution of N-(4-bromo-2-(2-hydroxy-2-phenylethoxy)phenyl)-2- fluorobenzenesulfonamide (0.373 g, 800 μηιοΐ, Eq: 1.00) and triphenylphosphine (0.47 g, 1.79 mmol, Eq: 2.24) in THF (10 mL). The reaction mixture was stirred at room temperature for 2 h, then MeOH ( 1 mL) was added and the reaction mixture was stirred for 12 h. The crude reaction mixture was concentrated in vacuo and triphenylphosphine oxide was removed by a quick flash column (silica gel, 40 g, 100% CH2C12) to give 7- bromo-4-(2-fluorophenylsulfonyl)-3-phenyl-3,4-dihydro-2H-ben zo[b][l,4]oxazine (0.27 g, 602 μηιοΐ, 75.3 % yield) as an oil.

Step 6 [3-(4-Chloro-phenyl)-4-(2-fluoro-benzenesulfonyl)-3 ,4-dihydro-2H- benzo[ 1 ,4]oxazin-7-yl]-(2-methoxy-ethyl)-amine

[353] 2-Methoxy-ethylamine (19 mg, 0.223 mmol), sodium tert-butoxide (21 mg,

0.223 mmol), bis(tri-t-butylphosphine)palladium(0) (19 mg, 0.037 mmol) and (N-(4- bromo-2-(2-hydroxy-2-phenylethoxy)phenyl)-2-fluorobenzenesul fonamide) (80 mg,

0.166 mmol) were added to dioxane (1.5 mL) in a reaction via. The vial was sealed and heated at 80°C for two hours with stirring, then cooled to room temperature and stirred for 18 hours. The reaction mixture was diluted with EtOAc (25 mL) and washed with sat. aqueous NaHC0 ₃ , and sat. aqueous NaCl, and then concentrated in vacuo. The crude material was purified by preparative TLC (silica gel, 1 : 1 Hexanes/EtOAc) to give 10 mg of [3-(4-chloro-phenyl)-4-(2-fluoro-benzenesulfonyl)-3,4-dihydr o-2H-benzo[l,4]oxazin- 7-yl]-(2-methoxy-ethyl)-amine, MS (M+H) = 478. Example 3

Formulations

[354] Pharmaceutical preparations for delivery by various routes are formulated as shown in the following Tables. "Active ingredient" or "Active compound" as used in the Tables means one or more of the Compounds of Formula I.

Com osition for Oral Administration

[356] The ingredients are combined and granulated using a solvent such as methanol. The formulation is then dried and formed into tablets (containing about 20 of active compound) with an appropriate tablet machine.

Composition for Oral Administration

Ingredient Amount

Active compound 1.0 g

Fumaric acid 0.5 g

Sodium chloride 2.0 g

Methyl paraben 0.15 g

Propyl paraben 0.05 g

Granulated sugar 25.5 g

Sorbitol (70% solution) 12.85 g

Veegum K (Vanderbilt Co.) l .O g

Flavoring 0.035 ml

Colorings 0.5 mg

Distilled water q.s. to 100 ml [357] The ingredients are mixed to form a suspension for oral administration.

Parenteral Formulation

[358] The active ingredient is dissolved in a portion of the water for injection. A sufficient quantity of sodium chloride is then added with stirring to make the solution isotonic. The solution is made up to weight with the remainder of the water for injection, filtered through a 0.2 micron membrane filter and packaged under sterile conditions.

[360] All of the ingredients, except water, are combined and heated to about

60°C with stirring. A sufficient quantity of water at about 60°C is then added with vigorous stirring to emulsify the ingredients, and water then added q.s. about 100 g.

Nasal Spray Formulations

[361] Several aqueous suspensions containing from about 0.025-0.5 percent active compound are prepared as nasal spray formulations. The formulations optionally contain inactive ingredients such as, for example, microcrystalline cellulose, sodium carboxymethylcellulose, dextrose, and the like. Hydrochloric acid may be added to adjust pH. The nasal spray formulations may be delivered via a nasal spray metered pump typically delivering about 50-100 microliters of formulation per actuation. A typical dosing schedule is 2-4 sprays every 4-12 hours.

Example 4

ASSAYS

[362] Compounds provided herein can be evaluated using cell-based assays such as electrophysiological assays, or can be evaluated in animal models ofautoimmune disease and immune-mediated responses and conditions. Examples of assays are described below.

[363] The compounds provided herein can be tested for antagonist activity at voltage-gated ion channels such as voltage-gated potassium channels by measuring their ability to affect channel opening or their ability to block permeation of ions through the channel pore in response to depolarizing voltage changes across the cell membrane. Functional tests of channel activity include but are not limited to ion flux resulting from channel opening measured by electrophysiological methods. These methods can be used to evaluate channel function when the relevant ion channel is heterologously expressed in a mammalian or amphibian cells. These methods can also be used to evaluate compounds provided herein in rodent primary lymphocytes and other mammalian primary cells and cell lines that endogenously express the channel of interest.

[364] Compounds can further be evaluated for their ability to bind the ion channel using biochemical approaches. Compounds can also be evaluated for their ability to modify lymphocyte signaling where the channels are known to have a role (e.g., cytokine production and cellular proliferation). Finally, compounds provided herein can be tested in vivo in animal models of immune -mediated processes and disorders and autoimmune diseases known to one skilled in the art, such as, for example, models of multiple sclerosis, arthritis, type I diabetes, and hypersensitivity in rodents or other mammals.

[365] The following biological examples are offered to illustrate the compounds, pharmaceutical compositions and methods provided herein and are not to be construed in any way as limiting the scope thereof. Cell Culture

[366] Chinese hamster ovary (CHO) cell lines stably expressing myc-epitope tagged K _v l .3 and flag-epitope tagged Kvl .5 were generated by transfection of the respective cDNA into the cells, subsequent selection exploiting the blastcidin marker and re-cloning to obtain a clonal cell line. Expression of K _v l .3 and K _v l .5 was followed by immunofluorescence analysis using the anti myc- (Kvl .3) or anti-flag (K _v 1.5)-antibody and subsequently by electrophysiological investigations (see below). Cells expressing the ion channels were maintained and cultivated twice weekly in MEM-alpha medium (Sigma) supplemented with 10% fetal calf serum (FCS), 1% glutamine (Sigma) and 1% Pen/Strep (Invitrogen). For the fluorometric assay cells were plated into Corning Costar 384 tissue culture plates 24 h prior to screening using a Multidrop (Labsystems). Cells were plated at 10,000 cells per well in a total volume of 75 μL and incubated overnight at 37°C and 5% C0 ₂ . For electrophysiological investigations using the manual patch clamping rig cells were plated the day before the analysis in 2cm-dishes at a density of 2.5E4 cells/ml. For tests on the planar patch clamping systems cells were harvested using trypsin/EDTA (Sigma), washed twice in PBS, and used at a density of approx 5E6/ml. Fluorometric assay

[367] Changes in membrane potential were detected through the use of a membrane -potential sensitive fluorescent dye (FMP; Molecular Devices). The buffer system used was carefully selected to optimize the cell membrane potential and favour conditions where the number of ion channels in the open state would be at a maximum. The FMP dye was re-suspended in 10 ml of assay buffer according to the supplier's protocol. Cell plates were washed using a standard 384 plate washer with assay buffer, incubated with dye, and diluted in assay buffer for 5 min before compound addition. Cell plates were subsequently incubated with compounds for 60min and different

concentrations, after which time the change in fluorescent signal was measured.

Fluorescent recordings were performed on the Tecan Saflre (Tecan, Crailsheim,

Germany) using excitation and emission filters of 540 and 555 nm, respectively, and incorporating a bandwidth of 5 nm (Journal of Biomolecular Screening 2006:57-64). Electrophysiology - manual patch clamping

[368] Patch-clamp experiments were performed in the voltage-clamp mode and whole-cell currents were recorded in (extracellular solution) NaCl 130mM, KC1 5.4mM, MgCl ₂ ImM, CaCl ₂ 1.8mM, HEPES 10 mM, and glucose 5 mM, pH 7.4, with aOH. Intracellular solution was KC1 130mM, HEPES lOmM, glucose 5 mM, EGTA 1 mM, ATP-Mg 2 mM, and MgCl ₂ 1 mM, pH 7.2, with KOH. For investigating effects and reversibility of a test compound on the potassium channels CHO-cells stably expressing the ion channels were clamped at a holding potential (HP) of-80 mV, hyperpolarized to -90 mV to determine the leak current, and depolarized for 200 ms to +40 mV. Intersweep interval was 20 sec. Pulse-cycling rate was 0.1 Hz. The area under curve (charge) during 200 ms of depolarization was analyzed. An initial period of 20 stimuli was recorded to determine a biexponential fit of endogenous current rundown. The total charge obtained is plotted against antagonist concentration on a logarithmic scale and IC50s are obtained using standard analysis protocols. After compound application, a washout period of approx. 5 min was recorded (Journal of Biomolecular Screening 2006:57-64).

Electrophysiology -patch clamping using the PatchLiner and the Port-a-Patch

[369] Recordings were made at room temperature using the Patchliner or Port-a-

Patch planar patch-clamp systems ( anion, Munchen, Germany) in whole-cell mode according to Nanion 's standard procedures. Prior to recordings cells were detached from culture flasks with 0.05% trypsin/EDTA and re-suspended at a density of approx. 5E6/ml cells per ml. The bath (external) solution contained (mM): NaCl 140, KC14, MgCl ₂ 1 , CaCl ₂ 2, Hepes 10, glucose 5; titrated to pH 7.4 with NaOH. The intracellular recording solution contained (mM): KC1 50, KF 60, NaCl 10, EGTA 20, HEPES 10; titrated to pH 7.2 with KOH. For investigating effects and reversibility of a test compound on the potassium channels CHO-cells stably expressing the ion channels were clamped at a holding potential (HP) of-100 mV, hyperpolarized to -120 mV to determine the leak current, and depolarized for 200 ms to +40 mV. Intersweep interval was 20 sec. The area under curve (charge) during 200 ms of depolarization was analyzed. An initial period of approx. 15 stimuli was recorded to determine an endogenous current rundown. In case of the Port-a-Patch the rundown was biexponentially fitted. In case of the PatchLiner the block of the compounds were referred to the charge obtained just prior to compound application. The total charge obtained is plotted against antagonist concentration on a logarithmic scale and IC50s are obtained using standard analysis protocols. After compound had been added a washout was recorded to analyse the reversibility of the block. Electrophysiological Experiments

Whole cell patch clamp:

[370] Whole cell recordings are made either manually using a Multiclamp 200B patch-clamp amplifier and Clampex acquisition program (Molecular Devices

Corporation) or by the automated 16 cell planar patch clamp instrument, QPatch

(Sophion Bioscience) or Whole-cell recordings are obtained from CHO cells stably or transiently transfected with cDNA of respective ion channels. For the manual patch- clamp recordings, solutions are either applied for periods of 1 to 3s by a gravity flow, 8- valve delivery system, or for periods of milliseconds using the quick-change Dynaflow perfusion system (Cellectricon Inc.). The internal pipette solution may include 60 mM Potassium-Chloride, 60 mM Potassium Flouride, 20 mM EGTA, and 5 mM Hepes at pH 7.2; normal external solution is 140 mM NaCl, 5 mM KC1, 1 mM CaCl _2, 2 mM MgCl ₂ , 25 mM Hepes, and 10 mM glucose. Concentration-response curves are obtained by recording currents in response to brief (200 msec) depolarizing voltage steps at 15 second intervals. A series of baseline current responses are obtained in the presence of regular external saline. To obtain inhibition curves, compounds are applied to the cells at increasing concentrations for a defined time period while obtaining current responses. The periods of compound application at each concentration and the number of voltage steps applied to the cell at each concentration are constant for the entire test concentration series. Currents are measured in cells that are voltage clamped at -90 millivolts.

Averaged steady- state current level at the end of the 3 minute compound application is measured and normalized to the maximum current response in control saline. These points are then plotted on a logarithmic scale and fitted by a Hill function. The IC ₅₀ is calculated from the resulting Hill fit.

[371] Compounds that inhibit K _v l .3 currents are tested for activity against other receptors to determine their selectivity for specific family members. The list of receptors assayed includes, but is not restricted to K _v l .1 , K _v l .5 , K _v l .4, K _v 3.1 , K _v 2.1 , hERG. The types of assay used for selectivity determination may include Electrophysiological determination of receptor inhibition in either mammalian cells stably expressing DNA encoding the channel of interest or inherently expressing the channel, including by using manual patch clamp or planar patch clamp system. Methods and data analysis will be similar to those described above for K _v l .3.

[372] Compounds of the invention were found to be active for Kvl .3. IC50 values for representative compounds are shown in Table 1.

In Vivo Assays:

[373] Various in vivo inflammatory, and visceral pain assays can be conducted in male Sprague-Dawley rats weighing 250-350 g. K _vl .3 modulators may be

administered prior to or post-induction of the inflammation model depending upon the specific model and the compound PK characteristics. The route of administration may include intraperitoneal, (i.p.), subcutaneous (s.c), oral (p.o.), intranvenous (i.v.), intrathecal (i. ), or intraplantar.

[374] Various models of autoimmune disease or immune -mediated disorders or diseases are available and may be suitable for testing compounds. The K _v l .3 modulators may be administered prior to or post-induction of the immune disease model depending upon the specific model and the compound PK characteristics. The route of

administration may include intraperitoneal, (i.p.), subcutaneous (s.c), oral (p.o.), intranvenous (i.v.), intrathecal (i.t.), or intraplantar. The endpoints for these studies may include mediation of immune reactions, inflammatory reactions, physical parameters, cellular parameters and will be appropriate for the model being utilized.

Multiple Sclerosis

EAE Model:

[375] Several animal models are utilized for testing compounds for effectiveness in treating MS. One immune -mediated model is the Experimental Allergic

Encephalomyelitis (EAE) model where injection of certain myelin components into genetically susceptible animals leads to T cell-mediated CNS demyelination. EAE is a T cell mediated autoimmune disease of the central nervous system (CNS). Disease is induced in susceptible strains of mice (SJL mice) by immunization with CNS myelin antigens or alternatively, disease is passively transferred to susceptible mice using antigen stimulated CD4+ T cells (Pettinelli, J. Immunol. 127, 1981, p. 1420). EAE is widely recognized as an acceptable animal model for multiple sclerosis in primates (Alvord et al. (eds.) 1984. Experimental allergic encephalomyelitis~A useful model for multiple sclerosis. Alan R. Liss, New York).

TMEV Model:

[376] Another experimental MS model is a viral model, whereby MS like disease is induced by Theiler's murine encephalomyelitis virus (TMEV) (Dal Canto, M.C., and Lipton, H.L., Am. J. Path., 88:497-500 (1977)). The TMEV picornavirus is inoculated intracerebrally in susceptible strains of mice and induces immune-mediated progressive CNS demyelination which is clinically and pathologically similar to MS. Intracerebral virus injection is performed on 4-6 week old animals that are lightly anesthetized with metofane. Virus is injected using a 27 gauge needle with a Hamilton syringe that delivers a 10 ul volume which contains 2xl0 ⁵ PFU of the Daniel's strain of TMEV. Intracerebral injection results in greater than 98% incidence of chronic viral infection with demyelination. Chronically infected animals for remyelination experiments are generally 6-8 months post-infection. Animals are sacrificed and spinal cord tissue is processed and morphological evaluation of the spinal cord of each animal is assessed histologically. Mice are anesthetized with pentobarbital and perfused by intracardiac administration of fixative (phosphate buffered 4% formaldehyde with 1% glutaraldehyde, pH 7.4). Spinal cords are removed and sectioned coronally into 1 mm blocks, postfixed with osmium, and embedded in araldite. One micron-thick cross- sections are cut from each block and stained with 4% paraphenyldiamine. Demyelination and remyelination are quantified using a Zeiss digital analysis system (ZIDAS) and camera lucida . For each mouse, ten spinal cord cross sections are examined which span the entire cord from the cervical to the proximal coccygeal spinal column regions. Areas of demyelination are characterized by large amounts of myelin debris, macrophages engulfing debris, cellular infiltration and naked axons. Oligodendrocyte remyelination is characterized by areas of axons with abnormally thin myelin sheaths and the absence of Schwann cells. Statistical comparison of the extent of demyelination and remyelination is performed using the Student's t test. Lysolecithin Induced Demyelination:

[377] 12 weeks old SJL/J mice are anesthetized with sodium pentobarbitol and a dorsal laminectomy is performed in the upper thoracic region of the spinal cord. A 34 guage needle attached to a Hamilton syringe is used to inject 1 ml of a 1% solution of lysolecithin directly into the dorsolateral aspect of the cord. The wound is sutured in two layers, and mice are allowed to recover. The day of lysolecithin injection was designated day 0. Seven days after lysolecithin injection, mice are treated with compound(s) (1 mg/injection each). Control mice are treated with bolus intraperitoneal injection of PBS or other appropriate control. Three weeks and five weeks after the lysolecithin injection, mice are sacrificed and one um thick sections are prepared. The araldite block showing the largest lysolecithin-induced demyelination lesion is used for quantitative analysis. The total area of the lesion is quantitated using a Zeiss interactive digital analysis system. The total number of remyelinated fibers is quantitated using a Nikon

microscope/computer analysis system. The data is expressed as number of remyelinated axons/mm of lesion.

Arthritis

[378] The collagen-induced arthritis model (CIA) is used as an autoimmune model for testing compounds (Myers, L.K. et al. Life Sci. (1997), 61(19): 1861 -1878). Immunization of genetically susceptible rodents or primates with Type II collagen (CII) in complete Freund's adjuvant leads to the development of a severe polyarticular arthritis after about 3 weeks. Synovitis and erosions of cartilage and bone are observed as the hallmarks of CIA and mimic RA. DBA 1 mice aged between 10 and 12 weeks are administered Hypnorm™ (0.1 ml, intraperitoneally) and shaved at the base of the tail. Bovine collagen type II (CII) is emulsified with complete Freund's adjuvant at a final concentration of 2 mg/ml, and a total of 0.1 ml is injected intradermally at three sites at the base of the tail. Twenty-one days later, a booster (0.1 ml) consisting of CII emusified with incomplete Freund's adjuvant (2 mg/ml) is injected intradermally across three sites at the tail base. A further 3 days later animals are injected with lipopolysaccharide (40 μg in 0.1 ml phosphate -buffered saline [PBS]; E. coli serotype 055:B5; Sigma- Aldrich Co. Ltd, Poole, UK) intraperitoneally to synchronize disease (Caccese R, et al (1992) Mediators Inflamm 1 :273-279) The development and progression of arthritis is monitored every 2-3 days and assigned a clinical score based on visual signs of arthritis (0.25 = swelling in a single digit; 0.5 = swelling in more than one digit; 1 = swelling and erythema of the paw; 2 = swelling of the paw and ankle; 3 = complete inflammation of the paw; the maximum score for each mouse was therefore 12) and the thickness of hind paws is measured using POCO 2T calipers (Krceplin Langenmesstechnik, Schluchtern, Germany) . Animals with a clinical score of 0.5 or above are used in compound assessment experiments and administered varying doses of test compound. Mice are monitored until 40 days after immunization, when they were killed and blood collected for serum and draining lymph nodes collected for assessment of stimulated cytokine secretion.

Diabetes

[379] The main clinical feature of IDDM is elevated blood glucose levels

(hyperglycemia) which is caused by auto-immune destruction of insulin-producing β- cells in the islets of Langerhans of the pancreas. A useful animal model in testing compounds for treating IDDM is the NOD mouse. The NOD mouse represents a model in which auto -immunity against beta-cells is the primary event in the development of IDDM. Diabetogenesis is mediated through a multi-factorial interaction between a unique MHC class II gene and multiple, unlinked, genetic loci, as in the human disease.

[380] Another animal model for studying the effects of compounds in IDDM is the streptozotocin (STZ) model (Hartner, A. et al. (2005), BMC Nephrol. 6(1):6). This model has been used extensively as an animal model to study the mechanisms involved in the destruction of pancreatic beta cells in IDDM. Diabetes is induced in rodents by the beta-cell toxin streptozotocin (STZ). STZ is taken up by the pancreatic beta cell through the glucose transporter GLUT-2. This substance decomposes intracellularly, and causes damage to DNA either by alkylation or by the generation of NO. The appearance of DNA strand breaks leads to the activation of the abundant nuclear enzyme poly(ADP-ribose) polymerase (PARP), which synthesizes large amounts of the (ADP-ribose) polymer, using NAD+ as a substrate. As a consequence of PARP activation, the cellular concentration of NAD+ may then decrease to very low levels, which is thought to abrogate the ability of the cell to generate sufficient energy and, finally, to lead to cell death. Delayed Type Hypersensitivity

[381] A pediatric animal model using the delayed type hypersensitivity (DTH) skin test is utilized to further evaluate compounds. Sprague Dawley rats (1 week old) are sensitized against keyhole limpet hemocyanin (KLH). Animals are then challenged 2 weeks later by an intradermal injection of KLH (0.3 mg) in sterile saline. Rats with positive DTH skin reactions at 24 and 48 hours after challenge (baseline) are divided randomly into control and compound treatment groups. Before each procedure (day 0) and on postoperative days 3 and 6, animals are again challenged intradermally with KLH (0.3 mg). DTH skin reaction is evaluated 24 and 48 hours later.

[382] Cobb et al have also described a model utilizing a hairless guinea pig for the determination of delayed-type hypersensitivity to tuberculin (Cobb A et al (2001) Int Immunopharmacology l(2):349-353). This DTH model uses hairless (IAF/HA-HO) guinea pigs, sensitized with complete Freund's adjuvant and repeatedly skin tested with tuberculin, purified protein derivative, (PPD). At 10 weeks after sensitization, intradermal PPD elicited minimal erythema at 6 h, which increased over the next 18 h to a maximum at 24 h, and declined by 48 h. The response could be quantified by bioassay using graded doses of PPD. Reactions at 24 h were characterized by predominantly mononuclear cell deep and superficial dermal infiltrate.

Graft Rejection

[383] Compounds are tested for prevention or alteration of human T-cell mediated skin graft rejection in a reconstituted mouse model of skin transplantation. Human T-cell mediated destruction of allogeneic diurnal microvessels is evaluated in a severe combined immunodeficient mouse in accordance with Murray et al (Murray, A.G. et al (1994) PNAS USA 91 :9146-9153).

[384] The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions and methods provided herein and are not to be construed in any way as limiting their scope. In the examples, all temperatures are in degrees Celsius (unless otherwise indicated). Compounds that can be prepared in accordance with the methods provided herein along with their biological activity data are presented in following Table. The syntheses of these representative compounds are or can be carried out in accordance with the methods set forth above. [385] While the present invention has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective spirit and scope of the present invention. All such modifications are intended to be within the scope of the claims appended hereto.