US007271274B2

(12) United States Patent (10) Patent N0.: US 7,271,274 B2 Meng et a]. (45) Date of Patent: Sep. 18, 2007

(54) PHENOLIC ANTIOXIDANTS FOR THE 6,881,860 B2 4/2005 Sikorski et al. TREATMENT OF DISORDERS INCLUDING 2002/0193446 A1 12/2002 Meng ARTHRITIS, ASTHMA AND CORONARY 2003/0064967 A1 4/2003 Luchoomun et al. ARTERY DISEASE 2004/0266879 A1 12/2004 Sikorski et al. 2005/0065121 A1 3/2005 Sikorski et al. (75) Inventors: Charles Q. Meng, Duluth, GA (US); i/(fmers M. David. WeIngarten,. Cummmg,. GA eng et a1. (Us) FOREIGN PATENT DOCUMENTS (73) Assignee: AhteroGenics, Inc., Alpharetta, GA EP 0 348 203 A1 12/1989 (Us) EP 0 405 788 A2 1/1991 EP 0 621 255 A1 10/1994 ( * ) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 (Continued) USC‘ 1540’) by 0 days‘ OTHER PUBLICATIONS (21) Appl. NO-Z 11/111,196 Barnhart, J.W., et al., “Chapter 10: The Synthesis, metabolism, and biological activity of probucol and its analogs,” Pharmacochem. (22) Filed: Apr. 20, 2005 Lib/1, in Antilipidemic Drugs: Medicinal, Chemical, and Biochemi cal Aspects, Witiak et al., Eds., (Elsevier Science: Amsterdam, (65) Prior Publication Data 1991), pp. 277-299. Baron, J .L., et al., “The pathogenesis of adoptive murine autoim US 2006/0020038 A1 Jan' 26> 2006 mune diabetes requires an interaction between a4-integrins and vascular cell adhesion molecule-1,” J. Clin. Invest, 93(4-6):1700 Related US. Application Data 1703 (1994), (60) Provisional application No. 60/600,029, ?led on Aug. (Continued) 9, 2004, prov1s1onal appl1cat1on No. 60/564,043, ?led on Apr' 20, 2004_ Primary ExamineriShailendra Kumar (74) Attorney, Agent, or F irmiKing & Spalding (51) Int. Cl. C07D 317/10 (2006.01) (57) ABSTRACT A61K 31/335 (2006.01) C07C 309/00 (2006.01) The invention relates to compounds, pharmaceutical com (52) US. Cl...... 549/449; 549/450; 549/453; positions comprising the compounds, and methods of using 514/467 the compounds, Wherein the compounds are of the following (58) Field of Classi?cation Search ...... 514/562, Formulas: 514/467, 618, 712; 564/162; 562/442; 568/47; 549/449, 453, 450 See application ?le for complete search history. I (56) References Cited S S U.S. PATENT DOCUMENTS 4,029,812 A 6/1977 Wagner et al. 4,076,841 A 2/1978 Wagner et al. HO I I.» 4,078,084 A 3/1978 Wagner et al. 4,752,616 A 6/1988 Hall et a1. | 4,954,514 A 9/1990 Kita et a1. RI 5,155,250 A 10/1992 Parker et a1. 5,206,247 A 4/1993 Regnier et a1. 5,262,439 A 11/1993 Parthasarathy et al. S 5,608,095 A 3/1997 Parker et a1. 5,627,205 A 5/1997 Regnier et a1. 5,750,351 A 5/1998 Medford et a1. 5,773,209 A 6/1998 Medford et a1. 5,773,231 A 6/1998 Medford et a1. 5,807,884 A 9/1998 Medford et a1. 5,811,449 A 9/1998 Medford et a1. 5,846,959 A 12/1998 Medford et a1. 6,121,319 A 9/2000 Somers et al. or a pharmaceutically acceptable salt, ester or prodrug 6,147,250 A 11/2000 Somers thereof, Wherein the substituents are de?ned in the applica 6,548,699 B1* 4/2003 Somers ...... 562/431 6,602,914 B2 8/2003 Meng tion. The invention further provides methods of treatment of 6,617,352 B2 9/2003 Somers in?ammatory disorders by administering the compounds. 6,828,447 B2 12/2004 Meng 6,852,878 B2* 2/2005 Meng et a1...... 562/42 25 Claims, No Drawings US 7,271,274 B2 Page 2

FOREIGN PATENT DOCUMENTS Lee, S.J., et al., “Adhesion molecule expression and regulation on cells of the central nervous system,” J. Neuroimmunol, 98177-88 EP 0 763 527 A1 3/1997 (1998). FR 2130975 A5 11/1972 Marx, N., et al., “Peroxisome proliferator-activated receptors FR 2133024 A5 11/1972 (PPARs) and their role in the vessel wall: possible mediators of FR 2134810 A5 12/1972 cardiovascular risk?” J. Cardiovasc. Risk, 8(4)1203-210 (Aug. FR 2140769 A5 1/1973 2001). FR 2140771 A5 1/1973 Meng, C.Q., et al., “NitrobenZene compounds inhibit expression of FR 2168137 A1 8/1973 VCAM-1,” Bioorganic & Medicinal Chemistry Letters, WO WO95/15760 A1 6/1995 11(14)11823-1827 (Jul. 23, 2001). WO WO95/30415 A1 11/1995 Meng, C.Q., et al., “Novel phenolic antioxidants as multifunctional WO WO97/15546 A1 5/1997 inhibitors of inducible VCAM-1 expression for use in atheroscle WO WO98/51289 A2 11/1998 rosis,” Bioorganic & Medicinal Chemistry Letters, 12:2545-2548 WO WO98/51662 A1 11/1998 (2002). W0 WO 00/26184 A1 5/2000 Meng, C.Q., “Probucol (Restenosis) Daiichi,” Curr Opin. W0 WO 01/07057 A2 9/2001 Cardiovasc. Pulm. Renal Invest. Drugs, 2(3)1294-298 (2000). OTHER PUBLICATIONS Morales-Ducret, J., et al., “014/51 integrin (VLA-4) ligands in arthritis: vascular cell adhesion molecule-1 expression in synovium Burkly, L.C., et al., “Protection against adoptive transfer of autoim and on ?broblast-like synoviocytes”, J. Immunol., 149(4)11424 mune diabetes mediated through very late antigen-4 integrin,” 1431 (Aug. 15, 1992). Diabetes, 431529-534 (1994). Nakao, H., et al., “An Inhibitor of VCAM-1 Expression and Its Carew, T.E., et al., “Antiatherogenic effect of probucol unrelated to Implication as a Novel Treatment of In?ammatory Disease,” J. its hypocholesterolemic effect: Evidene that antioxidants in vivo can Atheroscler Thromb., 41149-155 (1998). selectively inhibit low density lipoprotein degradation in macroph Nicolaou, K.C., et al., “Total Synthesis of Ionophore Antibiotic age-rich fatty streaks and slow the progression of atherosclerosis in X-14547A,”J. Org. Chem., 5011440-1456 (1985). the Watanabe heritable hyperlipidemic rabbit,” Proc. Natl. Acad. Oguchi, S., et al., “Monoclonal Antibody Against Vascular Cell Sci. USA, 8417725-7729 (Nov. 1987). Adhesion Molecule-1 Inhibits Neointimal Formation after De Meglio, P., et al. “New derivatives of clo?brate and probucol. Periadventitial Carotid Artery Injury in Genetically Preliminary studies on hypolipe-mic activity,” Farmaco, Ed. Sci., Hypercholesterolemic Mice,” Arterioscler. Thromb. Vasc. Biol., 40(11), 833-844 (1985). [In Italian; abstract provided in English: 2011729-1736 (2000). Chem Abstr. AN 1986128675, DN 104128675; & partial translation Ohkawara, Y., et al., “In situ expression of the cell adhesion in English. molecules in bronchial tissues form asthmatics with air ?ow limi Elovaara, I., et al., “Adhesion Moleules in Multiple Sclerosis,” tation: In vivo evidence of VCAM-1/VLA-4 interaction in selective Arch. Neurol., 571546-551 (2000). eosinophil in?ltration,” Am. J Respir Cell Mol. Biol., 1214-12 Folkman, J. and Shing, Y., “Angiogenesis,” J. Biol. Chem., 267(16)110931-10934 (Jun. 5, 1992). (1995). Foster, C.A., et al., “Novel Inhibitor of Endothelial Cell-Associated Pilewski, J .M., et al., “Cell Adhesion Molecules is Asthma: Hom VCAM-l Expression,” Skin Pharmacol., 91149 (from 12th Annual ing, Activation, and Airway Remodeling,”Am. J Respir Cell. Mol. Meeting at pp. 141-168) (1996). Biol., 1211-3 (1995). Haddad, J .J ., et al., “Redox regulation of TNF-alpha biosynthesis: Rabb, H.A., et al., “The Role of the Leukocyte Adhesion Molecules augmentation by irreversible inhibition of gamma-glutamylcysteine VLA-4, LFA-l, and Mac-1 in Allergic Airway Responses in the synthetase and the involvement of an IkappaB-alpha/NF-kappa.B Rat,” Am. J. Respir Care. Med, 149:1186-1191 (1994). independent pathway in alveolar epithelial cells,” Cell Signal, Rival, Y., et al., “PPARalpha and PPARdelta activators inhibit 14(3)1211-218 (Mar. 2002). cytokine-induced nuclear translocation of NF-kappaB and expres Kallmann, B.A., et al., “Cytokine-induced modulation of cellular sion ofVCAM-l in EAhy926 endothelial cells,”Eur. J. Pharmacol., adhesion to human cerebral endothelial cells is mediated by soluble 435(2-3)143-151 (Jan. 25, 2002). vascular cell adhesion molecule-1,” Brain, 123:687-697 (2000). Schreiner, E.P., et al., “Inhibitors of vascular cell adhesion mol Koch, A.E., et al., “ImmunolocaliZation of endothelial and ecule-1 expression,” Expert Opin. Ther Patents, 13(2):149-166 leukocyte adhesion molecules in human rheumatoid and (2003). osteoarthritic synovial tissues”, Lab. Investig., 64(3):313-320 Tardif, J.C., et al., “Effects of AGI-1067 and probucol after (1991). percutaneous coronary interventions,” Circulation, 107(4)1552-558 Koch, A.E., et al., “Angiogenesis mediated by soluble forms of (Feb. 4, 2003). E-selectin and vascular cell adhesion molecule-1,” Nature, Yang, X.D., et al., “Inhibition of insulitis and prevention of diabetes 376(6540)1517-519 (Aug. 10, 1995). in nonobese diabetic mice by blocking L-selectin and very late KudlacZ, E., et al., “Pulmonary eosinophilia in a murine model of antigen 4 adhesion receptors,” Proc. Natl. Acad. Sci. USA, allergic in?ammation is attenuated by small molecule alpha4beta1 90(22)110494-10498 (Nov. 15, 1993). antagonists,” J. Pharmacol. Exp. Ther., 301(2)1747-752 (May 2002). * cited by examiner US 7,271,274 B2 1 2 PHENOLIC ANTIOXIDANTS FOR THE and dose dependent expression of VCAM-1 and release of TREATMENT OF DISORDERS INCLUDING soluble VCAM-l Were detected in cultures of human cere ARTHRITIS, ASTHMA AND CORONARY bral endothelial cells induced by TNF-alpha, but not in ARTERY DISEASE peripheral blood mononuclear cells (Kallmann et al., Brain 12316874697, 2000). Clinical data also shoW that adhesion CROSS REFERENCE TO RELATED molecules in blood and cerebrospinal ?uid are up-regulated APPLICATIONS throughout the clinical spectrum of multiple sclerosis, fur ther supporting the belief that multiple sclerosis can be suppressed by interfering With cell adhesion molecules such This application claims priority to US. Provisional Patent as VCAM-l (Elovaara et al., Arch. Neurol. 57:546*551, Application Ser. Nos. 60/564,043 ?led Apr. 20, 2004 and 2000). 60/600,029 ?led Aug. 9, 2004. Avariety of agents have been reported as potent inhibitors of VCAM-1 expression. (Schreiner et al., Expert Opi. Ther. FIELD OF THE INVENTION Patents 2003, 13, 1494166; Meng et al., Bioorg. Med. Chem. Lett. 2001, 11, 182341827.) A cyclic depsipeptide effec This invention is phenolic antioxidants that can be used tively inhibited VCAM-1 expression and reduced in?am for the treatment of disorders in Which redox-sensitive mation in a dermal model of in?ammation (Foster et al., Skin pro-in?ammatory genes are involved including VCAM-l, Pharmacol. 1996, 9, 149). A monoclonal antibody against such as arthritis, asthma and coronary artery disease. VCAM-l inhibited neointimal formation in a murine model of arterial Wall injury. (Oguchi et al., Arterioscler. Thromb. BACKGROUND OF THE INVENTION 20 Vasc. Biol. 2000, 20, 172941736.) A disubstituted 1,4 diaZepine diminished the increase in paW sWelling in a VCAM-l is also a mediator of chronic in?ammatory mouse model of collagen-induced arthritis. (Nakao et al., J. disorders such as asthma, rheumatoid arthritis, autoimmune Atheroscler Thromb. 1998, 4, 1494155.) Some VLA-4 diabetes and multiple sclerosis. For example, it is knoWn antagonists have shoWn e?icacy in animal models of dis that the expression of VCAM-1 and ICAM-1 are increased 25 ease. CP-664511, a small-molecule VLA-4 antagonist in in asthmatics (PileWski, J. M., et al. Am. J. Respir. Cell Mol. clinical trials in asthmatic patients, inhibited airWay eosino Biol. 12, 143 (1995); OhkaWara, Y., et al.,Am. J. Respir. Cell phil in?ltration in a murine model of allergic pulmonary Mol. Biol. 12, 4412 (1995)). Additionally, blocking the in?ammation. (KudlacZ et al., J Pharmacol. Exp. Ther. integrin receptors for VCAM-l and ICAM-1 (VLA-4 and 2002, 301, 7474752.) Several activators of peroxisome LFA-l, respectively) suppressed both early and late phase proliferator-activated receptors (PPARs) inhibited the responses in an ovalbumin-sensitiZed rat model of allergic expression of VCAM-1 on endothelial cells, suggesting a airWay responses (Rabb, H. A., et al., Am. J Respir. Care role of VCAM-1 in the anti-in?ammatory response of PPAR Med. 149, 118641191 (1994)). There is also increased activation. (Marx et al., J. Cardiovasc. Risk 2001, 8, expression of endothelial adhesion molecules, including 2034210; and Rival et al., Eur. J. Pharmacol. 2002, 435, VCAM-1, in the microvasculature of rheumatoid synovium 1434151.) (Koch, A. E. et al., Lab. Invest. 64, 3134322 (1991); 35 Probucol has been shoWn to possess potent antioxidant Morales-Ducret, J. et al., Immunol. 149, 142141431 (1992)). properties and to block oxidative modi?cation of LDL. Neutralizing antibodies directed against VCAM-l or its Consistent With these ?ndings, probucol has been shoWn to counter receptor, VLA-4, can delay the onset of diabetes in actually sloW the progression of atherosclerosis in LDL a mouse model (NOD mice) Which spontaneously develops receptor-de?cient rabbits. (CareW et al. Proc. Natl. Acad. the disease (Yang, X. D. et al., Proc. Natl. Acad. Sci. U.S.A. 40 Sci. U.S.A. 841772547729 (1987); Meng, C. Q. Probucol 90, 10494410498 (1993); Burkly, L. C. et al., Diabetes 43, (Restenosis). Curr Opin. Cardiovasc. Pulm. Renal Invest. 5234534 (1994); Baron, J. L. et al., J. Clin. Invest. 93, Drugs 2000, 2, 2944298; Barnhart et al., The Synthesis, 170041708 (1994)). Metabolism, and Biological Activity of Probucol and Its VCAM-l is expressed by cells both as a membrane bound Analogs. In Antilipidemic Drugs: Medicinal, Chemical, and form and as a soluble form. The soluble form of VCAM-1 45 Biochemical Aspects, Witiak et al., Eds., Elsevier Science: has been shoWn to induce chemotaxis of vascular endothe Amsterdam, 1991, pp 2774299.) lial cells in vitro and stimulate an angiogenic response in rat Probucol is chemically related to the Widely used food cornea (Koch, A. F. et al., Nature 376, 5174519 (1995)). additives 2,(3)-tert-butyl-4-hydroxyanisole (BHA) and 2,6 Inhibitors of the expression of soluble VCAM-l have poten di-tert-butyl-4-methyl phenol (BHT). It is a thioketal having tial therapeutic value in treating diseases With a strong a chemical name of 4,4'-(isopropylidenedithio) bis(2,6-di angiogenic component, including tumor groWth and 50 tert-butylphenol) and has the folloWing chemical structure: metastasis (Folkman, J. and Shing, Y., Biol. Chem. 10931410934 (1992)). VCAM-l is expressed in cultured human vascular endot helial cells after activation by lipopolysaccharide (LPS) and S S cytokines such as interleukin-1 (IL-1) and tumor necrosis 55 factor (TNF-alpha). X It has been documented that VCAM-l is expressed on HO brain microvessel endothelial cells in active lesions of multiple sclerosis brain. Multiple sclerosis is a common demyelinating disorder of the central nervous system, caus 60 ing patches of sclerosis (plaques) in the brain and spinal cord. It occurs in young adults and has protean clinical manifestations. Experimental therapy using antibodies for While probucol is a potent chemical anti-oxitant, there is VCAM-1 in autoimmune encephalomyelitis, Which is an little data to indicate it can be used in in?ammatory diseases animal model for multiple sclerosis, has shoWn that adhe 65 that do not depend on changing lipid levels such as sheu sion molecules play a role in the pathogenesis of the disease matoid arthritis, asthma and COPD. It is used primarily to (Benveniste et al., J. Neuroimmunol. 98z77i88, 1999). Time loWer serum cholesterol levels in hypercholesterolemic US 7,271,274 B2 3 4 patients. Probucol is commonly administered in the form of corresponds to European Patent Application No. 621 255) tablets available under the trademark LorelcoTM. and European Patent Application No. 763 527. US. Pat. No. 5,262,439 to Parthasarathy discloses ana WO 97/15546 to Nippon Shinyaku Co. Ltd. discloses logs of probucol With increased Water solubility in Which carboxylic acid derivatives for the treatment of arterial one or both of the hydroxyl groups are replaced With ester groups. sclerosis, ischemic heart diseases, cerebral infarction and Certain probucol ester derivatives have been described as post PTCA restenosis. being hypocholesterolemic and hypolipidemic agents: Fr The DoW Chemical Company is the assignee of patents to 2168137 (bis 4-hydroxyphenylthioalkane esters); Fr hypolipidemic 2-(3,5-di-tert-butyl-4-hydroxyphenyl)thio 2140771 (tetralinyl phenoxy alkanoic esters of probucol); Fr carboxamides. For example, US. Pat. Nos. 4,029,812, 2140769 (benZofuryloxyalkanoic acid derivatives of probu 4,076,841 and 4,078,084 to Wagner, et al., disclose these col); Fr 2134810 (bis-(3-alkyl-5-t-alkyl-4-thiaZole-5-car compounds for reducing blood serum lipids, especially boxy)phenyl-thio)alkanes; FR 2133024 (bis-(4-nicotinoy cholesterol and triglyceride levels. loxyphenylthio)propanes; and Fr 2130975 (bis(4 PCT WO 98/51289, ?led by Emory University and listing (phenoxyalkanoyloxy)-phenylthio)alkanes). as inventors Russell M. Medford and Patricia K. Somers, De Meglio et al. have described several ethers of sym claims priority to provisional patent application U.S. Ser. metrical molecules for the treatment of hyperlipidemia. No. 60/047,020, ?led on May 14, 1997. This application These molecules contain tWo phenyl rings attached to each discloses that monoesters of probucol inhibit the expression other through a iS4C(CH3)2iSi bridge. In contrast to of VCAM-1, and may also exhibit the composite pro?le of probucol, the phenyl groups do not have t-butyl as substitu 20 loWering LDL and reducing cholesterol. ents. (De Meglio et al., New Derivatives of Clo?brate and Recent reports demonstrated that mono-esters potently probucol: Preliminary Studies ofHypolipemic Activity; Far inhibited cytokine-induced VCAM-1 and MCP-1 expression maco, Ed. Sci (1985), 40 (11), 833444). and smooth muscle cell proliferation in vitro, and progres WO 00/26184 discloses a large genus of compounds With sion of atherosclerosis in experimental animals. (Meng et a general formula of phenyl-S-alkylene-S-phenyl, in Which 25 al., Bioorg. Med. Chem. Lett. 2002, 12, 2545*2548. In one or both phenyl rings can be substituted at any position. clinical trials, AGI-1067 did not cause QTc prolongation, These compounds Were disclosed as lubricants. While probucol did. Tardif et al., Circulation 2003, 107, US. Pat. Nos. 5,750,351; 5,807,884; 5,811,449; 5,846, 959; 5,773,231, and 5,773,209 to Medford, et al. (assigned 5524558.) PCT WO 98/51662 and US. Pat. Nos. 6,147,250, 6,548, to Emory University), as Well as the corresponding WO95/ 30 30415 to Emory University indicate that polyunsaturated 699, 6,617,352 and 6,602,914 describe mono-esters of fatty acids (“PUFAs”) and their hydroperoxides (“ox-PU probucol for the treatment of VCAM-1 mediated diseases FAs”), Which are important components of oxidatively including cardiovascular and in?ammatory diseases. PCT modi?ed loW density lipoprotein (LDL), induce the expres US 01/09049 discloses thioketals and thioethers for the treatment of VCAM-1 mediated diseases including in?am sion of VCAM-1, but not intercellular adhesion molecule-1 35 (ICAM-1) or E-selectin in human aortic endothelial cells. matory disorders. US. Pat. No. 5,155,250 to Parker et al. discloses that There is a need for neW phenolic compounds that can be 2,6-dialkyl-4-silylphenols are antiatherosclerotic agents. used in the treatment of a variety of disorders. The same compounds are disclosed as serum cholesterol loWering agents in PCT Publication No. W0 95/ 15760, 40 SUMMARY OF THE INVENTION published on Jun. 15, 1995. US. Pat. No. 5,608,095 to Parker et al. discloses that alkylated-4-silyl-phenols inhibit In one embodiment, antioxidant compounds are provided the peroxidation of LDL, loWer plasma cholesterol, and that inhibit the expression of redox-sensitive pro-in?amma inhibit the expression of VCAM-1, and thus are useful in the tory genes, including VCAM-l, and can be used to treat a treatment of atherosclerosis. 45 patient With a disorder in Which VCAM-1 expression is A series of European patent applications of Shionogi involved. Since VCAM-1 can be a mediator or marker of Seiyaku Kabushiki Kaisha disclose phenolic thioethers for chronic in?ammatory disorders, the compounds, composi use in treating arteriosclerosis. European Patent Application tions and methods of the invention can be used to inhibit the No. 348 203 discloses phenolic thioethers Which inhibit the expression of VCAM-1 and to treat chronic in?ammatory denaturation of LDL and the incorporation of LDL by 50 disorders including cardiovascular and in?ammatory dis macrophages. The compounds are useful as anti-arterioscle eases. rosis agents. Hydroxamic acid derivatives of these com Compounds of the present invention include those of pounds are disclosed in European Patent Application No. Formula I 405 788 and are useful for the treatment of arteriosclerosis, ulcer, in?ammation and allergy. Carbamoyl and cyano 55 derivatives of the phenolic thioethers are disclosed in US. Pat. No. 4,954,514 to Kita et al. US. Pat. No. 4,752,616 to Hall et al. discloses arylthio alkylphenylcarboxylic acids for the treatment of thrombotic disease. The compounds disclosed are useful as platelet 60 aggregation inhibitors for the treatment of coronary or cerebral thromboses and the inhibition of bronchoconstric HO tion, among others. A series of patents to Adir et Compagnie disclose substi R1 tuted phenoxyisobutyric acids and esters useful as antioxi 65 dants and hypolipaemic agents. This series includes US. Pat. Nos. 5,206,247 and 5,627,205 to Regnier, et al. (Which US 7,271,274 B2 5 6 or a pharmaceutically acceptable salt or ester thereof, wherein -continued R1 is independently hydrogen or optionally substituted CFC4 alkyl; and X is independently C liC4 alkyl, option ally substituted by one or more hydroxyl or C(O)OH Wherein the one or more hydroxyl or C(O)OH groups are optionally protected With a protecting group. In yet another embodiment of the invention, a compound is provided of Formula II above Wherein Y is selected from In another embodiment, a compound of Formula II is provided: the group consisting of:

20 OH; and Olin" H OH

Examples of in?ammatory disorders in Which VCAM-1 Wherein Y is selected from the group consisting of: expression is involved and that can be treated or prophy lactically treated, as disclosed herein include, but are not limited to arthritis, rheumatoid arthritis, asthma, dermatitis, cystic ?brosis, post transplantation late and chronic solid HO organ rejection, multiple sclerosis, systemic lupus erythe 30 matosis, in?ammatory boWel diseases, autoimmune diabe tes, diabetic retinopathy, diabetic nephropathy, diabetic vas Mom culopathy, rhinitis, allergic rhinitis, ocular in?ammation, uveitis, ischemia-reperfusion injury, stenosis, restenosis, post-angioplasty restenosis, chronic obstructive pulmonary disease (COPD), glomerulonephritis, Graves disease, gas yolk 35 trointestinal allergies, conjunctivitis, atherosclerosis, coro nary artery disease, angina and small artery disease. In a particular embodiment, compounds, compositions and methods are provided for the treatment rheumatoid

40 arthritis. The compounds and compositions of the invention are also suitable as disease modifying anti-rheumatoid arthritis drugs (DMARDs). The compounds, methods and compositions disclosed herein also can be used for the treatment of ocular in?ammation, including uveitis. 45 In a particular embodiment, compounds, compositions and methods are provided for the treatment asthma, or other pulmonary in?ammatory diseases. The compounds and compositions of the invention are also suitable as disease modifying anti-asthma drugs. In another embodiment, the 50 compounds and compositions of the invention can be used to treat chronic obstructive pulmonary disease. In another embodiment, the compounds described herein are useful in both the primary and adjunctive medical treatment of cardiovascular disease. The compounds can be 55 used in primary treatment of, for example, coronary disease states including atherosclerosis, post-angioplasty restenosis, coronary artery diseases and angina. The compounds can be administered to treat small vessel disease that is not treatable by surgery or angioplasty, or other vessel disease in Which 60 surgery is not an option. The compounds can also be used to stabiliZe patients prior to revasculariZation therapy. Com pounds, methods and compositions of the invention can be used to inhibit the progression of atherosclerosis. The compounds, compositions and methods disclosed 65 herein can also be used in the treatment of in?ammatory skin OH; and diseases as Well as human endothelial disorders, Which include, but are not limited to psoriasis, dermatitis, including US 7,271,274 B2 7 8 ecZematous dermatitis, Kaposi’s sarcoma, multiple sclero As used herein, the term “compound substantially free of” sis, as Well as proliferative disorders of smooth muscle cells. or “compound substantially in the absence of” refers to a In yet another embodiment, the compounds, methods and form of a compound that is in an admixture With no more compositions disclosed herein can be selected to treat anti than 15%, or no more than 10% by Weight, or no more than in?ammatory conditions that are mediated by mononuclear 5%, 2%, 1% or 0% by Weight, of other enantiomeric or leucocytes. Further, they can be used to treat in?ammatory diastereomeric or other stereoisomeric forms of that com diseases by modulating the expression of pro-in?ammatory pound. genes such as TNFot, IL1[3, MCP-l and IL6. In one embodiment, in the methods, compositions and In one embodiment, the compounds, methods and com compounds of this invention, When stereochemistry is des positions disclosed herein are used in the prevention or ignated, the compounds are substantially free of their enan treatment of tissue or organ transplant rejection. Treatment and prevention of organ or tissue transplant rejection tiomers or other stereoisomeric forms. includes, but is not limited to the treatment of recipients of Similarly, the term “isolated” refers to a compound that heart, lung, combined heart-lung, liver, kidney, pancreatic, includes at least 85%, 90%, 95%, 98%, 99%, or 100% by skin, spleen, small boWel, or corneal transplants. The com Weight, of the compound, the remainder comprising other pounds, methods and compositions disclosed herein can also compounds. be used in the prevention or treatment of graft-versus-host The term “pharmaceutically acceptable salt” refer to a salt disease, such as sometimes occurs folloWing bone marroW or complex that retains the desired biological activity of a transplantation. compound of the present invention and exhibits minimal The compounds, methods and compositions can be used 20 undesired toxicological elfects. Nonlimiting examples of alone or as adjunct or combination therapy simultaneously such salts are (a) acid addition salts formed With inorganic or in series. acids (for example, hydrochloric acid, hydrobromic acid, Also provided are pharmaceutical compositions compris sulfuric acid, phosphoric acid, nitric acid, and the like), and ing a compound disclosed herein in a form suitable for oral, salts formed With organic acids such as acetic acid, oxalic parenteral, intravenous, intradermal, transdermal, subcuta 25 neous or topical administration. The pharmaceutical com acid, tartaric acid, succinic acid, malic acid, ascorbic acid, position is, e.g., in the form of a tablet or capsule. The benZoic acid, tannic acid, pamoic acid, alginic acid, poly compounds may be substantially free of other stereoisomer glutamic acid, naphthalenesulfonic acid, naphthalenedisul forms, and essentially enantiamerically pure. fonic acid, and polygalcturonic acid; (b) base addition salts 30 formed With metal cations such as Zinc, calcium, bismuth, Also Within the scope of the invention is the use of a barium, magnesium, aluminum, copper, cobalt, nickel, cad compound disclosed herein, or a pharmaceutically accept mium, sodium, potassium, and the like, or With a cation able salt or ester thereof, or the use in the manufacture of a formed from meglumine, ammonia, N,N-dibenZylethylene medicament, optionally With a pharmaceutically acceptable diamine, D-glucosamine, tetraethylammonium, or ethylene carrier or diluent, for the treatment of an in?ammatory 35 diamine; or (c) combinations of (a) and (b); e.g., a Zinc disease in a host, optionally in combination and/or alterna tannate salt or the like. Also included in this de?nition are tion With one or more other therapeutic agents such as pharmaceutically acceptable quaternary salts knoWn by anti-in?ammatory agents. those skilled in the art, Which speci?cally include the quaternary ammonium salt comprising a counterion, includ DETAILED DESCRIPTION OF THE 40 ing chloride, bromide, iodide, 4O-alkyl, toluenesulfonate, INVENTION methylsulfonate, sulfonate, phosphate, or carboxylate (such as benZoate, succinate, acetate, glycolate, maleate, malate, I. De?nitions citrate, tartrate, ascorbate, benZoate, cinnamoate, mande The term “alkyl”, as used herein, unless otherWise speci loate, benZyloate, and diphenylacetate). Other examples of ?ed, includes a saturated straight, branched, or cyclic, pri 45 pharmaceutically acceptable salts are organic acid addition mary, secondary, or tertiary hydrocarbon of typically C1 to salts formed With acids Which form a physiological accept C4, and speci?cally includes methyl, ethyl, propyl, isopro able anion, for example, arginine, lysine, tosylate, methane pyl, cyclopropyl, butyl, isobutyl, secbutyl, and t-butyl. The sulfonate, citrate, ot-ketoglutarate, and ot-glycerophosphate. alkyl is optionally substituted. Substituted alkyl groups Suitable inorganic salts may also be formed, including, include halogenated alkyl groups, including ?uorinated 50 sulfate, nitrate, bicarbonate, and carbonate salts. alkyl. groups. The alkyl group may be optionally substituted As used herein, the term “prodrug” includes a compound With a moiety such as halo (chloro, ?uoro, bromo or iodo), that, When administered to a subject, is converted under haloalkyl, hydroxyl, carboxyl, acyl, aryl, acyloxy, amino, physiological conditions to a compound of the invention. amido, carboxyl derivatives, alkylamino, dialkylamino, ary As used herein, the term “patient” refers to a Warm lamino, alkoxy, aryloxy, nitro, cyano, carboxylic acid, and 55 blooded animal or mammal, and in particular a human, in carbamate, either unprotected, or protected as necessary, as need of therapy. The term “host”, as used herein, refers to a knoWn to those skilled in the art, for example, as taught in unicellular or multicellular organism, including cell lines Greene et al., Protective Groups in Organic Synthesis, John and animals, including a human. Wiley & Sons, Second Edition, 1991. Examples include CF3 and CH2CF3. 60 II. Description Whenever a range of is referred to herein, it includes It has been discovered that the phenolic antioxidant independently and separately every member of the range. As compounds of the invention inhibit the expression of a nonlimiting example, the term “Cl£4 alkyl” (or Cl_4 VCAM-1, and thus can be used to treat a patient With a alkyl) is considered to include, independently, each member disorder in Which VCAM-1 expression is involved and/or of the group, such that, for example, CFC4 alkyl includes 65 certain redox-sensitive pro-in?ammatory genes are involved straight, branched and Where appropriate cyclic C1, C2, C3 such as TNFot, IL1[3, MCP-l and IL6. These compounds can and C4 alkyl functionalities. be administered to a host as monotherapy, or if desired, in US 7,271,274 B2 9 1 0 combination With another compound of the invention or [(2-{2,6-Di-tert-butyl-4-[l -(3 ,5 -di-tert-butyl-4-hydroxyphe another biologically active agent, as described in more detail nylsulfanyl)- l -methylethylsulfanyl]phenoxy}acetyl)me beloW. thylamino] acetic acid; A. Compounds In one embodiment, there is provided a compound of Formula 1: .0 X

3 -(2- {2, 6-Di-tert-butyl-4- [l -(3 ,5 -di-tert-butyl-4 -hydrox yphenylsulfanyl) -l -methylethylsulfanyl] HO phenoxy}acetylamino)propionic acid; and

s s OH 0 wherein HO o\)]\ OH R1 is independently hydrogen or CFC4 alkyl; and N H X is independently CFC4 alkyl, optionally substituted by 25 one or more hydroxyl or C(O)OH, Which are optionally protected. 2-{2,6-Di-tert-butyl-4-[ l -(3 ,5 -di-tert-butyl-4-hydroxyphe In one embodiment, the one or more hydroxyl or C(O)OH nylsulfanyl)- 1 -methyl -ethylsulfanyl]phenoxy} -N- (2 -hy groups are protected. droXy-l -hydroxymethyl-ethyl)ac etamide. Optionally, X is CFC4 alkyl substituted by tWo or more hydroxyl groups. In one embodiment, there is provided a compound of Formula 11: Optionally, X is C liC4 alkyl substituted by three or more hydroxyl groups. Optionally, R1 is substituted Cl£4 alkyl, e.g., substituted With a halogen, such as ?uoro. X is, e.g., a C1, C2, C3 or C4 alkyl. Speci?c embodiments include:

wherein Y is selected from the group consisting of: ?g .X. (FM0 (2- {2, 6-Di-tert-butyl-4- [l -(3, 5 -di-tert-butyl-4 -hydroxyphe nylsulfanyl)- l -methylethylsulfanyl] phenoxy}acetylamino)acetic acid; 1‘ OH; OH; 55 kw HO QH /; / ; 60 W0 E 0 .X. 0 OH 6H OH

OH; i? 95*“? 65 OH US 7,271,274 B2 11 1 2 4-{2,6-Di-tert-butyl-4-[ 1 -(3 ,5 -di-tert-butyl-4-hydroxyphe -continued nylsulfanyl)-1-methylethylsulfanyl]-phenoXy}butane-1 ,2 (S),3(R)-triol; and

m

H0

4-{2,6-Di-tert-butyl-4-[1-(3,5-di-tert-butyl-4-hydroxyphe nylsulfanyl)-1-methylethylsulfanyl]-phenoXy}butane-1 ,2 (R),3(S)-triol. In yet another embodiment of the invention, a compound is provided of Formula 11 above Wherein Y is selected from the group consisting of: Mom

25

OH.

OH

A further embodiment includes the intermediates used to make the ?nal compounds of the invention. Said interme diates are useful as starting materials for making the com pounds of the invention as Well as having pharmaceutical activity alone. 35 Another embodiment of the invention includes the pro cess for making both the intermediates as Well as the ?nal compounds. B. Synthesis of Active Compounds 4-{2,6-Di-tert-butyl-4-[ 1 -(3 ,5 -di-tert-butyl-4-hydroxyphe 40 nylsulfanyl)- 1 -methylethylsulfanyl]phenoxy}butane- 1 ,2 The compounds of the present invention can be prepared (S),3(S)-triol; by those skilled in the art of organic synthesis using the methods disclosed herein and techniques knoWn in the art, many of Which are described by J. March, in Advanced 45 Organic Chemistry, 4”’ Edition (Wiley-lnterscience, NeW York, 1992), incorporated herein by reference. Speci?c means of preparing the compounds of the inven SXS OH tion are schematically displayed beloW. SCHEMEL 50

4-{2,6-Di-tert-butyl-4-[ 1 -(3 ,5 -di-tert-butyl-4-hydroxyphe nylsulfanyl)- 1 -methylethylsulfanyl] -phenoXy}butane-1 ,2 55 HO OH (R),3(R)-triol;

60 8X8 IIIO

65 Mitsunobu Reaction US 7,271,274 B2 13 14

-continued -continued

S S S S X 0 O \/ 5 X 0 O \/ HO 0\/E\ HO 0\/'\ 0‘“y\0H 0 >/\OH 10 lR—X, Base lR—X, Base

S S X 0 O \/ HO 0\)\ 20 O>/\OR 1. Acid 2. Base 1. Acid 2. Base 25

S S X @H S S HO 0 _ \/:\/\; OR 30 X OH 5 HO O 6H OR

1. Acid OH 2. Base 35 1. Acid 2. Base

S S X @H S S —> 3 HO O X OH —> HO O OH

R is alkyl and X is halide OH 45 R is alkyl and X is halide

SQHENLELZ 50 SQHEMEl

S7

O O V 60 O

O Base (such as KF Mitsunobu Reaction 65 on alumina) US 7,271,274 B2 15 16

-continued -continued

s s X 0 m no 0% 0/ R 0% N/ x H

Base hydrolysis wherein X:Alkyl substituted by one or more iOH or %OOH C. Stereoisomerism and Polymorphism It is appreciated that compounds of the present invention having a chiral center may exist in and be isolated in optically active and racemic forms. Some compounds may exhibit polymorphism. It is to be understood that the present jig X or. 20 invention encompasses any racemic, optically-active, dias tereomeric, polymorphic, or stereoisomeric form, or mix tures thereof, of a compound of the invention, Which possess O the useful properties described herein, it being Well knoWn in the art hoW to prepare optically active forms (for example, 25 by resolution of the racemic form by recrystallization tech R” / n 0/ niques, by synthesis from optically-active starting materials, Amide formation (e.g., using EDCl) by chiral synthesis, or by chromatographic separation using a chiral stationary phase). Examples of methods to obtain optically active materials 30 are knoWn in the art, and include at least the following: i) physical separation of crystalsia technique Whereby 8% macroscopic crystals of the individual enantiomers are manually separated. This technique can be used if crystals of the separate enantiomers exist, i.e., the material is a con 35 glomerate, and the crystals are visually distinct; ii) simultaneous crystallizationia technique Whereby the Basic hydrolysis individual enantiomers are separately crystallized from a solution of the racemate, possible only if the latter is a conglomerate in the solid state; 40 s s iii) enzymatic resolutionsia technique Whereby partial X 0 or complete separation of a racemate by virtue of differing 0 rates of reaction for the enantiomers With an enzyme; HO 0 iv) enzymatic asymmetric synthesisia synthetic tech N / n on 45 nique Whereby at least one step of the synthesis uses an R” enzymatic reaction to obtain an enantiomerically pure or enriched synthetic precursor of the desired enantiomer; v) chemical asymmetric synthesisia synthetic technique In Scheme 3, X:halide; n:li5; R:alkyl; R':alkyl; R":H or Whereby the desired enantiomer is synthesized from an alkyl 50 achiral precursor under conditions that produce asymmetry (i.e., chirality) in the product, Which may be achieved using chiral catalysts or chiral auxiliaries; vi) diastereomer separationsia technique Whereby a racemic compound is reacted With an enantiomerically pure reagent (the chiral auxiliary) that converts the individual enantiomers to diastereomers. The resulting diastereomers are then separated by chromatography or crystallization by virtue of their noW more distinct structural differences and the chiral auxiliary later removed to obtain the desired enantiomer; vii) ?rst- and second-order asymmetric transforma tionsia technique Whereby diastereomers from the race mate equilibrate to yield a preponderance in solution of the diastereomer from the desired enantiomer or Where prefer Amide formation (e.g., using EDCl) 65 ential crystallization of the diastereomer from the desired enantiomer perturbs the equilibrium such that eventually in principle all the material is converted to the crystalline US 7,271 ,274 B2 17 18 diastereomer from the desired enantiomer. The desired enan naphthalenedisulfonic acid, and polygalcturonic acid; (b) tiomer is then released from the diastereomer; base addition salts formed With metal cations such as Zinc, viii) kinetic resolutionsithis technique refers to the calcium, bismuth, barium, magnesium, aluminum, copper, achievement of partial or complete resolution of a racemate cobalt, nickel, cadmium, sodium, potassium, and the like, or (or of a further resolution of a partially resolved compound) With a cation formed from ammonia, N,N-dibenZylethylene by virtue of unequal reaction rates of the enantiomers With diamine, D-glucosamine, tetraethylammonium, ethylenedi a chiral, non-racemic reagent or catalyst under kinetic con amine meglumine, arginine, or lysine; or (c) combinations of ditions; (a) and (b); e.g., a Zinc tannate salt or the like. Also included ix) enantiospeci?c synthesis from non-racemic precur in this de?nition are pharmaceutically acceptable quaternary sorsia synthetic technique Whereby the desired enantiomer salts knoWn by those skilled in the art, Which speci?cally is obtained from non-chiral starting materials and Where the include the quaternary ammonium salt of the formulaiNR+ stereochemical integrity is not or is only minimally com A‘, Wherein R is as de?ned above and A is a counterion, promised over the course of the synthesis; including chloride, bromide, iodide, 4O-alkyl, toluene x) chiral liquid chromatographyia technique Whereby sulfonate, methylsulfonate, sulfonate, phosphate, or car the enantiomers of a racemate are separated in a liquid boxylate (such as benZoate, succinate, acetate, glycolate, mobile phase by virtue of their differing interactions With a maleate, malate, citrate, tartrate, ascorbate, benZoate, cinna stationary phase. The stationary phase can be made of chiral moate, mandeloate, benZyloate, and diphenylacetate). material or the mobile phase can contain an additional chiral Particular FDA-approved salts can be conveniently material to provoke the differing interactions; divided betWeen anions and cations (Approved Drug Prod xi) chiral gas chromatographyia technique Whereby the 20 ucts With Therapeutic Equivalence Evaluations (1994) US. racemate is volatiliZed and enantiomers are separated by Department of Health and Human Services, Public Health virtue of their differing interactions in the gaseous mobile Service, FDA, Center for Drug Evaluation and Research, phase With a column containing a ?xed non-racemic chiral Rockville, Md.; L. D. Bighley, S. M. Berge and D. C. adsorbent phase; Monkhouse, Salt Forms of Drugs and Absorption, Encyclo xii) extraction With chiral solventsia technique Whereby 25 pedia ofPharmaceuZical Technology, Vol. 13, J. SWarbridk the enantiomers are separated by virtue of preferential and J. Boylan, eds., Marcel Dekker, N.Y. (1996)). Among dissolution of one enantiomer into a particular chiral sol the approved anions include aceglumate, acephyllinate, vent; acetamidobenZoate, acetate, acetylasparaginate, acetylaspar xiii) transport across chiral membranesia technique tate, adipate, aminosalicylate, anhydromethylenecitrate, Whereby a racemate is placed in contact With a thin mem 30 ascorbate, aspartate, benZoate, besylate, bicarbonate, bisul brane barrier. The barrier typically separates tWo miscible fate, bitartrate, borate, bromide, camphorate, camsylate, ?uids, one containing the racemate, and a driving force such carbonate, chloride, chlorophenoxyacetate, citrate, closy as concentration or pressure differential causes preferential late, cromesilate, cyclamate, dehydrocholate, dihydrochlo transport across the membrane barrier. Separation occurs as ride, dimalonate, edentate, edisylate, estolate, esylate, eth a result of the non-racemic chiral nature of the membrane 35 ylbromide, ethylsulfate, fendiZoate, fosfatex, fumarate, Which alloWs only one enantiomer of the racemate to pass gluceptate, gluconate, glucuronate, glutamate, glycerophos through. phate, glysinate, glycollylarsinilate, glycyrrhiZate, hippu rate, hemisulfate, hexylresorcinate, hybenZate, hydrobro D. Pharmaceutically Acceptable Salt Formulations mide, hydrochloride, hydroiodid, hydroxybenZenesulfonate, In cases Where compounds are suf?ciently basic or acidic 40 hydroxybenZoate, hydroxynaphthoate, hyclate, iodide, to form stable nontoxic acid or base salts, administration of isethionate, lactate, lactobionate, lysine, malate, maleate, the compound as a pharmaceutically acceptable salt may be mesylate, methylbromide, methyliodide, methylnitrate, appropriate. The term “pharmaceutically acceptable salts” or methylsulfate, monophosadenine, mucate, napadisylate, “complexes” refers to salts or complexes that retain the napsylate, nicotinate, nitrate, oleate, orotate, oxalate, oxo desired biological activity of the compounds of the present 45 glurate, pamoate, pantothenate, pectinate, phenylethylbarbi invention and exhibit minimal undesired toxicological turate, phosphate, pacrate, plicrilix, polistirex, polygalactu elfects. ronate, propionate, pyridoxylphosphate, saccharinate, Examples of pharmaceutically acceptable salts are salicylate, stearate, succinate, stearylsulfate, subacetate, suc organic acid addition salts formed With acids, Which form a cinate, sulfate, sulfosalicylate, tannate, tartrate, teprosilate, physiological acceptable anion, for example, tosylate, meth 50 terephthalate, teoclate, thiocyante, tidiacicate, timonacicate, anesulfonate, acetate, citrate, malonate, tartarate, succinate, tosylate, triethiodide, triethiodide, undecanoate, and xin benZoate, ascorbate, ot-ketoglutarate and ot-glycerophos afoate. The approved cations include ammonium, beneth phate. Suitable inorganic salts may also be formed, includ amine, benZathine, betaine, calcium, carnitine, clemiZole, ing, sulfate, nitrate, bicarbonate and carbonate salts. Alter chlorcycliZine, choline, dibenylamine, diethanolamine, natively, the pharmaceutically acceptable salts may be made 55 diethylamine, diethylammonium diolamine, eglumine, erbu With suf?ciently basic compounds such as an amine With a mine, ethylenediamine, heptaminol, hydrabamine, hydroxy suitable acid affording a physiologically acceptable anion. ethylpyrrolidone, imadaZole, meglumine, olamine, pipera Alkali metal (for example, sodium, potassium or lithium) or Zine, 4-phenylcyclohexylamine, procaine, pyridoxine, alkaline earth metal (for example calcium) salts of carboxy triethanolamine, and tromethamine. Metallic cations lic acids can also be made. 60 include, aluminum, bismuth, calcium lithium, magnesium, Nonlimiting examples of such salts are (a) acid addition neodymium, potassium, rubidium, sodium, strontium and salts formed With inorganic acids (for example, hydrochloric Zinc. acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric A particular class of salts can be classi?ed as organic acid, and the like), and salts formed With organic acids such amine salts. The organic amines used to form these salts can as acetic acid, oxalic acid, tartaric acid, succinic acid, malic 65 be primary amines, secondary amines or tertiary amines, and acid, ascorbic acid, benZoic acid, tannic acid, pamoic acid, the substituents on the amine can be straight, branched or alginic acid, polyglutamic acid, naphthalenesulfonic acid, cyclic groups, including ringed structures formed by attach US 7,271,274 B2 19 20 ment of tWo or more of the amine substituents. Of particular In a particular embodiment, compounds are used for interest are organic amines that are substituted by one or treatment of arthritis or rheumatoid arthritis. Nonlimiting more hydroxyalkyl groups, including alditol or carbohydrate examples of arthritis include rheumatoid (such as soft-tissue moieties. These hydroxy substituted organic amines can be rheumatism and non-articular rheumatism, ?bromyalgia, cyclic or acyclic, both classes of Which can be primary ?brositis, muscular rheumatism, myofascil pain, humeral amines, secondary amines or tertiary amines. A common epicondylitis, frozen shoulder, Tietze’s syndrome, fascitis, class of cyclic hydroxy substituted amines are the amino tendinitis, tenosynovitis, bursitis), juvenile chronic, spondy sugars. loarthropaties (ankylosing spondylitis), osteoarthritis, hype A particular class of acyclic organic amines are repre ruricemia and arthritis associated With acute gout, chronic sented by the formula gout and systemic lupus erythematosus. The compounds and compositions of the invention are also suitable as disease modifying anti-rheumatoid arthritis drugs (DMARDs). The compounds and compositions also can be used for the treatment of ocular in?ammation, including uveitis. In one embodiment, a method is provided for the treatment of arthritis or an arthritis related disorder including adminis tering to a host in need thereof a compound of the invention Wherein Y and Z are independently hydrogen or loWer alkyl or a pharmaceutically acceptable salt or ester thereof. or, may be taken together to form a ring, R is hydrogen, alkyl In a particular embodiment, compounds and compositions or hydroxyloWeralkyl, and n is l, 2, 3, 4, or 5. Among these 20 are provided for the treatment asthma, or other pulmonary hydroxylamines are a particular class characterized When n in?ammatory diseases. The compounds and compositions of is 4. A representative of this group is meglumine, repre the invention are also suitable as disease modifying anti sented When Y is hydrogen, Z is methyl and R is methoxy. asthma drugs. In another embodiment, the compounds and Meglumine is also knoWn in the art as N-methylglucamine, compositions of the invention can be used to treat chronic N-MG, and l-deoxy-l -(methylamino)-D-glucitol. 25 obstructive pulmonary disease. In one aspect of the inven tion, a patient in need of treatment for asthma or other Pharmaceutically Acceptable Prodrugs pulmonary in?ammatory disease is provided, including The invention also includes pharmaceutically acceptable administering to the patient an effective amount of a com prodrugs of the compounds. Pharmaceutically acceptable pound of the invention or a pharmaceutically acceptable salt prodrugs refer to a compound that is metabolized, for 30 or ester thereof. example hydrolyzed or oxidized, in the host to form the In another embodiment, the compounds described herein compound of the present invention. Typical examples of are useful in both the primary and adjunctive medical prodrugs include compounds that have biologically labile treatment of cardiovascular disease. The compounds can be protecting groups on a functional moiety of the active used in primary treatment of, for example, coronary disease compound. Prodrugs include compounds that can be oxi 35 states including atherosclerosis, post-angioplasty restenosis, dized, reduced, aminated, deaminated, hydroxylated, dehy coronary artery diseases and angina. The compounds can be droxylated, hydrolyzed, dehydrolyzed, alkylated, dealky administered to treat small vessel disease that is not treatable lated, acylated, deacylated, phosphorylated, by surgery or angioplasty, or other vessel disease in Which dephosphorylated to produce the active compound. surgery is not an option. The compounds can also be used to Any of the compounds described herein can be adminis 40 stabilize patients prior to revascularization therapy. Com tered as a prodrug to increase the activity, bioavailability, pounds and compositions of the invention can be used to stability or otherWise alter the properties of the compound. inhibit the progression of atherosclerosis. In one aspect of A number of prodrug ligands are knoWn. In general, alky the invention, a method is provided for treating cardiovas lation, acylation or other lipophilic modi?cation of the cular disorders in a patient in need thereof comprising compound Will increase its stability. 45 administering to said patient an effective amount of a compound of the invention or a pharmaceutically acceptable E. Treatment of Disorders salt thereof. The compounds of the present invention can be used to The compounds disclosed herein can be used in the treat any disorder in Which VCAM-1 expression is involved treatment of in?ammatory skin diseases and in particular, and/or certain redox-sensitive pro-in?ammatory genes are 50 human endothelial disorders, Which include, but are not involved such as TNFot, ILlB, MCP-l and IL6. Such limited to, psoriasis, dermatitis, including eczematous der disorders include, but are not limited to arthritis, asthma, matitis, and Kaposi’s sarcoma, as Well as proliferative dermatitis, psoriasis, cystic ?brosis, post transplantation late disorders of smooth muscle cells. and chronic solid organ rejection, multiple sclerosis, sys In yet another embodiment, the compounds disclosed temic lupus erythematosis, in?ammatory boWel diseases, 55 herein can be selected to treat anti-in?ammatory conditions autoimmune diabetes, diabetic retinopathy, diabetic nephr in Which mononuclear leucocytes are involved. Further, they opathy, diabetic vasculopathy, ocular in?ammation, uveitis, can be used to treat in?ammatory diseases by modulating the rhinitis, ischemia-reperfusion injury, post-angioplasty rest expression of pro-in?ammatory genes such as TNFot, ILlB, enosis, chronic obstructive pulmonary disease (COPD), IL6 and MCP-l. glomerulonephritis, Graves disease, gastrointestinal aller 60 In addition to inhibiting the expression of VCAM-l, some gies, conjunctivitis, atherosclerosis, coronary artery disease, of the compounds of the invention have the additional angina and small artery disease. properties of inhibiting monocyte chemoattractant protein-l In one aspect of the invention, a method is provided for (MCP-l) and/or smooth muscle cell proliferation. MCP-l is treating an in?ammatory disease or disorder comprising a chemoattractant protein produced by endothelial cells, administering to a patient an effective amount of a com 65 smooth muscle cells as Well as macrophages. MCP-l pro pound of the invention or a pharmaceutically acceptable salt motes integrin activation on endothelial cells thereby facili or ester thereof. tating adhesion of leukocytes to VCAM-l, and MCP-l is a US 7,271 ,274 B2 21 22 chemoattractant for monocytes. MCP-l has been shown to nation therapy is typically preferred over alternation therapy play a role in leukocyte recruitment in a number of chronic because it induces multiple simultaneous stresses on the in?ammatory diseases including atherosclerosis, rheumatoid condition. arthritis, and asthma. Its expression is upregulated in these Any method of alternation can be used that provides diseases and as such inhibition of MCP-l expression rep treatment to the patient. Nonlimiting examples of alternation resents a desirable property of anti-in?ammatory therapeu patterns include li6 Weeks of administration of an effective tics. Furthermore, smooth muscle cell hyperplasia and amount of one agent folloWed by li6 Weeks of administra resulting tissue remodeling and decreased organ function is tion of an effective amount of a second agent. The altema yet another characteristic of many chronic in?ammatory tion schedule can include periods of no treatment. Combi diseases including atherosclerosis, chronic transplant rejec nation therapy generally includes the simultaneous tion and asthma. Inhibition of the hyperproliferation of administration of an effective ratio of dosages of tWo or smooth muscle cells is another desirable property for thera more active agents. peutic compounds. Illustrative examples of speci?c agents that can be used in Cytokines are extracellular signaling proteins produced combination or alternation With the compounds of the by many cell types playing a central role in human immune present invention are described beloW in regard to asthma response, and can be categorized as either pro-in?ammatory and arthritis. The agents set out beloW or others can alter or anti-in?ammatory in action. TNF-(X, IL-l[3 and IL-6 are natively be used to treat a host suffering from any of the major pro-in?ammatory cytokines implicated in the patho other disorders listed above or that involve VCAM-l or genesis of numerous diseases. The expression of these MCP-l. Illustrative second biologically active agents for the proin?ammatory cytokines is also redox-regulated (Haddad, 20 treatment of cardiovascular disease are also provided beloW. J . 1.; Saade, N. E.; Sa?eh-Garabedian, B. Redox regulation of TNF-(x Biosynthesis: Augmentation by Irreversible Inhi i) Asthma bition of y-Glutamylcysteine Synthetase and the Involve In one embodiment, the compounds of the present inven ment of an IKB-ot/NF-KB-independent Pathway in Alveolar tion are administered in combination or alternation With Epithelial Cells. Cell Signal. 2002, 14, 2lli2l8). 25 heparin, frusemide, ranitidine, an agent that effects respira In another embodiment, the compounds of the present tory function, such as DNAase, or immunosuppressive invention can be selected for the prevention or treatment of agents, IV gamma globulin, troleandomycin, cyclosporin tissue or organ transplant rejection. Treatment and preven (Neoral), methotrexate, FK-506, gold compounds such as tion of organ or tissue transplant rejection includes, but are Myochrysine (gold sodium thiomalate), platelet activating not limited to treatment of recipients of heart, lung, com 30 factor (PAF) antagonists such as thromboxane inhibitors, bined heart-lung, liver, kidney, pancreatic, skin, spleen, leukotriene-D4-receptor antagonists such as Accolate small boWel, or corneal transplants. They are also indicated (Za?rlukast), Zi?o (Zileuton), leukotriene C1 or C2 antago for the prevention or treatment of graft-versus-host disease, nists and inhibitors of leukotriene synthesis such as Zileuton Which sometimes occurs folloWing bone marroW transplan for the treatment of asthma, or an inducible nitric oxide tation. 35 synthase inhibitor. In another aspect of the invention, the compounds can be In another embodiment, the active compound is admin used in compositions, including pharmaceutical composi istered in combination or alternation With one or more other tions or a pharmaceutically acceptable salt thereof and/or a prophylactic agent(s). Examples of prophylactic agents that pharmaceutically acceptable carrier. can be used in alternation or combination therapy include The compounds, methods and compositions can be used 40 but are not limited to sodium cromoglycate, Intal (cromolyn alone or as adjunct or combination therapy simultaneously sodium, Nasalcrom, Opticrom, Crolom, Ophthalmic Cro or in series. lom), Tilade (nedocromil, nedocromil sodium) and keto tifen. F. Combination and Alternation Therapy In another embodiment, the active compound is admin Any of the compounds disclosed herein can be adminis 45 istered in combination or alternation With one or more other tered in combination or alternation With a second biologi [32-adrenergic agonist(s) ([3 agonists). Examples of [32-adr cally active agent to increase its effectiveness against the energic agonists ([3 agonists) that can be used in alternation target disorder. or combination therapy include but are not limited to In combination therapy, effective dosages of tWo or more albuterol (salbutamol, Proventil, Ventolin), terbutaline, agents are administered together, Whereas during alternation 50 Maxair (pirbuterol), Serevent (salmeterol), epinephrine, therapy an effective dosage of each agent is administered metaproterenol (Alupent, Metaprel), Brethine (Bricanyl, serially. The dosages Will depend on absorption, inactivation Brethaire, terbutaline sulfate), Tomalate (bitolterol), isopre and excretion rates of the drug as Well as other factors naline, ipratropium bromide, bambuterol hydrochloride, knoWn to those of skill in the art. It is to be noted that dosage bitolterol meslyate, broxaterol, carbuterol hydrochloride, values Will also vary With the severity of the condition to be 55 clenbuterol hydrochloride, clorprenaline hydrochloride, e?r alleviated. It is to be further understood that for any par moterol fumarate, ephedra (source of alkaloids), ephedrine ticular subject, speci?c dosage regimens and schedules (ephedrine hydrochloride, ephedrine sulfate), etafedrine should be adjusted over time according to the individual hydrochloride, ethylnoradrenaline hydrochloride, fenoterol need and the professional judgment of the person adminis hydrochloride, hexoprenaline hydrochloride, isoetharine tering or supervising the administration of the compositions. 60 hydrochloride, isoprenaline, mabuterol, methoxyphenamine The e?icacy of a drug can be prolonged, augmented, or hydrochloride, methylephedrine hydrochloride, orciprena restored by administering the compound in combination or line sulphate, phenylephrine acid tartrate, phenylpropanola alternation With a second, and perhaps third, agent that mine (phenylpropanolamine polistirex, phenylpropanola induces a different biological pathWay from that caused by mine sulphate), pirbuterol acetate, procaterol hydrochloride, the principle drug. Alternatively, the pharmacokinetics, bio 65 protokylol hydrochloride, pseudoephedrine (pseudoephe distribution or other parameter of the drug can be altered by drine polixtirex, pseudoephedrine tannate, pseudoephedrine such combination or alternation therapy. In general, combi hydrochloride, pseudoephedrine sulphate), reproterol hydro US 7,271,274 B2 23 24 chloride, rimiterol hydrobromide, ritodrine hydrochloride, (loratadine), Seldane (terfenadine), Periactin (cyprohepta salmeterol xinafoate, terbutaline sulphate, tretoquinol dine), Nolamine (phenindamine, Nolahist), Phenameth hydrate and tulobuterol hydrochloride. (promethaZine, Phenergan), Tacaryl (methdilaZine) and In another embodiment, the active compound is admin Temaril (trimepraZine). istered in combination or alternation With one or more other Alternatively, the compound of the present invention may corticosteriod(s). Examples of corticosteriods that can be be administered in combination or alternation With used in alternation or combination therapy include but are (a) xanthines and methylxanthines, such as Theo-24 (theo not limited to glucocorticoids (GC), Aerobid (Aerobid-M, phylline, Slo-Phylline, Uniphyllin, Slobid, Theo-Dur), ?unisolide), AZmacort (triamcinolone acetonide), Beclovet Choledyl (oxitriphylline), aminophylline; (Vanceril, beclomethasone dipropionate), Flovent (?utica (b) anticholinergic agents (antimuscarinic agents) such as sone), Pulmicort (budesonide), prednisolone, hydrocorti belladonna alkaloids, Atrovent (ipratropium bromide), sone, adrenaline, Alclometasone Dipropionate, Aldosterone, atropine, oxitropium bromide; Amcinonide, Beclomethasone Dipropionate, Bendacort, (c) phosphodiesterase inhibitors, including phosphodi Betamethasone (Betamethasone Acetate, Betamethasone esterase IV inhibitors such as Zardaverine; BenZoate, Betamethasone Dipropionate, Betamethasone (d) calcium antagonists such as nifedipine; Sodium Phosphate, Betamethasone Valerate), Budesonide, (e) potassium activators such as cromakalim for the treat Ciclomethasone, Ciprocinonide, Clobetasol Propionate, ment of asthma; Clobetasone Butyrate, Clocortolone Pivalate, Cloprednol, (f) B-eotaxin chemokine receptor, CCR3, antagonists; or Cortisone Acetate, CortivaZol, De?aZacort, Deoxycortone (g) IL-5 antibodies, IL-l3 antibodies, IL-l3 antagonists, Acetate (Deoxycortone Pivalate), Deprodone, Desonide, 20 IL-4 receptor antagonists, and IgE antibodies @(olair). Desoxymethasone, Dexamethasone (Dexamethasone Acetate, Dexamethasone Isonicotinate, Dexamethasone ii) Arthritic disorders Phosphate, Dexamethasone Sodium MetasulphobenZoate, In one embodiment, the compound of the present inven Dexamethasone Sodium Phosphate), Dichlorisone Acetate, tion can also be administered in combination or alternation Di?orasone Diacetate, Di?ucortolone Valerate, Di?upred 25 With apaZone, amitriptyline, chymopapain, collegenase, nate, Domoprednate, Endrysone, FluaZacort, Fluclorolone cyclobenZaprine, diaZepam, ?uoxetine, pyridoxine, ademe Acetonide, Fludrocortisone Acetate, Flumethasone (Flu tionine, diacerein, glucosamine, hylan (hyaluronate), miso methasone Pivalate), Flunisolide, Fluocinolone Acetonide, prostol, paracetamol, superoxide dismutase mimics, IL-1 Fluocinonide, Fluocortin Butyl, Fluocortolone (Fluocor receptor antagonists, IL-2 receptor antagonists, IL-6 recep tolone Hexanoate, Fluocortolone Pivalate), Fluo 30 tor antagonists, TNFO. receptor antagonists, TNFO. antibod rometholone (Fluorometholone Acetate), Fluprednidene ies, P38 MAP Kinase inhibitors, tricyclic antidepressents, Acetate, Fluprednisolone, Flurandrenolone, Fluticasone cJun kinase inhibitors or immunosuppressive agents, IV Propionate, Formocortal, Halcinonide, Halobetasol Propi gamma globulin, troleandomycin, cyclosporin (Neoral), onate, Halometasone, Hydrocortamate Hydrochloride, methotrexate, FK-506, gold compounds such as Myo Hydrocortisone (Hydrocortisone Acetate, Hydrocortisone 35 chrysine (gold sodium thiomalate), platelet activating factor Butyrate, Hydrocortisone Cypionate, Hydrocortisone (PAF) antagonists such as thromboxane inhibitors, MAP Hemisuccinate, Hydrocortisone Sodium Phosphate, Hydro KAPK2 (MK2) Kinase inhibitors, Chemokine Receptor cortisone Sodium Succinate, Hydrocortisone Valerate), Antagonists such as CCR5 Receptor antagonists, Interleukin Medrysone, Meprednisone, Methylprednisolone (Methyl Converting Enzyme (ICE) inhibitors, IKB Kinase (IKKI, prednisolone Acetate, Methylprednisolone, Hemisuccinate, 40 IKK2) inhibitors, TNF-ot Convertase EnZyme (TACE) Methylprednisolone Sodium Succinate), Mometasone inhibitors, ICK Kinase inhibitors, Janus Kinase 3 (JAK3) Furoate, Paramethasone Acetate, Prednicarbate, Prednisola inhibitors, Kinase insert domain-containing Receptor (KdR) mate Hydrochloride, Prednisolone (Prednisolone Acetate, Kinase inhibitors, and inducible nitric oxide sythase (iNOS) Prednisolone Hemisuccinate, Prednisolone Hexanoate, inhibitors. Prednisolone Pivalate, Prednisolone Sodium Metasul 45 In another embodiment, the active compound is admin phobenZoate, Prednisolone Sodium Phosphate, Predniso istered in combination or alternation With one or more other lone Sodium Succinate, Prednisolone Steaglate, Predniso corticosteriod(s). Examples of corticosteriods that can be lone Tebutate), Prednisone (Prednisone Acetate), used in alternation or combination therapy include but are Prednylidene, Procinonide, Rimexolone, Suprarenal Cortex, not limited to glucocorticoids (GC), Aerobid (Aerobid-M, Tixocortol Pivalate, Triamcinolone (Triamcinolone 50 ?unisolide), AZmacort (triamcinolone acetonide), Beclovet Acetonide, Triamcinolone Diacetate and Triamcinolone (Vanceril, beclomethasone dipropionate), Flovent (?utica Hexacetonide). sone), Pulmicort (budesonide), prednisolone, hydrocorti In another embodiment, the active compound is admin sone, adrenaline, Alclometasone Dipropionate, Aldo sterone, istered in combination or alternation With one or more other Amcinonide, Beclomethasone Dipropionate, Bendacort, antihistimine(s) (H 1 receptor antagonists). Examples of anti 55 Betamethasone (Betamethasone Acetate, Betamethasone histimines (H1 receptor antagonists) that can be used in BenZoate, Betamethasone Dipropionate, Betamethasone alternation or combination therapy include alkylamines, Sodium Phosphate, Betamethasone Valerate), Budesonide, ethanolamines ethylenediamines, piperaZines, piperidines or Ciclomethasone, Ciprocinonide, Clobetasol Propionate, phenothiaZines. Some non-limiting examples of antihistimes Clobetasone Butyrate, Clocortolone Pivalate, Cloprednol, are Chlortrimeton (Teldrin, chlorpheniramine), Atrohist 60 Cortisone Acetate, CortivaZol, De?aZacort, Deoxycortone (brompheniramine, Bromarest, Bromfed, Dimetane), Actidil Acetate (Deoxycortone Pivalate), Deprodone, Desonide, (triprolidine), Dexchlor (Poladex, Polaramine, dexchlorphe Desoxymethasone, Dexamethasone (Dexamethasone niramine), Benadryl (diphen-hydramine), Tavist (clemas Acetate, Dexamethasone Isonicotinate, Dexamethasone tine), Dimetabs (dimenhydrinate, Dramamine, Marmine), Phosphate, Dexamethasone Sodium MetasulphobenZoate, PBZ (tripelennamine), pyrilamine, MareZine (cycliZine), 65 Dexamethasone Sodium Phosphate), Dichlorisone Acetate, Zyrtec (cetiriZine), hydroxyZine, Antivert (mecliZine, Bon Di?orasone Diacetate, Di?ucortolone Valerate, Di?upred ine), Allegra (fexofenadine), Hismanal (astemiZole), Claritin nate, Domoprednate, Endrysone, FluaZacort, Fluclorolone US 7,271,274 B2 25 26 Acetonide, Fludrocortisone Acetate, Flumethasone (Flu Diethylamine Salicylate, Diethylsalicylamide, Difenpira methasone Pivalate), Flunisolide, Fluocinolone Acetonide, mide, Di?unisal, Dipyrone, Droxicam, EpiriZole, EtenZa Fluocinonide, Fluocortin Butyl, Fluocortolone (Fluocor mide, Etersalate, Ethyl Salicylate, Etodolac, Etofenamate, tolone Hexanoate, Fluocortolone Pivalate), Fluo Felbinac, Fenbufen, Fenclofenac, Fenoprofen Calcium, Fen rometholone (Fluorometholone Acetate), Fluprednidene tiaZac, Fepradinol, FepraZone, Floctafenine, Flufenamic, Acetate, Fluprednisolone, Flurandrenolone, Fluticasone Flunoxaprofen, Flurbiprofen (Flurbiprofen Sodium), Fos Propionate, Formocortal, Halcinonide, Halobetasol Propi fosal, Furprofen, Glafenine, Glucametacin, Glycol Salicy onate, Halometasone, Hydrocortamate Hydrochloride, late, Gold Keratinate, Harpagophytum Procumbens, Hydrocortisone (Hydrocortisone Acetate, Hydrocortisone Butyrate, Hydrocortisone Cypionate, Hydrocortisone Ibufenac, Ibuprofen, Ibuproxam, ImidaZole Salicylate, Hemisuccinate, Hydrocortisone Sodium Phosphate, Hydro Indomethacin (Indomethacin Sodium), Indoprofen, Isami cortisone Sodium Succinate, Hydrocortisone Valerate), faZone, Isonixin, Isoxicam, KebuZone, Ketoprofen, Ketoro Medrysone, Meprednisone, Methylprednisolone (Methyl lac Trometamol, Lithium Salicylate, LonaZolac Calcium, prednisolone Acetate, Methylprednisolone, Hemisuccinate, Lomoxicam, Loxoprofen Sodium, Lysine Aspirin, Magne Methylprednisolone Sodium Succinate), Mometasone sium Salicylate, Meclofenamae Sodium, Mefenamic Acid, Furoate, Paramethasone Acetate, Prednicarbate, Prednisola Meloxicam, Methyl Butetisalicylate, Methyl Gentisate, mate Hydrochloride, Prednisolone (Prednisolone Acetate, Methyl Salicylate, MetiaZinic Acid, MetifenaZone, Mofeb Prednisolone Hemisuccinate, Prednisolone Hexanoate, utaZone, MofeZolac, MoraZone Hydrochloride, Momi?u Prednisolone Pivalate, Prednisolone Sodium Metasul mate, Morpholine Salicylate, Nabumetone, Naproxen phobenZoate, Prednisolone Sodium Phosphate, Predniso 20 (Naproxen Sodium), NifenaZone, Ni?umic Acid, Nime lone Sodium Succinate, Prednisolone Steaglate, Predniso sulide, Oxametacin, OxaproZin, Oxindanac, Oxyphenbuta lone Tebutate), Prednisone (Prednisone Acetate), Zone, Parsalmide, PhenybutaZone, Phenyramidol Hydro Prednylidene, Procinonide, Rimexolone, Suprarenal Cortex, chloride, Picenadol Hydrochloride, Picolamine Salicylate, Tixocortol Pivalate, Triamcinolone (Triamcinolone Piketoprofen, PiraZolac, Piroxicam, Pirprofen, Pranoprofen, Acetonide, Triamcinolone Diacetate and Triamcinolone 25 Pranosal, Proglumetacin Maleate, ProquaZone, ProtiZinic Hexacetonide). Acid, RamifenaZone, Salacetamide, Salamidacetic Acid, In another embodiment, the active compound is admin Salicylamide, Salix, Salol, Salsalate, Sodium Aurothioma istered in combination or alternation With one or more other late, Sodium Gentisate, Sodium Salicylate, Sodium non-steroidal anti-in?ammatory drug(s) (NSAIDS). Thiosalicylate, Sulindac, Superoxide Dismutase (Orgotein, Examples of NSAIDS that can be used in alternation or 30 Pegorgotein, Sudismase), Suprofen, SuxibuZone, Tenidap combination therapy are carboxylic acids, propionic acids, Sodium, Tenoxicam, Tetrydamine, Thurfyl Salicylate, fenamates, acetic acids, pyrazolones, oxicans, alkanones, Tiaprofenic, Tiaramide Hydrochloride, Tinoridine Hydro gold compounds and others that inhibit prostaglandin syn chloride, Tolfenamic Acid, Tometin Sodium, Triethanola thesis, preferably by selectively inhibiting cylcooxyge mine Salicylate, Ufenamate, Zaltoprofen, Zidometacin and nase-2 (COX-2). Some nonlimiting examples of COX-2 35 Zomepirac Sodium. inhibitors are Celebrex (celecoxib), Bextra (valdecoxib), iii) Cardiovascular Disease Dynastat (parecoxib sodium) and Vioxx (rofacoxib). Some non-limiting examples of NSAIDS are aspirin (acetylsali Compounds useful for combining With the compounds of cylic acid), Dolobid (di?unisal), Disalcid (salsalate, salicyl the present invention for the treatment of cardiovascular disease encompass a Wide range of therapeutic compounds. salicylate), Trisilate (choline magnesium trisalicylate), 40 sodium salicylate, Cuprimine (penicillamine), Tolectin (tol Ileal bile acid transporter (IBAT) inhibitors, for example, metin), ibuprofen (Motrin, Advil, Nuprin Rufen), Naprosyn are useful in the present invention, and are disclosed in (naproxen, Anaprox, naproxen sodium), Nalfon (fenopro patent application no. PCT/US95/10863, herein incorpo fen), Orudis (ketoprofen), Ansaid (?urbiprofen), Daypro rated by reference. More IBAT inhibitors are described in (oxaproZin), meclofenamate (meclofanamic acid, 45 PCT/US97/04076, herein incorporated by reference. Still Meclomen), mefenamic acid, Indocin (indomethacin), Cli further IBAT inhibitors useful in the present invention are noril (sulindac), tolmetin, Voltaren (diclofenac), Lodine described in US. application Ser. No. 08/816,065, herein (etodolac), ketorolac, ButaZolidin (phenylbutaZone), Tan incorporated by reference. More IBAT inhibitor compounds dearil (oxyphenbutaZone), piroxicam (Feldene), Relafen useful in the present invention are described in WO (nabumetone), Myochrysine (gold sodium thiomalate), 50 98/40375, and WO 00/38725, herein incorporated by refer Ridaura (aurano?n), Solganal (aurothioglucose), acetami ence. Additional IBAT inhibitor compounds useful in the nophen, colchicine, Zyloprim (allopurinol), Benemid present invention are described in US. application Ser. No. (probenecid), Anturane (su?npyriZone), Plaquenil (hydroxy 08/816,065, herein incorporated by reference. chloroquine), Aceclofenac, Acemetacin, Acetanilide, Act In another aspect, the second biologically active agent is arit, Alclofenac, Ahninoprofen, Aloxiprin, Aluminium Aspi 55 a statin. Statins loWer cholesterol by inhibiting of 3-hy rin, Amfenac Sodium, Amidopyrine, Aminopropylone, droxy-3-methylglutaryl coenZyme A (HMG CoA) reductase, Ammonium Salicylate, Ampiroxicam, Amyl Salicylate, a key enZyme in the cholesterol biosynthetic pathWay. The Anirolac, Aspirin, Aurano?n, Aurothioglucose, Aurotioprol, statins decrease liver cholesterol biosynthesis, Which AZapropaZone, BendaZac (BendaZac Lysine), Benorylate, increases the production of LDL receptors thereby decreas Benoxaprofen, BenZpiperylone, BenZydamine, Hydrochlo 60 ing plasma total and LDL cholesterol (Grundy, S. M. New ride, Bomyl Salicylate, Bromfenac Sodium, Bufexamac, Engl. J. Med. 319, 24 (1988); Endo,A. J. LipidRes. 33, 1569 BumadiZone Calcium, Butibufen Sodium, Capsaicin, Car (1992)). Depending on the agent and the dose used, statins baspirin Calcium, Carprofen, ChlorthenoxaZin, Choline may decrease plasma triglyceride levels and may increase Magnesium Trisalicylate, Choline Salicylate, Cinmetacin, HDLc. Currently the statins on the market are lovastatin Clofexamide, ClofeZone, Clometacin, Clonixin, Cloraceta 65 (Merck), simvastatin (Merck), pravastatin (Sankyo and dol, Cymene, Diacerein, Diclofenac (Diclofenac Diethylam Squibb) and ?uvastatin (SandoZ). A ?fth statin, atorvastatin monium Salt, Diclofenac Potassium, Diclofenac Sodium), (Parke-Davis/P?Zer), is the most recent entrant into the US 7,271,274 B2 27 28 statin market. Any of these statins can be used in combina as US. Pat. Nos. 6,313,142, 6,310,075, 6,197,786, 6,147, tion With the compounds of the present invention. 090, 6,147,089, 6,140,343, and 6,140,343, the disclosures of MTP inhibitor compounds useful in the combinations and Which is herein incorporated by reference. methods of the present invention comprise a Wide variety of In another aspect, the second biologically active agent can structures and functionalities. Some of the MTP inhibitor be a ?bric acid derivative. Fibric acid derivatives useful in compounds of particular interest for use in the present the combinations and methods of the present invention invention are disclosed in WO 00/ 38725, the disclosure from comprise a Wide variety of structures and functionalities Which is incorporated by reference. Descriptions of these Which have been reported and published in the art. therapeutic compounds can be found in Science, 282, 23 The compounds of the present invention may also be used October 1998, pp. 7514754, herein incorporated by refer in combination or alternation therapy With PPAR agonists ence. including PPARa/y dual agonists, PPARO. agonists, and Cholesterol absorption antagonist compounds useful in PPARY agonists. the combinations and methods of the present invention In another embodiment the present invention encom comprise a Wide variety of structures and functionalities. Some of the cholesterol absorption antagonist compounds of passes a therapeutic combination of a compound of the present invention and an antihypertensive agent. Hyperten particular interest for use in the present invention are sion is de?ned as persistently high blood pressure. In another described in US. Pat. No. 5,767,115, herein incorporated by embodiment, the compound is administered in combination reference. Further cholesterol absorption antagonist com With an ACE inhibitor, a beta andrenergic blocker, alpha pounds of particular interest for use in the present invention, andrenergic blocker, angiotensin II receptor antagonist, and methods for making such cholesterol absorption antago 20 vasodilator and diuretic. nist compounds are described in US. Pat. No. 5,631,365, herein incorporated by reference. G. Pharmaceutical Compositions A number of phytosterols suitable for the combination Any host organism, including a patient, mammal, and therapies of the present invention are described by Ling and speci?cally a human, su?fering from any of the above Jones in “Dietary Phytosterols: A RevieW of Metabolism, 25 described conditions can be treated by the administration of Bene?ts and Side E?fects,” Life Sciences, 57 (3), 1954206 a composition comprising an e?‘ective amount of the com (1995). Without limitation, some phytosterols of particular pound of the invention or a pharmaceutically acceptable salt use in the combination of the present invention are Clo? thereof, optionally in a pharmaceutically acceptable carrier brate, Feno?brate, Cipro?brate, BeZa?brate, Gem?broZil. or diluent. The structures of the foregoing compounds can be found in 30 WO 00/38725. The composition can be administered in any desired Phytosterols are also referred to generally by Nes (Physi manner, including oral, topical, parenteral, intravenous, ology and Biochemistry ofSterols, American Oil Chemists’ intradermal, intra-articular, intra-synovial, intrathecal, intra Society, Champaign, Ill., 1991, Table 7-2). Especially pre arterial, intracardiac, intramuscular, subcutaneous, intraor ferred among the phyto sterols for use in the combinations of 35 bital, intracapsular, intraspinal, intrastemal, topical, trans the present invention are saturated phytosterols or stanols. dermal patch, via rectal, vaginal or urethral suppository, Additional stanols are also described by Nes (Id.) and are peritoneal, percutaneous, nasal spray, surgical implant, inter useful in the combination of the present invention. In the nal surgical paint, infusion pump, or via catheter. In one combination of the present invention, the phytosterol pref embodiment, the agent and carrier are administered in a sloW release formulation such as an implant, bolus, microparticle, erably comprises a stanol. In one preferred embodiment the 40 stanol is campestanol. In another preferred embodiment the microsphere, nanoparticle or nanosphere. For standard infor stanol is cholestanol. In another preferred embodiment the mation on pharmaceutical formulations, see Ansel, et al., stanol is clionastanol. In another preferred embodiment the Pharmaceutical Dosage Farms and Drug Delivery Systems, stanol is coprostanol. In another preferred embodiment the Sixth Edition, Williams & Wilkins (1995). stanol is 22,23-dihydrobrassicastanol. In another embodi 45 An e?‘ective dose for any of the herein described condi ment the stanol is epicholestanol. In another preferred tions can be readily determined by the use of conventional embodiment the stanol is fucostanol. In another preferred techniques and by observing results obtained under analo embodiment the stanol is stigmastanol. gous circumstances. In determining the e?‘ective dose, a Another embodiment the present invention encompasses number of factors are considered including, but not limited a therapeutic combination of a compound of the present 50 to: the species of patient; its siZe, age, and general health; the invention and an HDLc elevating agent. In one aspect, the speci?c disease involved; the degree of involvement or the second HDLc elevating agent can be a CETP inhibitor such severity of the disease; the response of the individual as P?Zer’s Torcetrapib or a combination of a CETP inhibitor patient; the particular compound administered; the mode of and a statin, i.e., Torcetrapib and Atorvastatin. Individual administration; the bioavailability characteristics of the CETP inhibitor compounds useful in the present invention 55 preparation administered; the dose regimen selected; and the are separately described in WO 00/38725, the disclosure of use of concomitant medication. Typical systemic dosages for Which is herein incorporated by reference. Other individual all of the herein described conditions are those ranging from CETP inhibitor compounds useful in the present invention 0.1 mg/kg to 500 mg/kg of body Weight per day as a single are separately described in WO 99/ 14174, EP818448, WO daily dose or divided daily doses. Preferred dosages for the 99/15504, WO 99/14215, WO 98/04528, and WO 00/17166, 60 described conditions range from 541500 mg per day. Amore the disclosures of Which are herein incorporated by refer particularly preferred dosage for the desired conditions ence. Other individual CETP inhibitor compounds useful in ranges from 254750 mg per day. Typical dosages for topical the present invention are separately described in WO application are those ranging from 0.001 to 100% by Weight 00/18724, WO 00/18723, and WO 00/18721, the disclosures of the active compound. of Which are herein incorporated by reference. Other indi 65 The compound is administered for a suf?cient time period vidual CETP inhibitor compounds useful in the present to alleviate the undesired symptoms and the clinical signs invention are separately described in WO 98/35937 as Well associated With the condition being treated. US 7,271,274 B2 29 30 The active compound is included in the pharmaceutically agents such as ethylenediaminetetraacetic acid; buffers such acceptable carrier or diluent in an amount su?icient to as acetates, citrates or phosphates and agents for the adjust deliver to a patient a therapeutic amount of compound in ment of tonicity such as sodium chloride or dextrose. pH can vivo in the absence of serious toxic effects. be adjusted With acids or bases, such as hydrochloric acid or The concentration of active compound in the drug com sodium hydroxide. The parenteral preparation can be position Will depend on absorption, inactivation, and excre enclosed in ampoules, disposable syringes or multiple dose tion rates of the drug as Well as other factors knoWn to those vials made of glass or plastic. of skill in the art. It is to be noted that dosage values Will also If administered intravenously, preferred carriers are vary With the severity of the condition to be alleviated. It is physiological saline, bacteriostatic Water, Cremophor ELTM to be further understood that for any particular subject, (BASF, Parsippany, N.J.) or phosphate buffered saline speci?c dosage regimens should be adjusted over time (PBS). according to the individual need and the professional judg In one form, the active compounds are prepared With ment of the person administering or supervising the admin carriers that Will protect the compound against rapid elimi istration of the compositions, and that the dosage ranges set nation from the body, such as a controlled release formula forth herein are exemplary only and are not intended to limit tion, including implants and microencapsulated delivery the scope or practice of the claimed composition. The active systems. Biodegradable, biocompatible polymers can be ingredient may be administered at once, or may be divided used, such as ethylene vinyl acetate, polyanhydrides, polyg into a number of smaller doses to be administered at varying lycolic acid, collagen, polyorthoesters and polylactic acid. intervals of time. Methods for preparation of such formulations Will be appar A mode of administration of the active compound for 20 ent to those skilled in the art. The materials can also be systemic delivery is oral. Oral compositions Will generally obtained commercially from AlZa Corporation and Nova include an inert diluent or an edible carrier. They may be Pharmaceuticals, Inc. Liposomal suspensions (including enclosed in gelatin capsules or compressed into tablets. For liposomes targeted to infected cells With monoclonal anti the purpose of oral therapeutic administration, the active bodies to viral antigens) are also preferred as pharmaceuti compound can be incorporated With excipients and used in 25 cally acceptable carriers. These may be prepared according the form of tablets, troches or capsules. Pharmaceutically to methods knoWn to those skilled in the art, for example, as compatible binding agents, and/or adjuvant materials can be described in US. Pat. No. 4,522,811 (Which is incorporated included as part of the composition. herein by reference in its entirety). For example, liposome The tablets, pills, capsules, troches and the like can formulations may be prepared by dissolving appropriate contain any of the folloWing ingredients, or compounds of a 30 lipid(s) (such as stearoyl phosphatidyl ethanolamine, similar nature: a binder such as microcrystalline cellulose, stearoyl phosphatidyl choline, arachadoyl phosphatidyl cho gum tragacanth or gelatin; an excipient such as starch or line, and cholesterol) in an inorganic solvent that is then lactose, a disintegrating agent such as alginic acid, Primogel, evaporated, leaving behind a thin ?lm of dried lipid on the or corn starch; a lubricant such as magnesium stearate or surface of the container. An aqueous solution of the com Sterotes; a glidant such as colloidal silicon dioxide; a 35 pound is then introduced into the container. The container is sWeetening agent such as sucrose or saccharin; or a ?avoring then sWirled by hand to free lipid material from the sides of agent such as peppermint, methyl salicylate, or orange the container and to disperse lipid aggregates, thereby form ?avoring. ing the liposomal suspension. When the dosage unit form is a capsule, it can contain, in Suitable vehicles or carriers for topical application can be addition to material of the above type, a liquid carrier such 40 as a fatty oil. In addition, dosage unit forms can contain prepared by conventional techniques, such as lotions, sus various other materials Which modify the physical form of pensions, ointments, creams, gels, tinctures, sprays, poW the dosage unit, for example, coatings of sugar, shellac, or ders, pastes, sloW-release transdermal patches, suppositories other enteric agents. for application to rectal, vaginal, nasal or oral mucosa. In addition to the other materials listed above for systemic For example, the compound can be in the form of a dosage 45 unit including about 0.5*1000 mg of compound, e.g., in a administration, thickening agents, emollients and stabiliZers tablet or capsule. can be used to prepare topical compositions. Examples of The compound or its salts can be administered as a thickening agents include petrolatum, beesWax, xanthan component of an elixir, suspension, syrup, Wafer, cheWing gum, or polyethylene, humectants such as sorbitol, emol lients such as mineral oil, lanolin and its derivatives, or gum or the like. A syrup may contain, in addition to the 50 active compounds, sucrose as a sWeetening agent and certain squalene. preservatives, dyes and colorings and ?avors. Any of the compounds described herein for combination The compound can also be mixed With other active or alternation therapy can be administered as any derivative materials that do not impair the desired action, or With that upon administration to the recipient, is capable of materials that supplement the desired action. The com 55 providing directly or indirectly, the parent compound, or that pounds can also be administered in combination With non exhibits activity itself. Nonlimiting examples are the phar steroidal antiin?ammatories such as ibuprofen, indometha maceutically acceptable salts (alternatively referred to as cin, fenoprofen, mefenamic acid, ?ufenamic acid, sulindac. “physiologically acceptable salts”), and a compound Which The compound can also be administered With corticosteri has been alkylated or acylated at an appropriate position. ods. 60 The modi?cations can affect the biological activity of the Solutions or suspensions used for parenteral, intradermal, compound, in some cases increasing the activity over the subcutaneous, or topical application can include the folloW parent compound. This can easily be assessed by preparing ing components: a sterile diluent such as Water for injection, the derivative and testing its anti-in?ammatory activity saline solution, ?xed oils, polyethylene glycols, glycerine, according to knoWn methods. propylene glycol or other synthetic solvents; antibacterial 65 The folloWing examples are understood to be illustrative agents such as benZyl alcohol or methyl parabens; antioxi only and are not intended to limit the scope of the present dants such as ascorbic acid or sodium bisul?te; chelating invention in any Way. US 7,271,274 B2 31 32 EXAMPLES g) Was added. The mixture Was stirred at room temperature for 1 hour and then poured into 1 N HCl solution (100 mL). Intermediates and ?nal products Were characterized by It Was extracted With dichloromethane (2x100 mL), dried conventional proton NMR, mass spectral analyses and stan over magnesium sulfate, and evaporated. Silica gel chroma dard analytical methods knoWn to those skilled in the art. tography (dichloromethane/ethyl acetate 4:1) gave 4-{2,6 Di-tert-butyl-4- [1 -(3 ,5 -di-tert-butyl-4 -hydroxyphenylsulfa Example 1 nyl)-1-methylethyl-sulfanyl]phenoxy}butane-1,2(S),-3 (s) triol as a White poWder (0.45 g, 66%), mp 694710 C. lH-NMR (CDCl3) 6 7.56_(s, 2H, Ph-H), 7.45 (s, 2H, Ph-H), 10 s s 5.36 (s, 1H, PhOH), 4.264.34 (m, 1H, PhOCHZCH), 4.01 (dd,_ 1:9, 10, 1H, CHiOi), 3.723.86 (m, 4H, , CHiOi), X @H 2.692.77 (m, 2H, OH), 2.15 (br. t, 1H, OH), 1.421.47 (m, 42H, S,S'-isopropylidene, tert-butyls). MS m/Z: 643 ([M+ 2 OH Na]+, 100%). Anal. calcd. for C35H256O5S2: C67.70, H9.09, 15 S10.33; found: C67.33, H9.02, S10.03. Ollu H Example 2 4-{2,6-Di-tert-butyl-4-[1-(3,5-di-tert-butyl-4-hy droxyphenylsulfanyl)- 1 -methylethyl-sulfanyl] phenoxy}butane-1,2(S),3(S)-triol 20

Ex-1A HO <0 25 4- {2,6-Di-tert-butyl-4- [1 -(3 ,5 -di-tert-butyl-4-hy "HO droxyphenylsulfanyl)- 1 -methylethylsulfanyl] - do 30 phenoxy }butane- 1 ,2 (R), 3 (R)-triol

The title compound Was prepared using the same proce OH dure as described in Ex-1 starting from probucol and 2-ethoxy-1,3-dioxolane-4(R),5(R)-dimethanol. White solid, To a solution of probucol (5.16 g, 10 mmol) in THF (20 35 mp 694710 C. lH-NMR (CDCl3) 6 7.56 (s, 2H, Ph-H), 7.45 mL) cooled to 0° C. Were added triphenylphosphine (1.3 g, (s, 2H, Ph-H), 5.38 (s, 1H, PhOH), 4.254.32 (m,~ 1H, 5 mmol), diethyl aZodicarboxylate (0.8 mL g, 5 mmol) and 2-ethoxy-1,3-dioxolane-4(S),5(S)-dimethanol (0.89 g, 5 PhOCH2CH), 3.96 (dd, E9, 10, 1H, CHADi), 3.723.86 (m, 4H, , CHiOi), 3.003.40 (m, 3H, OH), 1.46 (s, 6H, mmol; Nicolaou, K. C. et al.: J. Org. Chem. 1985, 50, S,S'-isopropylidene), 1.45 (s, 18H, tert-butyls), 1.43 (s, 18H, 144041456). The resultant mixture Was stirred at re?ux for 40 3 hours and then evaporated. Silica gel chromatography tert-butyls). MS m/Z: 643 ([M+Na]+, 100%).Anal. calcd. for (hexanes/ethyl acetate 4: 1) gave 2,6-di-tert-butyl-4-{1-[3,5 C35H56O5S2.1/3H2O: C67.05, H9.11, S, 10.23; found: di-tert-butyl-4-(2 -ethoxy-5 (S) -hydroxymethyl[ 1 ,3] diox C67.03, H9.06, S10.01. olan-4(S)-yl-methoxy)-phenylsulfanyl]-1-methyl-ethylsul fanyl}-phenol in three parts: diasteroisomer A (0.36 g), Example 3 diastereoisomer B (0.22 g) and a mixture of the tWo (0.72 g) all as viscous residues (total yield: 39%). Diastereoisomer A: lH-NMR (CDCl3) 6 7.56 (s, 2H, Ph-H), 7.44 (s,_ 2H, Ph-H), 5.89 [s, 1H, CH(O) 3], 5.37 (s, 1H, PhOH), 4.734.81 (m, 1H, PhOCH2CH), 4.194.25 (m, 1H, CHCH2OH), 3.8 04.01 (m, 3H, CH2O), 3.623.96 (m, 3H, CH2O), 2.80 (dd, 50 1:9, 3, 1H, OH), 1.45 (s, 6H, S,S'-isopropylidene), 1.44 (s, .X. 0 18H, tert-butyls), 1.43 (s, 18H, tert-butyls), 1.25 (t, 3H, OCH2CH3). Diasteroisomer B: lH-NMR (CDCl3) 6 7.56 (s, 2H, Ph-H), 7.44 (s, 2H, Ph-H), 5.92 [s, 1H, CH(O) 3], 5.36 55 in? gmm (s, 1H, PhOH), 4.504.59 (m, 1H, PhOCHZCH), 4.134.30 (m, 3H, CHCH2OH, CH2O), 3.894.00 (m, 3H, CH2O), 3.61 (quad, 1:7, 2H, OCH2CH3), 1.86 (dd, 1:6, 8, 1H, OH), 1.4 11.46 (m, 42H, S,S'-isopropylidene, tert-butyls), 1.21 (t, 3H, (2-{2,6-Di-tert-butyl-4-[ 1 -(3 ,5 -di-tert-butyl-4-hy OCH2CH3). droxyphenylsulfanyl)- 1 -methylethylsulfanyl] The mixture obtained from Ex-1A (0.72 g, 1.1 mmol) Was phenoxy}acetylamino)acetic acid dissolved in methanol (25 mL). Acetic acid (2 mL) and Water (1 mL) Were added and the resultant mixture Was Example 3A stirred at re?ux for 3 hours and then evaporated to about 10 mL. It Was poured into Water (100 mL) and extracted With To a solution of probucol (0.5 g, 0.97 mmol) in dimeth dichloromethane (2><100 mL). The organic phase Was dried 65 ylformamide (1.5 mL) Were added ethyl iodoacetate (0.31 g, over magnesium sulfate and evaporated. The residue Was 1.45 mmol) and 40% potassium ?uoride on alumina (0.7 g, dissolved in methanol (15 mL) and potassium carbonate (0.5 4.8 mmol). The mixture Was stirred for 24 hours and then US 7,271,274 B2 33 34 diluted With ether (25 mL), ?ltered and Washed With Water Example 4 (2><5 mL). The ether layer Was dried over MgSO4, ?ltered and concentrated. Silica gel chromatography (hexanes/ ether 5:95) yielded 160 mg of {2,6-di-tert-butyl-4-[1-(3,5-di-tert butyl-4 -hydroxy-phenylsulfanyl)-1 -methyl-ethylsulfanyl] - phenoxy}-acetic acid ethyl ester Which Was used directly for the next step of reaction. .X. O

Example 3B {2,6-Di-tert-butyl-4-[1-(3,5-di-tert-butyl-4-hydroxy-phe i? 95*)“ nylsulfanyl)-1 -methyl-ethylsulfanyl] -phenoxy} -acetic acid ethyl ester (Ex-3A, 160 mg) Was dissolved in THF/HZO/ [(2-{2,6-Di-tert-butyl-4-[ 1 -(3 ,5 -di-tert-butyl-4-hy MeOH (4:1:1) (4 mL) and lithium hydroxide hydrate (50 droxyphenylsulfanyl)- 1 -methylethylsulfanyl] mg) Was added. The resultant mixture Was stirred for 1 h and phenoxy}acetyl)methylamino]acetic acid then neutraliZed With 1N HCl. It Was extracted With ether (2><10 mL), dried over MgSO4, ?ltered, and concentrated. The title compound Was prepared in a similar procedure Silica gel chromatography (hexanes/ ether 50:50) gave acetic 20 as described in Ex-3 starting from acetic acid, [4-[[1-[[3,5 acid, [4-[[1-[[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenyl] bis(1 ,1 -dimethylethyl) -4 -hydroxyphenyl]thio] -1 -methyl ethyl]thio]-2,6-bis(1, 1 -dimethylethyl)phenoxy] - (Ex-3B) thio]-1-methylethyl]thio]-2,6-bis(1,1-dimethylethyl)-phe and sarcosine ethyl ester. Off-White solid, mp 1154119o C. noxy]as a White solid (90 mg, 16%), mp 1644165o C. lH-NMR (CDCl3) 6 7.58 (s, 2H, Ph-H), 7.45 (s, 2H, Ph-H), lH-NMR (CDCl3) 6 7.55 (s, 2H, Ph-H), 7.40 (s, 2H, Ph-H), 5.37 (s, 1H, Ph-OH), 4.52 (s, 2H, PhOCHZ), 4.24 (s, 2H, 5.35 (s, 1H, Ph-OH), 4.40 (s, 2H, OCH2COOH), 1.43 (s, 6H, 25 NCH2), 3.03 (s, 3H, NCH3), 1.47 (s, 6H, S,S'-isopropy S,S'-isopropylidene), 1.41 (s, 9H, tert-butyl), 1.39 (s, 9H, lidene), 1.44 (s, 18H, tert-butyls), 1.43 (s, 18H, tert-butyls). tert-butyl). MS m/Z: 613 ([M+K]+, 60%), 159 (100%). Anal. MS m/Z: 668 ([M+Na]+, 100%). Anal. calcd. for C36H55NO5S2: C66.94, H9.58, N2.17, S9.93; found: calcd. for C33H5OO4S2: C68.95, H8.77, S11.15, found: C67.13, H8.58, N2.20, S9.79. C68.67, H8.72, S11.09. 30 Example 5 Example 3C

To a solution of acetic acid, [4-[[1-[[3,5-bis(1,1-dimeth ylethyl)-4-hydroxyphenyl]thio] -1 -methylethyl]thio] -2,6-bis 35 (1,1-dimethylethyl)phenoxy]-(Ex-3B, 50 mg, 0.087 mmol) in methylene chloride (0.87 mL) Were added glycine ethyl ester hydrochloride (15.8 mg, 0.11 mmol), 1-(3-dimethy .0 X laminopropyl-3-ethyl carbodiimide hydrochloride (22 mg, 0.11 mmol) and 4-dimethylaminopyridine (28 mg, 0.23 40 mmol). The reaction mixture Was stirred overnight and then the methylene chloride evaporated. The reaction Was diluted With ether (10 mL) and Washed With Water (2><3 mL), dried over MgSO4, ?ltered, and concentrated. The crude mixture 3 -(2- {2, 6-Di-tert-butyl-4- [1 -(3, 5-di-tert-butyl-4 Was puri?ed by silica gel chromatography (ether/hexanes hydroxyphenylsulfanyl)- 1 -methyl ethyl sulfanyl] 50:50) to give 50 mg of (2-{2,6-di-tert-butyl-4-[1-(3,5-di phenoxy}acetylamino)propionic acid tert-butyl-4 -hydroxy -phenylsulfanyl)-1 -methyl -ethyl sulfa nyl]-phenoxy}-acetylamino)-acetic acid ethyl ester Which as Example 5A used directly for the next step of reaction. 50 3-(2-{2,6-Di-tert-butyl-4-[1-(3,5-di-tert-butyl-4-hydrox (2-{2,6-Di-tert-butyl-4-[1-(3,5-di-tert-butyl-4-hydroxy yphenyl sulfanyl) -1 -methylethylsulfanyl] phenylsulfanyl)-1 -methyl -ethyl -sulfanyl] -phenoxy} -acety phenoxy}acetylamino)propionic acid ethyl ester Was pre lamino)-acetic acid ethyl ester (Ex-3C, 50 mg) Was dis pared in a similar procedure as described in Ex-3C starting solved in THF/HZO/MeOH (2:1:1, 1 mL), lithium hydroxide from acetic acid, [4-[[1-[[3,5-bis(1,1-dimethylethyl)-4-hy monohydrate (15 mg, 0.36 mmol) Was added, and the 55 droxyphenyl]thio]-1-methylethyl]thio]-2,6-bis(1, 1 -dimeth reaction stirred for 1 h. The reaction Was neutraliZed With 1N ylethyl)-phenoxy]-(Ex-3B) and ethyl 3-aminobutyrate. Off HCl and extracted With ether (2><10 mL), dried over MgSO4, White solid, mp 1164117o C. lH-NMR (CDCl3) 6 7.56 (s, ?ltered, and concentrated to give the title product as a 2H, Ph-H), 7.43 (s, 2H, Ph-H), 7.31 (t, 1:6, 1H, NH), 5.37 viscous residue (25 mg, 45%) Which solidi?ed, mp 914940 (s, 1H, Ph-OH), 4.24 (s, 2H, PhOCHz), 4.17 (quad, 1:7, 2H, 60 OCH2CH3), 3.69 (quad, 1:6, 2H, NHCH2), 2.61 (t, 1:6, C. lH-NMR (CDCl3) 6 7.56 (s, 2H, Ph-H), 7.42 (s, 2H, NHCH2CH2), 1.45 (s, 6H, S,S'-isopropylidene), 1.44 (s, Ph-H), 7.28 (t, 1H, NH), 5.39 (br s, 1H, Ph-OH), 4.31 [s, 2H, 18H, tert-butyls), 1.39 (s, 18H, tert-butyls), 1.28 (t, 1:7, 3H, OCH2C(O)], 4.22 (d, 1:52 HZ, 2H, NHCHziCOOH), 1.44 OCH2CH3). MS m/Z: 696 ([M+Na]+, 100%). Anal. calcd. (s, 6H, S,S'-isopropylidene), 1.42 (s, 18H, tert-butyls), 1.39 for C38H59NO5S2: C67.72, H8.82, N2.08; found: 67.67, (s, 18H, tert-butyls). MS m/Z: 654 ([M+Na]+, 100%). Anal. 65 H8.83, N2.04. calcd. for C35H53NO5S2: C66.52, H8.45, N2.22, S10.14; The title compound Was prepared in a similar procedure found: C66.23, H8.30, N2.23, S9.98. as described in Ex-3 by hydrolyZing 3-(2-{2,6-di-tert-butyl US 7,271,274 B2 35 3 6 4-[ 1 -(3 ,5 -di-tert-butyl-4-hydroxyphenylsulfanyl)-1 -methyl 4- {2,6-Di-tert-butyl-4- [1 -(3 ,5 -di-tert-butyl-4-hy ethylsulfanyl]phenoxy}acetylamino)propionic acid ethyl droxyphenylsulfanyl)- 1 -methylethylsulfanyl] - ester (Ex-5A). Off-White solid, mp 1834184o C. lH-NMR phenoxy}butane-1,2(S),3(R)-triol (CDCl3) 6 7.56 (s, 2H, Ph-H), 7.43 (s, 2H, Ph-H), 7.33 (t, 1:6, 1H, NH), 5.37 (s, 1H, Ph-OH), 4.25 (s, 2H, PhOCH2), 5 4.69 (quad, 1:6, 2H, NHCH2), 2.68 (t, 1:6, NHCH2CH2), 1.45 (s, 6H, S,S'-isopropylidene), 1.44 (s, 18H, tert-butyls), 1.38 (s, 18H, tert-butyls). MS m/Z: 668 ([M+Na]+, 100%). Anal. calcd. for C36H55NO5S2: C66.94, H8.58, N2.17; 10 found: 67.10, H8.62, N2.03. Example 6 OIII

4- {2,6-Di-tert-butyl-4- [1 -(3 ,5 -di-tert-butyl-4-hy droxyphenylsulfanyl)- 1 -methylethylsulfanyl] - s s OH phenoxy}butane-1,2(R),3(S)-triol 0 HO 0% OH The title compounds can be prepared using the same N procedure as described in Ex-1 starting from probucol and H 2-ethoxy-1,3-dioxolane-4(S),5(R)-dimethanol and perform ing a chiral separation. 25 Example 9 2-{2,6-Di-tert-butyl-4-[1-(3,5-di-tert-butyl-4-hy droxyphenylsulfanyl)- 1 -methyl -ethylsulfanyl]phe noxy} -N- (2 -hydroxy- 1 -hydroxymethyl-ethyl)aceta mide

To a solution of acetic acid, [4-[[1-[[3,5-bis(1,1-dimeth ylethyl)-4-hydroxyphenyl]-thio]-1-methylethyl]thio]-2,6 bis(1,1-dimethylethyl)phenoxy]-(Ex-3B, 4.87 g, 8.49 mmol) in dichloromethane (200 mL) Was added serinol (0.77 g, 8.49 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodi imide hydrochloride (3.26 g, 17 mmol), and the mixture Was stirred at It overnight. It Was poured into Water (100 ml) and 2, 6-Di-tert-butyl-4-{ 1 - [3 ,5 -di-tert-butyl-4-(2 -ethoxy the organic phase Was dried over magnesium sulfate and 5 (R) -hydroxymethyl-[ 1 ,3]dioxolan-4 (R)-yl methoxy)-phenyl sulfanyl] -1 -methyl-ethylsulfanyl} - evaporated. Silica gel chromatography (hexanes/ethyl phenol acetate 1:2) gave the title compound as a White solid (2.95 g, 57%), mp 1634164o C. lH-NMR (CDCl3) 6 7.57 (s, 2H, The title compound Was prepared using the same proce Ph-H), 7.49 (d, 1:7, 1H, NH), 7.44 (s, 2H, Ph-H), 5.37 (s, dure as described in Ex-1A starting from probucol and 1H, Ph-OH), 4.28 (s, 2H, PhOCHZ), 4.104.29 (m, 1H, 2-ethoxy-1,3-dioxolane-4(R),5(R)-dimethanol as a White NHCH), 3.824.00 (m, 4H, CHZOH), 1.45 (s, 6H, S,S' solid (10.3 g, 35%), mp 6(L62o C. 1H-NMR (300 MHZ, isopropylidene), 2.44 (dd, 1:6, 11, 2H, OH), 1.44 (s, 18H, CDCl3) 6 7.56 (s, 2H), 7.44 (s, 2H), 5.89 (s, 1H), 5.36 (s, tert-butyls), 1.41 (s, 18H, tert-butyls). MS m/Z: 670 ([M+ 1H), 4.754.78 (m, 1H), 4.194424 (m, 2H), 3.924399 (m, Na]+, 100%). Anal. calcd. for C36H57NO5S2: C66.73, 2H), 3.84 (dd, 1H, 1:100, 4.2 HZ), 36043.74 (m, 2H), 2.79 H8.87, N2.16 S9.90; found: C66.37, H8.90, N2.24, S9.92. (dd, 1H, 1:91, 3.3 HZ). 14341.45 (m, 42H), 1.28 (t, 3H, J:7.2 HZ). HRMS (ESI) calcd for C38H6OO6S2 (M+Na), Example 7 and Example 8 699.3729; found, 699.3756.Anal. calcd. for C38H6OO6S2: C, 67.42, H, 8.93, S, 9.47; found: C, 67.04, H, 9.00, S, 8.86. 55 Example 10

HO HO