WO2011017448A1

WO2011017448A1 - Use of histone deacetylase inhibitors for treatment of autoimmune diseases

Info

Publication number: WO2011017448A1
Application number: PCT/US2010/044443
Authority: WO
Inventors: Alain H. Rook
Original assignee: The Trustees Of The University Of Pennsylvania
Priority date: 2009-08-05
Filing date: 2010-08-04
Publication date: 2011-02-10

Abstract

The invention includes a method of treating, ameliorating or preventing an autoimmune disease or disorder in a subject, comprising administering to the subject an effective amount of a composition comprising a histone deacetylase (HDAC) inhibitor.

Description

TITLE OF THE INVENTION

Use of Histone Deacetylase Inhibitors for Treatment of Autoimmune Diseases

BACKGROUND OF THE INVENTION

Histone deacetylases (HDACs) are enzymes that catalyze the removal of acetyl groups from lysine residues in the amino terminal tails of the nucleosomal core histones. Histone tails are normally positively charged due to amine groups present on their lysine and arginine amino acids. These positive charges allow the histone tails to interact with and bind to the negatively charged phosphate groups on the DNA backbone. Acetylation, which occurs normally in a cell, neutralizes the positive charges on the histone by changing amines into amides, and decreases the ability of the histones to bind to DNA. This decreased binding allows chromatin expansion and permits genetic transcription to take place. On the other hand, histone deacetylases remove those acetyl groups, increasing the positive charge of histone tails and encouraging high-affinity binding between the histones and DNA backbone. The increased DNA binding condenses DNA structure, preventing transcription.

As such, HDACs together with histone acetyl transferases (HATs) regulate the acetylation status of histones. HDACs may be divided into three classes based on structural homology. Class I HDACs (HDACs 1, 2, 3 and 8) bear similarity to the yeast RPD3 protein, are located in the nucleus and are found in complexes associated with transcriptional co-repressors. Class II HDACs (HDACs 4, 5, 6, 7 and 9) are similar to the yeast HDAl protein, and have both nuclear and cytoplasmic sub- cellular localization. Both Class I and II HDACs are inhibited by hydroxamic acid- based HDAC inhibitors, such as SAHA. Class III HDACs form a structurally distinct class of NAD-dependent enzymes that are related to the yeast SIR2 proteins and are not inhibited by hydroxamic acid-based HDAC inhibitors. Inhibitors of HDACs, such as the hydroxamic acid-based hybrid polar compound suberoylanilide hydroxamic acid (SAHA), induce growth arrest, differentiation and/or apoptosis of transformed cells in vitro and inhibit tumor growth in vivo.

Autoimmunity is the failure of an organism to recognize its own constituent parts as self, which allows an immune response against its own cells and tissues. Any disease that results from such an aberrant immune response is termed an autoimmune disease. Prominent examples include coeliac disease, diabetes mellitus type 1 (IDDM), systemic lupus erythematosus (SLE), Sjogren's syndrome, Churg- Strauss Syndrome, Hashimoto's thyroiditis, Graves' disease, idiopathic

thrombocytopenic purpura, and rheumatoid arthritis (RA).

Several mechanisms are thought to be operative in the pathogenesis of autoimmune diseases, against a backdrop of genetic predisposition and environmental modulation. Some of the important mechanisms are: T-cell bypass; T-cell-B-cell discordance; aberrant B cell receptor-mediated feedback; molecular mimicry;

idiotype cross-reaction; cytokine dysregulation; dendritic cell apoptosis; and epitope spreading or epitope drift.

Autoimmune diseases may be broadly divided into systemic and organ-specific (or localized) autoimmune disorders, depending on the principal clinico-pathologic features of each disease.

(a) Systemic autoimmune diseases include systemic lupus erythematosus, Sjogren's syndrome, scleroderma, rheumatoid arthritis, and dermatomyositis. These conditions tend to be associated with auto-antibodies to antigens that are not tissue- specific. Thus, although polymyositis is more or less tissue-specific in presentation, it may be included in this group because the auto-antigens are often ubiquitous t-RNA synthetases.

(b) Local syndromes, which affect a specific organ or tissue include endocrinologic (diabetes mellitus type 1, Hashimoto's thyroiditis, Addison's disease); gastrointestinal (coeliac disease, pernicious anaemia); dermatologic: (pemphigus vulgaris, vitiligo); haematologic (auto-immune haemolytic anaemia, idiopathic thrombocytopenic purpura); and neurological (myasthenia gravis).

Using the traditional "organ specific" and "non-organ specific" classification scheme, many diseases have been lumped together under the

autoimmune disease umbrella. However, many chronic inflammatory human disorders lack the telltale associations of B and T cell driven immunopathology. In the last decade it has been firmly established that tissue "inflammation against self does not necessarily rely on abnormal T and B cell responses. This has led to the recent proposal that the spectrum of autoimmunity should be viewed along an

"immunological disease continuum," with classical autoimmune diseases at one extreme and diseases driven by the innate immune system at the other extreme.

Within this scheme, the full spectrum of autoimmunity may be included. Many common human autoimmune diseases may be seen to have a substantial innate immune mediated immunopathology using this new scheme.

Treatments for autoimmune disease have traditionally been immunosuppressive, anti-inflammatory, or palliative. Non-immunological therapies, such as hormone replacement in Hashimoto's thyroiditis or Type 1 diabetes mellitus, treat outcomes of the autoaggressive response and are palliative treatments. Dietary manipulation limits the severity of celiac disease. Steroidal or NSAID treatment limits inflammatory symptoms of many diseases. IVIG is used for chronic inflammatory polyneuropathy and Guilain-Barre syndrome. Specific

immunomodulatory therapies, such as the TNF-α antagonists (e.g. etanercept), the B- cell depleting agent rituximab, the anti-IL-6 receptor tocilizumab and the

costimulation blocker abatacept have been shown to be useful in treating rheumatoid arthritis. Some of these immunotherapies may be associated with increased risk of adverse effects, such as susceptibility to infection.

Autoimmune skin diseases pose a serious health threat due to life- threatening infection. In addition, autoimmune skin diseases may cause symptoms such as skin blisters, rashes, inflammation, lesions, and disfigurement. Pemphigus is a group of rare autoimmune skin disorders characterized by the development of blisters in the outer layer of the skin (epidermis) and mucous membranes (thin moist layers that line the body's internal surfaces). The location and type of blisters vary according to the type of pemphigus. If left untreated, pemphigus may be a serious illness.

Blisters in the outer layer of the skin are common to all types of pemphigus. Blisters develop due to the destruction of the "cement" that holds cells together (epidermal acantholysis), resulting in the separation of cells from one another. Soft (flaccid) blisters generally occur on the neck, scalp, mucous membranes, and/or underarm (axillary) and groin areas (inguinal). Most patients with pemphigus have deposits of IgG around the blistered areas in the epidermal cells called keratinocytes. Anti-epidermal antibodies directed against skin cells are typically present in the fluid of the blisters. The diagnosis of pemphigus requires microscopic examination of cells in the blisters, as well as detection of the IgG antibodies that characterize this disease.

Bullous pemphigoid is a rare, chronic condition in which fluid-filled blisters (bullae) erupt on the surface of the skin, usually the arms, legs or trunk of the affected individual. The cause of bullous pemphigoid is related to the deposition of an IgG antibody directed towards the bullous pemphigoid antigen within the skin.

The goal of bullous pemphigoid treatment is usually to reduce inflammation— thereby easing the symptoms— and suppressing the autoimmune response in skin. Treatment usually involves a multidrug approach, including the use of corticosteroids to relieve inflammation, immunosuppressants, such as azathioprine (Imuran) and mycophenolate mofetil (CellCept), to attenuate the autoimmune response in skin, which in turn reduces inflammation, and corticosteroid-sparing agents. Drugs that help reduce the dosage or the need for continued use of corticosteroids are referred to as "corticosteroid-sparing." Examples include dapsone and tetracycline medications, as well as immunosuppressant drugs in more severe cases. Left untreated, bullous pemphigoid may take as long as six to ten years to disappear completely. Even with therapy, bullous pemphigoid usually takes one and a half to five years to resolve.

There is an urgent need in the art for effective pharmacological approaches to suppress or control auto-immunity as a way to treat or ameliorate autoimmune diseases. There is also an urgent need in the art for effective

pharmacological approaches to treat, ameliorate or prevent autoimmune skin diseases, such as bullous pemphigoid. The present invention fills these needs.

BRIEF SUMMARY OF THE INVENTION

In one aspect, the invention includes a method of treating or ameliorating an autoimmune disease or disorder in a subject in need thereof. The method comprises administering to the subject a therapeutically effective amount of a composition comprising a pharmaceutically acceptable carrier and a compound selected from the group consisting of a histone deacetylase (HDAC) inhibitor and a pharmaceutically acceptable salt or hydrate thereof.

In one embodiment, the autoimmune disease or disorder is selected from the group consisting of coeliac disease, diabetes mellitus type 1 (IDDM), systemic lupus erythematosus (SLE), Sjogren's syndrome, Churg-Strauss Syndrome, Hashimoto's thyroiditis, Graves' disease, idiopathic thrombocytopenic purpura, rheumatoid arthritis (RA), polymyositis, ulcerative colitis, Crohn's disease, autoimmune carditis, Wegener's granulomatosis, autoimmune hemolytic anemia, polyarteritis nodosa, psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris.

In one embodiment, the autoimmune disease or disorder is a skin autoimmune disease or disorder. In another embodiment, the skin autoimmune disease or disorder is selected from the group consisting of psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris. In yet another embodiment, the skin autoimmune disease or disorder is bullous pemphigoid. In yet another embodiment, the HDAC inhibitor is vorinostat. In yet another embodiment, the subject is a mammal. In yet another embodiment, the mammal is a human. In yet another embodiment, the dose of the HDAC inhibitor in the composition is about 400 mg per day.

In another aspect, the invention includes a method of treating a subject at risk of developing an autoimmune disease or disorder. The method comprises administering to the subject a therapeutically effective amount of a composition comprising a histone deacetylase (HDAC) inhibitor, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

In one embodiment, the autoimmune disease or disorder is selected from the list consisting of coeliac disease, diabetes mellitus type 1 (IDDM), systemic lupus erythematosus (SLE), Sjogren's syndrome, Churg-Strauss Syndrome,

Hashimoto's thyroiditis, Graves' disease, idiopathic thrombocytopenic purpura, rheumatoid arthritis (RA), polymyositis, ulcerative colitis, Crohn's disease, autoimmune carditis, Wegener's granulomatosis, autoimmune hemolytic anemia, polyarteritis nodosa, psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris.

In yet another embodiment, the autoimmune disease or disorder is a skin autoimmune disease or disorder. In another embodiment, the skin autoimmune disease or disorder is selected from the group consisting of psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris. In yet another embodiment, the skin autoimmune disease or disorder is bullous pemphigoid. In yet another embodiment, the HDAC inhibitor is vorinostat. In yet another embodiment, the subject is a mammal. In yet another embodiment, the mammal is a human. In yet another embodiment, the dose of the HDAC inhibitor in the composition is about 400 mg per day.

BRIEF DESCRIPTION OF THE DRAWINGS

For the purpose of illustrating the invention, there are depicted in the drawings certain embodiments of the invention. However, the invention is not limited to the precise arrangements and instrumentalities of the embodiments depicted in the drawings.

Figure 1 is a bar graph illustrating the percentage of K562 cells that was lysed upon treatment with various compounds, for PBMC NK cells isolated from CTCL patients. Effector-to-target ratio was 50:1.

Figure 2 is a bar graph illustrating the percentage of K562 cells that was lysed upon treatment with various molecules, for purified NK cells. Effector-to- target ratio was 2: 1.

Figure 3, comprising Figures 3 A and 3B, is a set of bar graphs illustrating the inhibition of dendritic cell-dependent cytokine production upon treatment with various molecules. Figure 3 A illustrates the effect of vorinostat on INF-α production. Figure 3 B illustrates the effect of vorinostat on IL- 12 production.

Figure 4 is a bar graph illustrating the effect of vorinostat on activation of antigen presenting cells (APCs).

DETAILED DESCRIPTION OF THE INVENTION

In one aspect, the present invention relates to the unexpected discovery that histone deacetylase (HDCA) inhibitors are useful in treating, ameliorating or preventing an autoimmune disease or disorder in a subject. In another aspect, the present invention relates to the unexpected discovery that HDCA inhibitors are useful in treating, ameliorating or preventing an autoimmune skin disease or disorder in a subject.

The present invention provides methods of treating a subject that is afflicted with or at risk of developing an autoimmune disease or disorder by administering a pharmaceutical compositions comprising a HDAC inhibitor to the subject in need thereof. In another aspect, the present invention provides methods of treating a subject that is afflicted with or at risk of developing an autoimmune skin disease or disorder by administering a pharmaceutical compositions comprising a HDAC inhibitor to the subject in need thereof.

Definitions.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although any methods and materials similar or equivalent to those described herein may be used in the practice for testing of the present invention, the preferred materials and methods are described herein. In describing and claiming the present invention, the following terminology will be used.

It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

The articles "a" and "an" are used herein to refer to one or to more than one (i.e. to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

The term "about" will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term "about" is meant to encompass variations of ± 20% or ± 10%, more preferably ± 5%, even more preferably ± 1 %, and still more preferably ± 0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

As used herein, the term "APC" refers to antigen presenting cell.

As used herein, the term "immunoglobulin" or "Ig" is defined as a class of proteins that function as antibodies. The five members included in this class of proteins are IgA, IgG, IgM, IgD, and IgE. IgA is the primary antibody that is present in body secretions, such as saliva, tears, breast milk, gastrointestinal secretions and mucus secretions of the respiratory and genitor-urinary tracts. IgG is the most common circulating antibody. IgM is the main immunoglobulin produced in the primary immune response in most mammals. It is the most efficient

immunoglobulin in agglutination, complement fixation, and other antibody responses, and is important in defense against bacteria and viruses. IgD is the immunoglobulin that has no known antibody function, but may serve as an antigen receptor. IgE is the immunoglobulin that mediates immediate hypersensitivity by causing release of mediators from mast cells and basophils upon exposure to allergen.

As used herein, the term "vorinostat" refers to TV-hydroxy-iV-phenyl- octane-l,8-diamide, and the term "SAHA" refers to suberoylanilide hydroxamic acid. The term "vorinostat" and "SAHA" refer to the same compound, marketed under the name Zolinza® (Patheon, Mississauga, Ontario, CA, & Merck & Co., White House Station, NJ).

The phrase "autoimmune disease or disorder," as used herein, refers to an aberrant reaction of the host adaptive or innate immune systems such that the host immune system is activated against substances (such as proteins or antigens), cells, tissues, or organs normally present in the body.

The phrase "autoimmune skin disease or disorder," as used herein, refers to an aberrant reaction of the host adaptive or innate immune systems such that the host immune system is activated against substances (such as proteins or antigens), cells, or tissues normally present in the skin.

As used herein, the term "ameliorate" or "alleviate" refers to the lessening, decrease, or diminishing of a symptom, state, or condition. In one aspect, a symptom of a disease is ameliorated or alleviated when the symptom decreases in severity of occurrence or effect in a patient. In another aspect, a symptom of a disease is ameliorated or alleviated when the symptom is completely eradicated or eliminated from the patient.

As used herein, a "subject" of diagnosis or treatment is a mammal, including a human. Non -human animals subject to diagnosis or treatment include, for example, primates, mice, rats, cattle, sheep, goats, horses, canines, felines and the like.

As used herein, a "therapeutically effective amount" of a histone deacetylase (HDAC) inhibitor is one wherein the HDAC inhibitor produces a therapeutically beneficial effect in the subject.

As used herein, a "therapeutically beneficial effect" is any clinical improvement experienced by the subject, such as a reversal in at least one symptom, an elimination of at least one symptom, or a reduction in the severity, duration, and/or frequency of symptoms caused by the autoimmune disease or disorder.

"Pharmaceutically acceptable" refers to those properties and/or substances that are acceptable to the patient from a pharmacological/toxicological point of view and to the manufacturing pharmaceutical chemist from a

physical/chemical point of view regarding composition, formulation, stability, patient acceptance and bioavailability.

As used herein, the language "pharmaceutically acceptable salt" refers to a salt of the administered compounds prepared from pharmaceutically acceptable non-toxic acids, including inorganic acids, organic acids, solvates, hydrates, or clathrates thereof.

As used herein, the term "composition," "pharmaceutical composition" or "pharmaceutically acceptable composition" refers to a mixture of at least one compound or molecule useful within the invention with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound or molecule to a patient. Multiple techniques of administering a compound or molecule exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary and topical administration.

"Pharmaceutically acceptable carrier" refers to a medium that does not interfere with the effectiveness of the biological activity of the active ingredient(s) and is not toxic to the host to which it is administered. As used herein, the language "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical

administration. Supplementary active compounds may also be incorporated into the compositions. The term "pharmaceutically acceptable carrier" also means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound or molecule useful within the invention within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the invention, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein, "pharmaceutically acceptable carrier" also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the invention, and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions. The "pharmaceutically acceptable carrier" may further include a pharmaceutically acceptable salt of the compound or molecule useful within the invention. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the invention are known in the art and described, for example in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

The term "treating" in its various grammatical forms in relation to the present invention refers to preventing, (i.e., chemoprevention ), curing, reversing, attenuating, alleviating, minimizing, suppressing or halting the deleterious effects of a disease state, disease progression, disease causative agent (e.g., bacteria or viruses) or other abnormal condition. For example, treatment may involve alleviating a symptom (i.e., not necessary all symptoms) of a disease or attenuating the progression of a disease. Because some of the inventive methods involve the physical removal of the etiological agent, the artisan will recognize that they are equally effective in situations where the inventive compound is administered prior to, or simultaneous with, exposure to the etiological agent (prophylactic treatment) and situations where the inventive compounds are administered after (even well after) exposure to the etiological agent.

Description

An autoimmune disease or disorder comprises any disease or disorder that arises as a result of a host's immune system directing an immune response against components of the host's own tissues. Non-limiting examples of autoimmune diseases or disorder are coeliac disease, diabetes mellitus type 1 (IDDM), systemic lupus erythematosus (SLE), Sjogren's syndrome, Churg-Strauss Syndrome,

A skin autoimmune disease or disorder comprises any disease or disorder that arises as a result of a host's immune system directing an immune response against components of the host's skin. Examples of autoimmune skin diseases or disorders include but are not limited to psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, as well as the pemphigus group of disorders including bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris.

Although the methods of the present invention may be practiced in vitro, it is contemplated that the preferred embodiment for the methods of selectively treating autoimmune diseases or disorders will comprise contacting cells in vivo in the subject, i.e., by administering a compound to a subject in need of treatment of an autoimmune disease or disorder. The methods of the present invention are intended for the treatment of any subject afflicted with an autoimmune disease or disorder. In one embodiment, the autoimmune disease or disorder is an autoimmune skin disease or disorder.

In one embodiment of the invention, the subject is a mammal. In another embodiment of the invention, the subject is a human.

Compounds Useful Within the Methods of the Invention

Histone deacetylase (HDAC) inhibitors are useful within the methods of the invention. HDAC inhibitors, as that term is used herein, are compounds that are capable of inhibiting the activity of at least one histone deacetylase in vivo and/or in vitro. This inhibition results in an accumulation of acetylated histone. Procedures that may assay for the accumulation of acetylated histones may thus be used as a suitable biological marker to determine the HDAC inhibitory activity of compounds of interest. It is also to be understood that the compounds of the present invention are capable of inhibiting any of the histone deacetylases set forth herein, or any other histone deacetylases.

The present invention includes within its scope compositions comprising HDAC inhibitors, which are: 1) hydroxamic acid derivatives; 2) short- chain fatty acids (SCFAs); 3) cyclic tetrapeptides; 4) benzamides; 5) electrophilic ketones; and/or any other class of compounds capable of inhibiting histone deacetylases, for use in inhibiting histone deacetylase, inducing terminal

differentiation, cell growth arrest and/or apoptosis in neoplastic cells, and/or inducing differentiation, cell growth arrest and/or apoptosis of tumor cells in a tumor.

Non-limiting examples of such HDAC inhibitors are set forth herein. It is understood that the present invention includes any salts, crystal structures, amorphous structures, hydrates, derivatives, metabolites, stereoisomers, structural isomers, polymorphs and prodrugs of the HDAC inhibitors described herein.

(A) Hydroxamic Acid Derivatives such as suberoylanilide hydroxamic acid

(SAHA or vorinostat) (Richon et al., 1998, Proc. Natl. Acad. Sci. USA 95:3003-07); m-carboxycinnamic acid bishydroxamide (CBHA) (Richon et al., 1998, Proc. Natl. Acad. Sci. USA 95:3003-07); pyroxamide; trichostatin analogues such as trichostatin A (TSA) and trichostatin C (Koghe et al., 1998, Biochem. Pharmacol. 56:1359-64); salicylhydroxamic acid (Andrews et al., 2000, Intl. J. Parasitology 30:761-68);

suberoyl bishydroxamic acid (SBHA) (U.S. Patent No. 5,608,108); azelaic bishydroxamic acid (ABHA) (Andrews et al., 2000, Intl. J. Parasitology 30:761-68); azelaic- l-hydroxamate-9-anilide (AAHA) (Qiu et al., 2000, MoI. Biol. Cell

11 :2069-83); 6-(3-chlorophenylureido) carpoic hydroxamic acid (3C1-UCHA);

oxamflatin [(2E)-5-[3-[(phenylsufonyl) amino]phenyl]-pent-2-en-4-ynohydroxamic acid] (Kim et al., 1999, Oncogene 18:2461-70); A-161906, Scriptaid (Su et al., 2000, Cancer Research 60:3137-3142); PXD-101 (Prolifix); LAQ-824; CHAP; MW2796 (Andrews et al., 2000, Intl. J. Parasitology 30:761-68); MW2996 (Andrews et al., 2000, Intl. J. Parasitology 30:761-68); or any of the hydroxamic acids disclosed in U.S. Patent Nos. 5,369,108, 5,932,616, 5,700,811, 6,087,367 or 6,511,990.

(B) Cyclic Tetrapeptides such as trapoxin A (TPX)-cyclic tetrapeptide (cyclo- (L-phenylalanyl-L-phenylalanyl-D-pipecolinyl-L-2-amino-8-oxo-9,10-epoxy decanoyl)) (Kijima et al., 1993, J. Biol. Chem. 268:22429-35); FR901228 (FK 228, depsipeptide) (Nakajima et al., 1998, Ex. Cell Res. 241 :126-33); FR225497 cyclic tetrapeptide (Mori et al., PCT Application WO 00/08048); apicidin cyclic tetrapeptide [cyclo(N-O-methyl-L-tryptophanyl-L-isoleucinyl-D-pipecolinyl-L-2-amino-8- oxodecanoyl)] (Darkin-Rattray et al., 1996, Proc. Natl. Acad. Sci. USA 93:13143-47); apicidin Ia, apicidin Ib, apicidin Ic, apicidin Ia, and apicidin lib (P. Dulski et al., PCT Application WO 97/1 1366); CHAP, HC-toxin cyclic tetrapeptide (Bosch et al., 1995, Plant Cell 7:1941-50); WF27082 cyclic tetrapeptide (PCT Application WO

98/48825); and chlamydocin (Bosch et al., 1995, Plant Cell 7:1941-50).

(C) Short chain fatty acid (SCFA) derivatives such as: sodium butyrate (Cousens et al., 1979, J. Biol. Chem. 254: 1716-23); isovalerate (McBain et al., 1997, Biochem. Pharm. 53 : 1357-68); valerate (McBain et al., 1997, Biochem. Pharm.

53:1357-68); 4-phenylbutyrate (4-PBA) (Lea & Tulsyan, 1995, Anticancer Res.

15:879-73); phenylbutyrate (PB) (Wang et al., 1999, Cancer Res. 59:2766-99);

propionate (McBain et al., 1997, Biochem. Pharm. 53:1357-68); butyramide (Lea &

Tulsyan, 1995, Anticancer Res. 15:879-73); isobutyramide (Lea & Tulsyan, 1995, Anticancer Res. 15:879-73); phenylacetate (Lea & Tulsyan, 1995, Anticancer Res.

15:879-73); 3-bromopropionate (Lea & Tulsyan, 1995, Anticancer Res. 15:879-73); tributyrin (Guan et al., 2000, Cancer Research, 60:749-55); valproic acid valproate and Pivanex™

(D) Benzamide derivatives such as CI-994; MS-27-275 [N-(2-aminophenyl)- 4-[N-(pyridin-3-yl methoxycarbonyl)aminomethyl]benzamide] (Saito et al., Proc.

Natl. Acad. Sci. USA 96, 4592-4597 (1999)); and 3 '-amino derivative of MS-27-275 (Saito et al., supra).

(E) Electrophilic ketone derivatives such as trifluoromethyl ketones (Frey et al, 2002, Bioorg. & Med. Chem. Lett. 12:3443-47; U.S. Patent No. 6,511,990) and α-keto amides such as N-methyl-α-ketoamides.

(F) Other HDAC Inhibitors such as natural products psammaplins and depudecin (Kwon et al., 1998, PNAS 95:3356-3361).

Preferred hydroxamic acid based HDAC inhibitors are suberoylanilide hydroxamic acid (SAHA or vorinostat), m-carboxycinnamic acid bishydroxamate (CBHA) and pyroxamide. SAHA has been shown to bind directly in the catalytic pocket of the HDAC enzyme. SAHA induces cell cycle arrest, differentiation and/or apoptosis of transformed cells in culture and inhibits tumor growth in rodents. SAHA is effective at inducing these effects in both solid tumors and hematological cancers. It has been shown that SAHA is effective at inhibiting tumor growth in animals with no toxicity to the animal. The SAHA-induced inhibition of tumor growth is associated with an accumulation of acetylated histones in the tumor. SAHA is effective at inhibiting the development and continued growth of carcinogen-induced (N-methylnitrosourea) mammary tumors in rats. SAHA was administered to the rats in their diet over the 130 days of the study. SAHA is thus a nontoxic, orally active antitumor agent which mechanism of action involves the inhibition of histone deacetylase activity.

HDAC inhibitors may be found in U.S. Patent No. 5,369,108, issued on Nov. 29, 1994, U.S. Patent. No. 5,700,811, issued on Dec. 23, 1997, U.S. Patent No. 5,773,474, issued on Jun. 30, 1998, U.S. Patent No. 5,932,616, issued on

Aug. 3, 1999 and U.S. Patent No. 6,511,990, issued Jan. 28, 2003, all to Breslow et al.; U.S. Patent No. 5,055,608, issued on Oct. 8, 1991, U.S. Patent No. 5,175,191, issued on Dec. 29, 1992 and U.S. Patent No. 5,608,108, issued on Mar. 4, 1997, all to Marks et al.; as well as Yoshida et al., 1995, Bioassays 17:423-30; Saito et al., 1999, PNAS USA 96:4592-97; Furamai et al., 2001, PNAS USA 98(l):87-92; Komatsu et al., 2001, Cancer Res. 61(1 1):4459-66; Su et al., 2000, Cancer Res. 60:3137-42; Lee et al., Cancer Res. 61(3):931-934; Suzuki et al., 1999, J. Med. Chem. 42(15):3001- 03; published PCT Application WO 01/18171 published on Mar. 15, 2001 to Sloan- Kettering Institute for Cancer Research and The Trustees of Columbia University; published PCT Application WO02/246144 to Hoffmann-La Roche; published PCT Application WO02/22577 to Novartis; published PCT Application WO02/30879 to Prolifix; published PCT Applications WO 01/38322 (published May 31, 2001), WO 01/70675 (published on Sep. 27, 2001) and WO 00/71703 (published on Nov. 30, 2000) all to Methylgene, Inc.; published PCT Application WO 00/21979 published on Oct. 8, 1999 to Fujisawa Pharmaceutical Co., Ltd.; published PCT Application WO 98/40080 published on Mar. 11, 1998 to Beacon Laboratories, L.L.C.; and Curtin M. (Current patent status of histone deacetylase inhibitors Expert Opin. Ther. Patents 2002, 12(9): 1375-84 and references cited therein).

In a preferred embodiment, the HDAC inhibitor comprises vorinostat. Any of the HDACs may be synthesized according to the methods outlined in the Experimental Details Section, or according to the method set forth in U.S. Patent Nos. 5,369,108, 5,700,811, 5,932,616 and 6,51 1,990, or according to any other method known to a person skilled in the art. The invention, in addition to the above listed HDAC inhibitors, is intended to encompass the use of homologs and analogs of such compounds. In this context, homologs are molecules having substantial structural similarities to the above-described compounds and analogs are molecules having substantial biological similarities regardless of structural similarities.

Salts of the Compounds Useful Within the Methods of the Invention

The compounds useful within the invention may form salts with acids or bases, and such salts are included in the present invention. In one embodiment, the salts are pharmaceutically-acceptable salts. The term "salts" embraces addition salts of free acids or free bases that are compounds useful within the invention. The term "pharmaceutically acceptable salt" refers to salts that possess toxicity profiles within a range that affords utility in pharmaceutical applications. Pharmaceutically

unacceptable salts may nonetheless possess properties such as high crystallinity, which have utility in the practice of the present invention, such as for example utility in process of synthesis, purification or formulation of compounds useful within the invention.

Suitable pharmaceutically-acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric, and phosphoric acids. Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic, 2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric, salicylic, galactaric and galacturonic acid.

Examples of pharmaceutically unacceptable acid addition salts include, for example, perchlorates and tetrafluoroborates.

Suitable pharmaceutically acceptable base addition salts of compounds useful within the invention include, for example, metallic salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N'-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine,

ethylenediamine, meglumine (N-methylglucamine) and procaine. Examples of pharmaceutically unacceptable base addition salts include lithium salts and cyanate salts. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound.

Methods of the Invention

In one aspect, the present invention includes a method of treating or ameliorating an autoimmune disease or disorder in a subject. The method comprises administering a therapeutically effective amount of a composition comprising a pharmaceutically acceptable carrier and a compound selected from the group consisting of a histone deacetylase (HDAC) inhibitor and a pharmaceutically acceptable salt or hydrate thereof. In one embodiment, the autoimmune disease or disorder is selected from the group consisting of coeliac disease, diabetes mellitus type 1 (IDDM), systemic lupus erythematosus (SLE), Sjogren's syndrome, Churg- Strauss Syndrome, Hashimoto's thyroiditis, Graves' disease, idiopathic

thrombocytopenic purpura, rheumatoid arthritis (RA), polymyositis, ulcerative colitis, Crohn's disease, autoimmune carditis, Wegener's granulomatosis, autoimmune hemolytic anemia, polyarteritis nodosa, psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris.

In one embodiment, the autoimmune disease or disorder is a skin autoimmune disease or disorder. In another embodiment, the skin autoimmune disease or disorder is selected from the group consisting of psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris. In yet another embodiment, the skin autoimmune disease or disorder is bullous pemphigoid. In yet another embodiment, the HDAC inhibitor is vorinostat. In yet another embodiment, the subject is a mammal. In yet another embodiment, the mammal is a human. In yet another embodiment, the dose of the HDAC inhibitor in the composition is about 400 mg per day. In another aspect, the present invention includes a method of treating a subject at risk of developing an autoimmune disease or disorder. The method comprises administering to the subject a therapeutically effective amount of a composition comprising a pharmaceutically acceptable carrier and a compound selected from the group consisting of a histone deacetylase (HDAC) inhibitor and a pharmaceutically acceptable salt or hydrate thereof.

In yet another aspect, the invention includes a method of treating a subject afflicted with a skin autoimmune disease or disorder. The method comprises administering to the subject in need thereof a therapeutically effective amount of a composition comprising a pharmaceutically acceptable carrier and a compound selected from the group consisting of a histone deacetylase (HDAC) inhibitor and a pharmaceutically acceptable salt or hydrate thereof.

In one embodiment, the skin autoimmune disease or disorder is selected from the group consisting of psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris. In another embodiment, the skin autoimmune disease or disorder is bullous pemphigoid. In yet another embodiment, the HDAC inhibitor is vorinostat. In yet another embodiment, the subject is a mammal. In yet another embodiment, the mammal is a human. In yet another embodiment, the dose of the HDAC inhibitor is about 400 mg per day.

In yet another aspect, the invention includes a method of treating a subject at risk of developing a skin autoimmune disease or disorder. The method comprises administering to the subject in need thereof a therapeutically effective amount of a composition comprising a pharmaceutically acceptable carrier and a compound selected from the group consisting of a histone deacetylase (HDAC) inhibitor and a pharmaceutically acceptable salt or hydrate thereof.

In one embodiment, the skin autoimmune disease or disorder is selected from the list consisting of psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris. In another embodiment, the skin autoimmune disease or disorder is bullous pemphigoid. In yet another embodiment, the HDAC inhibitor is vorinostat. In yet another embodiment, the subject is a mammal. In yet another embodiment, the mammal is a human. In yet another embodiment, the dose of the HDAC inhibitor is about 400 mg per day.

Pharmaceutical Compositions & Formulations

In one aspect, the pharmaceutical compositions useful within the methods of the invention comprise an HDAC inhibitor. In another aspect, the pharmaceutical compositions useful within the methods of the invention comprise a pharmaceutically acceptable salt of a HDAC inhibitor. In yet another aspect, the pharmaceutical compositions useful within the methods of the invention comprise a hydrate of a HDAC inhibitor. The term "hydrate" includes but is not limited to hemihydrate, monohydrate, dihydrate, trihydrate and the like.

In one aspect, the pharmaceutical compositions useful within the methods of the invention comprise any solid or liquid physical form of vorinostat or suberoylanilide hydroxamic acid (SAHA), or any of the other HDAC inhibitors. For example, the HDAC inhibitors may be in a crystalline form, in amorphous form, and have any particle size. The HDAC inhibitor particles may be micronized, or may be agglomerated, particulate granules, powders, oils, oily suspensions or any other form of solid or liquid physical form.

Compositions comprising an HDAC inhibitor may be incorporated into pharmaceutical compositions suitable for administration to a subject, preferably a mammal, more preferably a human. A formulated composition comprising an HDAC inhibitor may assume a variety of states. Generally, the composition is formulated in a manner that is compatible with the intended method of administration.

A composition of the instant invention may be formulated in combination with another agent, e.g., another therapeutic agent or an agent that stabilizes the composition comprising an HDAC inhibitor.

Pharmaceutical compositions of the invention include a pharmaceutical carrier that may contain a variety of components that provide a variety of functions, including regulation of drug concentration, regulation of solubility, chemical stabilization, regulation of viscosity, absorption enhancement, regulation of pH, and the like. The pharmaceutical carrier may comprise a suitable liquid vehicle or excipient and an optional auxiliary additive or additives. The liquid vehicles and excipients are conventional and commercially available. Illustrative examples thereof are distilled water, physiological saline, aqueous solutions of dextrose, and the like. For water soluble formulations, the pharmaceutical composition preferably includes a buffer such as a phosphate buffer, or other organic acid salt, preferably at a pH of between about 7 and 8. Other components may include antioxidants, such as ascorbic acid, hydrophilic polymers, such as monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, dextrins, chelating agents, such as EDTA, and like components well known to those in the pharmaceutical sciences, e.g., Remington's Pharmaceutical Science, latest edition (Mack Publishing Company, Easton, PA).

Any inert excipient that is commonly used as a carrier or diluent may be used in the formulations of the present invention, such as for example, a gum, a starch, a sugar, a cellulosic material, an acrylate, or mixtures thereof. A preferred diluent is microcrystalline cellulose. The compositions may further comprise a disintegrating agent (e.g., sodium croscarmellose) and a lubricant (e.g., magnesium stearate), and in addition may comprise one or more additives selected from a binder, a buffer, a protease inhibitor, a surfactant, a solubilizing agent, a plasticizer, an emulsifier, a stabilizing agent, a viscosity increasing agent, a sweetener, a film forming agent, or any combination thereof.

Furthermore, the compositions of the present invention may be in the form of controlled release or immediate release formulations. Controlled- or sustained-release formulations of a pharmaceutical composition of the invention may be made using conventional technology.

Although the description of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions that are suitable for ethical administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts.

Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist may design and perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the pharmaceutical compositions of the invention is contemplated include, but are not limited to, humans and other primates, mammals including commercially relevant mammals such as non-human primates, cattle, pigs, horses, sheep, cats, and dogs.

A composition of the invention may be administered into a recipient in a wide variety of ways. Preferred modes of administration are parenteral,

intravenous, intra-arterial, intramuscular, surgical implant, infusion pump, via catheter, or topically.

Pharmaceutical compositions that are useful in the methods of the invention may be prepared, packaged, or sold in formulations suitable for ophthalmic, oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, or another route of administration. Other contemplated formulations include projected nanoparticles, liposomal preparations, resealed erythrocytes containing the active ingredient, and immunologically-based formulations.

A pharmaceutical composition of the invention may be prepared, packaged, or sold in bulk, as a single unit dose, or as a plurality of single unit doses. As used herein, a "unit dose" is discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient that would be administered to a subject or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.

The relative amounts of the active ingredient, the pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100% (w/w) active ingredient.

In addition to the active ingredient, a pharmaceutical composition of the invention may further comprise one or more additional pharmaceutically active agents. Other active agents useful in the treatment of fibrosis include antiinflammatories, including corticosteroids, and immunosuppressants.

A formulation of a pharmaceutical composition of the invention suitable for oral administration may be prepared, packaged, or sold in the form of a discrete solid dose unit including, but not limited to, a tablet, a hard or soft capsule, a cachet, a troche, a dispersion or a lozenge, each containing a predetermined amount of the active ingredient. Other formulations suitable for oral administration include, but are not limited to, a powdered or granular formulation, an aqueous or oily suspension, an aqueous or oily solution, or an emulsion. In a preferred embodiment, the composition is in the form of a gelatin capsule.

As used herein, an "oily" liquid is one that comprises a carbon- containing molecule and which exhibits a less polar character than water.

A tablet comprising the active ingredient may, for example, be made by compressing or molding the active ingredient, optionally with one or more additional ingredients. Compressed tablets may be prepared by compressing, in a suitable device, the active ingredient in a free-flowing form such as a powder or granular preparation, optionally mixed with one or more of a binder, a lubricant, an excipient, a surface active agent, and a dispersing agent. Molded tablets may be made by molding, in a suitable device, a mixture of the active ingredient, a

pharmaceutically acceptable carrier, and at least sufficient liquid to moisten the mixture. Pharmaceutically acceptable excipients used in the manufacture of tablets include, but are not limited to, inert diluents, granulating and disintegrating agents, binding agents, and lubricating agents. Known dispersing agents include, but are not limited to, potato starch and sodium starch glycollate. Known surface active agents include, but are not limited to, sodium lauryl sulphate. Known diluents include, but are not limited to, calcium carbonate, sodium carbonate, lactose, microcrystalline cellulose, calcium phosphate, calcium hydrogen phosphate, and sodium phosphate. Known granulating and disintegrating agents include, but are not limited to, corn starch and alginic acid. Known binding agents include, but are not limited to, gelatin, acacia, pre-gelatinized maize starch, polyvinylpyrrolidone, and hydroxypropyl methylcellulose. Known lubricating agents include, but are not limited to, magnesium stearate, stearic acid, silica, and talc.

Tablets may be non-coated or they may be coated using known methods to achieve delayed disintegration in the gastrointestinal tract of a subject, thereby providing sustained release and absorption of the active ingredient. By way of example, a material such as glyceryl monostearate or glyceryl distearate may be used to coat tablets. Further by way of example, tablets may be coated using methods described in U.S. Patents numbers 4,256,108; 4,160,452; and 4,265,874 to form osmotically-controlled release tablets. Tablets may further comprise a sweetening agent, a flavoring agent, a coloring agent, a preservative, or some combination of these in order to provide pharmaceutically elegant and palatable preparation.

Hard capsules comprising the active ingredient may be made using a physiologically degradable composition, such as gelatin. Such hard capsules comprise the active ingredient, and may further comprise additional ingredients including, for example, an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin.

Soft gelatin capsules comprising the active ingredient may be made using a physiologically degradable composition, such as gelatin. Such soft capsules comprise the active ingredient, which may be mixed with water or an oil medium such as peanut oil, liquid paraffin, or olive oil.

Liquid formulations of a pharmaceutical composition of the invention which are suitable for oral administration may be prepared, packaged, and sold either in liquid form or in the form of a dry product intended for reconstitution with water or another suitable vehicle prior to use.

Liquid suspensions may be prepared using conventional methods to achieve suspension of the active ingredient in an aqueous or oily vehicle. Aqueous vehicles include, for example, water and isotonic saline. Oily vehicles include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis, olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. Liquid suspensions may further comprise one or more additional ingredients including, but not limited to, suspending agents, dispersing or wetting agents, emulsifying agents, demulcents, preservatives, buffers, salts, flavorings, coloring agents, and sweetening agents. Oily suspensions may further comprise a thickening agent. Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fats, sodium alginate, polyvinylpyrrolidone, gum tragacanth, gum acacia, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose. Known dispersing or wetting agents include, but are not limited to, naturally-occurring phosphatides such as lecithin, condensation products of an alkylene oxide with a fatty acid, with a long chain aliphatic alcohol, with a partial ester derived from a fatty acid and a hexitol, or with a partial ester derived from a fatty acid and a hexitol anhydride (e.g. polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, and polyoxyethylene sorbitan monooleate, respectively). Known emulsifying agents include, but are not limited to, lecithin and acacia. Known preservatives include, but are not limited to, methyl, ethyl, or n-propyl-para-hydroxybenzoates, ascorbic acid, and sorbic acid. Known sweetening agents include, for example, glycerol, propylene glycol, sorbitol, sucrose, and saccharin. Known thickening agents for oily

suspensions include, for example, beeswax, hard paraffin, and cetyl alcohol.

Liquid solutions of the active ingredient in aqueous or oily solvents may be prepared in substantially the same manner as liquid suspensions, the primary difference being that the active ingredient is dissolved, rather than suspended in the solvent. Liquid solutions of the pharmaceutical composition of the invention may comprise each of the components described with regard to liquid suspensions, it being understood that suspending agents will not necessarily aid dissolution of the active ingredient in the solvent. Aqueous solvents include, for example, water and isotonic saline. Oily solvents include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis, olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin.

Powdered and granular formulations of a pharmaceutical preparation of the invention may be prepared using known methods. Such formulations may be administered directly to a subject, used, for example, to form tablets, to fill capsules, or to prepare an aqueous or oily suspension or solution by addition of an aqueous or oily vehicle thereto. Each of these formulations may further comprise one or more of dispersing or wetting agent, a suspending agent, and a preservative. Additional excipients, such as fillers and sweetening, flavoring, or coloring agents, may also be included in these formulations.

A pharmaceutical composition of the invention may also be prepared, packaged, or sold in the form of oil-in- water emulsion or a water-in-oil emulsion. The oily phase may be a vegetable oil such as olive or arachis oil, a mineral oil such as liquid paraffin, or a combination of these. Such compositions may further comprise one or more emulsifying agents such as naturally occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soybean or lecithin phosphatide, esters or partial esters derived from combinations of fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of such partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. These emulsions may also contain additional ingredients including, for example, sweetening or flavoring agents.

As used herein, "parenteral administration" of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subject and administration of the pharmaceutical composition through the breach in the tissue. Parenteral administration thus includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration is contemplated to include, but is not limited to, intraocular, intravitreal, subcutaneous, intraperitoneal, intramuscular, intrasternal injection, intratumoral, and kidney dialytic infusion techniques.

Formulations of a pharmaceutical composition suitable for parenteral administration comprise the active ingredient combined with a pharmaceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending, stabilizing, or dispersing agents. In one embodiment of a formulation for parenteral administration, the active ingredient is provided in dry (i.e. powder or granular) form for reconstitution with a suitable vehicle (e.g. sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

The pharmaceutical compositions may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to the known art, and may comprise, in addition to the active ingredient, additional ingredients such as the dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations may be prepared using a non-toxic parenterally-acceptable diluent or solvent, such as water or 1,3-butanediol, for example. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or di-glycerides. Other parentally-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer systems. Compositions for sustained release or implantation may comprise pharmaceutically acceptable polymeric or hydrophobic materials such as an emulsion, an ion exchange resin, a sparingly soluble polymer, or a sparingly soluble salt.

A pharmaceutical composition of the invention may be prepared, packaged, or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 nanometers, and preferably from about 1 to about 6 nanometers. Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant may be directed to disperse the powder or using a self-propelling solvent/powder-dispensing container such as a device comprising the active ingredient dissolved or suspended in a low- boiling propellant in a sealed container. Preferably, such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than

0.5 nanometers and at least 95% of the particles by number have a diameter less than 7 nanometers. More preferably, at least 95% of the particles by weight have a diameter greater than 1 nanometer and at least 90% of the particles by number have a diameter less than 6 nanometers. Dry powder compositions preferably include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form.

Low boiling propellants generally include liquid propellants having a boiling point of below 65°F at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w/w) of the composition, and the active ingredient may constitute 0.1 to 20% (w/w) of the composition. The propellant may further comprise additional ingredients such as a liquid non-ionic or solid anionic surfactant or a solid diluent (preferably having a particle size of the same order as particles comprising the active ingredient).

Pharmaceutical compositions of the invention formulated for pulmonary delivery may also provide the active ingredient in the form of droplets of a solution or suspension. Such formulations may be prepared, packaged, or sold as aqueous or dilute alcoholic solutions or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization or atomization device. Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, or a preservative such as methylhydroxybenzoate. The droplets provided by this route of administration preferably have an average diameter in the range from about 0.1 to about 200 nanometers.

The formulations described herein as being useful for pulmonary delivery are also useful for intranasal delivery of a pharmaceutical composition of the invention.

Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle from about 0.2 to 500 micrometers. Such a formulation is administered in the manner in which snuff is taken i.e. by rapid inhalation through the nasal passage from a container of the powder held close to the nares. Formulations suitable for nasal administration may, for example, comprise from about as little as 0.1% (w/w) and as much as 100% (w/w) of the active ingredient, and may further comprise one or more of the additional ingredients described herein.

A pharmaceutical composition of the invention may be prepared, packaged, or sold in a formulation suitable for buccal administration. Such formulations may, for example, be in the form of tablets or lozenges made using conventional methods, and may, for example, 0.1 to 20% (w/w) active ingredient, the balance comprising an orally dissolvable or degradable composition and, optionally, one or more of the additional ingredients described herein. Alternately, formulations suitable for buccal administration may comprise a powder or an aerosolized or atomized solution or suspension comprising the active ingredient. Such powdered, aerosolized, or aerosolized formulations, when dispersed, preferably have an average particle or droplet size in the range from about 0.1 to about 200 nanometers, and may further comprise one or more of the additional ingredients described herein.

As used herein, "additional ingredients" include, but are not limited to, one or more of the following: excipients; surface active agents; dispersing agents; inert diluents; granulating and disintegrating agents; binding agents; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives;

physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents, demulcents; buffers; salts; thickening agents; fillers; emulsifying agents; antioxidants; antibiotics; antifungal agents; stabilizing agents; and

pharmaceutically acceptable polymeric or hydrophobic materials. Other "additional ingredients" which may be included in the pharmaceutical compositions of the invention are known in the art and described, for example in Remington's

Pharmaceutical Sciences (1985, Genaro, ed., Mack Publishing Co., Easton, PA), which is incorporated herein by reference.

Formulations suitable for topical administration include, but are not limited to, liquid or semi-liquid preparations such as liniments, lotions, oil-in-water or water-in-oil emulsions such as creams, ointments or pastes, and solutions or suspensions. Topically-administrable formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of the active ingredient may be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further comprise one or more of the additional ingredients described herein.

Dosing

The compound may be administered to a subject as frequently as several times daily, or it may be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less. The frequency of the dose will be readily apparent to the skilled artisan and will depend upon any number of factors, such as, but not limited to, the type and severity of the disease being treated, the type and age of the subject, etc. The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or one or more other accessory ingredients, and then, if necessary or desirable, shaping or packaging the product into a desired single- or multi-dose unit.

A composition useful within the methods of the invention may be administered to a subject in need thereof once daily (QD), or divided into multiple daily doses such as twice daily (BID), and three times daily (TID). The composition inhibitor may be administered to a subject at a total daily dosage of up to about 4000 mg, e.g., about 200 mg, about 400 mg, about 600 mg, about 800 mg, about 1000 mg, about 2000 mg, or about 4000 mg, which may be administered in one daily dose or may be divided into multiple daily doses as described above, and in any whole or partial increments therebetween.

In addition, the administration may be continuous, i.e., every day, or intermittently. The terms "intermittent" or "intermittently" as used herein means stopping and starting at either regular or irregular intervals. For example, intermittent administration of a composition may be administration one to six days per week or it may mean daily administration for two consecutive weeks, then no administration for one week or it may mean administration on alternate days.

Any of the HDAC inhibitors are administered to the subject at a total daily dosage of between about 25-4000 mg. A currently preferred treatment protocol comprises continuous administration (i.e., every day), once, twice or three times daily at a total daily dose in the range of about 200 mg to about 600 mg.

Another currently preferred treatment protocol comprises intermittent administration to a subject of between three to five days a week, once, twice or three times daily at a total daily dose in the range of about 200 mg to about 600 mg.

In one particular embodiment, the HDAC inhibitor is administered to a subject continuously once daily at a dose of about 400 mg or twice daily at a dose of about 200 mg. In another particular embodiment, the HDAC inhibitor is administered to a subject intermittently three days a week, once daily at a dose of about 400 mg or twice daily at a dose of about 200 mg.

In another particular embodiment, the HDAC inhibitor is administered to a subject intermittently four days a week, once daily at a dose of about 400 mg or twice daily at a dose of about 200 mg.

In another particular embodiment, the HDAC inhibitor is administered to a subject intermittently five days a week, once daily at a dose of about 400 mg or twice daily at a dose of about 200 mg.

In one particular embodiment, the HDAC inhibitor is administered to a subject continuously once daily at a dose of about 600 mg, twice daily at a dose of about 300 mg, or three times daily at a dose of about 200 mg.

In another particular embodiment, the HDAC inhibitor is administered to a subject intermittently three days a week, once daily at a dose of about 600 mg, twice daily at a dose of about 300 mg, or three times daily at a dose of about 200 mg.

In another particular embodiment, the HDAC inhibitor is administered to a subject intermittently four days a week, once daily at a dose of about 600 mg, twice daily at a dose of about 300 mg, or three times daily at a dose of about 200 mg.

In another particular embodiment, the HDAC inhibitor is administered to a subject intermittently five days a week, once daily at a dose of about 600 mg, twice daily at a dose of about 300 mg, or three times daily at a dose of about 200 mg.

In addition, the HDAC inhibitor may be administered to a subject according to any of the schedules described above, consecutively for a few weeks, followed by a rest period. For example, the HDAC inhibitor may be administered to a subject according to any one of the schedules described above from two to eight weeks, followed by a rest period of one week, or twice daily at a dose of 300 mg for three to five days a week. In another particular embodiment, the HDAC inhibitor is administered to a subject three times daily for two consecutive weeks, followed by one week of rest.

It should be apparent to a person skilled in the art that the various dosages and dosing schedules described herein merely set forth specific embodiments and should not be construed as limiting the broad scope of the invention. Any permutations, variations and combinations of the dosages and dosing schedules are included within the scope of the present invention. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents were considered to be within the scope of this invention and covered by the claims appended hereto. For example, it should be understood, that modifications in reaction conditions, including but not limited to reaction times, reaction size/volume, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions, e.g., nitrogen atmosphere, and reducing/oxidizing agents, with art-recognized alternatives and using no more than routine experimentation, are within the scope of the present application.

It is to be understood that wherever values and ranges are provided herein, all values and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present invention. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application.

The following examples further illustrate aspects of the present invention. However, they are in no way a limitation of the teachings or disclosure of the present invention as set forth herein. EXAMPLES

The invention is now described with reference to the following Examples. These Examples are provided for the purpose of illustration only and the invention should in no way be construed as being limited to these Examples, but rather should be construed to encompass any and all variations that become evident as a result of the teaching provided herein.

Example 1:

A 59-year-old woman, was diagnosed as having mycosis fungoides/cutaneous T-cell lymphoma (MF/CTCL), stage IB, with a medical history of varicella zoster, gastritis, and osteopenia. She also had a history of sulfa allergy.

Despite multiple treatments including psoralen UV-A phototherapy, topical chemotherapy, topical corticosteroids, oral bexarotene, chemotherapy, autologous stem cell transplant, denileukin diftitox, and interferon gamma, the patient experienced disease progression to stage IVA with skin tumors, nodal involvement with large cell transformation, and peripheral blood involvement (6% CD4+CD7- cells and a discrete population of 38% CD4+CD26- cells). She subsequently underwent total skin electron beam radiation (TSEB) therapy and started treatment with extracorporeal photopheresis (ECP) in conjunction with interferon gamma.

Three months later, the patient was admitted to the Hospital of the

University of Pennsylvania, Philadelphia, with a 2-month history of progressive skin pain, itching, blisters on their buttocks and feet, oral and vaginal erosions,

odynophagia, and lower extremity edema for which furosemide was previously prescribed. Initially, the concern was for radiation dermatitis vs disseminated herpetic infection. Despite aggressive skin care, pain control, and negative testing for herpes simplex and varicella zoster virus, the patient's skin condition progressed to erythroderma, erosions, and tense bullae over the arms, abdomen, chest, back, legs, and mouth (sparing the eyes), raising the possibility of Stevens-Johnson

syndrome/toxic epidermal necrolysis (TEN) or bullous drug hypersensitivity reaction. However, findings from multiple skin biopsy specimens were inconclusive for TEN.

Hematoxylin-eosin staining demonstrated a subepidermal blistering disorder. Although findings were consistent with bullous pemphigoid (BP), other diagnoses could not be completely ruled out based on histologic results at that time, including radiation recall dermatitis, drug-induced blistering reaction, and erythema multiforme. The inflammatory infiltrate was especially sparse in biopsied bulla, suggesting that the cutaneous T-cell lymphoma (CTCL) was not playing a role in the bullous process. Direct immunofluorescence at that time was inconclusive, although repeated biopsies of specimens of both skin and buccal mucosa demonstrated focal linear IgG and complement factor 3 deposition at the mucosal/submucosal interface.

The patient was transferred to the medical intensive care unit, started on intravenous (IV) methylprednisolone and IV immunoglobulin and had several drugs discontinued. Her oral lesions and skin improved with decreased erythema and bullae formation. She was discharged while prescribed a tapered dose of prednisone, and treatment with low-dose interferon gamma was restarted.

Over the next month, as the dose of corticosteroids was slowly tapered, the patient developed recurrent intense generalized pruritus, new bullae, and oral erosions. Indirect immunofluorescence examination of her serum performed on salt- split skin at Johns Hopkins was positive and diagnostic for BP without evidence for paraneoplastic pemphigus. In addition, assays for circulating antibodies against the BP 180 and BP230 antigens were positive. Treatment with a high dose of prednisone was restarted.

Over the next 3 months, the patient was treated with monthly IV immunoglobins (2 g/kg for 3 days) as a corticosteroid and immunosuppressive drug- sparing agent owing to the fact that adding additional immunosuppressive therapies would put her at risk for CTCL progression. Despite continuing low-dose interferon gamma, photopheresis, and topical corticosteroids, the patient developed multiple refractory CTCL plaques on her face, trunk, and extremities that required local radiation treatment. The IV immunoglobin treatment was discontinued because it was felt that may have been contributing to the worsening of the patient's CTCL. Once the prednisone taper approached 20 mg/d, the patient's BP flared and new, widespread skin blisters and oral erosions developed requiring increasing prednisone dose.

The patient with refractory aggressive MF/CTCL developed severe BP following TSEB therapy, which proved to be both a diagnostic and therapeutic challenge. Attempts at weaning the patient off systemic corticosteroids or instituting treatments for CTCL, including interferon gamma, resulted in flares of her bullous disease. However, treatments aimed at control of the BP resulted in progression of her known MF/CTCL.

Vorinostat, 400 mg/day, was added to the therapeutic regimen comprising interferon gamma, ECP, prednisone, and topical corticosteroids. Within 2 weeks of starting treatment with vorinostat, the patient noted a significant decrease in pruritus and blister formation. After 6 weeks on this regimen, most of the blisters had healed, and the patient was able to begin tapering her prednisone dose. Existing MF/ CTCL plaques responded to local radiation, and no new plaques appeared once treatment with vorinostat was initiated. Adverse effects included fatigue and thrombocytopenia, which improved on decreasing the dosage of vorinostat to 300 mg/day, 5 days a week. Simultaneously, the patient's prednisone dosage was tapered from 30 mg/day down to 15 mg/day. No new blister formation and a decrease in pruritus was observed. The patient remained blister-free for 2 months. However, after 4 months, owing to fatigue and thrombocytopenia, the dose of vorinostat was decreased to 200 mg/day, and the patient's CTCL progressed such that she required a change in therapeutic management. Multimodality therapy with vorinostat, interferon gamma, and ECP was discontinued. Gemcitabine hydrochloride was prescribed. It is worth noting that the patient's BP flared within 2 weeks of discontinuation of the vorinostat and her symptoms of skin fragility and pruritus worsened. Corticosteroids were increased to over 30 mg/day.

This is exemplary of the use of an HDAC inhibitor for the treatment of BP, an autoimmune blistering disease characterized by tense, subepidermal bullae secondary to IgG autoantibodies against the hemidesmosomal antigens, BP230 (BPAgI) and BP 180 (BPAg2), involved in dermoepidermal adhesion. Pruritus is common, and oral involvement occurs in approximately 20% of cases. Humoral immunity and autoreactive T cells, of both the TH2 and THl types, are thought to be important in pathogenesis. Treatments include topical and systemic corticosteroids, tetracycline hydrochloride, dapsone, azathioprine sodium, mycophenolate mofetil hydrochloride, methotrexate, cyclophosphamide, chorambucil, IV immunoglobins, cyclosporine, and rituximab. These therapeutic options are generally

immunosuppressive or are associated with long-term complications, as is the case with systemic corticosteroids.

Mycosis fungoides and Sezary syndrome are CTCL variants characterized by clonal malignant CD4+ T-lymphocytes. It is believed that an intact immune system is important in controlling the disease and preventing its progression. Commonly used multimodality treatment regimens for CTCL include interferon therapy in the form of either interferon alfa or interferon gamma. Interferons counteract several of the immune abnormalities of the skewed TH2 cytokine profile exhibited in CTCL. A well-known adverse effect of interferon therapy is an increased risk of autoimmune disorders in predisposed individuals.

Another accepted association is between autoimmunity and lymphoid neoplasms. Lymphocytic neoplasms are more common in the setting of autoimmune diseases, and autoimmunity occurs at increased frequencies in patients with lymphocytic malignant diseases. Bullous pemphigoid has been rarely reported in the setting of CTCL, namely, in the context of phototherapy treatment. It is worth noting that the pa tient described herein had a history of phototherapy which may have increased the risk for developing the bullous disorder. In addition, furosemide may have contributed to the development of BP.

Although HDAC inhibitors, such as vorinostat, are thought to be targeted therapy for the treatment of neoplasia, they affect a number of histone and nonhistone proteins. They result in the accumulation of acetylated proteins, leading to changes in gene expression and in the acetylation of proteins important in cell proliferation, apoptosis, cell motility, and angiogenesis, which are all important in tumor suppression. Anti-inflammatory effects of HDAC inhibitors have also been documented, including the reduction of proinflammatory cytokines (e.g. TNF, interleukin 1 β, IL-8), the down regulation of immune stimulators (e.g., IL-6), and the inhibition of production of nitric oxide, a contributor of many inflammatory disorders.

Impaired T-reg cells may contribute to autoimmune diseases. Histone deacetylase inhibitors were shown to generate T-reg cells, leading to enhanced T-reg sup pression with considerable T-reg-dependent effects on immune disease, including decreasing host injury in a murine model of autoimmune colitis in vivo. It is possible that anti-inflammatory effects may be the result of a specific suppression of THl responses by HDAC inhibitors. Another possibility is that these effects are mediated through the induction of T-reg cells.

In the patient described herein with concomitant CTCL and BP, vorinostat proved to be corticosteroid-sparing within several weeks of beginning therapy. Likewise, on cessation of therapy with vorinostat, the patient's BP rapidly flared, and again required increased doses of systemic corticosteroids in an attempt to control the blistering disorder. Taken together, these data suggest vorinostat is effective for the treatment of BP, and it is also a viable therapeutic option to treat autoimmune disease in the setting of malignant disease in an attempt to avoid other, more immunosuppressive, treatments.

Example 2:

Cutaneous T-cell lymphoma (CTCL) is a group of lymphoproliferative disorders caused by clonally derived, skin homing CD4+/CD26-/CD7-/CLA+/CCR4+ T-cells. Advanced CTCL disease is characterized by Th2 skewing, with decreased numbers of circulating cytotoxic T cells, natural killer (NK) cells, and dendritic cells (DCs). This immune dysregulation results in reduced antitumor responses and susceptibility to infectious agents.

Multimodality immunotherapy with biologic agents in an attempt to boost host antitumor response is a mainstay of therapy in advanced disease.

Vorinostat is an inhibitor of class I and II Zn-dependent HDACs. This compound has been approved by the FDA for treatment of CTCL in 2006 with a 29.7% overall response rate (ORR). Vorinostat causes selective apoptosis of malignant T cells by increasing expression of p21^WAF1, bax, decreasing Stat-6, and activating caspase-3 in CTCL cell lines and patients' peripheral blood lymphocytes (PBL).

Materials & Methods.

PBMCs were isolated from healthy donors and Sezary Syndrome (SS) patients. The cells were treated with 1 μM vorinostat alone, or in combination with stimulatory cytokines such as IFN-γ, IFN-α, IL-21, and TLR 7/8 agonist.

Inflammatory cytokine production was assessed by ELISA, and NK cytolytic activity was assessed by ⁵¹Cr assay with K562 cells. Flow cytometry was utilized to assess activation markers .

Results.

Peripheral blood mononuclear cells (PBMCs) isolated from healthy, age matched controls and CTCL patients were treated with vorinostat alone or in combination with immune stimulatory agents (IFN-γ, IFN-α, IL-21 or a TLR agonist), and malignant cell apoptosis and CMI were assessed (Figure 1). As illustrated in Figure 1, vorinostat inhibited NK cell cytotoxicity against K562 cells. Vorinostat greatly increased malignant CD4+ T-cell apoptosis but not CD8+ cells apoptosis.

Marked inhibition of immune response by vorinostat was associated with suppressed production of immune stimulatory cytokines by dendritic cells (IL- 12 and IFN-α; Figure 3) and NK cells (IFN-γ). This suppression was dose dependent and partially reversed by combining vorinostat with a TLR agonist or IFN-γ

(Figure 3A and Figure 3B). This effect was observed by overnight incubation with vorinostat at concentrations (1-2 μM) that approximate the serum concentrations among treated patients. As illustrated in Figure 3, vorinostat inhibited cytokine production by dendritic cells. Interestingly, both IL- 12 and IFN-α are thought to be important in the genesis of autoimmunity. IFN-γ from NK cells is also thought to play an important role in autoimmunity, and thus inhibition of production of this interferon by a HDAC inhibitor could play an important role in preventing

autoimmunity.

Vorinostat profoundly suppressed NK cell activity, which in experiments using purified NK cells appeared to be directly mediated on this population (Figure 2). This experiment suggested that vorinostat directly and profoundly inhibited NK cell activity. NK cell function with vorinostat was partially preserved by co exposure to IFN-α, IL-21 or both. As NK cells may mediate antibody-dependent cell-mediated cytotoxicity against tissues via the use of autoantibodies, inhibiting the activity of NK cells with a HDAC inhibitor could play an important role in suppressing autoimmunity in a subject. In one aspect, suppression of autoimmunity could take place in the skin of the subject.

Vorinostat also showed an effect on activation of antigen presenting cells (APCs) (Figure 4). Vorinostat inhibited the activation of APCs as evidenced by the inhibition of expression of the co-stimulatory molecule CD80.

Therefore, vorinostat profoundly suppressed TLR 7/8 agonist, IFN-α, and IL-21 -mediated NK cell cytotoxicity. Vorinostat also inhibited production of dendritic cell dependent cytokines IFN-α and IL- 12 by PBMCs. Furthermore, vorinostat blunted APC activation as evidenced by CD80 expression (an important co-stimulatory molecule). Blunting of the host immune response by vorinostat could be ameliorated by combining vorinostat with stimulatory cytokines such as IFN-γ,

IFN-α, or a TLR agonist, and this approach could be useful for treatment of refractory CTCL.

Since vorinostat potently suppressed multiple arms of the immune system, it may be combined with immunomodulating agents in the treatment of CTCL in order to minimize blunting of the host immune response and optimize host antitumor immunity. As illustrated herein, an HDAC inhibitor such as vorinostat may be effective in treatment of inflammatory disease and promotion of graft tolerance. An HDAC inhibitor such as vorinostat may further be used to treat, ameliorate or prevent autoimmune disorders or diseases in a subject.

While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety.

Claims

What is claimed: 1. A method of treating or ameliorating an autoimmune disease or disorder in a subject in need thereof, said method comprising administering to said subject a therapeutically effective amount of a composition comprising a

pharmaceutically acceptable carrier and a compound selected from the group consisting of a histone deacetylase (HDAC) inhibitor and a pharmaceutically acceptable salt or hydrate thereof.

2. The method of claim 1 , wherein said autoimmune disease or disorder is selected from the group consisting of coeliac disease, diabetes mellitus type 1 (IDDM), systemic lupus erythematosus (SLE), Sjogren's syndrome, Churg- Strauss Syndrome, Hashimoto's thyroiditis, Graves' disease, idiopathic

3. The method of claim 1 , wherein said autoimmune disease or disorder is a skin autoimmune disease or disorder.

4. The method of claim 3, wherein said skin autoimmune disease or disorder is selected from the group consisting of psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris.

5. The method of claim 4, wherein said skin autoimmune disease or disorder is bullous pemphigoid.

6. The method of claim 1 , wherein said HDAC inhibitor is vorinostat.

7. The method of claim 1 , wherein said subject is a mammal.

8. The method of claim 7, wherein said mammal is a human.

9. The method of claim 1 , wherein the dose of said HDAC inhibitor in said composition is about 400 mg per day.

10. A method of treating a subject at risk of developing an autoimmune disease or disorder, said method comprising administering to said subject a therapeutically effective amount of a composition comprising a pharmaceutically acceptable carrier and a compound selected from the group consisting of a histone deacetylase (HDAC) inhibitor and a pharmaceutically acceptable salt or hydrate thereof.

11. The method of claim 10, wherein said autoimmune disease or disorder is selected from the list consisting of coeliac disease, diabetes mellitus type 1 (IDDM), systemic lupus erythematosus (SLE), Sjogren's syndrome, Churg-Strauss Syndrome, Hashimoto's thyroiditis, Graves' disease, idiopathic thrombocytopenic purpura, rheumatoid arthritis (RA), polymyositis, ulcerative colitis, Crohn's disease, autoimmune carditis, Wegener's granulomatosis, autoimmune hemolytic anemia, polyarteritis nodosa, psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris.

12 The method of claim 10, wherein said autoimmune disease or disorder is a skin autoimmune disease or disorder.

13. The method of claim 12, wherein said skin autoimmune disease or disorder is selected from the group consisting of psoriasis, vitiligo, epidermolysis bullosa, scleroderma, alopecia areata, epidermolysis bullosa acquisita, bullous pemphigoid, pemphigus foliaceous, and pemphigus vulgaris.

14. The method of claim 13, wherein said skin autoimmune disease or disorder is bullous pemphigoid.

15. The method of claim 10, wherein said HDAC inhibitor is vorinostat.

16. The method of claim 10, wherein said subject is a mammal.

17. The method of claim 16, wherein said mammal is a human.

18. The method of claim 10, wherein the dose of said HDAC inhibitor in said composition is about 400 mg per day.