US008716252B2

(12) United States Patent (10) Patent No.: US 8,716.252 B2 Schafer et al. (45) Date of Patent: May 6, 2014

(54) (METHYLSULFONYL) ETHYL BENZENE (51) Int. Cl. ISONDOLINEDERVATIVES AND THEIR A613 L/70 (2006.01) PHARMACEUTICALUSES A613 L/40 (2006.01) C07H 17/02 (2006.01) (75) Inventors: Peter H. Schafer, Somerset, NJ (US); C07D 209/16 (2006.01) Anthony J. Frank, Easton, PA (US); (52) U.S. Cl. Hon-Wah Man, Princeton, NJ (US); Sai USPC ...... 514/27: 514/416:536/17.4: 548/472 L. Shankar, Chesterfield, NJ (US) (58) Field of Classification Search None (73) Assignee: Celgene Corporation, Summit, NJ (US) See application file for complete search history. (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 (56) References Cited U.S.C. 154(b) by 0 days. U.S. PATENT DOCUMENTS (21) Appl. No.: 13/518,843 6,667,316 B1* 12/2003 Man et al...... 514,323 (22) PCT Filed: Dec. 21, 2010 FOREIGN PATENT DOCUMENTS (86). PCT No.: PCT/US2O10/06142O WO WOO1,34606 5, 2001 S371 (c)(1), * cited by examiner (2), (4) Date: Aug. 21, 2012 Primary Examiner — Traviss C McIntosh, III (87) PCT Pub. No.: WO2011/079091 (74) Attorney, Agent, or Firm — Jones Day PCT Pub. Date: Jun. 30, 2011 (57) ABSTRACT (65) Prior Publication Data Provided are (methylsulfonyl)ethyl benzene isoindoline US 2013/O1162O4 A1 May 9, 2013 compounds, and pharmaceutically acceptable salts, Solvates, O O or stereoisomers thereof. Methods of use and pharmaceutical Related U.S. Application Data compositions of these compounds are also disclosed. (60) Provisional application No. 61/289,356, filed on Dec. 22, 2009. 13 Claims, No Drawings US 8,716,252 B2 1. 2 (METHYLSULFONYL) ETHYL BENZENE has been implicated in a number of diseases and medical ISONDOLINEDERVATIVES AND THEIR conditions including, but not limited to, ocular neovascular PHARMACEUTICALUSES diseases, choroidal neovascular diseases, retina neovascular diseases, rubeosis (neovascularization of the angle), viral dis This application is a S371 national stage application of 5 eases, genetic diseases, inflammatory diseases, allergic dis PCT/US2010/061420, filed Dec. 21, 2010, which claims pri eases, tumor growth and autoimmune diseases. Examples of ority to U.S. Provisional Application No. 61/289.356, filed Such diseases and conditions include, but are not limited to: Dec. 22, 2009, both of which are incorporated herein by diabetic retinopathy; retinopathy of prematurity; corneal reference in their entireties. graft rejection; neovascular glaucoma; retrolental fibroplasia; 10 arthritis; and proliferative vitreoretinopathy. 1. FIELD Accordingly, compounds that can control angiogenesis, Provided herein are (methylsulfonyl)ethylbenzene isoin inhibit PDE 4, and/or inhibit the production of certain cytok doline derivatives. Pharmaceutical compositions comprising ines, including TNFC, may be useful in the treatment and the compounds and methods for treating, preventing and 15 prevention of various diseases and conditions. managing various disorders are also provided. 3. SUMMARY 2. BACKGROUND Provided herein are (methylsulfonyl)ethylbenzene isoin Many cellular functions are mediated by levels of adenos- 20 doline analogs, and pharmaceutically acceptable salts, Sol ine 3',5'-cyclic monophosphate (cAMP). Such cellular func Vates (e.g., hydrates), prodrugs, clathrates, or stereoisomers tions can contribute to inflammatory conditions and diseases thereof. including asthma, inflammation, and other conditions (Lowe Also provided are methods of treating and managing vari and Cheng, Drugs of the Future, 17(9), 799-807, 1992). It has ous diseases or disorders. The methods comprise administer been shown that the elevation of cAMP in inflammatory 25 ing to a patient in need of such treatment or management, or leukocytes inhibits their activation and the Subsequent release having Such a disease or disorder, a therapeutically effective of inflammatory mediators, including TNFC. Increased levels amount of a compound provided herein, or a pharmaceuti of cAMP also lead to the relaxation of airway smooth muscle. cally acceptable salt, Solvate, prodrug, clathrate, or stereoiso In addition, excessive or unregulated TNFC. production has mer thereof. been implicated in a number of disease conditions including 30 Also provided herein are methods of preventing various but not limited to autoimmune and inflammatory diseases. diseases and disorders, which comprise administering to a Specific disease conditions include endotoxemia and/or toxic patient in need of such prevention, orata risk of such a disease shock syndrome Tracey, et al., Nature 330, 662-664 (1987) or disorder, a prophylactically effective amount of a com and Hinshaw, et al., Circ. Shock 30, 279-292 (1990), rheu pound provided herein, or a pharmaceutically acceptable salt, matoid arthritis, inflammatory bowel disease, cachexia De- 35 Solvate, prodrug, clathrate, or stereoisomer thereof. Zube, et al., Lancet, 335 (8690), 662 (1990), lupus and can Also provided herein are pharmaceutical compositions, C. single unit dosage forms, dosing regimens and kits which The primary cellular mechanism for the inactivation of comprise a compound provided herein, or a pharmaceutically cAMP is the breakdown of cAMP by a family of isoenzymes acceptable salt, Solvate, prodrug, clathrate, or stereoisomer referred to as cyclic nucleotide phosphodiesterases (PDE) 40 thereof. Beavo and Reitsnyder, Trends in Pharm., 11, 150-155, 1990. There are greater than ten known members of the 4. DETAILED DESCRIPTION family of PDEs. It is well documented that the inhibition of PDE type IV (PDE4) enzyme is particularly effective in both In one embodiment, provided are (methylsulfonyl)ethyl the inhibition of inflammatory mediator release and the relax- 45 benzene isoindoline compounds, and pharmaceutically ation of airway Smooth muscle (Verghese, et al., Journal of acceptable salts, Solvates, prodrugs, clathrate, and stereoiso Pharmacology and Experimental Therapeutics, 272(3), mers thereof. 1313-1320, 1995). In another embodiment, provided are methods of treating, Increasing cAMP levels (e.g., inhibiting PDE 4) and/or managing, and preventing various diseases and disorders, thus decreasing TNFC. levels constitutes a valuable therapeu- 50 which comprises administering to a patient a therapeutically tic strategy for the treatment of many inflammatory, infec or prophylactically effective amount of a compound provided tious, immunological, and malignant diseases. These include herein, or a pharmaceutically acceptable salt, Solvate, pro but are not restricted to: pulmonary diseases, septic shock, drug, clathrate, or stereoisomer thereof. Examples of diseases sepsis, endotoxic shock, hemodynamic shock and sepsis Syn and disorders are described herein. drome, post ischemic reperfusion injury, malaria, mycobac- 55 In other embodiments, a compound provided herein, or a terial infection, meningitis, many types of psoriasis and other pharmaceutically acceptable salt, Solvate, prodrug, clathrate, dermal diseases, congestive heart failure, fibrotic disease, or stereoisomer thereof, is administered in combination with cachexia, graft rejection, cancer, tumor growth, undesirable another drug (“second active agent') or treatment. Second angiogenesis, autoimmune disease, opportunistic infections active agents include Small molecules and large molecules in AIDS, rheumatoid arthritis, rheumatoid spondylitis, 60 (e.g., proteins and antibodies), examples of which are pro osteoarthritis, other arthritic conditions, inflammatory bowel vided herein, as well as stem cells. Methods, or therapies, that disease, Crohn's disease, ulcerative colitis, multiple Sclero can be used in combination with the administration of com sis, systemic lupus erythematosis, ENL in leprosy, radiation pounds provided herein include, but are not limited to, Sur damage, and hyperoxic alveolar injury. gery, blood transfusions, immunotherapy, biological therapy, In addition, a variety of other diseases and disorders are 65 radiation therapy, and other non-drug based therapies pres also associated with, or characterized by, undesired angio ently used to treat, prevent or manage various disorders genesis. For example, enhanced or unregulated angiogenesis described herein. US 8,716,252 B2 3 4 Also provided are pharmaceutical compositions (e.g., In another embodiment, R' is (cyclopropyl)-C(O) , single unit dosage forms) that can be used in the methods wherein cyclopropyl is substituted with an —OH, and R is provided herein. In one embodiment, pharmaceutical compo ethyl. In another embodiment where R is (OH-substituted sitions comprise a compound provided herein, or a pharma cyclopropyl)-C(O)— and R is ethyl, R is hydrogen. In ceutically acceptable salt, Solvate, prodrug, clathrate, or Ste another embodiment where R' is (OH-substituted cyclopro reoisomer thereof, and optionally a second active agent. pyl)-C(O)—and R is ethyl, R is gluc. 4.1 Compounds Specific compounds include, but are not limited to: In one embodiment, provided herein is a compound of formula (I): 10 O-gluc (I) OR2 NH2 O

15 NHR OR, 21 HO-H- N 21 N S CH R*-- N / S. vich, O NH2 O O-gluc, or a pharmaceutically acceptable salt, Solvate, or stereoiso mer thereof, wherein: 25 21 R" is hydrogen, glucuronide ("gluc”), or (cyclopropyl)-C HO-H- N / (O)—, wherein the cyclopropyl may be substituted with one N S 7 CH or more hydroxyl groups; R is hydrogen, methyl, or gluc; R is hydrogen, ethyl, or gluc; and 30 R" is hydrogen, hydroxyl, or -O-gluc; with the proviso that when R is methyl and R is ethyl, then R' cannot be unsub NH O stituted (cyclopropyl)-O-. In one embodiment, R is hydrogen. In another embodi 21 ment, R' is gluc. In another embodiment, R' is (cyclopropyl)- 35 gluc-O-- N C(O)—. In another embodiment, R' is (cyclopropyl)-C N (O)—, wherein the cyclopropyl is substituted with a hydroxyl group. In one embodiment, R is hydrogen. In another embodi ment, R is methyl. In another embodiment, R is gluc. 40 In one embodiment, R is hydrogen. In another embodi ment, R is ethyl. In another embodiment, R is gluc. In one embodiment, R is hydroxyl. In another embodi ment, R is -O-gluc. 45 Each of the combinations resulting from R'-R is also provided herein. In one embodiment, R' is hydrogen, R is methyl, and R is ethyl. In another embodiment, R' is hydrogen, R is gluc, and R' 50 is ethyl. In another embodiment, R is (cyclopropyl)-C(O)— wherein the cyclopropyl is substituted with an -OH, R is methyl, and R is ethyl. In another embodiment, R' is (cyclopropyl)-C(O)— and 55 R is ethyl. In another embodiment where R' is (cyclopropyl)- C(O)—and R is ethyl, R is hydrogen. In another embodi ment where R' is (cyclopropyl)-C(O)—and R is ethyl, R is gluc. In one embodiment, R and R are both hydrogen. In 60 another embodiment where R and Rare both hydrogen, R' is (cyclopropyl)-C(O)—. In another embodiment, R' is (cyclopropyl)-C(O)— and R’ is methyl. In another embodiment where R' is (cyclopro pyl)-C(O)— and R is methyl, R' is hydrogen. In another 65 embodiment where R' is (cyclopropyl)-C(O)— and R is methyl, R' is gluc. US 8,716,252 B2 5 6 -continued -continued O O OCH3

NH v's O Ho-X, O OCHs. N O M R CH3 10 O

As used herein, and unless otherwise specified, the term “pharmaceutically acceptable salt” refers to salts prepared 15 from pharmaceutically acceptable non-toxic acids, including inorganic acids and organic acids. Suitable non-toxic acids include inorganic and organic acids such as, but not limited to, acetic, alginic, anthranilic, benzenesulfonic, benzoic, cam phorsulfonic, citric, ethenesulfonic, formic, fumaric, furoic, gluconic, glutamic, glucorenic, galacturonic, glycidic, hydro bromic, hydrochloric, isethionic, lactic, maleic, malic, man delic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phenylacetic, propionic, phosphoric, Salicylic, Stearic, suc OH, cinic, Sulfanilic, Sulfuric, tartaric acid, p-toluenesulfonic and 25 the like. In one embodiment, suitable are hydrochloric, hydrobromic, phosphoric, and Sulfuric acids. As used herein, and unless otherwise specified, the term “solvate” means a compound that further includes a stoichio metric or non-stoichiometric amount of solvent bound by 30 non-covalent intermolecular forces. Where the solvent is water, the solvate is a hydrate. As used herein, and unless otherwise specified, the term “prodrug” means a derivative of a compound that can hydro lyze, oxidize, or otherwise react under biological conditions 35 (in vitro or in vivo) to provide the compound. Examples of prodrugs include, but are not limited to, compounds that comprise biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, O-gluc biohydrolyzable carbonates, biohydrolyzable ureides, and 40 biohydrolyzable phosphate analogues. Other examples of prodrugs include compounds that comprise —NO. —NO, OCHs. —ONO, or—ONO, moieties. Prodrugs can typically be pre pared using well-known methods, such as those described in Burger's Medicinal Chemistry and Drug Discovery, 172 45 178, 949-982 (Manfred E. Wolff ed., 5th ed. 1995), and Design of Prodrugs (H. Bundgaard ed., Elselvier, N.Y. 1985). As used herein, and unless otherwise specified, the terms “biohydrolyzable carbamate.” “biohydrolyzable carbonate.” O “biohydrolyzable ureide' and “biohydrolyzable phosphate' 50 mean a carbamate, carbonate, ureide and phosphate, respec tively, of a compound that either: 1) does not interfere with the Ho-3, biological activity of the compound but can confer upon that compound advantageous properties in vivo. Such as uptake, duration of action, or onset of action; or 2) is biologically 55 inactive but is converted in vivo to the biologically active compound. Examples of biohydrolyzable carbamates include, but are not limited to, carbamates that include lower O-gluc alkylamine, Substituted ethylenediamine, aminoacid, hydroxyalkylamine, heterocyclic and heteroaromatic amine, 60 and polyether amine moieties. y NH OCH5, and As used herein, and unless otherwise specified, the term HO 'stereoisomer encompasses all enantiomerically/stereo merically pure and enantiomerically/stereomerically enriched compounds provided herein. 65 As used herein and unless otherwise indicated, the term 'stereomerically pure” means a composition that comprises one stereoisomer of a compound and is Substantially free of US 8,716,252 B2 7 8 other stereoisomers of that compound. For example, a stereo 4.2 Methods of Treatment, Prevention and Management merically pure composition of a compound having one chiral Provided herein are methods of treating, preventing, and/or center will be substantially free of the opposite enantiomer of managing various diseases or disorders using a compound the compound. A stereomerically pure composition of a com provided herein, or a pharmaceutically acceptable salt, Sol pound having two chiral centers will be substantially free of 5 vate (e.g., hydrate), prodrug, clathrate, or Stereoisomer other diastereomers of the compound. A typical stereomeri thereof. Without being limited by a particular theory, com cally pure compound comprises greater than about 80% by pounds provided herein can inhibit PDE 4 enzymatic activity, weight of one stereoisomer of the compound and less than control angiogenesis or inhibit the production of certain about 20% by weight of other stereoisomers of the com cytokines including, but not limited to, TNF-C. INF-C. IP-10, 10 MIG, IL-1 B, IL-12, IL-18, GM-CSF, and/or IL-6. Without pound, greater than about 90% by weight of one stereoisomer being limited by a particular theory, compounds provided of the compound and less than about 10% by weight of the herein can stimulate the production of cetain other cytokines other stereoisomers of the compound, greater than about 95% including IL-10, and also act as inhibitors of T cell activation, by weight of one stereoisomer of the compound and less than resulting in decreased production of cytokines such as, but not about 5% by weight of the other stereoisomers of the com 15 limited to, IL-2 and/or IFN-y. In addition, compounds pro pound, or greater than about 97% by weight of one stereoi vided herein can inhibit the effects of NK cells. Further, somer of the compound and less than about 3% by weight of compounds provided herein may be immunomodulatory and/ the other Stereoisomers of the compound. or cytotoxic, and thus, may be useful as chemotherapeutic As used herein and unless otherwise indicated, the term agents, in particular against diseases such as, but not limited 'stereomerically enriched' means a composition that com to, chronic lymphocytic leukemia, acute lymphoblastic leu prises greater than about 55% by weight of one stereoisomer kemia, and diffuse large B cell lymphoma. Compounds pro of a compound, greater than about 60% by weight of one vided herein can also be anti-angiogenic, and thus, may be stereoisomer of a compound, greater than about 70% by useful for the treatment of cancer. Consequently, without weight, or greater than about 80% by weight of one stereoi being limited by a particular theory, some or all of Such Somer of a compound. 25 characteristics possessed by the compounds provided herein As used herein, and unless otherwise indicated, the term may render them useful in treating, managing, and/or pre “enantiomerically pure’ means a stereomerically pure com venting various diseases or disorders. position of a compound having one chiral center. Similarly, Examples of diseases or disorders include, but are not the term “enantiomerically enriched” means a stereomeri limited to: pulmonary disorders including, but not limited to, cally enriched composition of a compound having one chiral 30 asthma, COPD, scleroderma and idiopathic pulmonary fibro Center. sis; skin diseases including, but not limited to, cutaneous As used herein, and unless otherwise indicated, the term lupus erythmatosus, discoid lupus erythmatosus, dermato “alkyl refers to a saturated straight chain or branched hydro myositis, Systemic lupus erythmatosus, psoriasis, atomic der carbon having a number of carbon atoms as specified herein. matitis, sarcoidosis and cutaneous sarcoidosis; TNFC. related Representative saturated Straight chain alkyls include -me 35 disorders including, but not limited to, inflammatory dis thyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl, eases, autoimmune diseases, rheumatoid arthritis, psoriatic while saturated branched alkyls include -isopropyl, -sec-bu arthritis, ankylosing spondylitis, gouty arthritis, psoriasis, tyl, -isobutyl, -tert-butyl, -isopentyl, 2-methylbutyl, 3-meth Crohn's disease and ulcerative colitis; cancer; leukemia ylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, including chronic lymphocytic leukemia and acute lympho 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methyl 40 blastic leukemia; lymphoma including diffuse large B cell hexyl, 2,3-dimethylbutyl, and the like. The term “alkyl also lymphoma; disorders associated with angiogenesis Such as encompasses cycloalkyl. cancer or tumor growth; pain including, but not limited to, As used herein, and unless otherwise specified, the term Complex Regional Pain Syndrome (“CRPS); Macular “cycloalkyl means a specie of alkyl containing from 3 to 15 Degeneration (“MD) and uveitis and syndromes related carbonatoms, without alternating or resonating double bonds 45 thereof, asbestos-related disorders; parasitic diseases; immu between carbon atoms. It may contain from 1 to 4 rings. nodeficiency disorders; CNS disorders; CNS injury; athero Examples of unsubstituted cycloalkyls include, but are not Sclerosis and related disorders; dysfunctional sleep and limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, related disorders; hemoglobinopathy and related disorders and adamanty1. A cycloalkyl may be substituted with one or (e.g., anemia); and other various diseases and disorders. more of the substituents. 50 As used herein, and unless otherwise specified, the terms As used herein, the term “aryl' means a carbocyclic aro “treat,” “treating and “treatment” refer to the eradication or matic ring containing from 5 to 14 ring atoms. The ring atoms amelioration of a disease or disorder, or of one or more of a carbocyclic aryl group are all carbon atoms. Aryl ring symptoms associated with the disease or disorder. In certain structures include compounds having one or more ring struc embodiments, the terms refer to minimizing the spread or tures such as mono-, bi-, or tricyclic compounds as well as 55 worsening of the disease or disorder resulting from the benzo-fused carbocyclic moieties such as 5,6,7,8-tetrahy administration of one or more prophylactic or therapeutic dronaphthyl and the like. Specifically, the aryl group is a agents to a Subject with Such a disease or disorder. In some monocyclic ring or bicyclic ring. Representative aryl groups embodiments, the terms refer to the administration of a com include phenyl, anthracenyl, fluorenyl, indenyl, azulenyl, pound provided herein, with or without other additional phenanthrenyl and naphthyl. 60 active agent, after the onset of symptoms of the particular It should be noted that if there is a discrepancy between a disease. depicted structure and a name given that structure, the As used herein, and unless otherwise specified, the terms depicted structure is to be accorded more weight. In addition, “prevent,” “preventing and “prevention” refer to the preven if the stereochemistry of a structure or a portion of a structure tion of the onset, recurrence or spread of a disease or disorder, is not indicated with, for example, bold or dashed lines, the 65 or of one or more symptoms thereof. In certain embodiments, structure or portion of the structure is to be interpreted as the terms refer to the treatment with or administration of a encompassing all stereoisomers of it. compound provided herein, with or without other additional US 8,716,252 B2 10 active compound, prior to the onset of symptoms, particularly drugs and toxins such as fenfluramines, congenital heart dis to patients at risk of disease or disorders provided herein. The ease, pulmonary venous hypertension, chronic obstructive terms encompass the inhibition or reduction of a symptom of pulmonary disease, interstitial lung disease, sleep-disordered the particular disease. Patients with familial history of a dis breathing, alveolar hypoVentilation disorder, chronic expo ease in particular are candidates for preventive regimens in Sure to high altitude, neonatal lung disease, alveolar-capillary certain embodiments. In addition, patients who have a history dysplasia, sickle cell disease, other coagulation disorder, of reccurring symptoms are also potential candidates for the chronic thromboemboli, connective tissue disease, lupus prevention. In this regard, the term “prevention' may be inter including systemic and cutaneous lupus, Schistosomiasis, Sar changeably used with the term “prophylactic treatment.” coidosis or pulmonary capillary hemangiomatosis. As used herein, and unless otherwise specified, the terms 10 In one embodiment, provided herein are methods of treat “manage.” “managing” and “management” refer to prevent ing, preventing or managing asthma. In another embodiment, ing or slowing the progression, spread or worsening of a provided herein are methods of treating, preventing or man disease or disorder, or of one or more symptoms thereof. aging COPD. In another embodiment, provided herein are Often, the beneficial effects that a subject derives from a methods of treating, preventing or managing idiopathic pull prophylactic and/ortherapeutic agent do not resultina cure of 15 monary fibrosis. the disease or disorder. In this regard, the term “managing Examples of skin diseases include, but are not limited to, encompasses treating a patient who had suffered from the those described in U.S. publication no. 2005/0214328A1, particular disease in an attempt to prevent or minimize the published Sep. 29, 2005, which is incorporated herein by recurrence of the disease. reference. Specific examples include, but are not limited to, As used herein, and unless otherwise specified, a “thera keratoses and related symptoms, skin diseases or disorders peutically effective amount of a compound is an amount characterized with overgrowths of the epidermis, acne, sufficient to provide a therapeutic benefit in the treatment or wrinkles, and other skin diseases. management of a disease or disorder, or to delay or minimize As used herein, the term “keratosis” refers to any lesion on one or more symptoms associated with the disease or disor the epidermis marked by the presence of circumscribed over der. Atherapeutically effective amount of a compound means 25 growths of the horny layer, including but not limited to actinic an amount of therapeutic agent, alone or in combination with keratosis, seborrheic keratosis, keratoacanthoma, keratosis other therapies, which provides a therapeutic benefit in the follicularis (Darier disease), inverted follicular keratosis, pal treatment or management of the disease or disorder. The term moplantar keratoderma (PPK, keratosis palmaris et “therapeutically effective amount” can encompass an amount plantaris), keratosis pilaris, and stucco keratosis. The term that improves overall therapy, reduces or avoids symptoms or 30 “actinic keratosis' also refers to senile keratosis, keratosis causes of disease or disorder, or enhances the therapeutic senilis, Verruca senilis, plana Senilis, Solar keratosis, kerato efficacy of another therapeutic agent. derma or keratoma. The term "seborrheic keratosis' also As used herein, and unless otherwise specified, a “prophy refers to seborrheic wart, senile wart, or basal cell papilloma. lactically effective amount of a compound is an amount Keratosis is characterized by one or more of the following Sufficient to prevent a disease or disorder, or prevent its recur 35 symptoms: rough appearing, Scaly, erythematous papules, rence. A prophylactically effective amount of a compound plaques, spicules or nodules on exposed surfaces (e.g., face, means an amount of therapeutic agent, alone or in combina hands, ears, neck, legs and thorax), excrescences of keratin tion with other agents, which provides a prophylactic benefit referred to as cutaneous horns, hyperkeratosis, telangiecta in the prevention of the disease. The term “prophylactically sias, elastosis, pigmented lentigines, acanthosis, parakerato effective amount' can encompass an amount that improves 40 sis, dyskeratoses, papillomatosis, hyperpigmentation of the overall prophylaxis or enhances the prophylactic efficacy of basal cells, cellular atypia, mitotic figures, abnormal cell-cell another prophylactic agent. adhesion, dense inflammatory infiltrates and Small preva As used herein, and unless otherwise specified, the term lence of Squamous cell carcinomas. “administration' encompasses provision of the compounds Examples of skin diseases or disorders characterized with provided herein to a patient as they exist outside the body of 45 overgrowths of the epidermis include, but are not limited to, the patient. The term therefore encompasses provision of an any conditions, diseases or disorders marked by the presence isolated or purified compound to a patient. of overgrowths of the epidermis, including but not limited to, Examples of pulmonary disorders include, but are not lim infections associated with papilloma virus, arsenical kera ited to, those described in U.S. publication no. 2005/ toses, sign of Leser-Trélat, warty dyskeratoma (WD), tricho 0239842A1, published Oct. 27, 2005, which is incorporated 50 stasis spinulosa (TS), erythrokeratodermia variabilis (EKV), herein by reference. Specific examples include pulmonary ichthyosis fetalis (harlequin ichthyosis), knuckle pads, cuta hypertension and related disorders. Examples of pulmonary neous melanoacanthoma, porokeratosis, psoriasis, squamous hypertension and related disorders include, but are not limited cell carcinoma, confluent and reticulated papillomatosis to: asthma; chronic obstructive pulmonary disease (COPD); (CRP), acrochordons, cutaneous horn, cowden disease (mul primary pulmonary hypertension (PPH); secondary pulmo 55 tiple hamartoma syndrome), dermatosis papulosa nigra nary hypertension (SPH); familial PPH; sporadic PPH; pre (DPN), epidermal nevus syndrome (ENS), ichthyosis Vul capillary pulmonary hypertension; pulmonary arterial hyper garis, molluscum contagiosum, prurigo nodularis, and acan tension (PAH); pulmonary artery hypertension; idiopathic thosis nigricans (AN). pulmonary hypertension, idiopathic pulmonary fibrosis, Other skin diseases include, but are not limited to, cutane thrombotic pulmonary arteriopathy (TPA); plexogenic pull 60 ous lupus erythmatosus, discoid lupus erythmatosus, der monary arteriopathy; functional classes I to IV pulmonary matomyositis, systemic lupus erythmatosus, psoriasis and hypertension; and pulmonary hypertension associated with, cutaneous sarcoidosis. In one embodiment, provided herein related to, or secondary to, left Ventricular dysfunction, mitral are methods of treating, preventing or managing cutaneous valvular disease, constrictive pericarditis, aortic Stenosis, car lupus erythmatosus. In another embodiment, provided herein diomyopathy, mediastinal fibrosis, anomalous pulmonary 65 are methods of treating, preventing or managing discoid venous drainage, pulmonary venoocclusive disease, collagen lupus erythmatosus. In another embodiment, provided herein Vasular disease, congenital heart disease, HIV virus infection, are methods of treating, preventing or managing dermato US 8,716,252 B2 11 12 myositis. In another embodiment, provided herein are meth myelocytic leukemias. The compounds provided herein can ods of treating, preventing or managing systemic lupus eryth be used for treating, preventing or managing either primary or matosus. In another embodiment, provided herein are metastatic tumors. methods of treating, preventing or managing psoriasis. In Other specific cancers include, but are not limited to, another embodiment, provided herein are methods of treat advanced malignancy, amyloidosis, neuroblastoma, menin ing, preventing or managing cutaneous sarcoidosis. gioma, hemangiopericytoma, multiple brain metastase, glio Examples of elevated TNFC. related disorders include, but blastoma multiforms, glioblastoma, brain stem glioma, poor are not limited to, those described in WO98/03502 and WO prognosis malignant brain tumor, malignant glioma, recur 98/54170, both of which are incorporated herein in their rent malignant glioma, anaplastic astrocytoma, anaplastic oli 10 godendroglioma, neuroendocrine tumor, rectal adenocarci entireties by reference. Specific examples include, but are not noma, Dukes C & D colorectal cancer, unresectable limited to: endotoxemia or toxic shock syndrome; cachexia; colorectal carcinoma, metastatic hepatocellular carcinoma, adult respiratory distress syndrome; bone resorption diseases Kaposi's sarcoma, karotype acute myeloblastic leukemia, such as arthritis; hypercalcemia; Graft versus Host Reaction; chronic lymphocytic leukemia (CLL), Hodgkin’s lymphoma, cerebral malaria; inflammation; tumor growth; chronic pull 15 non-Hodgkin’s lymphoma, cutaneous T-Cell lymphoma, monary inflammatory diseases; reperfusion injury; myocar cutaneous B-Cell lymphoma, diffuse large B-Cell lym dial infarction; stroke; circulatory shock; rheumatoid arthri phoma, low grade follicular lymphoma, metastatic melanoma tis; Crohn's disease; HIV infection and AIDS; other disorders (localized melanoma, including, but not limited to, ocular Such as rheumatoid arthritis, gouty arthritis, rheumatoid melanoma), malignant mesothelioma, malignant pleural spondylitis, ankylosing spondylitis, osteoarthritis, psoriatic effusion mesothelioma syndrome, peritoneal carcinoma, pap arthritis and other arthritic conditions, septic shock, septis, illary serous carcinoma, gynecologic sarcoma, Soft tissue endotoxic shock, graft versus host disease, wasting, Crohn's sarcoma, Scleroderma, cutaneous vasculitis, Langerhans cell disease, ulcerative colitis, multiple Sclerosis, systemic lupus histiocytosis, leiomyosarcoma, fibrodysplasia ossificans pro erythromatosis, ENL in leprosy, HIV, AIDS, and opportunis gressive, hormone refractory prostate cancer, resected high tic infections in AIDS; disorders such as septic shock, sepsis, 25 risk soft tissue sarcoma, unrescectable hepatocellular carci endotoxic shock, hemodynamic shock and sepsis syndrome, noma, Waldenstrom's macroglobulinemia, Smoldering post ischemic reperfusion injury, malaria, mycobacterial myeloma, indolent myeloma, fallopian tube cancer, androgen infection, meningitis, psoriasis, congestive heart failure, independent prostate cancer, androgen dependent stage IV fibrotic disease, fever, cachexia, graft rejection, oncogenic or non-metastatic prostate cancer, hormone-insensitive prostate cancerous conditions, asthma, autoimmune disease, radiation 30 cancer, chemotherapy-insensitive prostate cancer, papillary damages, and hyperoxic alveolar injury; viral infections. Such thyroid carcinoma, follicular thyroid carcinoma, medullary as those caused by the herpesviruses: viral conjunctivitis; or thyroid carcinoma, and leiomyoma. In a specific embodi atopic dermatitis. ment, the cancer is metastatic. In another embodiment, the In one embodiment, provided herein are methods of treat cancer is refractory or resistance to chemotherapy or radia ing, preventing or managing rheumatoid arthritis. In another 35 tion. embodiment, provided herein are methods of treating, pre In one embodiment, provided herein are methods of treat venting or managing psoriatic arthritis. In another embodi ing, preventing or managing various forms of leukemias Such ment, provided herein are methods of treating, preventing or as chronic lymphocytic leukemia, chronic myelocytic leuke managing ankylosing spondylitis. In another embodiment, mia, acute lymphoblastic leukemia, acute myelogenous leu provided herein are methods of treating, preventing or man 40 kemia and acute myeloblastic leukemia, including leukemias aging gouty arthritis. In another embodiment, provided that are relapsed, refractory or resistant, as disclosed in U.S. herein are methods of treating, preventing or managing publication no. 2006/0030594, published Feb. 9, 2006, which Crohn's disease. In another embodiment, provided herein are is incorporated in its entirety by reference. methods of treating, preventing or managing ulcerative coli The term “leukemia' refers malignant neoplasms of the tis. In another embodiment, provided herein are methods of 45 blood-forming tissues. The leukemia includes, but is not lim treating, preventing or managing psoriasis. ited to, chronic lymphocytic leukemia, chronic myelocytic Examples of cancer and precancerous conditions include, leukemia, acute lymphoblastic leukemia, acute myelogenous but are not limited to, those described in U.S. Pat. Nos. 6,281, leukemia and acute myeloblastic leukemia. The leukemia can 230 and 5,635,517 to Muller et al., in various U.S. patent be relapsed, refractory or resistant to conventional therapy. publications to Zeldis, including publication nos. 2004/ 50 The term “relapsed” refers to a situation where patients who 022014.4A1, published Nov. 4, 2004 (Treatment of Myelod have had a remission of leukemia after therapy have a return ysplastic Syndrome); 2004/0029832A1, published Feb. 12, of leukemia cells in the marrow and a decrease in normal 2004 (Treatment of Various Types of Cancer); and 2004/ blood cells. The term “refractory or resistant” refers to a 0087546, published May 6, 2004 (Treatment of Myeloprolif circumstance where patients, even after intensive treatment, erative Diseases). Examples also include those described in 55 have residual leukemia cells in their marrow. In one embodi WO 2004/103274, published Dec. 2, 2004. All of these ref ment, provided herein are methods of treating, preventing or erences are incorporated herein in their entireties by refer managing chronic lymphocytic leukemia. In another embodi CCC. ment, provided herein are methods of treating, preventing or Specific examples of cancer include, but are not limited to, managing acute lymphoblastic leukemia. cancers of the skin, such as melanoma; lymph node; breast; 60 In another embodiment, provided herein are methods of cervix; uterus; gastrointestinal tract, lung; ovary; prostate; treating, preventing or managing various types of lympho colon; rectum; mouth; brain; head and neck; throat; testes; mas, including Non-Hodgkin’s lymphoma (NHL). The term kidney; pancreas; bone; spleen; liver, bladder, larynx, nasal “lymphoma’ refers a heterogenous group of neoplasms aris passages; and AIDS-related cancers. The compounds are also ing in the reticuloendothelial and lymphatic systems. “NHL useful for treating cancers of the blood and bone marrow, Such 65 refers to malignant monoclonal proliferation of lymphoid as multiple myeloma and acute and chronic leukemias, for cells in sites of the immune system, including lymph nodes, example, lymphoblastic, myelogenous, lymphocytic, and bone marrow, spleen, liver and gastrointestinal tract. US 8,716,252 B2 13 14 Examples of NHL include, but are not limited to, mantle cell include, but are not limited to, atrophic (dry) MD, exudative lymphoma (MCL), lymphocytic lymphoma of intermediate (wet) MD, age-related maculopathy (ARM), choroidal differentiation, intermediate lymphocytic lymphoma (ILL), neovascularisation (CNVM), retinal pigment epithelium diffuse poorly differentiated lymphocytic lymphoma (PDL), detachment (PED), and atrophy of retinal pigment epithelium diffuse large B cell lymphoma, centrocytic lymphoma, dif (RPE). In one embodiment, provided herein are methods of fuse small-cleaved cell lymphoma (DSCCL), follicular lym treating, preventing or managing uveitis. phoma, and any type of the mantle cell lymphomas that can be Examples of asbestos-related disorders include, but not seen under the microscope (nodular, diffuse, blastic and limited to, those described in U.S. publication no. 2005/ mentle Zone lymphoma). In one embodiment, provided 0100529, published May 12, 2005, which is incorporated herein are methods of treating, preventing or managing dif 10 herein by reference. Specific examples include, but are not fuse large B cell lymphoma. limited to, mesothelioma, asbestosis, malignant pleural effu Examples of diseases and disorders associated with, or Sion, benign exudative effusion, pleural plaques, pleural cal characterized by, undesired angiogenesis include, but are not cification, diffuse pleural thickening, rounded atelectasis, limited to, inflammatory diseases, autoimmune diseases, fibrotic masses, and lung cancer. viral diseases, genetic diseases, allergic diseases, bacterial 15 Examples of parasitic diseases include, but are not limited diseases, ocular neovascular diseases, choroidal neovascular to, those described in U.S. publication no. 2006/0154880, diseases, retina neovascular diseases, cancer and rubeosis published Jul. 13, 2006, which is incorporated herein by (neovascularization of the angle). Specific examples of the reference. Parasitic diseases include diseases and disorders diseases and disorders associated with, or characterized by, caused by human intracellular parasites Such as, but not lim undesired angiogenesis include, but are not limited to, arthri ited to, P falcifarium, P ovale, P. vivax, P malariae, L. tis, endometriosis, Crohn's disease, heart failure, advanced donovari, L. infantum, L. aethiopica, L. major; L. tropica, L. heart failure, renal impairment, endotoxemia, toxic shock mexicana, L. braziliensis, TGondii, B. microti, B. divergens, syndrome, osteoarthritis, retrovirus replication, wasting, B. coli, C. parvum, C. Cayetanensis, E. histolytica, I. belli, S. meningitis, silica-induced fibrosis, asbestos-induced fibrosis, mansonii, S. haematobium, Trypanosoma ssp., Toxoplasma Veterinary disorder, malignancy-associated hypercalcemia, 25 ssp., and O. volvulus. Other diseases and disorders caused by stroke, circulatory shock, periodontitis, gingivitis, macro non-human intracellular parasites such as, but not limited to, cytic anemia, refractory anemia, and 5q-deletion syndrome. Babesia bovis, Babesia Canis, Banesia Gibsoni, Besnoitia Examples of pain include, but are not limited to those darlingi, Cytauxzoon felis, Eimeria ssp., Hammondia ssp., described in U.S. patent publication no. 2005/0203142, pub and Theileria ssp., are also encompassed. Specific examples lished Sep. 15, 2005, which is incorporated herein by refer 30 include, but are not limited to, malaria, babesiosis, trypano ence. Specific types of pain include, but are not limited to, Somiasis, leishmaniasis, toxoplasmosis, meningoencephali nociceptive pain, neuropathic pain, mixed pain of nociceptive tis, keratitis, amebiasis, giardiasis, cryptosporidiosis, isospo and neuropathic pain, visceral pain, migraine, headache and riasis, cyclosporiasis, microsporidiosis, ascariasis, post-operative pain. trichuriasis, ancylostomiasis, strongyloidiasis, toxocariasis, Examples of nociceptive pain include, but are not limited 35 trichinosis, lymphatic filariasis, onchocerciasis, filariasis, to, pain associated with chemical orthermal burns, cuts of the Schistosomiasis, and dermatitis caused by animal Schisto skin, contusions of the skin, osteoarthritis, rheumatoid arthri SOS. tis, tendonitis, and myofascial pain. Examples of immunodeficiency disorders include, but are Examples of neuropathic pain include, but are not limited not limited to, those described in U.S. publication no. 2006/ to, CRPS type I, CRPS type II, reflex sympathetic dystrophy 40 0188475, published Aug. 24, 2006. Specific examples (RSD), reflex neurovascular dystrophy, reflex dystrophy, include, but not limited to, adenosine deaminase deficiency, sympathetically maintained pain syndrome, causalgia, antibody deficiency with normal or elevated Igs, ataxia-ten Sudeck atrophy of bone, algoneurodystrophy, shoulder hand langiectasia, bare lymphocyte syndrome, common variable syndrome, post-traumatic dystrophy, trigeminal neuralgia, immunodeficiency, Ig deficiency with hyper-IgM, Ig heavy post herpetic neuralgia, cancer related pain, phantom limb 45 chain deletions, IgA deficiency, immunodeficiency with thy pain, fibromyalgia, chronic fatigue syndrome, spinal cord moma, reticular dysgenesis, Nezelof syndrome, selective IgG injury pain, central post-stroke pain, radiculopathy, diabetic Subclass deficiency, transient hypogammaglobulinemia of neuropathy, post-stroke pain, luetic neuropathy, and other infancy, Wistcott-Aldrich syndrome, X-linked agamma painful neuropathic conditions such as those induced by globulinemia, X-linked severe combined immunodeficiency. drugs such as Vincristine and velcade. 50 Examples of CNS disorders include, but are not limited to, As used herein, the terms “complex regional pain syn those described in U.S. publication no. 2005/0143344, pub drome,” “CRPS and “CRPS and related syndromes” mean a lished Jun. 30, 2005, which is incorporated herein by refer chronic pain disorder characterized by one or more of the ence. Specific examples include, but are not limited to, following: pain, whether spontaneous or evoked, including include, but are not limited to, Amyotrophic Lateral Sclerosis, allodynia (painful response to a stimulus that is not usually 55 Alzheimer Disease, Parkinson Disease, Huntington's Dis painful) and hyperalgesia (exaggerated response to a stimulus ease, Multiple Sclerosis other neuroimmunological disorders that is usually only mildly painful); pain that is disproportion Such as Tourette Syndrome, delerium, or disturbances in con ate to the inciting event (e.g., years of severe pain after an sciousness that occur over a short period of time, and amnes ankle sprain); regional pain that is not limited to a single tic disorder, or discreet memory impairments that occur in the peripheral nerve distribution; and autonomic dysregulation 60 absence of other central nervous system impairments. (e.g., edema, alteration in blood flow and hyperhidrosis) asso Examples of CNS injuries and related syndromes include, ciated with trophic skin changes (hair and nail growth abnor but are not limited to, those described in U.S. publication no. malities and cutaneous ulceration). 2006/0122228, published Jun. 8, 2006, which is incorporated Examples of MD and uveitis and related syndromes herein by reference. Specific examples include, but are not include, but are not limited to, those described in U.S. patent 65 limited to, CNS injury/damage and related syndromes, publication no. 2004/0091455, published May 13, 2004, include, but are not limited to, primary brain injury, second which is incorporated herein by reference. Specific examples ary brain injury, traumatic brain injury, focal brain injury, US 8,716,252 B2 15 16 diffuse axonal injury, head injury, concussion, post-concus -continued sion syndrome, cerebral contusion and laceration, Subdural Peroneal Calf hematoma, epidermal hematoma, post-traumatic epilepsy, Popliteal Knee chronic vegetative state, complete SCI, incomplete SCI, acute Posterior tibial Calf SCI, Subacute SCI, chronic SCI, central cord syndrome, Pulmonary Lungs Brown-Sequard syndrome, anterior cord syndrome, conus Radial Forearm Renal Kidney medullaris syndrome, cauda equina Syndrome, neurogenic Splenic Stomach, pancreas, and spleen shock, spinal shock, altered level of consciousness, headache, Subclavian Shoulder nausea, emesis, memory loss, dizziness, diplopia, blurred Superior mesenteric Pancreas, Small intestine, ascending and transverse vision, emotional lability, sleep disturbances, irritability, 10 colon inability to concentrate, nervousness, behavioral impairment, Testicular Testes cognitive deficit, and seizure. Ulnar Forearm Other disease or disorders include, but not limited to, viral, genetic, allergic, and autoimmune diseases. Specific Examples of dysfunctional sleep and related syndromes examples include, but not limited to, HIV, hepatitis, adult 15 include, but are not limited to, those disclosed in U.S. publi respiratory distress syndrome, bone resorption diseases, cation no. 2005/0222209A1, published Oct. 6, 2005, which is chronic pulmonary inflammatory diseases, dermatitis, cystic incorporated herein by reference. Specific examples include, fibrosis, septic shock, sepsis, endotoxic shock, hemodynamic but are not limited to, Snoring, sleep apnea, insomnia, narco shock, sepsis syndrome, post ischemic reperfusion injury, lepsy, restless leg syndrome, sleep terrors, sleep walking meningitis, psoriasis, fibrotic disease, cachexia, graft versus sleep eating, and dysfunctional sleep associated with chronic host disease, graft rejection, auto-immune disease, rheuma neurological or inflammatory conditions. Chronic neurologi toid spondylitis, Crohn's disease, ulcerative colitis, inflam calorinflammatory conditions, include, but are not limited to, matory-bowel disease, multiple Sclerosis, systemic lupus Complex Regional Pain Syndrome, chronic low back pain, erythrematosus, ENL in leprosy, radiation damage, cancer, musculoskeletal pain, arthritis, radiculopathy, pain associ asthma, or hyperoxic alveolar injury. 25 ated with cancer, fibromyalgia, chronic fatigue syndrome, Examples of atherosclerosis and related conditions visceral pain, bladderpain, chronic pancreatitis, neuropathies include, but are not limited to, those disclosed in U.S. publi (diabetic, post-herpetic, traumatic or inflammatory), and neu cation no. 2002/0054899, published May 9, 2002, which is rodegenerative disorders such as Parkinson's Disease, Alzhe incorporated herein by reference. Specific examples include, imer's Disease, amyotrophic lateral Sclerosis, multiple scle but are not limited to, all forms of conditions involving ath 30 rosis, Huntington's Disease, bradykinesia; muscle rigidity; erosclerosis, including restenosis after vascular intervention parkinsonian tremor, parkinsonian gait; motion freezing; Such as angioplasty, Stenting, atherectomy and grafting. All depression; defective long-term memory, Rubinstein-Taybi forms of vascular intervention are contemplated herein, syndrome (RTS): dementia; postural instability; hypokinetic including diseases of the cardiovascular and renal system, disorders; synuclein disorders; multiple system atrophies; Such as, but not limited to, renal angioplasty, percutaneous 35 striatonigral degeneration; olivopontocerebellar atrophy; coronary intervention (PCI), percutaneous transluminal coro Shy-Drager syndrome; motor neuron disease with parkinso nary angioplasty (PTCA), carotid percutaneous transluminal nian features; Lewy body dementia; Tau pathology disorders; angioplasty (PTA), coronary by-pass grafting, angioplasty progressive Supranuclear palsy, corticobasal degeneration; with stent implantation, peripheral percutaneous translumi frontotemporal dementia; amyloid pathology disorders; mild nal intervention of the iliac, femoral or popliteal arteries, and 40 cognitive impairment; Alzheimer disease with parkinsonism; Surgical intervention using impregnated artificial grafts. The Wilson disease: Hallervorden-Spatz disease; Chediak-Ha following chart provides a listing of the major systemic arter gashi disease; SCA-3 spinocerebellar ataxia; X-linked dysto ies that may be in need of treatment, all of which are contem nia parkinsonism; prion disease; hyperkinetic disorders; cho plated herein: rea; ballismus; dystonia tremors; Amyotrophic Lateral 45 Sclerosis (ALS); CNS trauma and myoclonus. Examples of hemoglobinopathy and related disorders Artery Body Areas Supplied include, but are not limited to, those described in U.S. publi Axillary Shoulder and axilla cation no. 2005/0143420A1, published Jun. 30, 2005, which Brachial Upper arm is incorporated herein by reference. Specific examples Brachiocephalic Head, neck, and arm 50 include, but are not limited to, hemoglobinopathy, sickle cell Celiac Divides into left gastric, splenic, and hepatic arteries Common carotid Neck anemia, and any other disorders related to the differentiation Common iliac Divides into external and internal iliac arteries of CD34+ cells. Coronary Heart In other embodiments, the use of compounds provided Deep femoral Thigh herein in various immunological applications, in particular, Digital Fingers 55 as vaccine adjuvants, particularly anticancer vaccine adju Dorsalis pedis Foot External carotid Neck and external head regions vants, as disclosed in U.S. Publication No. 2007/0048327, External iliac Femoral artery published Mar. 1, 2007, which is incorporated herein in its Femoral Thigh entirety by reference, is also encompassed. These embodi Gastric Stomach Hepatic Liver, gallbladder, pancreas, and duodenum ments also relate to the uses of compounds provided herein in inferior mesenteric Descending colon, rectum, and pelvic wall 60 combination with vaccines to treat or prevent cancer or infec internal carotid Neck and internal head regions tious diseases, and other various uses of immunomodulatory internal iliac Rectum, urinary bladder, external genitalia, buttocks compounds such as reduction or desensitization of allergic muscles, uterus and vagina reactions. Left gastric Esophagus and stomach Middle sacral Sacrum Doses of a compound provided herein, or a pharmaceuti Ovarian Ovaries 65 cally acceptable salt, Solvate, clathrate, stereoisomer or pro Palmar arch Hand drug thereof, vary depending on factors such as: specific indication to be treated, prevented, or managed, age and con US 8,716,252 B2 17 18 dition of a patient; and amount of second active agent used, if lowering agents, thromboxane inhibitors, Steroids and other any. Generally, a compound provided herein, or a pharma therapeutics known to reduce pulmonary artery pressure. ceutically acceptable salt, Solvate, clathrate, Stereoisomer or Specific examples include, but are not limited to, warfarin prodrug thereof, may be used in an amount of from about 0.1 (CoumadinR), a diuretic, a cardiac glycoside, digoxin-oxy mg to about 500 mg per day, and can be adjusted in a con gen, diltiazem, nifedipine, a vasodilator Such as prostacyclin ventional fashion (e.g., the same amount administered each (e.g., prostaglandin I2 (PGI2), epoprostenol (EPO, Floran R), day of the treatment, prevention or management period), in treprostinil (Remodulin(R), nitric oxide (NO), bosentan (Tra cycles (e.g., one week on, one week off), or in an amount that cleer(R), amlodipine, epoprostenol (Floran R), treprostinil increases or decreases over the course of treatment, preven (Remodulin(R), prostacyclin, tadalafil (Cialis(R), simvastatin tion, or management. In other embodiments, the dose can be 10 (Zocor R), omapatrilat (Vanlev(R), irbesartan (AvaproR), from about 1 mg to about 300 mg, from about 0.1 mg to about pravastatin (PravacholR), digoxin, L-arginine, iloprost, 150 mg, from about 1 mg to about 200 mg, from about 10 mg betaprost, and sildenafil (ViagraR). to about 100 mg, from about 0.1 mg to about 50 mg, from In some embodiments, the second active agents include, about 1 mg to about 50 mg, from about 10 mg to about 50 mg. but are not limited to, keratolytics, retinoids, C-hydroxy from about 20 mg to about 30 mg, from about 1 mg to about 15 acids, antibiotics, collagen, botulinum toxin, interferon, Ste 30 mg. or from about 1 mg to about 20 mg. roids, and immunomodulatory agents. Specific examples 4.3 Second Active Agents include, but are not limited to, 5-fluorouracil, masoprocol, A compound provided herein, or a pharmaceutically trichloroacetic acid, Salicylic acid, lactic acid, ammonium acceptable salt, Solvate, prodrug, clathrate, or stereoisomer lactate, urea, tretinoin, isotretinoin, antibiotics, collagen, thereof, can be combined with other pharmacologically botulinum toxin, interferon, corticosteroid, transretinoic acid active compounds (“second active agents’) in methods and and collagens such as human placental collagen, animal pla compositions provided herein. Certain combinations may cental collagen, Dermalogen, Alloderm, Fascia, Cymetra, work synergistically in the treatment of particular types dis Autologen, Zyderm, Zyplast, Resoplast, and Isolagen. eases or disorders, and conditions and symptoms associated In other embodiments, the second active agents include, with Such diseases or disorders. A compound provided 25 but are not limited to: Semaxanib., cyclosporin; etanercept; herein, or a pharmaceutically acceptable salt, Solvate, clath doxycycline; bortezomib; acivicin; aclarubicin; acodazole rate, Stereoisomer or prodrug thereof, can also work to alle hydrochloride; acronine; adoZelesin; aldesleukin; altre viate adverse effects associated with certain second active tamine; ambomycin; ametantrone acetate; amsacrine; anas agents, and vice versa. trozole; anthramycin; asparaginase; asperlin; azacitidine; One or more second active ingredients or agents can be 30 aZetepa; azotomycin; batimastat; benzodepa; bicalutamide; used in the methods and compositions provided herein. Sec bisantrene hydrochloride; bisnafide dimesylate; bizelesin; ond active agents can be large molecules (e.g., proteins) or bleomycin sulfate; brequinar sodium; bropirimine; busulfan; Small molecules (e.g., synthetic inorganic, organometallic, or cactinomycin; calusterone; caracemide; carbetimer; carbopl organic molecules). atin: carmustine; carubicin hydrochloride; carzelesin: cedef Examples of large molecule active agents include, but are 35 ingol; celecoxib; chlorambucil; cirolemycin; cisplatin: not limited to, hematopoietic growth factors, cytokines, and cladribine; crisinatol mesylate; cyclophosphamide; cytara monoclonal and polyclonal antibodies. Specific examples of bine; dacarbazine; dactinomycin; daunorubicin hydrochlo the active agents are anti-CD40 monoclonal antibodies (such ride; decitabine; dexormaplatin; deZaguanine; deZaguanine as, for example, SGN-40); histone deacetylyase inhibitors mesylate; diaziquone; docetaxel, doxorubicin; doxorubicin (such as, for example, SAHA and LAQ 824); heat-shock 40 hydrochloride; droloxifene; droloxifene citrate; dromo protein-90 inhibitors (such as, for example, 17-AAG); insu stanolone propionate; duaZomycin; ediatrexate; eflornithine lin-like growth factor-1 receptor kinase inhibitors; vascular hydrochloride; elsamitrucin; enloplatin; enpromate; epipro endothelial growth factor receptor kinase inhibitors (such as, pidine; epirubicin hydrochloride; erbulozole; esorubicin for example, PTK787); insulin growth factor receptor inhibi hydrochloride; estramustine; estramustine phosphate tors; lysophosphatidic acid acyltransrerase inhibitors; IkB 45 Sodium; etanidazole; etoposide; etoposide phosphate: eto kinase inhibitors: p38 MAPK inhibitors: EGFR inhibitors prine; fadrozole hydrochloride; fazarabine; fenretinide; (such as, for example, gefitinib and erlotinib HCL); HER-2 floxuridine; fludarabine phosphate; fluorouracil; fluorocitab antibodies (such as, for example, trastuzumab (HerceptinR) ine; fosquidone; fostriecin Sodium; gemcitabine; gemcitab and pertuzumab (OmnitargTM)); VEGFR antibodies (such as, ine hydrochloride; hydroxyurea; idarubicin hydrochloride: for example, bevacizumab (AvastinTM)); VEGFR inhibitors 50 ifosfamide; ilmofosine, iproplatin; irinotecan; irinotecan (such as, for example, flk-1 specific kinase inhibitors, hydrochloride; lanreotide acetate; letrozole; leuprolide SU5416 and ptk787/zk222584); PI3K inhibitors (such as, for acetate; liaroZole hydrochloride; lometrexol Sodium, lomus example, wortmannin); C-Met inhibitors (such as, for tine; losoxantrone hydrochloride; masoprocol; maytansine; example, PHA-665752); monoclonal antibodies (such as, for mechlorethamine hydrochloride; megestrol acetate; example, rituximab (Rituxae), to situmomab (BeXXae), 55 melengestrol acetate; melphalan; menogaril; mercaptopu edrecolomab (Panorex) and G250); and anti-TNF-C. antibod rine; methotrexate; methotrexate Sodium; metoprine; meture ies. Examples of small molecule active agents include, but are depa; mitindomide; mitocarcin, mitocromin, mitogillin; not limited to, anticancer agents and antibiotics (e.g., mitomalcin, mitomycin; mitosper, mitotane; mitoxantrone clarithromycin). Specific second active compounds that can hydrochloride; mycophenolic acid; nocodazole; nogalamy be combined with compounds provided herein vary depend 60 cin; ormaplatin; OXisuran, paclitaxel, pegaspargase; peliomy ing on the specific indication to be treated, prevented or cin; pentamustine; peplomycin Sulfate; perfosfamide; pipo managed. broman; piposulfan, piroxantrone hydrochloride; In some embodiments, the second active agents include, plicamycin; plomestane; porfimer Sodium; porfiromycin; but are not limited to, anticoagulants, diuretics, cardiac gly prednimustine; procarbazine hydrochloride; puromycin; cosides, calcium channel blockers, vasodilators, prostacyclin 65 puromycin hydrochloride; pyrazofurin, riboprine; Safingol; analogues, endothelin antagonists, phosphodiesterase inhibi Safingol hydrochloride; Semustine; simtraZene; sparfosate tors (e.g., PDE V inhibitors), endopeptidase inhibitors, lipid Sodium; sparsomycin; Spirogermanium hydrochloride; spiro US 8,716,252 B2 19 20 mustine; spiroplatin; Streptonigrin, streptozocin, Sulofenur, factor; leukocyte alpha interferon; leuprolide+estrogen talisomycin; tecogalan Sodium; taxotere; tegafur, telox progesterone; leuprorelin; levamisole; liarozole; linear antrone hydrochloride; temoporfin, teniposide; teroxirone; polyamine analogue; lipophilic disaccharide peptide; lipo testolactone; thiamiprine; thioguanine; thiotepa, tiazofurin; philic platinum compounds; lissoclinamide 7: lobaplatin: tirapazamine; toremifene citrate; trestolone acetate; tricirib lombricine; lometrexol; lonidamine; losoxantrone; loxorib ine phosphate; trimetrexate; trimetrexate glucuronate; trip ine; lurtotecan; lutetium texaphyrin; lysofylline; lytic pep torelin; tubulozole hydrochloride: uracil mustard; uredepa; tides; maitansine; mannostatin A; marimastat; masoprocol; vapreotide; verteporfin; vinblastine sulfate; Vincristine sul maspin; matrilysin inhibitors; matrix metalloproteinase fate; Vindesine; Vindesine Sulfate; Vinepidine Sulfate; Vingly inhibitors; menogaril; merbarone; meterelin; methioninase; cinate sulfate; Vinleurosine sulfate; Vinorelbine tartrate; Vin 10 metoclopramide; MIF inhibitor; mifepristone; miltefosine; rosidine sulfate; Vinzolidine sulfate; Vorozole; Zeniplatin: Zinostatin; and Zorubicin hydrochloride. mirimostim; mitoguaZone; mitolactol; mitomycin analogues; Other second agents include, but are not limited to: 20-epi mitonafide; mitotoxin fibroblast growth factor-saporin; 1.25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone: mitoxantrone; mofarotene; molgramoStim; Erbitux, human aclarubicin; acylfulvene; adecypenol; adoZelesin; aldesleu 15 chorionic gonadotrophin, monophosphoryl lipid A+myobac kin; ALL-TK antagonists; altretamine; ambamustine; ami terium cell wall sk; mopidamol, mustard anticancer agent; doX, amifostine; aminolevulinic acid; amrubicin; amsacrine; my caperoxide B; mycobacterial cell wall extract; myriapor anagrelide; anastrozole; andrographolide; angiogenesis one; N-acetyldinaline; N-substituted benzamides; nafarelin; inhibitors; antagonist D; antagonist G, antarelix; anti-dorsal nagrestip; naloxone-pentazocine; napavin; naphterpin, nar izing morphogenetic protein-1, antiandrogen, prostatic car tograstim; nedaplatin; memorubicin; neridronic acid; niluta cinoma; antiestrogen; antineoplaston; antisense oligonucle mide; nisamycin; nitric oxide modulators; nitroxide antioxi otides; aphidicolin glycinate; apoptosis gene modulators; dant; nitrullyn; oblimersen (Genasense(R): apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA: O6-benzylguanine; octreotide; okicenone; oligonucleotides; arginine deaminase; asulacrine; atameStane; atrimustine; axi onapristone; ondansetron; ondansetron; oracin; oral cytokine nastatin 1; axinastatin 2; axinastatin 3; aZasetron; azatoxin; 25 inducer, ormaplatin: osaterone; oxaliplatin, oxaunomycin; aZatyrosine; baccatin III derivatives; balanol; batimastat; paclitaxel, paclitaxel analogues; paclitaxel derivatives; BCR/ABL antagonists; benzochlorins; benzoylstaurospo palauamine; palmitoylrhizoxin; pamidronic acid; panax rine; beta lactam derivatives; beta-alethine; betaclamycin B; ytriol; panomifene; parabactin; paZelliptine; pegaspargase; betulinic acid; bFGF inhibitor, bicalutamide; bisantrene; peldesine; pentosan polysulfate sodium; pentostatin: pentro bisaziridinylspermine; bisnafide; bistratene A: bizelesin; 30 Zole; perflubron; perfosfamide; perillyl alcohol; phenazino breflate; bropirimine; budotitane; buthionine sulfoximine; mycin; phenylacetate; phosphatase inhibitors; picibanil; pilo calcipotriol; calphostin C; camptothecin derivatives; capecit carpine hydrochloride; pirarubicin; piritrexim; placetin A; abine; carboxamide-amino-triazole; carboxyamidotriazole; placetin B; plasminogen activator inhibitor; platinum com CaRest M3; CARN 700; cartilage derived inhibitor; carze plex; platinum compounds; platinum-triamine complex; por lesin; casein kinase inhibitors (ICOS); castanospermine; 35 cecropin B; cetrorelix; chlorins; chloroquinoxaline Sulfona fimer Sodium; porfiromycin; prednisone; propyl bis-acri mide; cicaprost; cis-porphyrin, cladribine; clomifene ana done; prostaglandin J2, proteasome inhibitors; protein logues; clotrimazole; collismycin A; collismycin B; combre A-based immune modulator, protein kinase C inhibitor, pro tastatin A4, combretastatin analogue; conagenin: tein kinase C inhibitors, microalgal; protein tyrosine phos crambescidin 816; crisinatol; cryptophycin 8: cryptophycin A 40 phatase inhibitors; purine nucleoside phosphorylase inhibi derivatives; curacin A; cyclopentanthraquinones; cyclo tors; purpurins; pyrazoloacridine; pyridoxylated hemoglobin platam, cypemycin; cytarabine ocfosfate; cytolytic factor, polyoxyethylene conjugate; raf antagonists; raltitrexed: cytostatin: dacliximab; decitabine; dehydrodidemnin B; ramosetron; ras farnesyl protein transferase inhibitors; ras deslorelin; dexamethasone; dexifosfamide; dexraZOxane: inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhe deXVerapamil: diaziquone; didemnin B; didox; diethylnor 45 nium Re 186 etidronate; rhizoxin: ribozymes: RH retinamide: spermine; dihydro-5-azacytidine; dihydrotaxol. 9-; dioxamy rohitukine; romurtide; roquinimex: rubiginone B1; ruboxyl, cin; diphenyl spiromustine; docetaxel, docosanol; dolas Safingol; Saintopin; SarCNU; sarcophytol A. SargramoStim; etron: doxifluridine: doxorubicin; droloxifene; dronabinol; Sdi 1 mimetics; semustine; senescence derived inhibitor 1; duocarmycin SA; ebSelen; ecomustine, edelfosine; edrecolo sense oligonucleotides; signal transduction inhibitors; siZo mab; eflornithine; elemene; emitefur; epirubicin; epristeride; 50 furan; Sobuzoxane; sodium borocaptate; sodium phenylac estramustine analogue; estrogen agonists; estrogen antago etate; solverol; somatomedin binding protein; Sonermin; nists; etanidazole; etoposide phosphate; exemestane; fadro sparfosic acid; spicamycin D; spiromustine; splenopentin; Zole; fazarabine; fenretinide; filgrastim; finasteride; fla spongistatin 1; squalamine; stipiamide; stromelysin inhibi Vopiridol; flezelastine; fluasterone; fludarabine; tors; Sulfinosine; Superactive vasoactive intestinal peptide fluorodaunorunicin hydrochloride; forfenimex; formestane: 55 antagonist; Suradista; Suramin; Swainsonine; tallimustine; fostriecin, fotemustine; gadolinium texaphyrin; gallium tamoxifen methiodide; tauromustine; tazarotene; tecogalan nitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcit Sodium, tegafur, tellurapyrylium; telomerase inhibitors; abine; glutathione inhibitors; hepsulfam; heregulin; hexam temoporfin, teniposide; tetrachlorodecaoxide; tetraZomine; ethylene bisacetamide; hypericin; ibandronic acid; idarubi thaliblastine; thiocoraline; thrombopoietin; thrombopoietin cin; idoxifene; idramantone; ilmofosine; illomastat; imatinib 60 mimetic; thymalfasin; thymopoietin receptor agonist; thy (Gleevec.R.), imiquimod; immunostimulant peptides; insulin motrinan; thyroid stimulating hormone; tin ethyl etiopurpu like growth factor-1 receptor inhibitor; interferon agonists; rin; tirapazamine; titanocene bichloride; topsentin; interferons; interleukins; iobenguane; iododoxorubicin; toremifene; translation inhibitors; tretinoin; triacetyluridine: ipomeanol, 4-, iroplact; irsogladine; isobengaZole; isohomo triciribine; trimetrexate; triptorelin; tropisetron; turosteride; halicondrin B; itasetron; jasplakinolide; kahalalide F; lamel 65 tyrosine kinase inhibitors; tyrphostins; UBC inhibitors: ube larin-N triacetate; lanreotide; leinamycin; lenograstim; len nimex: urogenital sinus-derived growth inhibitory factor; tinan Sulfate; leptolstatin; letrozole; leukemia inhibiting urokinase receptor antagonists; vapreotide; variolin B; US 8,716,252 B2 21 22 velaresol; veramine; verdins; verteporfin; vinorelbine; vinx purly tin, an angiostatic steroid, rhuFab, interferon-2C. pen altine; vitaxin; Vorozole; Zanoterone; Zeniplatin: Zilascorb: toxifylline, tin etiopurpurin, motexafin, lucentis, lutetium, and Zinostatin stimalamer. 9-fluoro-1 1,21-dihydroxy-16,17-1-methylethylidinebis In some embodiments, the active agents include, but are (oxy)pregna-1,4-diene-320-dione, latanoprost (see U.S. Pat. not limited to, 2-methoxyestradiol, telomestatin, inducers of 5 No. 6,225.348), tetracycline and its derivatives, rifamycin apoptosis in multiple myeloma cells (such as, for example, and its derivatives, macrollides, metronidazole (U.S. Pat. Nos. TRAIL), statins, Semaxanib, cyclosporin, etanercept, doxy 6.218.369 and 6,015,803), genistein, genistin, 6'-O-Mal cycline, bortezomib, oblimersen (Genasense(R), remicade, genistin, 6'-O-Ac genistin, daidzein, daidzin, 6'-O-Mal docetaxel, celecoxib, melphalan, dexamethasone (Dec daidzin, 6'-O-Ac daidzin, glycitein, glycitin, 6'-O-Mal gly adroe), Steroids, gemcitabine, cisplatinum, temozolomide, 10 citin, biochanin A, formononetin (U.S. Pat. No. 6,001,368), etoposide, cyclophosphamide, temodar, carboplatin, procar triamcinolone acetomide, dexamethasone (U.S. Pat. No. bazine, gliadel, tamoxifen, topotecan, methotrexate, Arisa R, 5,770.589), thalidomide, glutathione (U.S. Pat. No. 5,632, taxol, taxotere, fluorouracil, leucovorin, irinotecan, Xeloda, 984), basic fibroblast growth factor (bFGF), transforming CPT-11, interferon alpha, pegylated interferon alpha (e.g., growth factor b (TGF-b), brain-derived neurotrophic factor PEG INTRON-A), capecitabine, cisplatin, thiotepa, fludara 15 (BDNF), plasminogen activator factor type 2 (PAI-2), bine, carboplatin, liposomal daunorubicin, cytarabine, dox EYE101 (Eyetech Pharmaceuticals), LY333531 (Eli Lilly), etaxol, pacilitaxel, vinblastine, IL-2, GM-CSF, dacarbazine, Miravant, and RETISERT implant (Bausch & Lomb). All of Vinorelbine, Zoledronic acid, palmitronate, biaxin, buSul the references cited herein are incorporated in their entireties phan, prednisone, bisphosphonate, arsenic trioxide, Vincris by reference. tine, doxorubicin (DOXil(R), paclitaxel, ganciclovir, adriamy In some embodiments, the second active agents include, cin, estramustine Sodium phosphate (EmcytR), Sulindac, but are not limited to, anthracycline, platinum, alkylating etoposide, and various cytotoxic and/or cytostatic agents. agent, oblimersen (Genasense(R), cisplatinum, cyclophos In some embodiments, examples of specific second agents phamide, temodar, carboplatin, procarbazine, gliadel, tamox according to the indications to be treated, prevented, or man ifen, topotecan, methotrexate, taxotere, irinotecan, capecit aged can be found in the following references, all of which are 25 abine, cisplatin, thiotepa, fludarabine, carboplatin, liposomal incorporated herein in their entireties: U.S. Pat. Nos. 6,281, daunorubicin, cytarabine, doxetaXol, pacilitaxel, vinblastine, 230 and 5,635,517; and U.S. publication nos. 2004/0220144, IL-2, GM-CSF, dacarbazine, vinorelbine, Zoledronic acid, 2004/0190609, 2004/0087546, 2005/0203142, 2004/ palmitronate, biaxin, buSulphan, prednisone, bisphospho 0091455, 2005/0100529, 2005/0214328, 2005/0239842, nate, arsenic trioxide, Vincristine, doxorubicin (Doxil R), 2006/0154880, 2006/0188475, 2006/0122228, and 2005/ 30 paclitaxel, ganciclovir, adriamycin, bleomycin, hyalu O143344 ronidase, mitomycin C, mepacrine, thiotepa, tetracycline and In some embodiments, the second active agents include, gemcitabine. but are not limited to, antidepressants, anticonvulsants, anti In some embodiments, the second active agents include, hypertensives, anxiolytics, calcium channel blockers, muscle but are not limited to, chloroquine, quinine, quinidine, relaxants, non-narcotic analgesics, opioid analgesics, anti 35 pyrimethamine, Sulfadiazine, doxycycline, clindamycin, inflammatories, cox-2 inhibitors, immunomodulatory agents, mefloquine, halofantrine, primaquine, hydroxychloroquine, alpha-adrenergic receptor agonists or antagonists, immuno proguanil, atovaquone, azithromycin, Suramin, pentamidine, Suppressive agents, corticosteroids, hyperbaric oxygen, ket melarsoprol, nifurtimox, benznidazole, amphotericin B, pen amine, other anesthetic agents, NMDA antagonists, NSAIDs tavalent antimony compounds (e.g., Sodium Stiboglucur and other therapeutics found, for example, in the Physician's 40 onate), interfereon gamma, itraconazole, a combination of Desk Reference 2003. Specific examples include, but are not dead promastigotes and BCG, leucovorin, corticosteroids, limited to, salicylic acid acetate (AspirinR), celecoxib (Cele Sulfonamide, spiramycin, IgG (serology), trimethoprim, and brexR), Enbrel(R), ketamine, ibuprofen, gabapentin (Neuron Sulfamethoxazole. tin R), phenyloin (DilantinR), carbamazepine (Tegretol R), In Some embodiments, the second active agents, but are not oxcarbazepine (Trileptal(R), valproic acid (DepakeneR), 45 limited to: antibiotics (therapeutic or prophylactic) Such as, morphine Sulfate, hydromorphone, prednisone, griseofulvin, but not limited to, amplicillin, tetracycline, penicillin, cepha penthonium, alendronate, dyphenhydramide, guanethidine, losporins, streptomycin, kanamycin, and erythromycin; anti ketorolac (Aculae), thyrocalcitonin, dimethylsulfoxide virals such as, but not limited to, amantadine, rimantadine, (DMSO), clonidine (Catapress(R), bretylium, ketanserin, acyclovir, and ribavirin; immunoglobulin; plasma; immuno reserpine, droperidol, atropine, phentolamine, bupivacaine, 50 logic enhancing drugs such as, but not limited to, levami sole lidocaine, acetaminophen, nortriptyline (PamelorR), ami and isoprinosine; biologics such as, but not limited to, gam triptyline (Elavil(R), imipramine (TofranilR), doxepin (Sine maglobulin, transfer factor, interleukins, and interferons; hor quan R), clomipramine (AnafranilR), fluoxetine (ProzacR), mones such as, but not limited to, thymic; and other immu sertraline (Zoloft(R), naproxen, nefazodone (SerZone(R), ven nologic agents such as, but not limited to, B cell stimulators lafaxine (EffexorR), trazodone (DesyrelR), bupropion (Well 55 (e.g., BAFF/BlyS), cytokines (e.g., IL-2, IL-4, and IL-5), butrinR), mexiletine, nifedipine, propranolol, tramadol, lam growth factors (e.g., TGF-C.), antibodies (e.g., anti-CD40 and otrigine, vioXX, Ziconotide, ketamine, dextromethorphan, IgM), oligonucleotides containing unmethylated CpG benzodiazepines, baclofen, tizanidine and phenoxyben motifs, and vaccines (e.g., viral and tumor peptide vaccines). Zamine. In some embodiments, the second active agents include, In some embodiments, the second active agents include, 60 but are not limited to: opioids; a dopamine agonist orantago but are not limited to, a steroid, a light sensitizer, an integrin, nist, such as, but not limited to, Levodopa, L-DOPA, cocaine, an antioxidant, an interferon, a Xanthine derivative, a growth C.-methyl-tyrosine, reserpine, tetrabenazine, benzotropine, hormone, a neutrotrophic factor, a regulator of neovascular pargyline, fenodolpam mesylate, cabergoline, pramipexole ization, an anti-VEGF antibody, a prostaglandin, an antibi dihydrochloride, ropinorole, amantadine hydrochloride, sel otic, a phytoestrogen, an anti-inflammatory compound or an 65 egiline hydrochloride, carbidopa, pergolide mesylate, antiangiogenesis compound, or a combination thereof. Spe Sinemet CR, and Symmetrel; a MAO inhibitor, such as, but cific examples include, but are not limited to, Verteporfin, not limited to, iproniazid, clorgyline, phenelZine and isocar US 8,716,252 B2 23 24 boxazid; a COMT inhibitor, such as, but not limited to, tol In some embodiments, the second active agents include, capone and entacapone; a cholinesterase inhibitor, Such as, but are not limited to, a tricyclic antidepressantagent, a selec but not limited to, physostigmine Saliclate, physostigmine tive serotonin reuptake inhibitor, an antiepileptic agent (gaba Sulfate, physostigmine bromide, meostigmine bromide, neo pentin, pregabalin, carbamazepine, oXcarbazepine, levitirac Stigmine methylsulfate, ambenonim chloride, edrophonium etam, topiramate), an antiaryhthmic agent, a sodium channel chloride, tacrine, pralidoxime chloride, obidoxime chloride, blocking agent, a selective inflammatory mediator inhibitor, trimedoxime bromide, diacetyl monoxim, endrophonium, an opioid agent, a second immunomodulatory compound, a pyridostigmine, and demecarium; an anti-inflammatory combination agent, and other known or conventional agents agent, Such as, but not limited to, naproxen Sodium, used in sleep therapy. Specific examples include, but are not diclofenac sodium, diclofenac potassium, celecoxib, Sulin 10 limited to, Neurontin, oxycontin, morphine, topiramate, dac, oxaprozin, diflunisal, etodolac, meloxicam, ibuprofen, amitryptiline, nortryptiline, carbamazepine, Levodopa, ketoprofen, nabumetone, refecoxib, methotrexate, lefluno L-DOPA, cocaine, C.-methyl-tyrosine, reserpine, tetrabena mide, Sulfasalazine, gold salts, Rho-D Immune Globulin, Zine, benzotropine, pargyline, fenodolpam mesylate, caber mycophenylate mofetil, cyclosporine, azathioprine, tacroli goline, pramipexole dihydrochloride, ropinorole, amantadine mus, basiliximab, daclizumab, Salicylic acid, acetylsalicylic 15 hydrochloride, selegiline hydrochloride, carbidopa, per acid, methyl salicylate, diflunisal, Salsalate, olsalazine, Sul golide mesylate, Sinemet CR, Symmetrel, iproniazid, clor fasalazine, acetaminophen, indomethacin, Sulindac, mefe gyline, phenelZine, isocarboxazid, tolcapone, entacapone, namic acid, meclofenamate Sodium, tolmetin, ketorolac, physostigmine saliclate, physostigmine Sulfate, physostig dichlofenac, flurbinprofen, oxaprozin, piroxicam, meloxi mine bromide, meostigmine bromide, neostigmine methyl cam, ampiroXicam, droxicam, pivoxicam, tenoxicam, phe Sulfate, ambenonim chloride, edrophonium chloride, tacrine, nylbutaZone, oxyphenbutaZone, antipyrine, aminopyrine, pralidoxime chloride, obidoxime chloride, trimedoxime bro apaZone, Zileuton, aurothioglucose, gold sodium thiomalate, mide, diacetyl monoxim, endrophonium, pyridostigmine, auranofin, methotrexate, colchicine, allopurinol, probenecid, demecarium, naproxen Sodium, diclofenac sodium, Sulfinpyrazone and benzbromarone or betamethasone and diclofenac potassium, celecoxib, Sulindac, oxaprozin, other glucocorticoids; and an antiemetic agent, such as, but 25 diflunisal, etodolac, meloxicam, ibuprofen, ketoprofen, nabu not limited to, metoclopromide, domperidone, prochlorpera metone, refecoxib, methotrexate, leflunomide, Sulfasalazine, Zine, promethazine, chlorpromazine, trimethobenzamide, gold salts, RHo-D Immune Globulin, mycophenylate mofetil, ondansetron, granisetron, hydroxy Zine, acetyleucine mono cyclosporine, azathioprine, tacrolimus, basiliximab, dacli ethanolamine, lalizapride, aZasetron, benzquinamide, biet Zumab, Salicylic acid, acetylsalicylic acid, methyl salicylate, anautine, bromopride, buclizine, clebopride, cyclizine, 30 diflunisal, Salsalate, olsalazine, SulfaSalazine, acetami dimenhydrinate, diphenidol, dolasetron, meclizine, methal nophen, indomethacin, Sulindac, mefenamic acid, meclofe latal, metopimazine, nabilone, oxyperndyl, pipamazine, sco namate sodium, tolmetin, ketorolac, dichlofenac, flurbinpro polamine, Sulpiride, tetrahydrocannabinol, thiethylperazine, fen, Oxaprozin, piroxicam, meloxicam, ampiroxicam, thioproperazine, tropisetron, and a mixture thereof. droxicam, pivoxicam, tenoxicam, phenylbutaZone, oxyphen In some embodiments, the second active agents include, 35 butaZone, antipyrine, aminopyrine, apaZone, Zileuton, but are not limited to, immunomodulatory agents, immuno aurothioglucose, gold sodium thiomalate, auranofin, meth Suppressive agents, antihypertensives, anticonvulsants, otrexate, colchicine, allopurinol, probenecid, Sulfinpyrazone, fibrinolytic agents, antiplatelet agents, antipsychotics, anti benzbromarone, betamethasone and other glucocorticoids, depressants, benzodiazepines, buspirone, amantadine, and metoclopromide, domperidone, prochlorperazine, promet other known or conventional agents used in patients with 40 hazine, chlorpromazine, trimethobenzamide, ondansetron, CNS injury/damage and related syndromes. Specific granisetron, hydroxy Zine, acetylleucine monoethanolamine, examples include, but are not limited to: Steroids (e.g., glu alizapride, aZasetron, benzquinamide, bietanautine, bro cocorticoids, such as, but not limited to, methylprednisolone, mopride, buclizine, clebopride, cyclizine, dimenhydrinate, dexamethasone and betamethasone); an anti-inflammatory diphenidol, dolasetron, meclizine, methallatal, metopi agent, including, but not limited to, naproxen Sodium, 45 mazine, nabilone, oxyperndyl, pipamazine, Scopolamine, diclofenac sodium, diclofenac potassium, celecoxib, Sulin Sulpiride, tetrahydrocannabinol, thiethylperazine, thioprop dac, oxaprozin, diflunisal, etodolac, meloxicam, ibuprofen, erazine, tropisetron, and a mixture thereof. ketoprofen, nabumetone, refecoxib, methotrexate, lefluno In some embodiments, the second active agents include, mide, sulfasalazine, gold salts, RHo-D Immune Globulin, but are not limited to: interleukins, such as IL-2 (including mycophenylate mofetil, cyclosporine, azathioprine, tacroli 50 recombinant IL-II (rIL2) and canarypox IL-2), IL-10, mus, basiliximab, daclizumab, Salicylic acid, acetylsalicylic IL-12, and IL-18; interferons, such as interferon alfa-2a, acid, methyl salicylate, diflunisal, Salsalate, olsalazine, Sul interferon alfa-2b, interferon alfa-nl, interferon alfa-n3, inter fasalazine, acetaminophen, indomethacin, Sulindac, mefe feron beta-Ia, and interferon gamma-Ib; and G-CSF; hydrox namic acid, meclofenamate Sodium, tolmetin, ketorolac, yurea; butyrates or butyrate derivatives; nitrous oxide: dichlofenac, flurbinprofen, oxaprozin, piroxicam, meloxi 55 hydroxy urea; HEMOXINTTM (NIPRISANTM; see U.S. Pat. cam, ampiroXicam, droxicam, pivoxicam, tenoxicam, phe No. 5,800.819); Gardos channel antagonists such as clotri nylbutaZone, oxyphenbutaZone, antipyrine, aminopyrine, mazole and triaryl methane derivatives; Deferoxamine; pro apaZone, Zileuton, aurothioglucose, gold sodium thiomalate, tein C; and transfusions of blood, or of a blood substitute such auranofin, methotrexate, colchicine, allopurinol, probenecid, as HemospanTM or HemospanTM PS (Sangart). Sulfinpyrazone and benzbromarone; a cAMP analog includ 60 Administration of a compound provided herein, or a phar ing, but not limited to, db-cAMP; an agent comprising a maceutically acceptable salt, Solvate, clathrate, Stereoisomer methylphenidate drug, which comprises 1-threo-meth or prodrug thereof, and the second active agents to a patient ylphenidate, d-threo-methylphenidate, dl-threo-meth can occur simultaneously or sequentially by the same or ylphenidate, 1-erythro-methylphenidate, d-erythro-meth different routes of administration. The suitability of a particu ylphenidate, dl-erythro-methylphenidate, and a mixture 65 lar route of administration employed for a particular active thereof, and a diuretic agent such as, but not limited to, agent will depend on the active agent itself (e.g., whether it mannitol, furosemide, glycerol, and urea. can be administered orally without decomposing prior to US 8,716,252 B2 25 26 entering the blood stream) and the disease being treated. One about one to about 24 cycles, from about two to about 16 of administration for compounds provided herein is oral. cycles, or from about four to about three cycles. Routes of administration for the second active agents or 4.5 Pharmaceutical Compositions and Dosage Forms ingredients are knownto those of ordinary skill in the art. See, Pharmaceutical compositions can be used in the prepara e.g., Physicians' Desk Reference (60" ed., 2006). tion of individual, single unit dosage forms. Pharmaceutical In one embodiment, the second active agent is adminis compositions and dosage forms provided herein comprise a tered intravenously or Subcutaneously and once or twice daily compound provided herein, or a pharmaceutically acceptable in an amount of from about 1 to about 1000 mg, from about 5 salt, Solvate, Stereoisomer, clathrate, or prodrug thereof. Phar to about 500 mg, from about 10 to about 350 mg. or from maceutical compositions and dosage forms can further com about 50 to about 200 mg. The specific amount of the second 10 prise one or more excipients. active agent will depend on the specific agent used, the type of Pharmaceutical compositions and dosage forms provided disease being treated or managed, the severity and stage of herein can also comprise one or more additional active ingre disease, and the amount(s) of compounds provided herein and dients. Examples of optional second, or additional, active any optional additional active agents concurrently adminis 15 ingredients are disclosed in Section 4.3, above. tered to the patient. Single unit dosage forms provided herein are suitable for As discussed elsewhere herein, also encompassed is a oral, mucosal (e.g., nasal, Sublingual, vaginal, buccal, or rec method of reducing, treating and/or preventing adverse or tal), parenteral (e.g., Subcutaneous, intravenous, bolus injec undesired effects associated with conventional therapy tion, intramuscular, or intraarterial), topical (e.g., eye drops or including, but not limited to, Surgery, chemotherapy, radia other ophthalmic preparations), transdermal or transcutane tion therapy, hormonal therapy, biological therapy and immu ous administration to a patient. Examples of dosage forms notherapy. Compounds provided herein and other active include, but are not limited to: tablets; caplets; capsules. Such ingredients can be administered to a patient prior to, during, as Soft elastic gelatin capsules; cachets; troches; lozenges; or after the occurrence of the adverse effect associated with dispersions; Suppositories; powders; aerosols (e.g., nasal conventional therapy. 25 sprays or inhalers); gels; liquid dosage forms suitable for oral 4.4 Cycling Therapy or mucosal administration to a patient, including Suspensions In certain embodiments, the prophylactic or therapeutic (e.g., aqueous or non-aqueous liquid Suspensions, oil-in-wa agents provided herein are cyclically administered to a ter emulsions, or a water-in-oil liquid emulsions), Solutions, patient. Cycling therapy involves the administration of an and elixirs, liquid dosage forms suitable for parenteral admin active agent for a period of time, followed by a rest (i.e., 30 istration to a patient; eye drops or other ophthalmic prepara discontinuation of the administration) for a period of time, tions Suitable for topical administration; and sterile Solids and repeating this sequential administration. Cycling therapy (e.g., crystalline or amorphous solids) that can be reconsti can reduce the development of resistance to one or more of the tuted to provide liquid dosage forms suitable for parenteral therapies, avoid or reduce the side effects of one of the thera administration to a patient. pies, and/or improve the efficacy of the treatment. 35 The composition, shape, and type of dosage forms will Consequently, in one embodiment, a compound provided typically vary depending on their use. For example, a dosage herein is administered daily in a single or divided doses in a form used in the acute treatment of a disease may contain four to six week cycle with a rest period of about a week or larger amounts of one or more of the active ingredients it two weeks. Cycling therapy further allows the frequency, comprises thana dosage form used in the chronic treatment of number, and length of dosing cycles to be increased. Thus, 40 the same disease. Similarly, a parenteral dosage form may another embodiment encompasses the administration of a contain Smaller amounts of one or more of the active ingre compound provided herein for more cycles than are typical dients it comprises than an oral dosage form used to treat the when it is administered alone. In yet another embodiment, a same disease. These and other ways in which specific dosage compound provided herein is administered for a greater num forms are used will vary from one another will be readily ber of cycles than would typically cause dose-limiting toxic 45 apparent to those skilled in the art. See, e.g., Remington's ity in a patient to whom a second active ingredient is not also Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton being administered. Pa. (1990). In one embodiment, a compound provided herein is admin In one embodiment, pharmaceutical compositions and istered daily and continuously for three or four weeks at a dosage forms comprise one or more excipients. Suitable dose of from about 0.1 mg to about 500 mg per day, followed 50 excipients are well known to those skilled in the art of phar by a rest of one or two weeks. In other embodiments, the dose macy, and non-limiting examples of Suitable excipients are can be from about 1 mg to about 300 mg, from about 0.1 mg provided herein. Whether a particular excipient is suitable for to about 150 mg, from about 1 mg to about 200 mg, from incorporation into a pharmaceutical composition or dosage about 10 mg to about 100 mg, from about 0.1 mg to about 50 form depends on a variety of factors well known in the art mg, from about 1 mg to about 50 mg, from about 10 mg to 55 including, but not limited to, the way in which the dosage about 50 mg, from about 20 mg to about 30 mg. or from about form will be administered to a patient. For example, oral 1 mg to about 20 mg, followed by a rest. dosage forms such as tablets may contain excipients not In one embodiment, a compound provided herein and a suited for use in parenteral dosage forms. The suitability of a second active ingredient are administered orally, with admin particular excipient may also depend on the specific active istration of the compound provided herein occurring 30 to 60 60 ingredients in the dosage form. For example, the decomposi minutes prior to the second active ingredient, during a cycle tion of Some active ingredients may be accelerated by some of four to six weeks. In another embodiment, the combination excipients such as lactose, or when exposed to water. Active of a compound provided herein and a second active ingredient ingredients that comprise primary or secondary amines are is administered by intravenous infusion over about 90 min particularly susceptible to Such accelerated decomposition. utes every cycle. 65 Consequently, provided are pharmaceutical compositions Typically, the number of cycles during which the combi and dosage forms that contain little, if any, lactose other nation treatment is administered to a patient will be from mono- or di-saccharides. As used herein, the term “lactose US 8,716,252 B2 27 28 free” means that the amount of lactose present, if any, is 4.5.1 Oral Dosage Forms insufficient to Substantially increase the degradation rate of an Pharmaceutical compositions that are suitable for oral active ingredient. administration can be provided as discrete dosage forms. Such Lactose-free compositions can comprise excipients that as, but not limited to, tablets (e.g., chewable tablets), caplets, are well known in the art and are listed, for example, in the capsules, and liquids (e.g., flavored syrups). Such dosage U.S. Pharmacopeia (USP) 25-NF20 (2002). In general, lac forms contain predetermined amounts of active ingredients, tose-free compositions comprise active ingredients, a binder/ and may be prepared by methods of pharmacy well known to filler, and a lubricant in pharmaceutically compatible and those skilled in the art. Seegenerally, Remington's Pharma pharmaceutically acceptable amounts. In one embodiment, ceutical Sciences, 18th ed., Mack Publishing, Easton Pa. lactose-free dosage forms comprise active ingredients, 10 (1990). microcrystalline cellulose, pre-gelatinized starch, and mag Oral dosage forms provided herein are prepared by com nesium Stearate. bining the active ingredients in an intimate admixture with at Also provided are anhydrous pharmaceutical compositions least one excipient according to conventional pharmaceutical and dosage forms comprising active ingredients, since water compounding techniques. Excipients can take a wide variety can facilitate the degradation of some compounds. For 15 of forms depending on the form of preparation desired for example, the addition of water (e.g., 5%) is widely accepted administration. For example, excipients suitable for use in in the pharmaceutical arts as a means of simulating long-term oral liquid or aerosol dosage forms include, but are not limited storage in order to determine characteristics such as shelf-life to, water, glycols, oils, alcohols, flavoring agents, preserva or the stability of formulations over time. See, e.g., Jens T. tives, and coloring agents. Examples of excipients suitable for Carstensen, Drug Stability: Principles & Practice, 2d. Ed., use in Solid oral dosage forms (e.g., powders, tablets, cap Marcel Dekker, NY, N.Y., 1995, pp. 379-80. In effect, water Sules, and caplets) include, but are not limited to, starches, and heat accelerate the decomposition of Some compounds. Sugars, micro-crystalline cellulose, diluents, granulating Thus, the effect of water on a formulation can be of great agents, lubricants, binders, and disintegrating agents. significance since moisture and/or humidity are commonly In one embodiment, oral dosage forms are tablets or cap encountered during manufacture, handling, packaging, Stor 25 Sules, in which case Solid excipients are employed. In another age, shipment, and use of formulations. embodiment, tablets can be coated by standard aqueous or Anhydrous pharmaceutical compositions and dosage nonaqueous techniques. Such dosage forms can be prepared forms can be prepared using anhydrous or low moisture con by any of the methods of pharmacy. In general, pharmaceu taining ingredients and low moisture or low humidity condi tical compositions and dosage forms are prepared by uni tions. Pharmaceutical compositions and dosage forms that 30 formly and intimately admixing the active ingredients with comprise lactose and at least one active ingredient that com liquid carriers, finely divided solid carriers, or both, and then prises a primary or secondary amine are preferably anhydrous shaping the product into the desired presentation if necessary. if Substantial contact with moisture and/or humidity during For example, a tablet can be prepared by compression or manufacturing, packaging, and/or storage is expected. molding. Compressed tablets can be prepared by compress An anhydrous pharmaceutical composition should be pre 35 ing in a suitable machine the active ingredients in a free pared and stored Such that its anhydrous nature is maintained. flowing form such as powder or granules, optionally mixed Accordingly, anhydrous compositions are, in one embodi with an excipient. Molded tablets can be made by molding in ment, packaged using materials known to prevent exposure to a suitable machine a mixture of the powdered compound water such that they can be included in suitable formulary moistened with an inert liquid diluent. kits. Examples of Suitable packaging include, but are not 40 Examples of excipients that can be used in oral dosage limited to, hermetically sealed foils, plastics, unit dose con forms provided herein include, but are not limited to, binders, tainers (e.g., vials), blister packs, and strip packs. fillers, disintegrants, and lubricants. Binders suitable for use Also provided are pharmaceutical compositions and dos in pharmaceutical compositions and dosage forms include, age forms that comprise one or more compounds that reduce but are not limited to, corn starch, potato starch, or other the rate by which an active ingredient will decompose. Such 45 starches, gelatin, natural and synthetic gums such as acacia, compounds, which are referred to herein as “stabilizers.” Sodium alginate, alginic acid, other alginates, powdered include, but are not limited to, antioxidants such as ascorbic tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl acid, pH buffers, or salt buffers. cellulose, cellulose acetate, carboxymethyl cellulose cal Like the amounts and types of excipients, the amounts and cium, Sodium carboxymethyl cellulose), polyvinyl pyrroli specific types of active ingredients in a dosage form may 50 done, methyl cellulose, pre-gelatinized Starch, hydroxypro differ depending on factors such as, but not limited to, the pyl methyl cellulose, (e.g., Nos. 2208, 2906, 2910), route by which it is to be administered to patients. In one microcrystalline cellulose, and mixtures thereof. embodiment, dosage forms comprise a compound provided Suitable forms of microcrystalline cellulose include, but herein in an amount of from about 0.10 to about 500 mg. In are not limited to, the materials sold as AVICEL-PH-101, other embodiments, dosage forms comprise a compound pro 55 AVICEL-PH-103 AVICELRC-581, AVICEL-PH-105 (avail vided herein in an amount of about 0.1, 1, 2, 5, 7.5, 10, 12.5, able from FMC Corporation, American Viscose Division, 15, 17.5, 20, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. An or 500 mg. specific binder is a mixture of microcrystalline cellulose and In other embodiments, dosage forms comprise the second sodium carboxymethyl cellulose sold as AVICEL RC-581. active ingredient in an amount of 1 to about 1000 mg, from 60 Suitable anhydrous or low moisture excipients or additives about 5 to about 500 mg, from about 10 to about 350 mg. or include AVICEL-PH-103TM and Starch 1500 LM. from about 50 to about 200 mg. Of course, the specific Examples of fillers suitable for use in the pharmaceutical amount of the second active agent will depend on the specific compositions and dosage forms provided herein include, but agent used, the diseases or disorders being treated or man are not limited to, talc, calcium carbonate (e.g., granules or aged, and the amount(s) of a compound provided herein, and 65 powder), microcrystalline cellulose, powdered cellulose, any optional additional active agents concurrently adminis dextrates, kaolin, mannitol, silicic acid, Sorbitol, starch, pre tered to the patient. gelatinized starch, and mixtures thereof. The binder or filler in US 8,716,252 B2 29 30 pharmaceutical compositions is, in one embodiment, present In one embodiment, controlled-release pharmaceutical in from about 50 to about 99 weight percent of the pharma products improve drug therapy over that achieved by their ceutical composition or dosage form. non-controlled counterparts. In another embodiment, the use Disintegrants may be used in the compositions to provide of a controlled-release preparation in medical treatment is tablets that disintegrate when exposed to an aqueous environ characterized by a minimum of drug Substance being ment. Tablets that contain too much disintegrant may disin employed to cure or control the condition in a minimum tegrate in storage, while those that contain too little may not amount of time. Advantages of controlled-release formula disintegrate at a desired rate or under the desired conditions. tions include extended activity of the drug, reduced dosage Thus, a Sufficient amount of disintegrant that is neither too frequency, and increased patient compliance. In addition, 10 controlled-release formulations can be used to affect the time much nor too little to detrimentally alter the release of the ofonset of action or other characteristics, such as blood levels active ingredients may be used to form Solid oral dosage of the drug, and can thus affect the occurrence of side (e.g., forms. The amount of disintegrant used varies based upon the adverse) effects. type of formulation, and is readily discernible to those of In another embodiment, the controlled-release formula ordinary skill in the art. In one embodiment, pharmaceutical 15 tions are designed to initially release an amount of drug compositions comprise from about 0.5 to about 15 weight (active ingredient) that promptly produces the desired thera percent of disintegrant, or from about 1 to about 5 weight peutic or prophylactic effect, and gradually and continually percent of disintegrant. release of other amounts of drug to maintain this level of Disintegrants that can be used in pharmaceutical compo therapeutic or prophylactic effect over an extended period of sitions and dosage forms include, but are not limited to, time. In one embodiment, in order to maintain a constant level agar-agar, alginic acid, calcium carbonate, microcrystalline of drug in the body, the drug can be released from the dosage cellulose, croScarmellose Sodium, crospovidone, polacrilin form at a rate that will replace the amount of drug being potassium, Sodium starch glycolate, potato or tapioca starch, metabolized and excreted from the body. Controlled-release other starches, pre-gelatinized starch, other starches, clays, of an active ingredient can be stimulated by various condi otheralgins, other celluloses, gums, and mixtures thereof. 25 tions including, but not limited to, pH, temperature, enzymes, Lubricants that can be used in pharmaceutical composi water, or other physiological conditions or compounds. tions and dosage forms include, but are not limited to, calcium 4.5.3 Parenteral Dosage Forms Stearate, magnesium Stearate, mineral oil, light mineral oil, Parenteral dosage forms can be administered to patients by glycerin, Sorbitol, mannitol, polyethylene glycol, other gly various routes including, but not limited to, Subcutaneous, cols, Stearic acid, sodium lauryl Sulfate, talc, hydrogenated 30 intravenous (including bolus injection), intramuscular, and Vegetable oil (e.g., peanut oil, cottonseed oil, Sunflower oil, intraarterial. In some embodiments, administration of a sesame oil, olive oil, corn oil, and soybean oil), Zinc stearate, parenteral dosage form bypasses patients natural defenses ethyl oleate, ethyl laureate, agar, and mixtures thereof. Addi against contaminants, and thus, in these embodiments, tional lubricants include, for example, a Syloid silica gel parenteral dosage forms are sterile or capable of being ster (AEROSIL200, manufactured by W.R. Grace Co. of Balti 35 ilized prior to administration to a patient. Examples of more, Md.), a coagulated aerosol of synthetic silica (marketed parenteral dosage forms include, but are not limited to, solu by Degussa Co. of Plano, Tex.), CAB-O-SIL (a pyrogenic tions ready for injection, dry products ready to be dissolved or silicon dioxide product sold by Cabot Co. of Boston, Mass.), Suspended in a pharmaceutically acceptable vehicle for injec and mixtures thereof. Ifused at all, lubricants may be used in tion, Suspensions ready for injection, and emulsions. an amount of less than about 1 weight percent of the pharma 40 Suitable vehicles that can be used to provide parenteral ceutical compositions or dosage forms into which they are dosage forms are well known to those skilled in the art. incorporated. Examples include, but are not limited to: Water for Injection In one embodiment, a solid oral dosage form comprises a USP; aqueous vehicles such as, but not limited to, Sodium compound provided herein, anhydrous lactose, microcrystal Chloride Injection, Ringer's Injection, Dextrose Injection, line cellulose, polyvinylpyrrolidone, Stearic acid, colloidal 45 Dextrose and Sodium Chloride Injection, and Lactated Ring anhydrous silica, and gelatin. er's Injection; water-miscible vehicles such as, but not limited 4.5.2 Controlled Release Dosage Forms to, ethyl alcohol, polyethylene glycol, and polypropylene Active ingredients provided herein can be administered by glycol; and non-aqueous vehicles such as, but not limited to, controlled release means or by delivery devices that are well corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, known to those of ordinary skill in the art. Examples include, 50 isopropyl myristate, and benzyl benzoate. but are not limited to, those described in U.S. Pat. Nos. 3,845, Compounds that increase the solubility of one or more of 770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719, 5,674, the active ingredients disclosed herein can also be incorpo 533, 5,059,595, 5,591,767, 5,120,548, 5,073,543, 5,639,476, rated into the parenteral dosage forms. For example, cyclo 5,354,556, and 5,733,566, each of which is incorporated dextrin and its derivatives can be used to increase the solubil herein by reference. Such dosage forms can be used to pro 55 ity of a compound provided herein. See, e.g., U.S. Pat. No. vide slow or controlled-release of one or more active ingre 5,134,127, which is incorporated herein by reference. dients using, for example, hydropropylmethyl cellulose, 4.5.4 Topical and Mucosal Dosage Forms other polymer matrices, gels, permeable membranes, osmotic Topical and mucosal dosage forms provided herein systems, multilayer coatings, microparticles, liposomes, include, but are not limited to, sprays, aerosols, Solutions, microspheres, or a combination thereof to provide the desired 60 emulsions, Suspensions, eye drops or other ophthalmic prepa release profile in varying proportions. Suitable controlled rations, or other forms known to one of skill in the art. See, release formulations known to those of ordinary skill in the e.g., Remington's Pharmaceutical Sciences, 16th and 18th art, including those described herein, can be readily selected eds. Mack Publishing, Easton Pa. (1980 & 1990); and Intro for use with the active agents provided herein. In one embodi duction to Pharmaceutical Dosage Forms, 4th ed., Lea & ment, provided are single unit dosage forms Suitable for oral 65 Febiger, Philadelphia (1985). Dosage forms suitable for treat administration Such as, but not limited to, tablets, capsules, ing mucosal tissues within the oral cavity can be formulated gelcaps, and caplets that are adapted for controlled-release. as mouthwashes or as oral gels. US 8,716,252 B2 31 32 Suitable excipients (e.g., carriers and diluents) and other Chloride Injection, Ringer's Injection, Dextrose Injection, materials that can be used to provide topical and mucosal Dextrose and Sodium Chloride Injection, and Lactated Ring dosage forms encompassed herein are well known to those er's Injection; water-miscible vehicles such as, but not limited skilled in the pharmaceutical arts, and depend on the particu to, ethyl alcohol, polyethylene glycol, and polypropylene lar tissue to which a given pharmaceutical composition or 5 glycol; and non-aqueous vehicles such as, but not limited to, dosage form will be applied. In one embodiment, excipients corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, include, but are not limited to, water, acetone, ethanol, ethyl isopropyl myristate, and benzyl benzoate. ene glycol, propylene glycol, butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil, and mixtures 5. EXAMPLES thereof to form solutions, emulsions or gels, which are non 10 toxic and pharmaceutically acceptable. Moisturizers or Certain embodiments of the invention are illustrated by the humectants can also be added to pharmaceutical composi following non-limiting examples. tions and dosage forms. Examples of additional ingredients are well known in the art. See, e.g., Remington's Pharmaceu 5.1 (2S,3S4S.5R.6R)-6-(2-(2-((S)-1-(3-ethoxy-4- 15 methoxyphenyl)-2-(methylsulfonyl)ethyl)-1,3-diox tical Sciences, 16th and 18th eds., Mack Publishing, Easton oisoindolin-4-yl-amino)-2-oxoethoxy)-3,4,5-trihy Pa. (1980 & 1990). droxytetrahydro-2H-pyran-2-carboxylic Acid The pH of a pharmaceutical composition or dosage form may also be adjusted to improve delivery of one or more active ingredients. Also, the polarity of a solvent carrier, its ionic strength, or tonicity can be adjusted to improve delivery. Compounds Such as Stearates can also be added to pharma ceutical compositions or dosage forms to alter the hydrophi licity or lipophilicity of one or more active ingredients so as to improve delivery. In other embodiments, Stearates can serve 25 as a lipid vehicle for the formulation, as an emulsifying agent or Surfactant, or as a delivery-enhancing or penetration-en O hancing agent. In other embodiments, salts, Solvates, pro drugs, clathrates, or stereoisomers of the active ingredients 30 can be used to further adjust the properties of the resulting NH O OS-CH composition. 4.6 Kits N (H In one embodiment, active ingredients provided herein are not administered to a patient at the same time or by the same 35 OR i FO route of administration. In another embodiment, provided are kits which can simplify the administration of appropriate CH amounts of active ingredients. In one embodiment, a kit comprises a dosage form of a 40 Step 1: A reaction mixture of (S) N-(2-(1-(3-ethoxy-4- compound provided herein. Kits can further comprise addi hydroxyphenyl)-2-(methylsulfonyl)ethyl)-3-oxoisoindolin tional active ingredients such as oblimersen (Genasense R), 4-yl)cyclopropanecarboxamide (0.38 g. 0.83 mmol) and 3.4. melphalan, G-CSF, GM-CSF, EPO, topotecan, dacarbazine, 5-triacetoxy-6-(2.2.2-trichloro-acetimidoyloxy)-tetrahydro irinotecan, taxotere, IFN, COX-2 inhibitor, pentoxifylline, pyran-2-carboxylic acid benzyl ester (0.46g, 0.83 mmol) in ciprofloxacin, dexamethasone, IL2, IL8, IL18, Ara-C, 45 dichloromethane (40 mL) was chilled to -15°C. to -20°C., Vinorelbine, isotretinoin, 13 cis-retinoic acid, or a pharmaco treated with BF.OEt (0.10 mL, 0.83 mmol), and stirred for logically active mutant orderivative thereof, or a combination 2 hours at -15° C. to -20° C. After 2 hours, the reaction thereof. Examples of the additional active ingredients mixture was then allowed to warm to room temperature and include, but are not limited to, those disclosed herein (see, stirred overnight. The reaction mixture was diluted with 50 EtOAc (10 vol) then washed with saturated aqueous NaHCO e.g., section 4.3). (5 Vol), 2% NaCl (5 Vol), dried (MgSO), and concentrated. In other embodiments, kits can further comprise devices The crude mixture was eluted through a 120 g ISCO column, that are used to administer the active ingredients. Examples of 5% to 20% EtOAc/CHC1. After concentration of the frac Such devices include, but are not limited to, Syringes, drip tions, the product was dried under Vacuum for 12 hours, bags, patches, and inhalers. 55 which afforded (2S,3S4S.5R,6S)-2-(benzyloxycarbonyl)-6- Kits can further comprise cells or blood for transplantation (4-((S)-1-(7-(cyclopropanecarboxamido)-1-oxoisoindolin as well as pharmaceutically acceptable vehicles that can be 2-yl)-2-(methylsulfonyl)ethyl)-2-ethoxyphenoxy)tetrahy used to administer one or more active ingredients. For dro-2H-pyran-3,4,5-triyl triacetate (250 mg, 56% yield) as a example, if an active ingredient is provided in a solid form light yellow solid: 'H-NMR (DMSO-d): 10.4 (s, 1H), 8.26 that must be reconstituted for parenteral administration, the 60 8.23 (d. J=8 Hz, 1H), 7.53-7.48 (t, J–7 Hz, 1H), 7.39-7.31 (m, 5H), 7.20-7.18 (d. J=7 Hz, 1H), 7.13-7.12 (d, J-2 Hz, 1H), kit can comprise a sealed container of a suitable vehicle in 7.09-7.07 (d. J=9 Hz, 1H), 6.95-6.92 (dd, J=1.8 Hz, 1H), which the active ingredient can be dissolved to form a par 5.9-5.88 (dd, J=4, 10 Hz, 1H), 5.44-5.37 (overlapping dandt, ticulate-free sterile solution that is suitable for parenteral J.8 Hz, J-10 Hz, 2H), 5.17-5.03 (m, 4H), 4.70-4.63 (m, administration. Examples of pharmaceutically acceptable 65 2H), 4.38-4.30 (m. 1H), 4.21-4.15 (d. J=18 Hz, 1H), 4.08 vehicles include, but are not limited to: Water for Injection 3.95 (m, 3H), 3.04 (s, 3H), 2.00 (s, 3H), 1.97 (s, 3H), 1.81 USP; aqueous vehicles such as, but not limited to, Sodium 1.76 (m, 4H), 1.32-1.27 (t, J=7 Hz, 3H), 0.89-0.86 (d. J=6 Hz,

US 8,716,252 B2 35 36 A mixture of (S)-7-amino-2-(1-(3-ethoxy-4-methoxyphe The solvent was removed and the crude residue purified using nyl)-2-(methylsulfonyl) ethyl)isoindolin-1-one (5 g, 12.4 column chromatography (MeOH/CH.Cl: 0-5% over 60 mmol) and neat TMSI (25 g, 125 mmol) was heated at 80°C. minutes) to give 4.7 g of crude, 68% yield. Further purifica for 16 hours. The mixture was cooled to room temperature tion was then carried out by prep-HPLC to give (S)- N-(2- and quenched with 5% aq. sodium sulfite. Ethyl acetate was (1-(3-hydroxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl)- added and the layers were separated. The organic layer was 3-oxoisoindolin-4-yl)cyclopropanecarboxamide as a white washed with water, brine and then dried over NaSO. The solid (0.3 g, 4.3% yield). HPLC: HPLC: Hypersil DBS C8 organic layer was concentrated, followed by a solvent Swap to column, 250x4.6 mm, 5um: 99/1 to 85/15 over 15 minutes THF. To the THF solution, was added cyclopropanecarbonyl Gradient CHCN/10 mM aq. KHPO, 1.0 mL/min, 35°C., chloride (1.2 mL, 13.2 mmol). The mixture was heated to 40° 10 run time 20 minutes: to 11.84 minutes. H-NMR (DMSO-d): C. for 0.5 hour then cooled to room temperature. The mixture 10.50 (1H, s), 9.11 (1H, s), 8.22-825 (1H, d), 748-7.53 (1H, was quenched with 1 Naq. NaHCO then extracted with ethyl t), 7.18-7.2 (1H, d), 6.84-6.95 (3H, m), 5.80-5.85 (1H, m), 4.60-6.66 (1H, d), 3.90-430 (3H, m), 3.75 (3H, s), 3.04 acetate. The organic was washed with 1 N aq. NaHCO (3H, s), 1.79-1.83 (1H, m), 0.88-0.90 (4H, m); LC-MS ES" water, then brine. The solvent was removed and the crude was 15 suspended in ACN at 40° C. for 1 hour. The product was (M+1) 445. collected by filtration to give 1 g of crude, 19% yield. Further 5.5 7-amino-2-(1S)-1-(3-ethoxy-4-methoxy-phe purification by prep-HPLC followed by treatment with 2 N nyl)-2-methanesulfonyl-ethyl-2,3-dihydro-isoindol aq. HCl gave (S)- N-(2-(1-(3,4-dihydroxyphenyl)-2-(meth 1-one ylsulfonyl)ethyl)-3-oxoisoindolin-4-yl)cyclopropanecar boxamide as a white solid (0.25g, 5% yield). HPLC: HPLC: Hypersil DBS C8 column, 250x4.6 mm, 5um: 99/1 to 85/15 over 15 minutes Gradient CHCN/10 mM aq. KHPO, 1.0 O mL/min, 35° C. run time 20 minutes: to 10.91 minutes. 'H-NMR (DMSO-d): 10.51 (1H, s), 9.02-9.07 (2H, d), 8.22 25 8.25 (1H, d), 7.48-7.53 (1H, t), 7.18-7.20 (1H, d), 6.70-6.78 o (3H, m), 5.77-5.81 (1H, m), 4.59–4.65 (1H,d), 3.86-4.26 (3H, m), 3.02 (3H, s), 1.76-1.83 (1H, m), 0.88-0.90 (4H, m). LC O MS ES" (M+1) 431. 30 N \l 5.4 (S) N-(2-(1-(3-hydroxy-4-methoxyphenyl)-2- N (methylsulfonyl)ethyl)-3-oxosoindolin-4-yl)cyclo O propanecarboxamide 35 Step 1: A solution of 2-bromomethyl-6-nitro-benzoic acid methyl ester (4.0 g, 14.6 mmol), (1S)-1-(3-ethoxy-4-meth oxy-phenyl)-2-methanesulfonyl-ethylamine (4.0 g, 14.6 mmol) and triethylamine (4.4 mL, 31.6 mmol) in DMF (40 NH mL) was heated at 80°C.-90° C. for 1 day. The solvent was 40 removed in vacuo to give an oil. The oil was extracted with ethyl acetate (150 mL) and HCl (2N, 50 mL). The organic layer was washed with HCl (2N, 2x50 mL), water (2x50 mL), brine (2x50 mL) and dried over MgSO. The solvent was removed in vacuo to give a yellow oil. The oil was slurried in 45 ether (50 mL) to give a Suspension. The Suspension was filtered and washed with ether to give 2-(1S)-1-(3-ethoxy-4- Step 1: A mixture of (S)-7-amino-2-(1-(3-ethoxy-4-meth methoxy-phenyl)-2-methanesulfonyl-ethyl-7-nitro-2,3-di oxyphenyl)-2-(methylsulfonyl)ethyl)isoindolin-1-one (10g, hydro-isoindol-1-one as a yellow solid (3.43 g, 54% yield): 24.7 mmol) and neat TMSI (50g, 250 mmol) was heated at "H NMR (CDC1) & 1.5 (t, J–7 Hz, 3H, CH,), 2.99 (s.3H, 80° C. for 1.5 to 2 hours. The mixture was cooled to 0°C.-5° 50 CH), 3.69 (dd, J=5, 15 Hz, 1H, CHH), 3.86 (s, 3H, CH), C. and quenched with 5% aq. sodium sulfite. Methylene 4.07 (q, J–7 Hz, 2H, CH), 4.27 (dd, J=11, 15 Hz, 1H, CHH), chloride was added, and the slurry stirred vigorously until all 4.30 (d. J=17 Hz, 1H, NCHH), 4.54 (d. J=17 Hz, 1H, NCHH), the solids dissolved. The CHCl layer was separated and 5.70 (dd, J=5, 10 Hz, 1H, NCH), 6.85 (d. J=8 Hz, 1H, Ar), washed with water and brine, then concentrated to a foam (9.6 6.94-7.00 (m, 2H, Ar), 7.58-7.76 (m, 3H, Ar). g, >99% yield). The crude material was purified by column 55 Step 2: A suspension of 2-(1S)-1-(3-ethoxy-4-methoxy chromatography (MeOH 0-5%/CHCl over 60 minutes). phenyl)-2-methanesulfonyl-ethyl-7-nitro-2,3-dihydro The desired fractions were combined and concentrated to isoindol-1-one (3.0 g. 6.9 mmol) and Pd/C (10%. 350 mg) in give (S)-7-amino-2-(1-(3-hydroxy-4-methoxyphenyl)-2- methanol/ethyl acetate (150 mL each) was shaken under (methylsulfonyl)ethyl)isoindolin-1-one as a yellow solid (6 hydrogen (50-60 psi) in a Parr for 20 hours. The suspension g, 65% yield). UPLC: Acquity UPLC BEHC8, 2.1x50mm, 60 was filtered thru a pad of Celite, and washed with acetone 1.7L, 99:1 to 15:85 over 2 minutes gradient, 0.1% aq. TFA/ (300 mL). The solvent was removed in vacuo to give a solid. MeCN, 0.8 mL/min, 35° C.: to 0.80 minutes. The solid was slurried in methanol (10 mL) for 3 hours. The Step 2: To a mixture of (S)-7-amino-2-(1-(3-hydroxy-4- Suspension was filtered and washed with methanol to give methoxyphenyl)-2-(methylsulfonyl)ethyl)isoindolin-1-one 7-amino-2-(1S)-1-(3-ethoxy-4-methoxy-phenyl)-2-meth (5.86 g. 15.6 mmol) in THF (60 mL), was added cyclopro 65 anesulfonyl-ethyl-2,3-dihydro-isoindol-1-one as a white panecarbonyl chloride (1.5 mL, 16.1 mmol). The mixture was solid (1.5g, 55% yield):mp, 135-137° C.; H NMR (CDC1) heated to 60° C. for 1 hour then cooled to room temperature. 81.44 (t, J–7 Hz, 3H, CH), 2.93 (s.3H, CH), 3.66 (dd, J=5, US 8,716,252 B2 37 38 15 Hz, 1H, CHH), 3.85 (s, 3H, CH), 4.06 (q, J=7 Hz, 2H, in the absence or presence of compounds. Compounds pro CH), 4.13 (d.J=16 Hz, 1H, CHH), 4.22 (dd, J=10, 15 Hz, 1H, vided herein are dissolved in DMSO (Sigma) and further CHH), 4.35 (d. J=17 Hz, 1H, CHH), 5.19 (brs, 2H, NH), 5.64 dilutions are done in culture medium immediately before use. (dd, J–5, 10 Hz, 1H, NCH), 6.54 (d. J=8 Hz, 1H, Ar), 6.61 (d. The final DMSO concentration in all assays can be about J–7 Hz, 1H, Ar), 6.83 (d. J=8 Hz, 1H, Ar), 6.90-6.96 (m, 2H, 0.25%. Compounds are added to cells 1 hour before LPS Ar), 7.20 (t, J=8 Hz, 1H, Ar); C NMR (CDC1) & 14.59, stimulation. Cells are then incubated for 18-20hours at 37°C. 41.28, 48.01, 51.81, 55.87, 56.14, 64.48, 110.98, 111.38, in 5% CO, and supernatants are then collected, diluted with 112.22, 113.41, 114.75, 119.23, 129.89, 133.08, 142.36, culture medium and assayed for TNFC. levels by ELISA (En 146.02, 148.72, 149.45, 170.57: Anal Calcd for dogen, Boston, Mass., USA). ICsos are calculated using non CHNOS: C, 59.39; H, 5.98: N, 6.93. Found: C, 59.04; 10 linear regression, sigmoidal dose-response, constraining the H, 6.04; N, 6.74. 5.6 Assays top to 100% and bottom to 0%, allowing variable slope 5.6.1 PDE 4 Inhibition Assay (GraphPad Prism v3.02). PDE 4 inhibition may be assays using any methods con 5.6.3 Various PDE 4 Inhibition Related Assays 5.6.3.1 PBMC Purification and Stimulation ventionally known in the art. For example, using a modifica 15 tion of the method of Hill and Mitchell Hill and Mitchell, Human leukocytes units from healthy blood donors (Blood FasebJ., 8, A217 (1994), U937 cells (a human promonocytic Center of New Jersey, East Orange, N.J.) are diluted 1:1 with cell line) are grown to 1x10 cells/mL and collected by cen sterile Hank's Balanced Salt Solution (HBSS) and centri trifugation. A cell pellet of 1x10 cells is washed in phosphate fuged over room temperature Ficoll-Paque Plus (GE Health buffered saline and then frozen at -70° C. for later purifica care) to yield peripheral blood mononuclear cells (PBMC). tion or immediately lysed in cold homogenization buffer (20 PBMC are washed in HBSS and resuspended in Roswell Park mM Tris-HCl, pH 7.1, 3 mM 2-mercaptoethanol, 1 mM mag Memorial Institute (RPMI) complete medium (RPMI 1640, nesium chloride, 0.1 mMethylene glycol-bis-(B-aminoethyl BioWhittaker, Walkersville, Md.), 5% human serum, 100 ether)-N,N,N',N'-tetraacetic acid (EGTA), 1 uM phenylm U/ml penicillin, 100 mg/ml streptomycin, 2 mM ethylsulfonyl fluoride (PMSF), and 1 g/mL leupeptin). Cells 25 L-glutamine) and counted. One hundred ul (2x10"/ml) of are homogenized with 20 strokes in a Dounce homogenizer PBMC are added to each well of a 96 well flat-bottom plate and the Supernatant containing the cytosolic fraction are (final cell count=2x10/well) and incubated at 37° C. for 1 obtained by centrifugation. The Supernatant is then loaded hour. Fifty ml (4x) compound is added to each test well and 50 onto a Sephacryl S-200 column equilibrated in homogeniza ml medium containing 1% dimethylsulfoxide (DMSO) is tion buffer. The crude phosphodiesterase type 4 enzyme is 30 eluted in homogenization buffer at a rate of approximately 0.5 added to each control well (DMSO, 0.25%). The plate is mL/min and fractions are assayed for phosphodiesterase incubated for 1 hour at 37° C. Cells are then stimulated with activity using rolipram. Fractions containing PDE 4 activity 50 ul of 4 ng/ml lipopolysaccharide (LPS) from Salmonella (rolipram sensitive) are pooled and aliquoted for later use. abortus equii (Sigma) (LPS), 1 ng/ml) and incubated for The phosphodiesterase assay is carried out based on the 35 18 hours at 37° C. For stimulation with superantigen, PBMC procedure described by Hill and Mitchell. The assay is carried are plated in 96-well tissue culture plates at 3x10 cells/well out in a total Volume of 100 ul containing various concentra in complete medium, pretreated with compounds at 37°C. for tion of the compounds of interest, 50 mM Tris-HCl, pH 7.5, 5 1 hour, then stimulated with 100 ng/ml Staphylococcal mM magnesium chloride and 1 uMcAMP of which 1% is H enterotoxin B (SEB) (Toxin Technology, Sarasota, Fla.) for cAMP. Reactions are incubated at 30° C. for 30 minutes and 40 18 hours. then terminated by boiling for 2 minutes. The amount of PDE 5.6.3.2 Quantitative Reverse Transcription-Polymerase 4 containing extract used for these experiments is predeter Chain Reaction mined Such that reactions are within the linear range and Cells are harvested and RNA isolation is performed consume less than 15% of the total substrate. Following ter according to manufacturers instructions (e.g., RNeasy, mination of reaction, samples are chilled at 4° C. and then 45 Qiagen, Valencia, Calif.). Reverse transcription is performed treated with 10 ul of 10 mg/mL Snake venom for 15 minutes converting 1 ug RNA to cDNA for each sample according to at 30°C. Unused substrate then is removed by adding 200 ul manufacturer's protocol (e.g., RT Kit, Applied BioSystems, of a quaternary ammonium ion exchange resin (AG1-X8. Foster City, Calif.). Quantitative real-time polymerase chain BioRad) for 15 minutes. Samples then are spun at 3000 rpm reaction (PCR) is performed for gene expression analysis for 5 minutes and 50 ul of the aqueous phase are taken for 50 using 50 ng cDNA per sample. Gene expression assays for counting. Each data point is carried out in duplicate and target genes and endogenous glyceraldehyde-3-phosphate activity is expressed as percentage of control. The ICsos of the dehydrogenase (GAPDH) control may be from, for example, compounds are then determined from dose responsecurves of Applied Biosystems. Expression is measured on a real-time a minimum of three independent experiments. PCR System 7500 (Applied Biosystems). Relative quantifi 5.6.2 TNFC. Inhibition Assay in PMBC 55 cations are calculated, for example, with SDS V.1.3.1 soft Peripheral blood mononuclear cells (PBMC) from normal Wa. donors are obtained by Ficoll Hypaque (Pharmacia, Piscat 5.6.3.3 Cytokine and Chemokine Protein Analysis away, N.J., USA) density centrifugation. Cells are cultured in A 50 ul supernatant from each well is transferred into new RPMI 1640 (Life Technologies, Grand Island, N.Y., USA) round-bottomed 96 well plates and stored at -20° C. for supplemented with 10% AB+human serum (Gemini Bio 60 cytokine analysis by cytometric bead array using a LumineX products, Woodland, Calif., USA), 2 mM L-glutamine, 100 IS100 instrument (Luminex Corporation, Austin, Tex.). Lin U/ml penicillin, and 100 ug/ml streptomycin (Life Technolo coPlex kits with antibody bound beads for Luminex XMAP gies). Technology (Millipore) are combined into multiplex format PBMC (2x10 cells) are plated in 96-well flat-bottom Cos prior to assay. Data analysis is performed using Upstate Bead tar tissue culture plates (Corning, N.Y., USA) in triplicate. 65 view software. IL-2 and IFN-Y levels from SEB stimulated Cells are stimulated with LPS (from Salmonella abortus equi, PBMC are measured by enzyme-linked immunoabsorbant Sigma cat. no. L-1887, St. Louis, Mo., USA) at 1 ng/ml final assay (ELISA) (R&D Systems). US 8,716,252 B2 39 40 5.6.3.4 PMN Isolation and Stimulation bation, the Supernatants are harvested and analysed for levels Polymorphonuclear cells (PMN) are isolated from human of TNF-ct, IFN-y, GM-CSF, and MIP-1C. by ELISA (R&D leukocytes by separation from PBMC using Ficoll gradient Systems). centrifugation to remove PBMC. Erythrocytes are removed 5.6.3.6 Keratinocyte Proliferation, TNF-C. Production and by sedimentation in 3% dextran followed by hypotonic lysis Viability in 0.2% saline. Finally, any contaminating monocytes are For proliferation studies, human neonatal foreskin epider depleted using HLA class II magnetic beads. PMN (3x10 mal keratinocytes (HEKncells) are obtained from Cell Appli cells/well) are pretreated with titrated compound for 1 hour, cations, Inc. (San Diego, Calif.) and plated at 3000 cells/well and then stimulated with Zymosan A particles (heat-killed S in 96-well flat bottom tissue culture plates for two days. Cell cerevisiae) at various doses. Polymyxin B sulfate (40 nM 10 final) is added to all samples to neutralize any contaminating proliferation is measured using the Cell Counting Kit LPS. After overnight incubation, supernatants are harvested (Dojindo Molecular Technologies, Inc, Gaithersburg, Md.). and assayed for IL-8 by ELISA. For TNF-C. production and viability studies, HEKncells are For LTB production, PMN are resuspended in phosphate obtained from Cascade Biologics (Portland, Oreg.) and are buffered saline without calcium or magnesium (BioWhit 15 grown in serum-free medium Supplemented with growth fac taker) containing 10 mM 4-(2-hydroxyethyl)-1-pipera tors. When cells reach 80% confluency, cells are trypsinized zineethanesulfonic acid (HEPES) (pH-7.2) and plated in and plated at 1x10 cells/well in 6-well dishes. Plates are 96-well tissue culture plates at a concentration of 1.7x10' incubated for 24 hours to allow cell adhesion. HEKncells are cells/well. Cells are treated with 50 uMthimerosal (Sigma)/1 treated with test compound or 0.1% DMSO as the vehicle mM CaC1/1 mM MgCl, for 15 minutes at 37° C. 5% CO, control for 1 hour before ultraviolet B (UVB) irradiation with then treated with compound in a final DMSO concentration of 50 ml/cm in a UV Stratalinker 2400 (Stratagene, La Jolla, 0.01% in duplicate for 10 minutes. PMN are stimulated with Calif.) calibrated with 312 nm UVB bulbs. Media and com 1 uM formyl-Met-Leu-Phe (fMLF, Sigma) for 30 minutes, pounds are replaced, and cells are incubated for 18 hours. then lysed by the addition of methanol (20% final concentra Supernatants are removed for testing in a TNF-C. ELISA tion) and frozen in a dry icefisopropanol bath for 10 minutes. 25 before 100 ul of adenosine triphosphate (ATP)-lite reagent Lysates are stored at -70° C. until the LTB content is mea (PerkinElmer Life and Analytical Sciences, Shelton, Conn.) sured by competitive LTB ELISA (R&D Systems). is added to each well to assay for viability. Lysates are trans For CD18/CD11b (Mac-1) expression, PMN are pretreated ferred to plates and shaken for 2 minutes before chemilumi with compound for 10 minutes and stimulated with nescence is read on a TopCount NXT Luminescence Counter N-formyl-Met-Leu-Phe (fMLF) for 30 minutes, then placed 30 (PerkinElmer Life and Analytical Sciences). on ice, stained with anti-CD18-FITC and anti-CD11b-PE and 5.6.4 Cell Proliferation Assay analysed by flow cytometry using a fluorescence-activated Cell lines Namalwa, MUTZ-5, and UT-7 are obtained from cell sorter (FACS) Calibur flow cytometer (BD Biosciences). the Deutsche Sammlung von Mikroorganismen and Zellkul To measure adhesion of neutrophils to endothelial cells, turen GmbH (Braunschweig, Germany). The cell line KG-1 is human umbilical vein endothelial cells (HUVEC, Anthrogen 35 obtained from the AmericanType Culture Collection (Manas esis Corporation, Cedar Knolls, N.J.) are plated out onto 96 sas, Va., USA). Cell proliferation as indicated by H-thymi well plates (5x10 cells per well) in medium containing 2% dine incorporation is measured in all cell lines as follows. FBS 4 days prior to the experiment to ensure adhesion of Cells are plated in 96-well plates at 6000 cells per well in HUVEC to the plate. On the day of the experiment, neutro media. The cells are pre-treated with compounds at about phils are isolated from human leukocytes and labeled with the 40 100, 10, 1, 0.1, 0.01, 0.001, 0.0001 and 0 uM in a final fluorescent dye Calcein-AM (Molecular Probes, Oregon) for concentration of about 0.25% DMSO in triplicate at 37° C. in 1 hour. Labeled neutrophils (2x10 per well) are added to the a humidified incubator at 5% CO, for 72 hours. One micro adhered HUVECs and pretreated with compound for 10 min curie of H-thymidine (Amersham) is then added to each utes at 37° C. in a humidified incubator at 5% CO. fMLF is well, and cells are incubated again at 37°C. in a humidified added to trigger neutrophiladhesion to HUVECs for 30 min 45 incubator at 5% CO for 6 hours. The cells are harvested onto utes. The cells are washed with PBS containing 2% glucose to UniFilter GF/C filter plates (Perkin Elmer) using a cell har remove non-adherent neutrophils, and the number of adher vester (Tomtec), and the plates are allowed to dry overnight. ent neutrophils was measured on a fluorimeter. Microscint 20 (Packard) (25ul/well) is added, and plates are For IL-8 production assays, PMN are plated in 96-well analyzed in TopCount NXT (Packard). Each well is counted tissue culture plates at 3x10 cells/well in complete medium, 50 for one minute. Percent inhibition of cell proliferation is treated with compound in duplicate in a final DMSO concen calculated by averaging all triplicates and normalizing to the tration of 0.1% for 1 hour at 37° C. 5% CO. PMN are then DMSO control (0% inhibition). Each compound is tested in stimulated with unopsonized, boiled Zymosan A (Sigma) at each cell line in three separate experiments. Final ICsos are 2.5x10 particles/well for 18 hours. Supernatants are har calculated using non-linear regression, sigmoidal dose-re vested and tested for IL-8 by ELISA (R&D Systems). sponse, constraining the top to 100% and bottom to 0%. 5.6.3.5 NK Cell Purification and Stimulation allowing variable slope. (GraphPad Prism v3.02). NK cells are isolated from leukocyte units from healthy 5.6.5 Cell Cycle Analysis blood donors by 30-minute incubation with RossetteSep Cells are treated with DMSO oran amount of a compound cocktail for NK cell enrichment by negative selection (Stem provided herein overnight. Propidium iodide staining for cell Cell Technologies, Inc.), followed by Ficoll-Hypaque density 60 cycle is performed using CycleTEST PLUS (Becton Dickin gradient centrifugation. CD56'NK cells are isolated to -85% son) according to manufacturer's protocol. Following stain purity as determined by flow cytometry. Flat-bottom plates ing, cells are analyzed by a FACSCalibur flow cytometer are coated with 100 ug/mL of human IgG (Sigma) overnight using ModFit LT software (Becton Dickinson). at 4°C. The unbound IgG is washed away. NK cells are plated 5.6.6 Apoptosis Analysis at 2x10 cells/well into the 96-well plates, and 10 ng/mL of 65 Cells are treated with DMSO oran amount of a compound IL-2 (R&D Systems, Minneapolis Minn.) is added. Test com provided herein at various time points, then washed with pound is then added to the plate wells. After a 48-hour incu annexin-V washbuffer (BDBiosciences). Cells are incubated US 8,716,252 B2 41 42 with annexin-V binding protein and propidium iodide (BD are considered to be within the scope of the invention and are Biosciences) for 10 minutes. Samples are analyzed using flow encompassed by the appended claims. cytometry. All of the patents, patent applications and publications 5.6.7. Anti-Proliferation Assay referred to herein are incorporated herein in their entireties. Day 1: The cells are seeded to 96-well plate with 50 ul/well Citation oridentification of any reference in this application is in 10% FBS RPMI (w/Glutamine, w/o pen-strip) for over not an admission that Such reference is available as prior art to night. The following cells are used: this invention. The full scope of the invention is better under Colorectal cancer cell: Colo 2053200 cells/well; positive stood with reference to the appended claims. control irinotecan What is claimed is: Pancreatic cancer cell: BXPC-3 1200 cells/well; positive 10 1. A pharmaceutical composition comprising a compound control gemcitabine of formula (I): Prostate cancer cell: PC3 1200 cells/well; positive control docetaxel Breast cancer cell: MDA-MB-231 2400 cells/well; posi (I) tive control paclitaxel 15 OR2 Day 2: The compounds are serially diluted from 0.00001 um-10 um (or 0.000001~1 uM) with 50 ul/well (of 2x) and added to the plates in duplicate with relative positive control. NHR OR, The plates were then incubated at 37° C. for 72 hours. Day 5: The results are detected by CellTiter Glo method. 21 100 ul/well of CellTiter Glo reagent is added to the plates and R*-- N / incubated for 10 minutes at room temperature, and then ana lyzed on the Top Count reader. The ICs of each compound is N vich, typically based on the result of two or more individually O experiments. 25 5.7 PDE 4 Inhibition or a pharmaceutically acceptable salt, Solvate, or Stereoi Abilities of certain compounds for inhibiting PDE 4 were Somer thereof, wherein: determined using procedures substantially similar to those R" is glucuronide (gluc”), or (cyclopropyl)-C(O) , described in Sections 5.6.1 and 5.6.3, above. wherein the cyclopropyl may be substituted with one or The tested compounds included: (S) N-(2-(1-(3-ethoxy 30 more hydroxyl groups; 4-hydroxyphenyl)-2-(methylsulfonyl)ethyl)-3-oxoisoindo R is hydrogen, methyl, or gluc; lin-4-yl)cyclopropanecarboxamide: (S)-N-(2-(1-(3,4-dihy R is hydrogen, ethyl, or gluc; and droxyphenyl)-2-(methylsulfonyl)ethyl)-3-oxoisoindolin-4- R" is hydrogen, hydroxyl, or -O-gluc; yl)cyclopropanecarboxamide: (S) N-(2-(1-(3-hydroxy-4- with the proviso that when R is methyl and R is ethyl, methoxyphenyl)-2-(methylsulfonyl)ethyl)-3-oxoisoindolin 35 then R' cannot be unsubstituted (cyclopropyl)-C(O). 4-yl)cyclopropanecarboxamide; and 7-Amino-2-(1S)-1-(3- 2. The composition of claim 1, wherein R' is (cyclopro ethoxy-4-methoxy-phenyl)-2-methanesulfonyl-ethyl-2,3- pyl)-C(O)—. dihydro-isoindol-1-one. From the tests, it was determined 3. The composition of claim 1, wherein R is hydrogen. that ICso values of the tested compounds were in the range of 4. The composition of claim 1, wherein R is methyl. about 2 to about 85 uM. 40 5. The composition of claim 1, wherein R is gluc. 5.8 TNFO Inhibition 6. The composition of claim 1, wherein R is hydrogen. Abilities of certain compounds for inhibiting TNFC. were 7. The composition of claim 1, wherein R is ethyl. determined using procedures substantially similar to those 8. The composition of claim 1, wherein R is hydroxyl. described in Section 5.6.2, above. 9. The composition of claim 1, wherein R is —O-gluc. The tested compounds included: (2S,3S4S.5R.6R)-6-(2- 45 10. A pharmaceutical composition comprising a com (2-((S)-1-(3-Ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) pound having the structure: ethyl)-1,3-dioxoisoindolin-4-ylamino)-2-oxoethoxy)-3,4,5- trihydroxytetrahydro-2H-pyran-2-carboxylic acid: (S) N (2-(1-(3-ethoxy-4-hydroxyphenyl)-2-(methylsulfonyl) O-gluc ethyl)-3-oxoisoindolin-4-yl)cyclopropanecarboxamide; 50 (S)- N-(2-(1-(3,4-dihydroxyphenyl)-2-(methylsulfonyl) ethyl)-3-oxoisoindolin-4-yl)cyclopropanecarboxamide; NH O OCHs. (S)- N-(2-(1-(3-hydroxy-4-methoxyphenyl)-2-(methylsul 21 fonyl)ethyl)-3-oxoisoindolin-4-yl)cyclopropanecarboxam HO-H N / ide; and 7-Amino-2-(1S)-1-(3-ethoxy-4-methoxy-phenyl)- 55 2-methanesulfonyl-ethyl-2,3-dihydro-isoindol-1-one. N R CH3 ICso values of the tested compounds were not determined, O except for one test compound whose ICso was determined to OCH be about 0.6LM. For other test compounds, percent inhibition at 10 uM was determined. About 30 to about 45 percent 60 NH2 O O-gluc, inhibition of TNFC. was observed when each of the com pounds was present at 10 uM. 21 The embodiments of the invention described above are HO-H- N / intended to be merely exemplary, and those skilled in the art will recognize, or will be able to ascertain using no more than 65 N v CH routine experimentation, numerous equivalents of specific O compounds, materials, and procedures. All Such equivalents US 8,716,252 B2 43 44 -continued or a pharmaceutically acceptable salt, Solvate, or stereoiso OCH mer thereof. 11. The composition of claim 1, wherein the compound is:

NH2 O O-C2H5, 5 21 O O-gluc gluc-O-- N / N v CH3 v- O OC2H5, O 10 N O OCH3 M

vO ch NH-gluc O-CHs. 15 O OH 21 HO-H- N / v- O OC2H5, N R CH3 O N O O-gluc M

vich,O NH2 O O-CHs. O OH 25 N O v's O OH, C vich,M O 30 N O O OCH C vM CH O NH O O-gluc, O OCH3

35 N O M NH O OH, v CH3 O N O O OH M 40 iv CH NH O 13 O OC2H5, OCH3 HO

N O 45 NH2 O O-CHs. M v CH3 O N O O O-gluc M 50 vich, NH O 13 O OCHs. HO or a pharmaceutically acceptable salt, Solvate, or stereoiso N O mer thereof. A. 55 12. A method of treating or managing a disease or disorder S-CHV comprising administering to a patient a composition of claim O 1, or a pharmaceutically acceptable salt, Solvate, or stereoi O OCH Somer thereof, wherein the disease or disorder is cancer, pain, arthritis, Crohn's disease, 5q-deletion syndrome, sickle cell NH 60 13 O OC2H5, anemia, cachexia, adult respiratory distress syndrome, macu HO lar degeneration, Graft versus Host Reaction, chronic pulmo nary inflammatory diseases, psoriatic arthritis, multiple scle N O M rosis, systemic lupus erythromatosis, ENL in leprosy, v CH3 65 psoriasis, fibrotic disease, oncogenic or cancerous condi tions, asthma, chronic obstructive pulmonary disease, or O atopic dermatitis. US 8,716,252 B2 45 46 13. A method of treating or managing a disease or disorder comprising administering to a patient a compound of claim 10, or a pharmaceutically acceptable salt, Solvate, or stereoi Somer thereof, wherein the disease or disorder is cancer, pain, arthritis, Crohn's disease, 5q-deletion syndrome, sickle cell anemia, cachexia, adult respiratory distress syndrome, macu lar degeneration, Graft versus Host Reaction, chronic pulmo nary inflammatory diseases, psoriatic arthritis, multiple scle rosis, systemic lupus erythromatosis, ENL in leprosy, psoriasis, fibrotic disease, oncogenic or cancerous condi 10 tions, asthma, chronic obstructive pulmonary disease, or atopic dermatitis.