COMPOSITIONS AND METHODS FOR TREATING LYMPHOMA

CROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 61/259,396, filed on November 9, 2009, and U.S. Provisional Application No. 61/266,783, filed on December 4, 2009, each of which is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

[0002] There is a need for new chemotherapeutic agents for the treatment of cancers

including lymphoma and leukemia.

SUMMARY OF THE INVENTION

[0003] Lymphoma is a type of cancer that affects lymphocytes and presents as solid tumors in the lymph nodes or in other lymph tissues. Lymphomas account for about five percent of all cases of cancer in the United States. Lymphocytic tumors may have areas of normoxia and areas of hypoxia. Provided herein are compounds, compositions and methods for treatment of lymphoma.

[0004] Provided herein, in some embodiments, are method of treating lymphoma in a

patient in need thereof comprising administration of a formulation comprising S-2- amino-3-[4'-N,N,-/? s(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride, metabolite, analog, or derivative thereof, to the patient in need thereof.

[0005] In some embodiments, the N-oxide, metabolite, analog, or derivative thereof, has the structure of Formula I:

Formula I;

wherein:

A is an aryl or heteroaryl ring; R ¹ and R ² are each, independently H, -Ci-Ce alkyl, -Ci-Ce heteroalkyl, -(L) _m -(C3-Cio cycloalkyl), -(L) _m -(C ₂ -C ₁ o heterocycloalkyl); and wherein each alkyl, heteroalkyl, cycloalkyl, and heterocycloalkyl, is independently, at each occurrence, optionally substituted with 1 -3 substituents selected from R ⁵ ;

or R ¹ and R ² taken together with the nitrogen atom to which they are attached form a C2-C10 heterocycloalkyl ring wherein the ring optionally comprises one or more moieties selected from O, C=0, S(0) _m , NR ⁸ S(0) _m , NR ⁸ (C=0) and -R ⁸ , and wherein the ring is optionally substituted with 1-3 substituents selected from R ^a ; or R ² is attached to an adjacent carbon atom on ring A to form a saturated, partially saturated or unsaturated 5-6 member ring that is fused to ring A, and wherein the 5-6 membered ring that is fused to ring A is optionally substituted with 1-3 substituents selected from R ⁵ ;

R ³ and R ⁴ are each independently, at each occurrence, H, -Ci-Ce alkyl, halo, or -OH; each R ⁵ is independently, at each occurrence, halo, -CN, -N0 ₂ , -(L) _m -OR ⁶ , -(L) _m - SR ⁷ , -(L) _m -S(=0)R ⁷ , -(L) _m -S(=0) ₂ R ⁷ , -(L) _m -NHS(=0) ₂ R ⁷ , -(L) _m -C(=0)R ⁷ , - (L) _m -OC(=0)R ⁷ , -(L) _m -C0 ₂ R ⁶ , -(L) _m -OC0 ₂ R ⁶ , -(L) _m -CH(R ⁶ ) ₂ , -(L) _m -N(R ⁶ ) ₂ , - (L) _m -C(=0)N(R ⁶ ) ₂ , -(L) _m -OC(=0)N(R ⁶ ) ₂ , -(L) _m -NHC(=0)NH(R ⁶ ), -(L) _m - NHC(=0)R ⁷ , -(L) _m -NHC(=0)OR ⁷ , -(L) _m -C(OH)(R ⁶ ) ₂ , -(L) _m -C(NH ₂ )(R ⁶ ) ₂ , -d- C ₆ alkyl, -Ci-C ₆ fluoroalkyl or -Ci-C ₆ heteroalkyl;

each R ⁶ is independently, at each occurrence, H, a Ci-C ₆ alkyl, a Ci-C ₆ fluoroalkyl, a Ci-C ₆ heteroalkyl, a C3-Ciocycloalkyl, a C ₂ -Cioheterocycloalkyl, an aryl, a heteroaryl, -Ci-C4alkyl-(C3-Ciocycloalkyl), -Ci-C4alkyl-(C ₂ - Cioheterocycloalkyl), -Ci-C4alkyl-(aryl), or -Ci-C4alkyl-(heteroaryl), and wherein each alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently, at each occurrence, optionally substituted with 1-3 substituents selected from R ^a ; or

two R ⁶ groups attached to the same N atom are taken together with the N atom to which they are attached to form a C ₂ -Cioheterocycloalkyl wherein the ring optionally comprises one or more moieties selected from O, C=0, S(0) _m , NR ⁸ S(0) _m , NR ⁸ (C=0) and N-R ⁸ , and wherein the ring is optionally substituted with 1 -3 substituents selected from R ^a ; each R ⁷ is independently, at each occurrence, a Ci-C ₆ alkyl, a Ci-C ₆ fluoroalkyl, a Ci- Ceheteroalkyl, a C3-Ciocycloalkyl, a C2-Cioheterocycloalkyl, an aryl, a heteroaryl, -Ci-C4alkyl-(C3-Ciocycloalkyl), -Ci-C4alkyl-(C ₂ - Cioheterocycloalkyl), -Ci-C4alkyl-(aryl), or -Ci-C4alkyl-(heteroaryl), and wherein each alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently, at each occurrence, optionally substituted with 1-3 substituents selected from R ^a ;

each R ⁸ is independently, at each occurrence, H, Ci-Ce alkyl, Ci-Ce heteroalkyl or - C(=0)-C ₁ -C6 alkyl, and wherein each alkyl, and heteroalkyl is independently, at each occurrence, optionally substituted with 1 -3 substituents selected from R ^a ; each R ^a is independently, at each occurrence, oxo, halo, -CN, -N0 ₂ , -(L) _m -0-(Ci- C ₃ alkyl), -(L) _m -S-(Ci-C ₃ alkyl), -(L) _m -S(=0) -(Ci-C ₃ alkyl), -(L) _m -S(=0) ₂ -(d- C ₃ alkyl), -(L) _m -NHS(=0) ₂ -(C ₁ -C ₃ alkyl), -(L) _m -C(=0) -(C ₁ -C ₃ alkyl), -(L) _m - OC(=0) -(C ₁ -C ₃ alkyl), -(L) _m -C0 ₂ -(Ci-C ₃ alkyl), -(LVOC0 ₂ -(Ci-C ₃ alkyl), - (L) _m -CH(C ₁ -C ₃ alkyl) ₂ , -(L) _m -N(C ₁ -C ₃ alkyl) ₂ , -(L) _m -C(=0)N(C ₁ -C ₃ alkyl) ₂ , -(L) _m - OC(=0)N(Ci-C ₃ alkyl) ₂ , -(L) _m -NHC(=0)NH(Ci-C ₃ alkyl), -(L) _m -NHC(=0) ) - (Ci-C ₃ alkyl), -(L) _m -NHC(=0)0-(C ₁ -C ₃ alkyl), -(L) _m -C(OH)(Ci-C ₃ alkyl) ₂ , -(L) _m - C( H ₂ )( Ci-C ₃ alkyl) ₂ , -Ci-C ₆ alkyl, -Ci-C ₆ fluoroalkyl, or -Ci-C ₆ heteroalkyl;

L is independently, at each occurrence, a bivalent radical selected from -(Ci-C ₃ alkylene) _m -, -(C ₃ -C ₇ cycloalkylene), -(Ci-C ₃ alkylene) _m -0-(Ci-C ₃ alkylene) _m -, and -(Ci-C ₃ alkylene) _m -NH-(Ci-C ₃ alkylene) _m -;

each m is independently, at each occurrence, 0, 1 or 2;

n is 1, 2 or 3; and

p is 0, 1, 2, or 3;

or pharmaceutically acceptable salt, N-oxide, solvate or prodrug thereof.

[0006] In some embodiments, the compound of Formula I is a compound having the

structure of Formula II:

Formula II.

[0007] In some embodiments, the compound of Formula II has the structure of Formula III:

Formula III;

wherein:

R ⁹ is CN, halo, -C1-C3 alkyl, -0-Ci-C ₃ alkyl, -O-C1-C3 fluoroalkyl or -C1-C3 fluoroalkyl; and

k is 1, 2 or 3.

[0008] In some embodiments, the compound of Formula III has the structure of Formula IIIA, Formula IIIB or Formula IIIC:

Formula I Formula IIIB

Formula IIIC.

[0009] In some embodiments, the compound of Fomula I has the structure of Formula IV:

Formula IV.

[0010] In some embodiments, n is 1.

[0011] In some embodiments, R ³ and R ⁴ are each, independently at each occurrence, H, - C1-C3 alkyl, or halo.

[0012] The method of claim 2, wherein p is 0.

[0013] In some embodiments, the compound of Formula I is selected from

[0014] In some embodiments, the compound is administered together with a

pharmaceutically acceptable carrier or diluent.

[0015] In some embodiments, said lymphoma is under normoxic conditions. In some

embodiments, said lymphoma is under hypoxic conditions. In some embodiments, said lymphoma is selected from a group consisting of Hodgkin's lymphoma, non- Hodgkin lymphoma, nodal marginal zone B cell lymphoma (NMZL), mantle cell lymphoma, diffuse large B cell lymphoma, plasma cell neoplasms, chronic lymphocytic leukemia, small lymphocytic lymphoma, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, splenic marginal zone lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, primary effusion lymphoma, T cell prolymphocytic leukemia, T cell large granular lymphocytic leukemia, aggressive NK cell leukemia, adult T cell

leukemia/lymphoma, extranodal NK/T cell lymphoma, enteropathy -type T cell lymphoma, hepatosplenic T cell lymphoma, blastic NK cell lymphoma, mycosis fungoides, sezary syndrome, primary cutaneous CD30-positive T cell

lymphoproliferative disorders, primary cutaneous anaplastic large cell lymphoma, primary CNS lymphoma, lymphomatoid papulosis, angioimmunoblastic T cell lymphoma, peripheral T cell lymphoma, follicular lymphoma, Burkitt's lymphoma, gastric lymphoma, cutaneous T Cell lymphoma, anaplastic large cell lymphoma, and MALT lymphoma.

[0016] In some embodiments, said lymphoma is selected from a group consisting of

Hodgkin's lymphoma, non-Hodgkin lymphoma, diffuse large B cell lymphoma, follicular lymphoma, Burkitt's lymphoma, mantle cell lymphoma, peripheral T-cell lymphoma, extranodal natural killer/T-cell lymphoma, nasal type, cutaneous T-cell lymphoma, enteropathy type T-cell lymphoma, angioimmunoblastic T-cell lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma, extranodal marginal zone B-cell, nodal marginal zone B-cell lymphoma, hairy cell leukemia, splenic marginal zone B-cell lymphoma, and mediastinal (thymic) large B-cell lymphoma.

[0017] In some embodiments, said lymphoma is selected from a group consisting of

Hodgkin's lymphoma, non-Hodgkin lymphoma, diffuse large B cell lymphoma, and follicular lymphoma. In some embodiments, said lymphoma is diffuse large B cell lymphoma.

[0018] In one aspect, methods of inhibiting angiogenic growth in a patient in need thereof comprised of administering a therapeutically effective amount of S-2-amino-3-[4'- N,N,-Ws(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride, wherein the angiogenic growth is associated with lymphoma are provided.

[0019] In another aspect, methods of inhibiting tumor formation in a patient in need thereof comprised of administering a therapeutically effective amount of S-2-amino-3-[4'- N,N,-Ws(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride, wherein the tumor formation is associated with lymphoma are provided.

[0020] In another aspect methods of treating lymphoma comprising administering a

therapeutically effective amount of S-2-amino-3-[4'-N,N,-/?«(2- chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride are provided.

[0021] In another aspect, methods of treating Hodgkin's lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-/?«(2-chloroethyl)amino]phenyl propionic acid N- oxide dihydrochloride are provided.

[0022] In another aspect, methods of treating diffuse large B-cell lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-/?«(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride are provided.

[0023] In another aspect, methods of treating follicular lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-/?«(2-chloroethyl)amino]phenyl propionic acid N- oxide dihydrochloride are provided. [0024] In another aspect, methods of treating non-Hodgkin lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-/?«(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride are provided.

[0025] In another aspect, methods of treating Burkitt's lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-/?«(2-chloroethyl)amino]phenyl propionic acid N- oxide dihydrochloride are provided.

INCORPORATION BY REFERENCE

[0026] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

[0028] FIG. 1 illustrates the chemical structure of melphalan N-oxide (PX-478).

[0029] FIG. 2 illustrates the effect of PX-478 on HIF-Ια in diffuse large B-cell lymphoma (DLBCL) cell lines with increasing concentrations of PX-478 under normoxic or hypoxic conditions.

[0030] FIG. 3 A-C illustrates the effect of PX-478 on cell viability of SUDHL4 (A),

SUDHL6 (B) and SUDHL10 (C) lymphoma cell lines. Cell growth was measured by MTT assay.

[0031] FIG. 4A illustrates inhibition of HIF-1 alpha expression in SUDHL-4 cells as

measured by flow cytometry under normoxia and hypoxia. FIG. 4B illustrates downregulation of HIF- 1 alpha protein upon pretreatment of SUDHL4 cells with PX-478 followed by 20 hr incubation in normoxia (21% oxygent) or hypoxia (1% oxygen). HIF-1 alpha and HIF-2 alpha protein was detected using specific antibodies.

[0032] FIG. 5 illustrates apoptosis of DLBCL cells in the presence of PX-478 as measured by annexin V/PI staining followed by flow cytometry. SUDHL4 cells were pretreated with PX-478 for 6 hours and were then incubated with the indicated concentrations of PX-478 under normoxia or hypoxia for 18 hours.

[0033] FIG. 6 illustrates inhibition of colony formation of SUDHL4 cells in the presence of PX-478. SUDHL4 cells were grown on soft agarose feeder layer (1 : 1 agarose to RPMI) and plated. After the feeder layer was solidified, SUDHL4 cells were plated at 10,000 cells/mL in a 1 :3 agarose to RPMI mixture. After solidification of the top layer, cells were placed in the incubator and RPMI media was added the same day. Media was changed every 3 days and colonies (>50 cells) were counted on day 11. Colony formation was examined and photomicroscopy of cell colonies was taken at 5x magnification.

[0034] FIG. 7A illustrates oxygen-dependent regulation of HIF- 1 alpha activity. Oxygen regulates the rate at which HIF-1 alpha protein is degraded. In normoxic conditions, oxygen dependent prolyl hydroxylase-domain protein 1-3 is required for the binding of the von Hippel-Lindau (VHL) tumor suppressor protein. VHL binding is also promoted by acetylation of lysine (K) residue 532 by the ARDI acetyltransferase. Oxygen also regulates the interaction of HIF- 1 alpha with trancriptional co- activators. Oxygen dependent hydroxylaton of asparagine (N) residue 803 in HIF-1 alpha by the enzyme FIH-1 (factor inhibiting HIF-1) blocks the bindiing of p300 and CBP to HIF-1 alpha and therefore inhibits HIF-1 mediated gene transcription.

Under hypoxic conditions the rate of aspargine and proline hydroxylation decreases therefore VHL cannot bind to HIF-1 alpha, resulting in decrease rate of HIF-1 alpha degradation. p300 and CBP can bind to HIF-1 alpha allowing transcriptional activaton of HIF-1 target genes. FIG. 7B illustrates the HIF-1 alpha pathway.

DETAILED DESCRIPTION OF THE INVENTION

[0035] Solid tumors with areas of hypoxia have long been recognized as the most

aggressive and difficult tumors to treat. Lymphoma is a type of cancer that affects lymphocytes and presents as solid tumors in the lymph nodes or in other lymph tissues. Lymphocytic tumors occur under both normoxic and hypoxic conditions. [0036] The cellular response to hypoxia includes increased glycolysis, inhibited apoptosis, and increased angiogenesis and metastasis, and is mediated through the hypoxia- inducible factor-1 (HIF-1) transcription factor. HIF-1 is a heterodimer of the oxygen regulated HIF- la or HIF-2a and constitutively expressed HIF-Ι β and it activates transcription of a wide variety of genes involved in glycolysis, erythropoiesis, resistance to apoptosis, and promotion of angiogenesis. There is constitutive stabilization of HIF- la in many non-Hodgkin lymphoma (NHL) cell lines and in a significant fraction of diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma patients. Since the levels of HIF- la have been shown to control the activity of HIF-1 and are over-expressed in a range of human tumors, HIF- la is an attractive target for chemotherapy.

[0037] HIF- 1 a protein is found in a wide variety of human primary tumors, but only at very low levels in normal tissue. HIF- la protein levels are regulated in response to oxygen tension. In normal cells during normoxia, HIF- la is minimally expressed because any expressed protein is immediately broken down by the ubiquitin- proteosome system. However, in tumors, tissue hypoxia and cell growth leads to increased HIF activity. Thus, HIF- la is activated in cancer cells, even under normoxic conditions by oncogene products or impaired activity of tumor suppressor genes.

[0038] Oxygen regulates the rate at which HIF- la protein is degraded. In normoxic

conditions, oxygen dependent hydroxylation of proline residues 402 and 564 in HIF- la by the enzymes PHD (prolyl hydroxylase-domain protein) 1-3 is required for the binding of the von Hippel-Lindau (VHL) tumor-suppressor protein. VHL binding is also promoted by acetylation of lysine (K) residue 532 by the ARD1

acetyltransferase. Oxygen also regulates the interaction of HIF- la with

transcriptional co-activators. Oxygen-dependent hydroxylation of aspargine (N) residue 803 in HIF- la by the enzyme FIH-1 (factor inhibiting HIF-1) blocks the binding of p300 and CBP to HIF- la and therefore inhibits HIF- la mediated gene transcription. Under hypoxic conditions, the rate of aspargine and proline hydroxylation decreases therefore VHL cannot bind to HIF- la, resulting in a decreased rate of HIF- la degradation. p300 and CBP can bind to HIF- la allowing transcriptional activation of HIF-1 target genes. [0039] Because of the role of HIF-Ια in regulating the response of growing tumors to hypoxia it is a very important target for anticancer drug development. The importance of HIF-Ια to lymphomas is demonstrated by the PX-478 mediated dose- dependent downregulation of HIF-Ια protein levels in DLBCL cells (FIG. 2). There was no change in HIF-2a protein levels in either hypoxic or normoxic conditions. Furthermore, lower concentrations of PX-478 were needed for effective HIF-la downregulation in normoxia. In normoxic conditions, lower concentrations of PX- 478 are needed to regulate HIF-la protein levels in lymphocytes than needed in hypoxic conditions.

[0040] Accordingly, provided herein are methods of treatment of lymphomas with PX-478, or salts, metabolites, analogs or derivatives thereof. In some embodiments, the methods provided herein lead to a decrease in HIF-la protein and/or subsequent trans activation of the HIF-1 complex in a patient in need thereof, leading to decreased levels of downstream targets. In some embodiments, the methods of treatment provided herein lead to inhibition of thioredoxin-reductase in

lymphocytes.

Certain terminology

[0041] Optical Isomers— Diastereomers— Geometric Isomers— Tautomers. Compounds described herein may contain an asymmetric center and may thus exist as enantiomers. Where the compounds described herein possess two or more asymmetric centers, they may additionally exist as diastereomers. Contemplated within the scope of embodiments presented herein are all such possible

stereoisomers as substantially pure resolved enantiomers, racemic mixtures thereof, as well as mixtures of diastereomers. The formulas are shown without a definitive stereochemistry at certain positions. Contemplated within the scope of embodiments presented herein are all stereoisomers of such formulas and pharmaceutically acceptable salts thereof. Diastereoisomeric pairs of enantiomers is separated by, for example, fractional crystallization from a suitable solvent, and the pair of enantiomers thus obtained is separated into individual stereoisomers by conventional means, for example by the use of an optically active acid or base as a resolving agent or on a chiral HPLC column. Further, any enantiomer or diastereomer of a compound of the general formula is obtained by stereospecific synthesis using optically pure starting materials or reagents of known configuration. [0042] It must also be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to a "fibroblast" is a reference to one or more fibroblasts and equivalents thereof known to those skilled in the art, and so forth.

[0043] As used herein, the term "about" means plus or minus 10% of the numerical value of the number with which it is being used. Therefore, about 50% means in the range of 45%-55%.

[0044] The term "inhibiting" includes the administration of a compound described herein to prevent the onset of the symptoms, alleviating the symptoms, or eliminating the disease, condition or disorder.

[0045] By "pharmaceutically acceptable", it is meant the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0046] As used herein, the term "therapeutic" means an agent utilized to treat, combat, ameliorate, prevent or improve an unwanted condition or disease of a patient. In some embodiments, a therapeutic agent provided herein is useful for the treatment of cancer or the decrease in proliferation of cells.

[0047] A "therapeutically effective amount" or "effective amount" of a composition is a predetermined amount calculated to achieve the desired effect, e.g., to inhibit, block, or reverse the activation, migration, or proliferation of cells. The activity contemplated by the present methods includes both medical therapeutic and/or prophylactic treatment, as appropriate. The specific dose of a compound administered described herein to obtain therapeutic and/or prophylactic effects will be determined by the particular circumstances surrounding the case, including, for example, the compound administered, the route of administration, and the condition being treated. Specific modes of administration will depend on the indication. The selection of the specific route of administration and the dose regimen is to be adjusted or titrated by the clinician according to methods known to the clinician in order to obtain the optimal clinical response. The amount of compound to be administered is that amount which is therapeutically effective. The dosage to be administered will depend on the characteristics of the subject being treated, e.g., the particular animal treated, age, weight, health, types of concurrent treatment, if any, and frequency of treatments, and can be easily determined by one of skill in the art (e.g., by the clinician).

[0048] The compounds are effective over a wide dosage range and, for example, dosages per day will normally fall within the range of from 0.001 to 10 mg/kg, more usually in the range of from 0.01 to 1 mg/kg. However, it will be understood that the effective amount administered will be determined by the physician in the light of the relevant circumstances including the condition to be treated, the choice of compound to be administered, and the chosen route of administration, and therefore the above dosage ranges are not intended to limit the scope of embodiments presented herein in any way. A therapeutically effective amount of a compound described herein is typically an amount such that when it is administered in a physiologically tolerable excipient composition, it is sufficient to achieve an effective systemic concentration or local concentration in the tissue.

[0049] The terms "treat," "treated," or "treating" as used herein, in some embodiments, refers to therapeutic treatment. In other embodiments, the terms "treat," "treated," or "treating" as used herein refer to prophylactic and/or preventative measures. In some embodiments, "treating" slows down (lessen) an undesired physiological condition, disorder or disease (e.g., by delaying metastasis), to obtain beneficial or desired clinical results. In some embodiments, "treating" allows for beneficial or desired clinical results including, but not limited to, alleviation of symptoms;

diminishment of the extent of the condition, disorder or disease; stabilization (i.e., not worsening) of the state of the condition, disorder or disease; delay in onset or slowing of the progression of the condition, disorder or disease; amelioration of the condition, disorder or disease state; and remission (whether partial or total), whether detectable or undetectable, or enhancement or improvement of the condition, disorder or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment.

[0050] As used herein, "lymphoma" includes, but is not limited to, Hodgkin's lymphoma,

Non-Hodgkin's lymphoma, nodal marginal zone B cell lymphoma (NMZL), mantle cell lymphoma, diffuse large B cell lymphoma, plasma cell neoplasms, chronic lymphocytic leukemia, small lymphocytic lymphoma, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, splenic marginal zone lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, primary effusion lymphoma, T cell prolymphocytic leukemia, T cell large granular lymphocytic leukemia, aggressive NK cell leukemia, adult T cell

leukemia/lymphoma, extranodal NK/T cell lymphoma, enteropathy -type T cell lymphoma, hepatosplenic T cell lymphoma, blastic NK cell lymphoma, mycosis fungoides, sezary syndrome, primary cutaneous CD30-positive T cell

lymphoproliferative disorders, primary cutaneous anaplastic large cell lymphoma, primary CNS lymphoma, lymphomatoid papulosis, angioimmunoblastic T cell lymphoma, peripheral T cell lymphoma, follicular lymphoma, Burkitt's lymphoma, gastric lymphoma, cutaneous T Cell lymphoma, anaplastic large cell lymphoma, and MALT lymphoma.

Methods

[0051] Provided herein are methods of treating lymphoma comprising administering a therapeutically effective amount of S-2-amino-3-[4'-N,N,-bis(2- chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride (PX-478) to a patient in need thereof. Further provided herein are methods of treating lymphoma comprising administering a therapeutically effective amount of S-2-amino-3-[4'- N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, to a patient in need thereof.

[0052] Provided herein are methods of inhibiting angiogenic growth in a patient in need thereof comprising administering a therapeutically effective amount of S-2-amino-3- [4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, to a patient in need thereof, wherein the angiogenic growth is associated with lymphoma and/or leukemia.

[0053] Provided herein are methods of inhibiting HIF-1 in a patient in need thereof

comprising administering a therapeutically effective amount of S-2-amino-3-[4'- N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, to the patient in need thereof. Provided herein are methods of treating a HIF-1 alpha - mediated disease in a patient in need thereof comprising administering a therapeutically effective amount of S-2-amino-3-[4'- N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, to the patient in need thereof. [0054] Provided herein are methods of inhibiting tumor formation in a patient in need thereof comprising administering a therapeutically effective amount of S-2-amino-3- [4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, to the patient in need thereof, wherein the tumor formation is associated with lymphoma and/or leukemia.

[0055] Provided herein are methods of stabilizing disease or slowing progression of disease (e.g., metastasis) in a patient in need thereof (e.g., a patient suffering from lymphoma) comprising administering a therapeutically effective amount of S-2- amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, to the patient in need thereof.

[0056] Provided herein are methods of treating Hodgkins lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N- oxide, or salts, metabolites, analogs or derivatives thereof.

[0057] Provided herein are methods of treating diffuse large B-cell lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof.

[0058] Provided herein are methods of treating follicular lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N- oxide, or salts, metabolites, analogs or derivatives thereof.

[0059] Provided herein are methods of treating non-Hodgkin lymphoma in a patient

comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof.

[0060] Provided herein are methods of treating Burkitt's lymphoma in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of S-2-amino-3-[4'-N,N,-/?«(2-chloroethyl)amino]phenyl propionic acid N- oxide, or salts, metabolites, analogs or derivatives thereof.

[0061] In certain embodiments, S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride (PX-478) is administered to a patient suffering from any lymphoma or leukemia described herein. In certain embodiments, a metabolite of PX-478 (e.g., 2-amino-3-(4-(2- chloroethylamino)phenyl)propanoic acid) is administered to a patient suffering from any lymphoma or leukemia as described herein. In certain embodiments, an analog or derivative of PX-478 (e.g., a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC, or Formula IV, as described herein) is administered to a patient suffering from lymphoma or leukemia as described herein.

[0062] In a still further embodiment, administration of S-2-amino-3-[4'-N,N,-bis(2- chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride (PX-478), or salts, metabolites, analog or derivatives thereof inhibits angiogenic growth or tumor formation in a patient suffering from lymphoma. In some of such embodiments, administration of a metabolite of PX-478 (e.g., 2-amino-3-(4-(2- chloroethylamino)phenyl)propanoic acid) inhibits angiogenic growth or tumor formation in a patient suffering from lymphoma.

[0063] In certain embodiments, a therapeutically effective dose of S-2-amino-3-[4'-N,N,- bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salt, metabolite, analog or derivative thereof, provides therapeutic benefit without affecting HIF-2a protein levels. In some instances, a lower dosage of S-2-amino-3-[4'-N,N,-bis(2- chloroethyl)amino]phenyl propionic acid N-oxide, or salt, metabolite, analog or derivative thereof, is needed to regulate HIF- la protein levels under normoxic conditions compared to the dosage needed in hypoxic conditions..

f 64} Provided herein are methods for treatment of lymphoma using pharmaceutical

formulations comprising S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof. The formulations comprise the compound together with one or more of a

pharmaceutically acceptable carrier, a diluent, an aqueous solution, an adjuvant, or any other pharmaceutically acceptable excipient as described herein.

[0065] In any of the foregoing embodiments, an active metabolite or active degradation product of PX-478 is utilized alone or in combination with PX-478 in the composition and methods disclosed herein.

[0066] In any of the embodiments described above, the lymphoma is selected from

Hodgkin's lymphoma, Non-Hodgkin's lymphoma, nodal marginal zone B cell lymphoma (NMZL), mantle cell lymphoma, diffuse large B cell lymphoma, plasma cell neoplasms, chronic lymphocytic leukemia, small lymphocytic lymphoma, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, splenic marginal zone lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, primary effusion lymphoma, T cell prolymphocytic leukemia, T cell large granular lymphocytic leukemia, aggressive NK cell leukemia, adult T cell leukemia/lymphoma, extranodal NK/T cell lymphoma, enteropathy -type T cell lymphoma, hepatosplenic T cell lymphoma, blastic NK cell lymphoma, mycosis fungoides, sezary syndrome, primary cutaneous CD30-positive T cell

lymphoproliferative disorders, primary cutaneous anaplastic large cell lymphoma, primary CNS lymphoma, lymphomatoid papulosis, angioimmunoblastic T cell lymphoma, peripheral T cell lymphoma, follicular lymphoma, Burkitt's lymphoma, gastric lymphoma, cutaneous T Cell lymphoma, anaplastic large cell lymphoma, and MALT lymphoma.

[0067] In any of the foregoing embodiments, the lymphoma, or tumor cells thereof, is under normoxic conditions. In any of the foregoing embodiments, the lymphoma, or tumor cells thereof, is under hypoxic conditions. As discussed herein, the therapeutically effective amount of PX-478, salts, metabolites, analogs or derivatives thereof, is dependent upon whether the lymphoma or tumor associated therewith is under normoxic or hypoxic conditions.

Compounds

[0068] The structure of S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride is:

[0069] Also contemplated within the scope of the embodiments presented herein are other salts, metabolites, analogs and/or derivatives of S-2-amino-3-[4'-N,N,-bis(2- chloroethyl)amino]phenyl propionic acid N-oxide. The acids which are used to prepare the salts of the aforementioned compounds are those which form non-toxic pharmaceutically acceptable salts, i.e., salts containing pharmacologically acceptable anions, such as the hydrochloride, acetate, trifluoroacetic acid, tosylate, picrate, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, ethanesulfonate,

benzenesulfonate, p-toluenesulfonate and pamoate salts.

[0070] Encompassed within the scope of embodiments presented herein are other physical forms of S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N- oxide, or salts, metabolites, analogs or derivatives thereof, including, but not limited to, crystals, polymorphs, clathrates, solvates, hydrates, amorphous forms, cocrystals, and anhydrous forms.

[0071] Provided herein, in some embodiments, are salts, metabolites, analogs or derivatives of S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, that are compounds of Formula I:

Formula I;

wherein:

A is an aryl or heteroaryl ring;

R ¹ and R ² are each, independently H, -Ci-Ce alkyl, -Ci-Ce heteroalkyl, -(L) _m -(C3-Cio cycloalkyl), -(L) _m -(C ₂ -C ₁ o heterocycloalkyl); and wherein each alkyl, heteroalkyl, cycloalkyl, and heterocycloalkyl, is independently, at each occurrence, optionally substituted with 1-3 substituents selected from R ⁵ ;

or R ¹ and R ² taken together with the nitrogen atom to which they are attached form a C ₂ -C ₁₀ heterocycloalkyl ring wherein the ring optionally comprises one or more moieties selected from O, C=0, S(0) _m , NR ⁸ S(0) _m , NR ⁸ (C=0) and -R ⁸ , and wherein the ring is optionally substituted with 1-3 substituents selected from R ^a ; or R ² is attached to an adjacent carbon atom on ring A to form a saturated, partially saturated or unsaturated 5-6 member ring that is fused to ring A, and wherein the 5-6 membered ring that is fused to ring A is optionally substituted with 1-3 substituents selected from R ⁵ ; R ³ and R ⁴ are each independently, at each occurrence, H, -Ci-Ce alkyl, halo, or -OH; each R ⁵ is independently, at each occurrence, halo, -CN, -N0 ₂ , -(L) _m -OR ⁶ , -(L) _m - SR ⁷ , -(L) _m -S(=0)R ⁷ , -(L) _m -S(=0) ₂ R ⁷ , -(L) _m -NHS(=0) ₂ R ⁷ , -(L) _m -C(=0)R ⁷ , - (L) _m -OC(=0)R ⁷ , -(L) _m -C0 ₂ R ⁶ , -(L) _m -OC0 ₂ R ⁶ , -(L) _m -CH(R ⁶ ) ₂ , -(L) _m -N(R ⁶ ) ₂ , - (L) _m -C(=0)N(R ⁶ ) ₂ , -(L) _m -OC(=0)N(R ⁶ ) ₂ , -(L) _m -NHC(=0)NH(R ⁶ ), -(L) _m - NHC(=0)R ⁷ , -(L) _m -NHC(=0)OR ⁷ , -(L) _m -C(OH)(R ⁶ ) ₂ , -(L) _m -C(NH ₂ )(R ⁶ ) ₂ , -d- C ₆ alkyl, -Ci-C ₆ fluoroalkyl or -Ci-C ₆ heteroalkyl;

each R ⁶ is independently, at each occurrence, H, a Ci-C ₆ alkyl, a Ci-C ₆ fluoroalkyl, a Ci-C ₆ heteroalkyl, a C3-Ciocycloalkyl, a C ₂ -Cioheterocycloalkyl, an aryl, a heteroaryl, -Ci-C4alkyl-(C3-Ciocycloalkyl), -Ci-C4alkyl-(C ₂ - Cioheterocycloalkyl), -Ci-C4alkyl-(aryl), or -Ci-C4alkyl-(heteroaryl), and wherein each alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently, at each occurrence, optionally substituted with 1-3 substituents selected from R ^a ; or

two R ⁶ groups attached to the same N atom are taken together with the N atom to which they are attached to form a C ₂ -Cioheterocycloalkyl wherein the ring optionally comprises one or more moieties selected from O, C=0, S(0) _m , NR ⁸ S(0) _m , NR ⁸ (C=0) and N-R ⁸ , and wherein the ring is optionally substituted with 1 -3 substituents selected from R ^a ;

each R ⁷ is independently, at each occurrence, a Ci-C ₆ alkyl, a Ci-C ₆ fluoroalkyl, a Ci- Ceheteroalkyl, a C3-Ciocycloalkyl, a C ₂ -Cioheterocycloalkyl, an aryl, a heteroaryl, -Ci-C4alkyl-(C3-Ciocycloalkyl), -Ci-C4alkyl-(C ₂ - Cioheterocycloalkyl), -Ci-C4alkyl-(aryl), or -Ci-C4alkyl-(heteroaryl), and wherein each alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently, at each occurrence, optionally substituted with 1-3 substituents selected from R ^a ;

each R ⁸ is independently, at each occurrence, H, Ci-Ce alkyl, Ci-Ce heteroalkyl or - C(=0)-C ₁ -C6 alkyl, and wherein each alkyl, and heteroalkyl is independently, at each occurrence, optionally substituted with 1 -3 substituents selected from R ^a ; each R ^a is independently, at each occurrence, oxo, halo, -CN, -N0 ₂ , -(L) _m -0-(Ci- C ₃ alkyl), -(L) _m -S-(C ₁ -C ₃ alkyl), -(L) _m -S(=0) -(C ₁ -C ₃ alkyl), -(L) _m -S(=0) ₂ -(d- C ₃ alkyl), -(L) _m -NHS(=0) ₂ -(C ₁ -C ₃ alkyl), -(L) _m -C(=0) -(Ci-C ₃ alkyl), -(L) _m - OC(=0) -(d-Csalkyl), -(L) _m -C0 ₂ -(Ci-C ₃ alkyl), -(LVOC0 ₂ -(Ci-C ₃ alkyl), - (L) _m -CH(C ₁ -C ₃ alkyl) ₂ , -(L) _m -N(C ₁ -C ₃ alkyl) ₂ , -(L) _m -C(=0)N(C ₁ -C ₃ alkyl) ₂ , -(L) _m - OC(=0)N(C ₁ -C ₃ alkyl) ₂ , -(L) _m -NHC(=0)NH(C ₁ -C ₃ alkyl), -(L) _m -NHC(=0) ) - (Ci-C ₃ alkyl), -(L) _m -NHC(=0)0-(C ₁ -C ₃ alkyl), -(L) _m -C(OH)(Ci-C ₃ alkyl) ₂ , -(L) _m - C(NH ₂ )( Ci-C ₃ alkyl) ₂ , -Ci-C ₆ alkyl, -Ci-C ₆ fluoroalkyl, or -Ci-C ₆ heteroalkyl;

L is independently, at each occurrence, a bivalent radical selected from -(Ci-C ₃ alkylene) _m -, -(Q-C7 cycloalkylene), -(Ci-C ₃ alkylene) _m -0-(Ci-C ₃ alkylene) _m -, and -(Ci-C ₃ alkylene) _m -NH-(Ci-C ₃ alkylene) _m -;

each m is independently, at each occurrence, 0, 1 or 2;

n is 1, 2 or 3; and

p is 0, 1, 2, or 3;

or pharmaceutically acceptable salt, N-oxide, solvate or prodrug thereof.

[0072] In some embodiments, a compound of Formula I is a compound having the structure of Formula II:

Formula II.

[0073] In some embodiments, a compound of Formula II has the structure of Formula III:

Formula III;

wherein:

R ⁹ is CN, halo, -d-C ₃ alkyl, -0-Ci-C ₃ alkyl, -0-d-C ₃ fluoroalkyl or -Ci-C ₃ fluoroalkyl; and

k is 1, 2 or 3. [0074] In some embodiments, the compound of Formula III has the structure of Formula IIIA, Formula IIIB or Formula IIIC:

Formula I Formula IIIB

Formula IIIC.

[0075] In some embodiments, the compound of Fomula I has the structure of Formula IV:

Formula IV.

[0076] In some embodiments, for any compound described above, n is 1. In some

embodiments, for any compound described above, n is 2.

[0077] In some embodiments, for any compound described above, R ³ and R ⁴ are each, independently at each occurrence, H, -C1-C ₃ alkyl, or halo. In some embodiments, for any compound described above, R ³ and R ⁴ are H.

[0078] In some embodiments, for any compound described above, p is 0. In some

embodiments, for any compound described above, p is 1. In some embodiments, for any compound described above, R ⁵ is selected from -OH, CN, halo, -C1-C ₃ alkyl, -

O-C1-C ₃ alkyl, and -O-C1-C ₃ fluoroalkyl.

[0079] In some embodiments, a compound of Formula I is selected from compounds in Table 1.

-23-

[0080] In some embodiments, any compound of Formula I, Formula II, Formula III,

Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is a metabolite, derivative or analog of S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride (PX-478).

Certain Terminology

[0081] Unless otherwise stated, the following terms used in this application, including the specification and claims, have the definitions given below. It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise. Unless otherwise indicated, conventional methods of mass spectroscopy, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology are employed. In this application, the use of "or" or "and" means "and/or" unless stated otherwise. Furthermore, use of the term "including" as well as other forms, such as "include", "includes," and "included," is not limiting. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0082] An "alkyl" refers to an aliphatic hydrocarbon. The alkyl may be saturated or

unsaturated. The alkyl, whether saturated or unsaturated, is a branched alkyl or straight chain alkyl. Typical alkyl groups include, but are in no way limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec -butyl, tertiary butyl, pentyl, neopentyl, hexyl, allyl, but-2-enyl, but-3-enyl, and the like.

[0083] The term "alkylene" refers to a divalent alkyl radical. Any of the above mentioned monovalent alkyl groups may be an alkylene by abstraction of a second hydrogen atom from the alkyl. In one aspect, an alkylene is a Ci-C ₆ alkylene. In another aspect, an alkylene is a Ci-C ₄ alkylene. Typical alkylene groups include, but are not limited to, straight chain alkylenes, e.g., methylene, ethylene, propylene and butylene.

Typical alkylene groups include branched chain alkylenes including, but not limited to, -CH ₂ -, -CH(CH ₃ )-, -C(CH ₃ ) ₂ -, -CH ₂ CH ₂ -, -CH ₂ CH(CH ₃ )-, -CH ₂ C(CH ₃ ) ₂ -, - CH ₂ CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ CH ₂ -, and the like.

[0084] The term "halo" or, alternatively, "halogen" or "halide" means fluoro, chloro, bromo or iodo.

[0085] The term "fluoroalkyl" refers to an alkyl in which one or more hydrogen atoms are replaced by a fluorine atom.

[0086] The term "heteroalkyl" refers to optionally substituted alkyl, alkenyl and alkynyl radicals which have one or more skeletal chain atoms selected from an atom other than carbon, e.g., oxygen, nitrogen (e.g. NH or Nalkyl), sulfur, phosphorus, silicon, or combinations thereof. In some embodiments, one aspect, heteroalkyl refers to an alkyl group in which one of the skeletal atoms of the alkyl is oxygen. In some embodiments, heteroalkyl refers to an alkyl group in which one of the skeletal atoms of the alkyl is NH or Nalkyl. In some embodiments, in another aspect, a heteroalkyl refers to an alkyl group in which one of the skeletal atoms of the alkyl is O or S. Exemplary heteroalkyls include, but are not limited to, -(CH ₂ ) _n O-CH ₃ , - (CH ₂ ) _n OCH(CH ₃ ) ₂ , -CH(CH ₃ )0-(CH ₂ ) _n -CH ₃ , -C(CH ₃ ) ₂ 0-CH ₃ , -(CH ₂ ) _n S-CH ₃ , - (CH ₂ ) _n SCH(CH ₃ ) ₂ , -CH(CH ₃ )S-(CH ₂ ) _n -CH ₃ , -CH(CH ₃ )S0 ₂ -(CH ₂ ) _n -CH ₃ , - C(CH ₃ ) ₂ S0 ₂ -CH ₃ , -CH ₂ NH-(Ci-C ₆ alkyl), -C(CH ₃ ) ₂ NH-(Ci-C ₆ alkyl), -CH(CH ₃ )NH- (Ci-C ₆ alkyl) ₂ , In certain embodiments, the heteroatom(s) is placed at any interior position of the heteroalkyl group. Examples include, but are not limited to, -CH ₂ -0- CH ₃ , -CH ₂ -CH ₂ -0-CH ₃ , -CH ₂ -NH-CH ₃ , -CH ₂ -CH ₂ -NH-CH ₃ , -CH ₂ -N(CH ₃ )-CH ₃ , - CH ₂ -CH ₂ -NH-CH ₃ , -CH ₂ -CH ₂ -N(CH ₃ )-CH ₃ , -0-CH ₂ -CH ₂ -N(CH ₃ )-CH ₃ , -CH ₂ -S- CH ₂ -CH ₃ , -CH ₂ -CH ₂ ,-S(0)-CH ₃ , -CH ₂ -CH ₂ -S(0) ₂ -CH ₃ , -0-CH ₂ -CH ₂ -S(0) ₂ -CH ₃ ,- CH=CH-0-CH ₃ , -Si(CH ₃ ) ₃ , -CH ₂ -CH=N-OCH ₃ , and -CH=CH-N(CH ₃ )-CH ₃ . In some embodiments, up to two heteroatoms are consecutive, such as, by way of example, -CH ₂ -NH-OCH ₃ and -CH ₂ -0-Si(CH ₃ ) ₃ .

[0087] The term "optionally substituted" or "substituted" means that the referenced group may be substituted with one or more additional group(s) individually and independently selected from R ⁵ as described herein. In some embodiments, an optional substituent is selected from oxo, halogen, -CN, -NH ₂ , -OH, -NH(CH ₃ ), - N(CH ₃ ) ₂ , alkyl (including straight chain, branched and/or unsaturated alkyl), cycloalkyl, heterocycloalkyl, fluoroalkyl, heteroalkyl, alkoxy, fluoroalkoxy, -S- alkyl,-S(=0) ₂ alkyl, -C(=0)NH(alkyl or phenyl), -C(=0)N(alkyl) ₂ , - OC(=0)N(alkyl) ₂ , -NHC(=0)NH(alkyl or phenyl), -NHC(=0)alkyl, - N(alkyl)C(=0)alkyl, -NHC(=0)0(alkyl), -C(OH)(alkyl) ₂ , and -C(NH ₂ )(alkyl) ₂ . In some embodiments, an optional substituent is selected from oxo, fluorine, chlorine, bromine, iodine, -CN, -NH ₂ , -OH, -NH(CH ₃ ), -N(CH ₃ ) ₂ , -CH ₃ , -CH ₂ CH ₃ , - CH(CH ₃ ) ₂ , -CF ₃ , -CH ₂ CF ₃ , -OCH ₃ , -OCH ₂ CH ₃ , -OCH(CH ₃ ) _2; -OCF ₃ , - OCH ₂ CF ₃ , - S(=0) ₂ -CH ₃ , -C(=0)NH ₂ , -C(=0)-NHCH ₃ , -NHC(=0)NHCH ₃ , -C(=0)CH ₃ , - C(=0)OH. In some embodiments, substituted groups are substituted with one, two or three of the preceding groups. In some embodiments, substituted groups are substituted with one or two of the preceding groups. In some embodiments, substituted groups are substituted with one of the preceding groups.

[0088] As used herein, the term "aryl" refers to an aromatic ring wherein each of the atoms forming the ring is a carbon atom. Aryl rings described herein include rings having five, six, seven, eight, nine, or more than nine carbon atoms. Aryl groups are optionally substituted. Examples of aryl groups include, but are not limited to phenyl, and naphthalenyl.

[0089] The terms "heteroaryl" or, alternatively, "heteroaromatic" refers to an aryl group that includes one or more ring heteroatoms selected from nitrogen, oxygen and sulfur. An N-containing "heteroaromatic" or "heteroaryl" moiety refers to an aromatic group in which at least one of the skeletal atoms of the ring is a nitrogen atom. In certain embodiments, heteroaryl groups are monocyclic or polycyclic. Illustrative examples of heteroaryl groups include the following moieties:

and the like.

[0090] The term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic radical, wherein each of the atoms forming the ring (i.e. skeletal atoms) is a carbon atom. In various embodiments, cycloalkyls are saturated, or partially unsaturated. In some embodiments, cycloalkyls are fused with an aromatic ring. Cycloalkyl groups include groups having from 3 to 10 ring atoms. Illustrative examples of cycloalkyl groups include, but are not limited to, the following moieties:

and the like. Monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Dicylclic cycloalkyls include, but are not limited to tetrahydronaphthyl, indanyl, tetrahydropentalene or the like. Polycyclic cycloalkyls include admantane, norbornane or the like. The term cycloalkyl includes "unsaturated nonaromatic carbocyclyl" or "nonaromatic unsaturated carbocyclyl" groups both of which refer to a nonaromatic carbocycle, as defined herein, that contains at least one carbon carbon double bond or one carbon carbon triple bond.

[0091] The term "heterocycloalkyl" refers to heteroalicyclic groups containing one to four ring heteroatoms each selected from O, S and N. In certain instances, each heterocycloalkyl group has from 4 to 10 atoms in its ring system, and with the proviso that the ring of said group does not contain two adjacent O or S atoms. In some embodiments, heterocycloalkyls are fused with an aromatic ring. An example of a 3-membered heterocycloalkyl group is aziridine. Examples of 4-membered heterocycloalkyl groups include azetidine, a beta lactam and the like. Examples of 5-membered heterocycloalkyl groups include pyrrolidine, oxazolidine,

thiazolidinedione and the like. Examples of 6-membered heterocycloalkyl groups include and are not limited to piperidine, morpholine, piperazine and the like. Other examples of heterocycloalkyl groups are

and the like.

Combination therapy

[0092] In some embodiments, any compound (e.g., a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV) and/or composition thereof described herein is administered to an individual in need thereof (e.g., an individual suffering from cancer) in combination with one or more therapeutic agents. In some embodiments, the one or more therapeutic agents is a chemotherapeutic agent (e.g., an alkylating agent, an antimetabolite, a cytotoxic agent, an apoptotic agent, and the like). In some embodiments, the one or more therapeutic agents is an antibody. In some embodiments, a compound described herein (e.g., a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV) is administered in combination with radiation therapy (e.g., involved-field radiation therapy, extended field radiation therapy, radiotherapy and the like). In some embodiments, a compound described herein (e.g., a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV) is administered in combination with stem cell therapy. In some embodiments, a compound described herein (e.g., a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV) is administered in combination with an antisense therapeutic agent.

[0093] In some embodiments, a compound described herein is administered in combination with a currently used therapeutic regimen. Examples of currently used therapeutic regimens include a CHOP regimen (cyclophosphamide (also called

Cytoxan/Neosar), Doxorubicin (or Adriamycin), Vincristine (Oncovin) and Prednisolone), a R-CHOP regimen (Rituxan-CHOP), a ABVD regimen (Adriamycin (doxorubicin), Blenoxane (bleomycin), Velban (vinblastine), DTIC (dacarbazine)), a BEACOPP regimen (Cytoxan/Neosar (Cyclophosphamide), Adriamycin/Rubex (Doxorubicin), VePesid (Etoposide), Matulane (Procarbazine), Prednisone, Blenoxane (Bleomycin), Oncovin (Vincristine)), a Stanford V regimen

(Adriamycin/Rubex (Doxorubicin), Velban (Vinblastine), Mustargen

(Mechloroethamine/ Nitrogen Mustard), VePesid (Etoposide), Blenoxane

(Bleomycin), Oncovin (Vincristine), Prednisone, and radiation), and the like. By way of example, in some embodiments, a compound described herein is used in combination with any one of the aforementioned regimens for treatment of individuals suffering from hematological cancers (e.g., lymphoma, leukemia, myeloma and the like).

[0094] Examples of antibodies suitable for use in combination with a compound described herein include and are not limited to Rituxan, Zevalin (Ibritumomab Tiuxetan),

Bexxar (Tositumomab), Gemtuzumab (Mylotarg), alemtuzumab, trastuzumab, bortezomib (velcade), nimotuzumab, cetuximab, bevacizumab, and the like. In some embodiments, antibodies are specific to epitopes on cancer cells. In some embodiments, antibodies are tagged with radioactive isotopes and deliver radiation at the targeted site.

[0095] Examples of chemotherapeutic agents suitable for use in combination with any

compound described herein (e.g., a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV) include and are not limited to methotrexate (RHEUMATREX®, Amethopterin) cyclophosphamide (CYTOXAN®), thalidomide (THALIDOMID®), acridine carboxamide, actimid®, actinomycin, 17-N-allylamino-17-demethoxygeldanamycin, aminopterin, amsacrine, anthracycline, antineoplastic, antineoplaston, 5-azacytidine, azathioprine, BL22, bendamustine, biricodar, bleomycin, bortezomib, bryostatin, busulfan, calyculin, camptothecin, capecitabine, carboplatin, chlorambucil, cisplatin, cladribine, clofarabine, cytarabine, dacarbazine, dasatinib, daunorubicin, decitabine, dichloroacetic acid, discodermolide, docetaxel, doxorubicin, epirubicin, epothilone, eribulin, estramustine, etoposide, exatecan, exisulind, ferruginol, floxuridine, fludarabine, fluorouracil, fosfestrol, fotemustine, ganciclovir, gemcitabine, hydroxyurea, IT- 101, idarubicin, ifosfamide, imiquimod, irinotecan, irofulven, ixabepilone, laniquidar, lapatinib, lenalidomide, lomustine, lurtotecan, mafosfamide, masoprocol, mechlorethamine, melphalan, mercaptopurine, mitomycin, mitotane, mitoxantrone, nelarabine, nilotinib, oblimersen, oxaliplatin, PAC-1, paclitaxel, pemetrexed, pentostatin, pipobroman, pixantrone, plicamycin, procarbazine, proteasome inhibitors (e.g., bortezomib), raltitrexed, rebeccamycin, revlimid®, rubitecan, SN-38, salinosporamide A, satraplatin, streptozotocin, swainsonine, tariquidar, taxane, tegafur-uracil, temozolomide, testolactone, thioTEPA, tioguanine, topotecan, trabectedin, tretinoin, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, zosuquidar, or the like.

[0096] Examples of antisense therapeutic agents include and are not limited to OGX-427 (ISIS), ISIS-EIF4E _Rx , LY2181308 (Lilly), OGX-Ol l/TV-1011 (ISIS), and the like.

Examples of Pharmaceutical Compositions and Methods of Administration

[0097] Pharmaceutical compositions provided herein are administered for therapeutic or prophylactic treatments. A therapeutic amount is an amount sufficient to remedy a disease state or symptoms, or otherwise prevent, hinder, retard, or reverse the progression of disease or any other undesirable symptoms in any way whatsoever. In prophylactic applications, pharmaceutical compositions are administered to a patient susceptible to or otherwise at risk of a particular disease. Hence, a prophylactically effective amount is an amount sufficient to prevent, hinder or retard a disease state or its symptoms.

[0098] Provided herein, in certain embodiments, are compositions comprising a

therapeutically effective amount of any compound described herein (e.g., a compound of any one of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV).

[0099] Provided herein are pharmaceutical compositions that include one or more

compounds of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, and a pharmaceutically acceptable diluent(s), excipient(s), or carrier(s). In addition, a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is optionally administered as pharmaceutical compositions in which it is mixed with other active ingredients, as in combination therapy. In some embodiments, the pharmaceutical compositions includes other medicinal or pharmaceutical agents, carriers, adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure, and/or buffers. In addition, the

pharmaceutical compositions also contain other therapeutically valuable substances.

[00100] A pharmaceutical composition, as used herein, refers to a mixture of a

compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB,

Formula IIIC or Formula IV with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and/or excipients. The pharmaceutical composition facilitates administration of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB,

Formula IIIC or Formula IV to an organism. In practicing the methods of treatment or use provided herein, therapeutically effective amounts of a compound of Formula

I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV are administered in a pharmaceutical composition to a mammal having a condition, disease, or disorder to be treated. Preferably, the mammal is a human. A

therapeutically effective amount varies depending on the severity and stage of the condition, the age and relative health of an individual, the potency of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV used and other factors. The compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is optionally used singly or in combination with one or more therapeutic agents as components of mixtures.

[00101] The compounds described herein are administered in the conventional

manner by any route where they are active, including but not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, buccal, or ocular routes, or intravaginally, by inhalation, by depot injections, or by implants. Thus, modes of administration for the compounds described herein (either alone or in combination with other pharmaceuticals) can be, but are not limited to, sublingual, injectable (including short-acting, depot, implant and pellet forms injected subcutaneously or intramuscularly), or by use of vaginal creams, suppositories, pessaries, vaginal rings, rectal suppositories, intrauterine devices, and transdermal forms such as patches and creams.

[00102] By way of example only, Example 3 a is describes a parenteral formulation,

Example 3 f describes a rectal formulation. The pharmaceutical formulations described herein include, but are not limited to, aqueous liquid dispersions, self- emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate and controlled release formulations.

[00103] The pharmaceutical compositions will include at least one compound of

Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or

Formula IV, as an active ingredient in free-acid or free-base form, or in a pharmaceutically acceptable salt form. In addition, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), as well as active metabolites of these compounds of

Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or

Formula IV having the same type of activity. In some situations, compounds of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV exist as tautomers. All tautomers are included within the scope of the compounds presented herein. Additionally, the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV exists in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. The solvated forms of the compounds of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV presented herein are also considered to be disclosed herein.

[00104] "Carrier materials" include any commonly used excipients in pharmaceutics and should be selected on the basis of compatibility with compounds disclosed herein, such as, a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, and the release profile properties of the desired dosage form. Exemplary carrier materials include, e.g., binders, suspending agents, disintegration agents, filling agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, diluents, and the like. Suitable pharmaceutical vehicles and/or carrier materials also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like, and may also contain wetting agents, emulsifying agents, and/or pH buffering agents. In addition, auxiliary, stabilizing, thickening, lubricating, and/or coloring agents are used. In certain embodiments, the pharmaceutical composition is in the form of a capsule, tablet or other orally administrable formulation.

[00105] In some embodiments, pharmaceutical compositions provided herein

comprise a therapeutically effective amount of S-2-amino-3-[4'-N,N,-bis(2- chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, and in certain embodiments, in purified form, together with a suitable amount of one or more pharmaceutically acceptable vehicles, so as to provide a composition suitable for administration to a patient with lymphoma.

[00106] Pharmaceutical compositions comprising S-2-amino-3-[4'-N,N,-bis(2- chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, are manufactured by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes. Pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers, diluents, excipients, or auxiliaries, which facilitate processing of S-2-amino-3-[4'-N,N,-bis(2- chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, and one or more pharmaceutically acceptable vehicles into formulations that can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. A summary of pharmaceutical

compositions is found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Ea hston, Pa.: Mack Publishing Company, 1995);

Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins, 1999).

[00107] Moreover, the pharmaceutical compositions described herein, which include a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, are formulated into any suitable dosage form, including but not limited to, aqueous oral dispersions, liquids, gels, syrups, elixirs, slurries, suspensions and the like. For oral ingestion by a patient to be treated, solid oral dosage forms, aerosols, controlled release formulations, fast melt formulations, effervescent formulations, lyophilized formulations, tablets, powders, pills, dragees, capsules, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate release and controlled release formulations are also contemplated.

[00108] In some embodiments, a formulation comprising a compound of Formula I,

Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is a solid drug dispersion. A solid dispersion is a dispersion of one or more active ingredients in an inert carrier or matrix at solid state prepared by the melting (or fusion), solvent, or melting-solvent methods. (Chiou and Riegelman, Journal of

Pharmaceutical Sciences, 60, 1281 (1971)). The dispersion of one or more active agents in a solid diluent is achieved without mechanical mixing. Solid dispersions are also called solid-state dispersions. In some embodiments, any compound described herein (e.g., a compound of any one of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV) is formulated as a spray dried dispersion (SDD). An SDD is a single phase amorphous molecular dispersion of a drug in a polymer matrix. It is a solid solution prepared by dissolving the drug and a polymer in a solvent (e.g., acetone, methanol or the like) and spray drying the solultion. The solvent rapidly evaporates from droplets which rapidly solidifies the polymer and drug mixture trapping the drug in amorphous form as an amorphous molecular dispersion. In some embodiments, such amorphous dispersions are filled in capsules and/or constituted into oral powders for reconstitution. Solubility of an SDD comprising a drug is higher than the solubility of a crystalline form of a drug or a non-SDD amorphous form of a drug. In some embodiments of the methods described herein, compounds of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV are administered as SDDs constituted into appropriate dosage forms described herein.

[00109] In certain embodiments, a pharmaceutical composition comprising S-2- amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salts, metabolites, analogs or derivatives thereof, is formulated for oral administration, and in certain embodiments for sustained release oral administration. Oral

pharmaceutical compositions provided herein are optionally in the form of solutions, suspensions, emulsion, tablets, pills, pellets, capsules, capsules containing liquids, powders, sustained-release formulations, suppositories, emulsions, aerosols, sprays, lozenges, aqueous or oily suspensions, granules, suspensions, emulsions, syrups, or elixirs, or any other form suitable for use. In certain embodiments, the formulation is enteric coated, to provide absorption of the compound in the intestine, rather than the stomach.

[00110] Orally administered pharmaceutical compositions may contain one or more optional agents, for example, sweetening agents such as fructose, aspartame or saccharin, flavoring agents such as peppermint, oil of wintergreen, or cherry coloring agents and preserving agents, to provide a pharmaceutically palatable preparation. Moreover, in tablet or pill forms, the pharmaceutical compositions are coated to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over an extended period of time. Selectively permeable membranes surrounding an osmotically active driving compound are also suitable for orally administered compounds and pharmaceutical compositions. In these latter platforms, fluid from the environment surrounding the capsule is imbibed by the driving compound, which swells to displace the agent or agent composition through an aperture. These delivery platforms can provide an essentially zero order delivery profile as opposed to the spiked profiles of immediate release formulations. A time delay material such as glycerol monostearate or glycerol stearate may also be used. Oral pharmaceutical compositions may include standard vehicles such as mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Such vehicles are of pharmaceutical grade.

[00111] For oral liquid preparations such as suspensions, elixirs and solutions, can include suitable carriers, excipients, or diluents include water, saline, alkylene glycols (e.g., propylene glycol), polyalkylene glycols (e.g., polyethylene glycol) oils, alcohols, slightly acidic buffers from about pH 4 to about pH 6 (e.g., acetate, citrate, ascorbate from about 5 mM to about 50 mM), etc. Additionally, flavoring agents, preservatives, coloring agents, bile salts, acylcarnitines, and the like are added.

[00112] S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N- oxide, or salt, metabolite, analog or derivative thereof is formulated in a number of different dosage forms, which can be optionally adapted to provide sustained release S-2-amino-3-[4'-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide, or salt, metabolite, analog or derivative thereof, upon oral administration.

[00113] Pharmaceutical formulations comprising the compounds described herein and a suitable carrier are, in some embodiments, solid dosage forms which include, but are not limited to, tablets, capsules, cachets, pellets, pills, powders and granules; topical dosage forms which include, but are not limited to, solutions, powders, fluid emulsions, fluid suspensions, semi-solids, ointments, pastes, creams, gels and jellies, and foams; and parenteral dosage forms which include, but are not limited to, solutions, suspensions, emulsions, and dry powder. The active ingredients in such formulations are formulated with pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, hydrophobic vehicles, water soluble vehicles, emulsifiers, buffers, humectants, moisturizers, solubilizers, preservatives and the like. The means and methods for administration are described in, for example, Modern Pharmaceutics, Banker & Rhodes, Marcel Dekker, Inc. (1979); and Goodman & Gilman's The Pharmaceutical Basis of Therapeutics, 6th Edition,

MacMillan Publishing Co., New York (1980) can be consulted. [00114] Pharmaceutical preparations for oral use are optionally obtained by mixing one or more solid excipient with a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include, for example, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. If desired, disintegrating agents are added, such as the cross linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[00115] Dragee cores are provided with suitable coatings. For this purpose,

concentrated sugar solutions are generally used, which optionally contain gum arabic, talc, polyvinylpyrrolidone, earbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.

Dyestuffs or pigments are optionally added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.

[00116] In some embodiments, the solid dosage forms disclosed herein are in the form of a tablet, (including a suspension tablet, a fast-melt tablet, a bite- disintegration tablet, a rapid-disintegration tablet, an effervescent tablet, or a caplet), a pill, a powder (including a sterile packaged powder, a dispensable powder, or an effervescent powder) a capsule (including both soft or hard capsules, e.g., capsules made from animal-derived gelatin or plant-derived HPMC, or "sprinkle capsules"), solid dispersion, solid solution, bioerodible dosage form, controlled release formulations, pulsatile release dosage forms, multiparticulate dosage forms, pellets, granules, or an aerosol. By way of example, Example 3b describes a solid dosage formulation that is a capsule. In other embodiments, the pharmaceutical formulation is in the form of a powder. In still other embodiments, the pharmaceutical formulation is in the form of a tablet, including but not limited to, a fast-melt tablet. Additionally, pharmaceutical formulations of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV are optionally administered as a single capsule or in multiple capsule dosage form. In some embodiments, the pharmaceutical formulation is administered in two, or three, or four, capsules or tablets.

[00117] In another aspect, dosage forms include microencapsulated formulations. In some embodiments, one or more other compatible materials are present in the microencapsulation material. Exemplary materials include, but are not limited to, pH modifiers, erosion facilitators, anti-foaming agents, antioxidants, flavoring agents, and carrier materials such as binders, suspending agents, disintegration agents, filling agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, and diluents.

[00118] Exemplary microencapsulation materials useful for delaying the release of the formulations including a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, include, but are not limited to, hydroxypropyl cellulose ethers (HPC) such as Klucel® or Nisso HPC, low-substituted hydroxypropyl cellulose ethers (L-HPC), hydroxypropyl methyl cellulose ethers (HPMC) such as Seppifilm-LC, Pharmacoat®, Metolose SR, Methocel®-E, Opadry YS, PrimaFlo, Benecel MP824, and Benecel MP843, methylcellulose polymers such as Methocel®-A, hydroxypropylmethylcellulose acetate stearate Aqoat (HF-LS, HF-LG,HF-MS) and Metolose®, Ethylcelluloses (EC) and mixtures thereof such as E461, Ethocel®, Aqualon®-EC, Surelease®, Polyvinyl alcohol (PVA) such as Opadry AMB, hydroxyethylcelluloses such as Natrosol®, carboxymethylcelluloses and salts of carboxymethylcelluloses (CMC) such as Aqualon®-CMC, polyvinyl alcohol and polyethylene glycol co-polymers such as Kollicoat IR®, monoglycerides (Myverol), triglycerides (KLX), polyethylene glycols, modified food starch, acrylic polymers and mixtures of acrylic polymers with cellulose ethers such as Eudragit® EPO, Eudragit® L30D-55, Eudragit® FS 30D Eudragit® L100-55, Eudragit® L100, Eudragit® S100, Eudragit® RD100, Eudragit® E100, Eudragit® L12.5, Eudragit® S 12.5, Eudragit® NE30D, and Eudragit® NE 40D, cellulose acetate phthalate, sepifilms such as mixtures of HPMC and stearic acid, cyclodextrins, and mixtures of these materials.

[00119] The pharmaceutical solid oral dosage forms including formulations described herein, which include a compound of Formula I, Formula II, Formula III, Formula

IIIA, Formula IIIB, Formula IIIC or Formula IV, are optionally further formulated to provide a controlled release of the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV . Controlled release refers to the release of the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV from a dosage form in which it is incorporated according to a desired profile over an extended period of time.

Controlled release profiles include, for example, sustained release, prolonged release, pulsatile release, and delayed release profiles. In contrast to immediate release compositions, controlled release compositions allow delivery of an agent to an individual over an extended period of time according to a predetermined profile. Such release rates provide therapeutically effective levels of agent for an extended period of time and thereby provide a longer period of pharmacologic response while minimizing side effects as compared to conventional rapid release dosage forms. Such longer periods of response provide for many inherent benefits that are not achieved with the corresponding short acting, immediate release preparations.

[00120] In other embodiments, the formulations described herein, which include a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, are delivered using a pulsatile dosage form. A pulsatile dosage form is capable of providing one or more immediate release pulses at predetermined time points after a controlled lag time or at specific sites. Pulsatile dosage forms including the formulations described herein, which include a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, are optionally administered using a variety of pulsatile formulations that include, but are not limited to, those described in U.S. Pat. Nos. 5,01 1,692, 5,017,381, 5,229,135, and 5,840,329. Other pulsatile release dosage forms suitable for use with the present formulations include, but are not limited to, for example, U.S. Pat. Nos. 4,871,549, 5,260,068, 5,260,069, 5,508,040, 5,567,441 and 5,837,284.

[00121] For oral administration, the compounds are optionally formulated by

combining compounds described herein with pharmaceutically acceptable carriers.

Such carriers allow for compounds described herein to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use are obtained by adding a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include, but are not limited to, fillers such as sugars, including, but not limited to, lactose, sucrose, mannitol, and sorbitol; cellulose preparations such as, but not limited to, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and

polyvinylpyrrolidone (PVP). If desired, disintegrating agents can be added, such as, but not limited to, the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[00122] Dragee cores can be provided with suitable coatings. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.

Dyestuffs or pigments can be added to the tablets or dragee coatings for

identification or to characterize different combinations of active compound doses.

[00123] Pharmaceutical preparations which can be used orally include, but are not limited to, push- fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push- fit capsules can contain the active ingredients in admixture with filler such as, e.g., lactose, binders such as, e.g., starches, and/or lubricants such as, e.g., talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers can be added. All formulations for oral administration should be in dosages suitable for such administration.

[00124] Liquid formulation dosage forms for oral administration are optionally

aqueous suspensions selected from the group including, but not limited to, pharmaceutically acceptable aqueous oral dispersions, emulsions, solutions, elixirs, gels, and syrups. See, e.g., Singh et al, Encyclopedia of Pharmaceutical

Technology, 2nd Ed., pp. 754-757 (2002). In addition to the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, the liquid dosage forms optionally include additives, such as: (a) disintegrating agents; (b) dispersing agents; (c) wetting agents; (d) at least one preservative, (e) viscosity enhancing agents, (f) at least one sweetening agent, and (g) at least one flavoring agent. In some embodiments, the aqueous dispersions further includes a crystal-forming inhibitor.

[00125] In some embodiments, the pharmaceutical formulations described herein are self-emulsifying drug delivery systems (SEDDS). Emulsions are dispersions of one immiscible phase in another, usually in the form of droplets. Generally, emulsions are created by vigorous mechanical dispersion. SEDDS, as opposed to emulsions or microemulsions, spontaneously form emulsions when added to an excess of water without any external mechanical dispersion or agitation. An advantage of SEDDS is that only gentle mixing is required to distribute the droplets throughout the solution. Additionally, water or the aqueous phase is optionally added just prior to administration, which ensures stability of an unstable or hydrophobic active ingredient. Thus, the SEDDS provides an effective delivery system for oral and parenteral delivery of hydrophobic active ingredients. In some embodiments, SEDDS provides improvements in the bioavailability of hydrophobic active ingredients. Methods of producing self-emulsifying dosage forms include, but are not limited to, for example, U.S. Pat. Nos. 5,858,401, 6,667,048, and 6,960,563.

[00126] For buccal administration, the compositions can take the form of, e.g., tablets or lozenges formulated in a conventional manner. Buccal formulations that include a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB,

Formula IIIC or Formula IV include, but are not limited to, formulations described in, for example, U.S. Pat. Nos. 4,229,447, 4,596,795, 4,755,386, and 5,739, 136. In addition, the buccal dosage forms described herein optionally further include a bioerodible (hydrolysable) polymeric carrier that also serves to adhere the dosage form to the buccal mucosa. The buccal dosage form is fabricated so as to erode gradually over a predetermined time period, wherein the delivery of the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or

Formula IV, is provided essentially throughout. Buccal drug delivery avoids the disadvantages encountered with oral drug administration, e.g., slow absorption, degradation of the active agent by fluids present in the gastrointestinal tract and/or first-pass inactivation in the liver. The bioerodible (hydrolysable) polymeric carrier generally comprises hydrophilic (water-soluble and water-swellable) polymers that adhere to the wet surface of the buccal mucosa. Examples of polymeric carriers useful herein include acrylic acid polymers and co, e.g., those known as "carbomers" (Carbopol®, which may be obtained from B.F. Goodrich, is one such polymer). Other components also be incorporated into the buccal dosage forms described herein include, but are not limited to, disintegrants, diluents, binders, lubricants, flavoring, colorants, preservatives, and the like. For buccal or sublingual administration, the compositions optionally take the form of tablets, lozenges, or gels formulated in a conventional manner. By way of example, Example 3c and Example 3d describe sublingual formulations.

[00127] Suitable intranasal formulations include those described in, for example, U.S.

Pat. Nos. 4,476, 116, 5, 1 16,817 and 6,391,452. Nasal dosage forms generally contain large amounts of water in addition to the active ingredient. Minor amounts of other ingredients such as pH adjusters, emulsifiers or dispersing agents, preservatives, surfactants, gelling agents, or buffering and other stabilizing and solubilizing agents are optionally present.

[00128] For administration by inhalation, the compound of Formula I, Formula II,

Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is optionally in a form as an aerosol, a mist or a powder. Pharmaceutical compositions described herein are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit is determined by providing a valve to deliver a metered amount. Capsules and cartridges of, such as, by way of example only, gelatin for use in an inhaler or insufflator are formulated containing a powder mix of the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV and a suitable powder base such as lactose or starch. By way of example, Example 3e describes an inhalation formulation.

[00129] In transdermal administration, the compounds of the present invention, for example, can be applied to a plaster, or can be applied by transdermal, therapeutic systems that are consequently supplied to the organism. Transdermal formulations of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB,

Formula IIIC or Formula IV are administered for example by those described in U.S.

Pat. Nos. 3,598, 122, 3,598, 123, 3,710,795, 3,731,683, 3,742,951, 3,814,097,

3,921,636, 3,972,995, 3,993,072, 3,993,073, 3,996,934, 4,031,894, 4,060,084, 4,069,307, 4,077,407, 4,201, 211, 4,230, 105, 4,292,299, 4,292,303, 5,336, 168, 5,665,378, 5,837,280, 5,869,090, 6,923,983, 6,929,801 and 6,946, 144. By way of example, Example 3g describes a topical formulation.

[00130] The transdermal formulations described herein include at least three

components: (1) a formulation of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV , (2) a penetration enhancer; and (3) an aqueous adjuvant. In addition, transdermal formulations include components such as, but not limited to, gelling agents, creams and ointment bases, and the like. In some embodiments, the transdermal formulation further includes a woven or non-woven backing material to enhance absorption and prevent the removal of the transdermal formulation from the skin. In other embodiments, the transdermal formulations described herein maintain a saturated or supersaturated state to promote diffusion into the skin.

[00131] In some embodiments, formulations suitable for transdermal administration of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB,

Formula IIIC or Formula IV employ transdermal delivery devices and transdermal delivery patches and are lipophilic emulsions or buffered, aqueous solutions, dissolved and/or dispersed in a polymer or an adhesive. Such patches are optionally constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents. Still further, transdermal delivery of a compound of Formula I, Formula II,

Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is optionally accomplished by means of iontophoretic patches and the like. Additionally, transdermal patches provide controlled delivery of a compound of Formula I,

Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV .

The rate of absorption is optionally slowed by using rate-controlling membranes or by trapping the compound of Formula I, Formula II, Formula III, Formula IIIA,

Formula IIIB, Formula IIIC or Formula IV within a polymer matrix or gel.

Conversely, absorption enhancers are used to increase absorption. An absorption enhancer or carrier includes absorbable pharmaceutically acceptable solvents to assist passage through the skin. For example, transdermal devices are in the form of a bandage comprising a backing member, a reservoir containing the compound of

Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or

Formula IV optionally with carriers, optionally a rate controlling barrier to deliver the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV to the skin of the host at a controlled and predetermined rate over a prolonged period of time, and means to secure the device to the skin.

[00132] Formulations that include a compound of Formula I, Formula II, Formula III,

Formula IIIA, Formula IIIB, Formula IIIC or Formula IV suitable for intramuscular, subcutaneous, or intravenous injection include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents, or vehicles including water, ethanol, polyols (propyleneglycol, polyethylene-glycol, glycerol, cremophor and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity is maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Formulations suitable for subcutaneous injection also contain optional additives such as preserving, wetting, emulsifying, and dispensing agents.

[00133] For intravenous injections, a compound of Formula I, Formula II, Formula

III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is optionally formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. For other parenteral injections, appropriate formulations include aqueous or nonaqueous solutions, preferably with physiologically compatible buffers or excipients.

[00134] The compounds described herein are optionally formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. The compounds can be administered by continuous infusion subcutaneously over a period of about 15 minutes to about 24 hours. Formulations for injection can be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents. In some embodiments, the pharmaceutical composition described herein are in a form suitable for parenteral injection as sterile suspensions, solutions or emulsions in oily or aqueous vehicles, and contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV in water soluble form. Additionally, suspensions of the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV are optionally prepared as appropriate oily injection suspensions.

[00135] In addition to the formulations described previously, the compounds of the present invention can also be formulated as a depot preparation. Such long acting formulations can be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection.

[00136] Depot injections can be administered at about 1 to about 6 months or longer intervals. Thus, for example, the compounds can be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

[00137] In some embodiments, the compound of Formula I, Formula II, Formula III,

Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is administered topically and formulated into a variety of topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams or ointments. Such pharmaceutical compositions optionally contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.

[00138] The compound of Formula I, Formula II, Formula III, Formula IIIA,

Formula IIIB, Formula IIIC or Formula IV is also optionally formulated in rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas, containing conventional suppository bases such as cocoa butter or other glycerides, as well as synthetic polymers such as polyvinylpyrrolidone, PEG, and the like.

[00139] Pharmaceutical compositions of the compounds also can comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as, e.g., polyethylene glycols.

Examples of Methods of Dosing and Treatment Regimens

[00140] A compound of Formula I, Formula II, Formula III, Formula IIIA, Formula

IIIB, Formula IIIC or Formula IV is optionally used in the preparation of medicaments for the prophylactic and/or therapeutic treatment of diseases mediated by HIF-1. In addition, a method for treating any of the diseases or conditions described herein in an individual in need of such treatment, involves administration of pharmaceutical compositions containing at least one compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV described herein, or a pharmaceutically acceptable salt, pharmaceutically acceptable N-oxide, pharmaceutically active metabolite, pharmaceutically acceptable prodrug, or pharmaceutically acceptable solvate thereof, in therapeutically effective amounts to said individual.

[00141] In the case wherein the patient's condition does not improve, upon the

doctor's discretion the administration of the a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is optionally administered chronically and/or at a higher dose, including throughout the duration of the patient's life in order to ameliorate or otherwise control or limit the disease progression.

[00142] In the case wherein the patient's status does improve, upon the doctor's discretion the administration of the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is optionally given continuously; alternatively, the dose of drug being administered is temporarily reduced or temporarily suspended for a certain length of time (i.e., a "drug holiday"). The length of the drug holiday optionally varies between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. The dose reduction during a drug holiday includes from 10%- 100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. [00143] Once improvement of the patient's conditions has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, is reduced, as a function of improvement in a patient, to a level at which the improved disease is retained. In some embodiments, patients require intermittent treatment on a long-term basis upon any recurrence of symptoms and/or condition.

[00144] In some embodiments, the pharmaceutical compositions described herein are in unit dosage forms suitable for single administration of precise dosages. In unit dosage form, the formulation is divided into unit doses containing appropriate quantities of one or more compounds of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV . In some embodiments, the unit dosage is in the form of a package containing discrete quantities of the formulation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. In some embodiments, aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multiple-dose reclosable containers are used, in which case it is typical to include a preservative in the composition. By way of example only, formulations for parenteral injection are presented in unit dosage form, which include, but are not limited to ampoules, or in multi dose containers, with an added preservative.

[00145] The daily dosages appropriate for the a compound of Formula I, Formula II,

Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV are from about 0.01 to 2.5 mg/kg per body weight. An indicated daily dosage in the larger mammal, including, but not limited to, humans, is in the range from about 0.5 mg to about 1000 mg, conveniently administered in divided doses, including, but not limited to, up to four times a day or in extended release form. Suitable unit dosage forms for oral administration include from about 1 to 500 mg active ingredient, from about 1 to 250 mg of active ingredient, or from about 1 to about 100 mg active ingredient. The foregoing ranges are merely suggestive, as the number of variables in regard to an individual treatment regime is large, and considerable excursions from these recommended values are not uncommon. Such dosages are optionally altered depending on a number of variables, not limited to the activity of the compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB,

Formula IIIC or Formula IV used, the condition being treated, the mode of administration, the requirements of an individual, the severity of the disease or condition being treated, and the judgment of the practitioner.

[00146] Toxicity and therapeutic efficacy of such therapeutic regimens are optionally determined in cell cultures or experimental animals, including, but not limited to, the determination of the LD50 (the dose lethal to 50% of the population) and the ED50

(the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index, which is expressed as the ratio between LD50 and ED50. Compounds of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV exhibiting high therapeutic indices are preferred. The data obtained from cell culture assays and animal studies is optionally used in formulating a range of dosage for use in human. The dosage of such a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV lies preferably within a range of circulating concentrations that include the ED50 with minimal toxicity. The dosage optionally varies within this range depending upon the dosage form employed and the route of administration utilized.

Screening assays

[00147] The compounds described herein (e.g., compounds of Formula I, Formula II,

Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV) are screened for activity using suitable cancer cell lines. Typically, the screening assays involve incubation of cells in the presence of test compounds in a suitable culture medium under normoxic (-21 % oxygen) or hypoxic (~ 1.5% oxygen) conditions. The anticancer activity of test compounds is determined by

(1) measuring cell viability in a MTT assay;

(2) measuring expression of HIF-Ια and HIF-2a by Western blotting and/or flow cytometry;

(3) measuring apoptosis by annexin V/PI staining followed by flow cytometry; or any combination thereof.

[00148] Suitable cancer cell lines include and are not limited to the NCI60 human tumor cell line panel (See, Nature Reviews Cancer 2006, 6, 813-823) that includes leukemia, non-small cell lung cancer, colon, CNS, Melanoma, ovarian, renal, prostate and breast cancer cell lines. Examples of cell lines in the NCI60 human tumor cell line include and are not limited to A549, K-562, HT29, SF-268, M14, IGROV1, TK-10, PC-3, MCF7 and the like.

[00149] This invention and embodiments illustrating the method and materials used is further understood by reference to the following non-limiting examples.

EXAMPLE 1

[00150] HIF-1 is a transcription factor that serves as a master regulator of cellular responses to hypoxia and regulates genes required for adaptation to hypoxia.

Although the expression of HIF-la subunit is constitutive, HIF-la protein levels are regulated in response to oxygen tension. Under normoxic conditions, HIF-la is degraded by the proteasome, and HIF-1 remains inactive. In hypoxia, HIF-la is stabilized and forms a complex with HIF- 1 β that allows HIF- 1 to function as a transcription factor. Thus, HIF-la is activated only during hypoxia under normal physiologic conditions. By contrast, HIF-la is frequently activated in cancer cells, including under normoxic conditions by oncogene products or impaired activity of tumor suppressor genes. There is constitutive stabilization of HIF-la in many non- Hodgkin lymphoma (NHL) cell lines as well as among a significant fraction of diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma patients.

Constitutive expression of HIF-la enhances vascularization, increases glucose metabolism, and induces the expression of anti-apoptotic proteins in cancer tissues. HIF-la is thought to be one of the most important molecular targets in the treatment of cancer.

[00151] Methods: We treated several DLBCL cell lines (SUDHL4, SUDHL6, and

SUDHL10) with increasing concentrations (2-25μΜ) of PX-478 for 4 hours followed by 20-hour incubation under normoxic (21% oxygen) or hypoxic (1.5% oxygen) conditions. Cell viability was assessed by MTT assay. Expression of HIF- la and HIF -2a were measured by Western blotting and flow cytometry after preincubation of cells with 5-25μΜ PX-478 followed by 20-hour incubation under normoxia or hypoxia. Apoptosis was measured by annexin V/PI staining followed by flow cytometry.

[00152] Results: Under hypoxic conditions, dose-dependent downregulation of HIF- la protein levels were noted with complete absence of HIF-la by 20μΜ (See FIG. 2) Interestingly, lower concentrations of PX-478 were needed for effective HIF-la downregulation in normoxia. Of note, there was no change in HIF-2a protein levels observed in either normoxic or hypoxic conditions (See Fig. 4). In cell viability studies, time- and dose-dependent cell death of PX-478 was documented in all cell lines with an associated IC50 of 15-20μΜ (See FIG. 3 A, B and C).

[00153] Conclusion: Results support that PX-478 effectively downregulates HIF-la protein at low concentrations and induces cell death in DLBCL cells.

[00154] Compounds 1-34, shown in Table 1, will be tested using the above protocol.

EXAMPLE 2

[00155] Methods: We grew SUDHL4 cells on soft agar and a feeder layer (1 : 1 agarose to RPMI) was plated. After the feeder layer was solidified, SUDHL4 cells were plated at 10,000 cells/mL in a 1 :3 agarose to RPMI mixture. After solidification of the top layer, cells were placed in the incubator and RPMI media was added the same day. Media was changed every 3 days, and colonies (>50 cells) were counted on day 1 1. Colony formation was examined and photomicroscopy of cell colonies was taken at 5x magnification. For results, see FIG. 4.

[00156] Conclusion: Results support that PX-478 inhibits colony formation under hypoxia and normoxia. The fact that PX-478 inhibits cell growth and the proliferation of lymphoma cells under hypoxia as well as normoxia indicates that there is oxygen independent upregulation of HIF-1 a in lymphoma cells.

EXAMPLE 3

Example 3a: Parenteral Composition

[00157] To prepare a parenteral pharmaceutical composition suitable for

administration by injection, 100 mg of a water-soluble salt of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is dissolved in DMSO and then mixed with 10 mL of 0.9% sterile saline. The mixture is incorporated into a dosage unit form suitable for

administration by injection.

Example 3b: Oral Composition

[00158] To prepare a pharmaceutical composition for oral delivery, 100 mg of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB,

Formula IIIC or Formula IV is mixed with 750 mg of starch. The mixture is incorporated into an oral dosage unit for, e.g., a hard gelatin capsule, which is suitable for oral administration.

Example 3c: Sublingual (Hard Lozenge) Composition

[00159] To prepare a pharmaceutical composition for buccal delivery, such as a hard lozenge, mix 100 mg of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV with 420 mg of powdered sugar mixed, with 1.6 mL of light corn syrup, 2.4 mL distilled water, and 0.42 mL mint extract. The mixture is gently blended and poured into a mold to form a lozenge suitable for buccal administration.

Example 3d: Fast-Disintegrating Sublingual Tablet

[00160] A fast-disintegrating sublingual tablet is prepared by mixing 48.5% by weigh of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV, 44.5% by weight of microcrystalline cellulose (KG- 802), 5% by weight of low-substituted hydroxypropyl cellulose (50 μιη), and 2% by weight of magnesium stearate. Tablets are prepared by direct compression (AAPS PharmSciTech. 2006;7(2):E41). The total weight of the compressed tablets is maintained at 150 mg. The formulation is prepared by mixing the amount of compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV with the total quantity of microcrystalline cellulose (MCC) and two-thirds of the quantity of low-substituted hydroxypropyl cellulose (L-HPC) by using a three dimensional manual mixer (lnversina ®, Bioengineering AG, Switzerland) for 4.5 minutes. All of the magnesium stearate (MS) and the remaining one-third of the quantity of L-HPC are added 30 seconds before the end of mixing.

Example 3e: Inhalation Composition

[00161] To prepare a pharmaceutical composition for inhalation delivery, 20 mg of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is mixed with 50 mg of anhydrous citric acid and 100 mL of 0.9% sodium chloride solution. The mixture is incorporated into an inhalation delivery unit, such as a nebulizer, which is suitable for inhalation administration. Example 3f: Rectal Gel Composition

[00162] To prepare a pharmaceutical composition for rectal delivery, 100 mg of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is mixed with 2.5 g of methylcelluose (1500 mPa), 100 mg of methylparapen, 5 g of glycerin and 100 mL of purified water. The resulting gel mixture is then incorporated into rectal delivery units, such as syringes, which are suitable for rectal administration.

Example 3g: Topical Gel Composition

[00163] To prepare a pharmaceutical topical gel composition, 100 mg of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is mixed with 1.75 g of hydroxypropyl celluose, 10 mL of propylene glycol, 10 mL of isopropyl myristate and 100 mL of purified alcohol USP. The resulting gel mixture is then incorporated into containers, such as tubes, which are suitable for topical administration.

Example 3h: Ophthalmic Solution Composition

[00164] To prepare a pharmaceutical opthalmic solution composition, 100 mg of a compound of Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is mixed with 0.9 g of NaCl in 100 mL of purified water and filterd using a 0.2 micron filter. The resulting isotonic solution is then incorporated into ophthalmic delivery units, such as eye drop containers, which are suitable for ophthalmic administration.

Example 3i: Nasal spray solution

[00165] To prepare a pharmaceutical nasal spray solution, 10 g of a compound of

Formula I, Formula II, Formula III, Formula IIIA, Formula IIIB, Formula IIIC or Formula IV is mixed with 30 mL of a 0.05M phosphate buffer solution (pH 4.4). The solution is placed in a nasal administrator designed to deliver 100 μΐ of spray for each application.

EXAMPLE 4

[00166] This study is adapted from an animal model reported by Bittner, C. et al,

Lab Invest 2000, 80: 1523-1531. A murine anaplastic lymphoid cell line TS1G6 (about 10 ⁴ cells) is injected into nude or immunocompetent C57B1/6 mice. Mice develop tumors within 4-8 weeks. Mice are subjected to daily oral treatment with PX-478 or Compound 1 or vehicle for 5 days.

[00167] Tumor weight, tumor size and the ratio of total tumor to total body weight is determined in the treatment group and control group to determine therapeutic efficacy of PX-478 and Compound 1. EXAMPLE 5

[00168] This study will determine the efficacy and safety of Compound 1 in

treatment of relapsed or refractory mantle cell lymphoma, diffuse large B cell lymphoma and multiple myeloma. Patients will be administered Compound 1 orally or intravenously.

[00169] Disease will be assessed every 6 weeks using International Working Group

Response Criteria for Non-Hodgkins Lymphoma in the mantle cell and diffuse large B cell lymphoma patients and the International Myeloma Working Group Response Criteria for multiple myeloma patients. Patients will continue to be treated and disease assessed until disease progression or unacceptable toxicity.

[00170] Duration of response will be determined if disease progression is

documented in patients who have an objective response.

[00171] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein is employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.