WO2010062372A2

WO2010062372A2 - Methods for modulating nf-kb using gibberellins

Info

Publication number: WO2010062372A2
Application number: PCT/US2009/005938
Authority: WO
Inventors: Angela N. Koehler
Original assignee: President And Fellows Of Harvard College
Priority date: 2008-11-03
Filing date: 2009-11-03
Publication date: 2010-06-03
Also published as: WO2010062372A3

Abstract

The present invention provides methods for modulating the NF-κB pathway in a cell using gibberellins such as gibberellic acid. The invention also provides methods for using gibberellins in treating proliferative diseases (e.g., cancer), neurodegenerative diseases, autoimmune diseases, inflammatory diseases, and infectious diseases.

Description

METHODS FOR MODULATING NF-KB USING GIBBERELLINS

Related Applications

[0001] The present application claims priority under 35 U.S. C. § 119(e) to U.S. provisional application, U.S. S.N. 61/110,728, filed November 3, 2008, which is incorporated herein by reference.

Government Support

[0002] This invention was made with U.S. Government support under NOl-CO-12400 awarded by the National Cancer Institute's Initiative for Chemical Genetics. The U.S. Government has certain rights in the invention.

Background of the Invention

[0003] Transcription facts that become overactive in cancers are promising yet untested targets for cancer therapeutics. These proteins mediate the excessive transcription of genes whose products are required for tumor growth and metastasis (Darnell, Natl. Rev. Cancer (2002) 2:740-749; incorporated herein by reference). Directly inhibiting or activating the function of a transcription factor requires specific disruption or recruitment of DNA-protein or protein-protein interactions. The discovery or design of small molecules that specifically disrupt or promote these interactions has thus far proven to be a significant challenge, and this protein class is often perceived to be "undruggable." Transcription factors are cogent targets because many more signaling proteins are affected in cancer than transcription factors, and the myriad of protein-protein interactions in transcriptional complexes may provide selective sites for targeting by small molecules. Compelling arguments have been presented for targeting latent cytoplasmic transcription factors that have increased activity in most human cancers such as the STATs, β-catenin, and NF-κB.

[0004] The mammalian nuclear transcription factor NF-κB is the master immunomodulatory transcription factor, and aberrant NF-κB activity is associated with various cancers as well as inflammatory and autoimmune diseases. NF -KB is a multisubunit complex involved in the activation of gene transcription, including the regulation of apoptosis (Baldwin, Ann. Rev. Immunol. (1996) 14: 649-683). NF-κB exists mainly as a homodimer (p50/p50) or heterodimer (p50/p65) in the cytoplasm in the form of an inactive complex with the inhibitory IKB protein. Many cellular stimuli including antineoplastic agents, viruses (e.g., HIV-I), inflammatory cytokines (e.g., TNF-α, IL-I), phorbol esters, bacterial products (LPS), and oxidative stress result in the IKK-mediated phosphorylation of serines 32 and 36 of IKB, followed by IKB 's ubiquitination and subsequent degradation by the 26S proteasome (Baeuerle and Henkel, Ann. Rev. Immunol. (1994) 12:141-179). Degradation of IKB leads to the release of NF-κB. Upon release, NF-κB translocates into the nucleus where the subunits bind specific DNA control elements and initiate gene transcription. Inhibition of NF-κB- mediated gene transcription can be accomplished through inhibition of phosphorylation of the inhibitory protein IKB, inhibition of IKB ubiquitination, inhibition of IKB degradation, inhibition of NF-κB (p50/p65) nuclear translocation, or the inhibition of NF-κB-DNA binding or NF-κB-mediated DNA transcription. Genes regulated by NF-κB activation include a number of cytokines (e.g., TNF-α, IL-I, IL-2, IL-6, IL-8, iNOS), chemokines, cell adhesion molecules, acute phase proteins, immunoregulatory proteins, eicosanoid metabolizing enzymes, and anti-apoptotic genes.

[0005] NF-κB has been shown to play an important role in many disorders, including cancer and other proliferative disorders, immune disorders (e.g., autoimmune disease and immunodeficiency), inflammatory disorders, neurodegenerative diseases, and sepsis. NF-κB is also implicated in cancers resistant to chemotherapy. There remains a need for compounds that modulate the NF-κB pathway, particularly compounds that directly target NF-κB.

Summary of the Invention

[0006] The present invention stems from the discovery that gibberellins (such as gibberellic acid (GA₃), gibberellin A4 (GA₄), gibberellin A9 methyl ester (GA₉ME), allogibberic acid (AGA), and an ethylene glycol derivative of AGA) are useful as modulators of NF-κB activity and therefore may be useful in treating NF-κB-mediated disorders or studying the NF-κB pathway. A general approach to identifying small molecules that modulate the activity of transcription factors using small-molecule microarrays and functional assays (Koehler et al, J. Am. Chem. Soc. (2003) 125:8420-8241; Vegas et al, Chem. Soc. Rev. (2008) 37: 1385-94; each of which is incorporated herein by reference) was applied to discovering ligands of NF-κB. Using this approach and confirmed by surface plasmon resonance (SPR), gibberellins were discovered to be binders of NF-κB. Gibberellins were also found to inhibit TNFα-stimulated transcription. Therefore, gibberellins may be used as probes of transcription factor function, imaging tools, therapeutic agents, or therapeutic leads. In certain embodiments, the gibberellins directly target the transcription factor itself. The gibberellins as described herein may be used to treat any disease associated with aberrant NF-κB activity, including proliferative diseases such as cancer and benign neoplasms, autoimmune diseases, immunodeficiency disorders, inflammatory diseases, neurodegenerative diseases, infectious diseases, and sepsis. A gibberellin may be combined with other known chemotherapeutic agents (e.g., kinase inhibitors, proteasome inhibitors, HDAC inhibitors) in the treatment of proliferative diseases. Gibberellins may also be used as screening tools, imaging agents, or biological probes. For example, the gibberellins may be used to inhibit the proliferation of cells or modulate the NF-κB pathway in cells including cells in culture as well as cells in a subject (e.g., human, mouse, dog, rat). [0007] In certain embodiments, the invention provides a method for modulating the

NF-κB pathway with a compound of formula I:

wherein

R¹ is hydrogen or a group -OR²⁰, where R²⁰ is hydrogen, a glycoside, Cj_-6 alkyl group, or R¹ together with R² or R¹⁰ forms a bond (Cj-C₂ or C]-Ci₀ double bond, respectively);

R² is hydrogen or a group -OR²¹, where R²¹ is hydrogen, a glycoside; or R² together with R⁴ forms a lactone; or R² together with R¹ or R³ forms a bond (Ci-C₂ or C₂-C₃ double bond, respectively); or R² together with R¹ or R³ and the intervening atoms forms an epoxide ring;

R³ is hydrogen, =0, or -OR² , where R ² is hydrogen or a glycoside; or R³ together with R forms a bond (C₂-C₃ double bond); or R together with R and the intervening atoms forms an epoxide ring;

R⁴ is -OH, or -OR²³, where R²³ is unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, arylalkyl, amidine, -NR²⁴R²⁵ or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur; R²⁴ and R²⁵ may or may not be the same, are hydrogen, or Ci_-20 alkyl, allyl, aryl, arylalkyl or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; or R⁴ together with R²¹ or R²⁸ forms a lactone;

R is hydrogen, a glycoside, unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, or arylalkyl;

R⁶ is hydrogen or -OH; or R⁶ together with R⁷ forms a bond (Cn-Ci₂ double bond); or R⁶ together with R⁷ and the intervening atoms forms an epoxide ring;

R⁷ is hydrogen, =0, or -OR²⁶, where R²⁶ is hydrogen or a glycoside; or R⁷ together with R⁶ forms a bond (Cn-Ci₂ double bond); or R⁷ together with R⁶ and the intervening atoms forms an epoxide ring;

R⁸ is hydrogen, hydroxyl, mercaptan, or halogen, amino, azido, -NR²⁴R²⁵, unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, or arylalkyl, or —OR²⁷, where R²⁷ is hydrogen or a glycoside;

R⁹ is hydrogen or hydroxyl; or R⁹ together with R¹⁵ forms a bond (C₉-C₁₅ double bond); or R together with R¹⁵ and the intervening atoms forms an epoxide ring;

R¹⁰ is hydrogen, methyl, -CHO, -CO₂H, or a glycoside ester of said -CO₂H, -CH₂OR²⁸ or -OR²⁸, where R²⁸ is hydrogen or together with R⁴ forms a lactone; or R¹⁰ together with R¹ forms a bond (Ci-Ci₀ double bond); or R¹⁰ together with R¹ and the intervening atoms forms an epoxide ring;

R¹ ' is hydrogen or hydroxyl or is absent;

R¹² is methyl, -CH₂OH, -CO₂H, or a glycoside ester of said -CO₂H;

R¹³ is methylene, or a divalent heteroatom, or -NR²⁹, where R²⁹ is -NHR³⁰ or -OR³⁰ where R³⁰ is hydrogen, or Ci_-20 alkyl, aryl, alkylaryl; and a double bond is present between Ci₆ and R¹³ when R¹¹ is absent; or R¹³ is hydrogen, hydroxyl, methyl, -CHO, -CH₂X, where X is halogen, -CH₂NR²⁹, where R²⁹ is -NR³⁰ or -OR³⁰, where R³⁰ is hydrogen, or C_-20 alkyl, aryl, or alkylaryl when R¹ ' is hydrogen or hydroxyl;

R¹⁴ is hydrogen or hydroxyl;

R¹⁵ is hydrogen, or R¹⁵ together with R⁹ forms a bond (C₉-Ci₅ double bond); or R¹⁵ together with R⁹ and the intervening atoms forms an epoxide ring; or a pharmaceutically acceptable salt thereof. The compound typically directly affects NF-κB. In certain embodiments, the compound is gibberellic acid (GA₃) of the formula:

[0008] In one aspect, compounds of formula I may be used to inhibit expression of a eukaryotic gene whose transcription is regulated by NF -KB. In another aspect, a compound of formula I may be used to inhibit the expression of a viral gene whose transcription is regulated by NF-κB in a eukaryotic cell. The compounds of formula I are useful in the treatment of cancer and benign neoplasms, autoimmune diseases, immunodeficiency disorders, inflammatory diseases, neurodegenerative diseases, infectious diseases, and sepsis. Pharmaceutical compositions with a therapeutically effective amount of an inventive compound of formula I are also provided by the present invention. Inventive pharmaceutical compositions may optionally include a pharmaceutically acceptable excipient and/or an additional therapeutic agents (e.g., chemotherapeutic agent, anti-inflammatory agent). [0009] In another aspect, the present invention provides methods of screening for compounds that modulate the NF-κB pathway using small molecule microarrays. For example, compounds identified from screening may inhibit, modulate, or bind the p50 subunit, the p65 subunit, or both. In another aspect, the present invention provides methods for identifying inhibitors of NF-κB by competition experiments using gibberellic acid or another gibberellin. The binding of the identified compound to NF -KB may be confirmed by other assays including surface plasmon resonance, a TNFα-stimulated transcription assay, cell viability assays, nuclear translocation assays, DNA-binding assays, and protein-protein interaction assays.

[0010] All patents, published patent applications, and journal articles cited in this application are incorporated herein by reference.

Definitions

[0011] Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75^th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March March 's Advanced Organic Chemistry, 5^th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3^rd Edition, Cambridge University Press, Cambridge, 1987. [0012] The compounds of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)- isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention.

[0013] Where an isomer/enantiomer is preferred, it may, in some embodiments, be provided substantially free of the corresponding enantiomer, and may also be referred to as "optically enriched." "Optically enriched," as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments the compound of the present invention is made up of at least about 90% by weight of a preferred enantiomer. In other embodiments the compound is made up of at least about 95%, 98%, or 99% by weight of a preferred enantiomer. Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts or prepared by asymmetric syntheses. See, for example, Jacques et ah, Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et ah, Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, Tables of Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972).

[0014] It will be appreciated that the compounds of the present invention, as described herein, may be substituted with any number of substituents or functional moieties. In general, the term "substituted" whether preceded by the term "optionally" or not, and substituents contained in formulas of this invention, refer to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. When more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. As used herein, the term "substituted" is contemplated to include substitution with all permissible substituents of organic compounds, any of the substituents described herein (for example, aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, etc.), and any combination thereof (for example, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like) that results in the formation of a stable moiety. The present invention contemplates any and all such combinations in order to arrive at a stable substituent/moiety. Additional examples of generally applicable substitutents are illustrated by the specific embodiments shown in the Examples, which are described herein. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.

[0015] As used herein, substituent names which end in the suffix "-ene" refer to a biradical derived from the removal of two hydrogen atoms from the substitutent. Thus, for example, acyl is acylene; alkyl is alkylene; alkeneyl is alkenylene; alkynyl is alkynylene; heteroalkyl is heteroalkylene, heteroalkenyl is heteroalkenylene, heteroalkynyl is heteroalkynylene, aryl is arylene, and heteroaryl is heteroarylene.

[0016] The term "acyl," as used herein, refers to a group having the general formula -

C(=O)R^X1, -C(=O)OR^X1, -C(=O)-O-C(=O)R^X1, -C(=O)SR^X1, -C(=O)N(R^X1)₂, -C(=S)R^X1, -C(=S)N(R^X1)₂, and -C(=S)S(R^X1), -C(=NR^X1)R^X1, -C(=NR^X1)OR^X1, -C(=NR^X1)SR^X1, and -C(=NR^X1)N(R^X1)₂, wherein R^XI is hydrogen; halogen; substituted or unsubstituted hydroxyl; substituted or unsubstituted thiol; substituted or unsubstituted amino; substituted or unsubstituted acyl, cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched alkyl; cyclic or acyclic, substituted or unsubstituted, branched or unbranched alkenyl; substituted or unsubstituted alkynyl; substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, mono- or di- aliphaticamino, mono— or di- heteroaliphaticamino, mono— or di- alkylamino, mono- or di- heteroalkylamino, mono- or di- arylamino, or mono- or di- heteroarylamino; or two R^xl groups taken together form a 5- to 6- membered heterocyclic ring. Exemplary acyl groups include aldehydes (— CHO), carboxylic acids (-CO₂H), ketones, acyl halides, esters, amides, imines, carbonates, carbamates, and ureas. Acyl substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted). [0017] The term "acyloxy" refers to a "substituted hydroxyl" of the formula (-OR'), wherein R' is an optionally substituted acyl group, as defined herein, and the oxygen moiety is directly attached to the parent molecule.

[0018] The term "aliphatic," as used herein, includes both saturated and unsaturated, nonaromatic, straight chain (i.e., unbranched), branched, acyclic, and cyclic (i.e., carbocyclic) hydrocarbons, which are optionally substituted with one or more functional groups. As will be appreciated by one of ordinary skill in the art, "aliphatic" is intended herein to include, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties. Thus, as used herein, the term "alkyl" includes straight, branched and cyclic alkyl groups. An analogous convention applies to other generic terms such as "alkenyl", "alkynyl", and the like. Furthermore, as used herein, the terms "alkyl", "alkenyl", "alkynyl", and the like encompass both substituted and unsubstituted groups. In certain embodiments, as used herein, "aliphatic" is used to indicate those aliphatic groups (cyclic, acyclic, substituted, unsubstituted, branched or unbranched) having 1-20 carbon atoms. Aliphatic group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted).

[0019] The term "alkyl," as used herein, refers to saturated, straight- or branched- chain hydrocarbon radicals derived from a hydrocarbon moiety containing between one and twenty carbon atoms by removal of a single hydrogen atom. In some embodiments, the alkyl group employed in the invention contains 1-20 carbon atoms. In another embodiment, the alkyl group employed contains 1-15 carbon atoms. In another embodiment, the alkyl group employed contains 1-10 carbon atoms. In another embodiment, the alkyl group employed contains 1—8 carbon atoms. In another embodiment, the alkyl group employed contains 1-5 carbon atoms. Examples of alkyl radicals include, but are not limited to, methyl, ethyl, n- propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, sec-pentyl, iso-pentyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, and the like, which may bear one or more sustitutents. Alkyl group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted).

[0020] The term "alkenyl," as used herein, denotes a monovalent group derived from a straight- or branched-chain hydrocarbon moiety having at least one carbon-carbon double bond by the removal of a single hydrogen atom. In certain embodiments, the alkenyl group employed in the invention contains 2-20 carbon atoms. In some embodiments, the alkenyl group employed in the invention contains 2-15 carbon atoms. In another embodiment, the alkenyl group employed contains 2-10 carbon atoms. In still other embodiments, the alkenyl group contains 2-8 carbon atoms. In yet other embodiments, the alkenyl group contains 2-5 carbons. Alkenyl groups include, for example, ethenyl, propenyl, butenyl, l-methyl-2- buten-1-yl, and the like, which may bear one or more substituents. Alkenyl group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted).

[0021] The term "alkynyl," as used herein, refers to a monovalent group derived from a straight- or branched-chain hydrocarbon having at least one carbon-carbon triple bond by the removal of a single hydrogen atom, hi certain embodiments, the alkynyl group employed in the invention contains 2-20 carbon atoms. In some embodiments, the alkynyl group employed in the invention contains 2-15 carbon atoms. In another embodiment, the alkynyl group employed contains 2-10 carbon atoms. In still other embodiments, the alkynyl group contains 2-8 carbon atoms. In still other embodiments, the alkynyl group contains 2-5 carbon atoms. Representative alkynyl groups include, but are not limited to, ethynyl, 2- propynyl (propargyl), 1-propynyl, and the like, which may bear one or more substituents. Alkynyl group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted).

[0022] The term "amino," as used herein, refers to a group of the formula (-NH₂). A

"substituted amino" refers either to a mono-substituted amine (— NHR^h) of a disubstitued amine (-NR^h ₂), wherein the R^h substituent is any substitutent as described herein that results in the formation of a stable moiety (e.g., a suitable amino protecting group; aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, amino, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted). In certain embodiments, the R^h substituents of the di- substituted amino group (-NR^h ₂) form a 5- to 6- membered hetereocyclic ring. [0023] The term "alkoxy" refers to a "substituted hydroxyl" of the formula (-OR¹), wherein R¹ is an optionally substituted alkyl group, as defined herein, and the oxygen moiety is directly attached to the parent molecule.

[0024] The term "alkylthioxy" refers to a "substituted thiol" of the formula (-SR^r), wherein R^r is an optionally substituted alkyl group, as defined herein, and the sulfur moiety is directly attached to the parent molecule. [0025] The term "alkylamino" refers to a "substituted amino" of the formula (-NR^h ₂), wherein R is, independently, a hydrogen or an optionally subsituted alkyl group, as defined herein, and the nitrogen moiety is directly attached to the parent molecule. [0026] The term "aryl," as used herein, refer to stable aromatic mono- or polycyclic ring system having 3-20 ring atoms, of which all the ring atoms are carbon, and which may be substituted or unsubstituted. In certain embodiments of the present invention, "aryl" refers to a mono, bi, or tricyclic C₄-C₂₀ aromatic ring system having one, two, or three aromatic rings which include, but not limited to, phenyl, biphenyl, naphthyl, and the like, which may bear one or more substituents. Aryl substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety {e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted). [0027] The term "arylalkyl," as used herein, refers to an aryl substituted alkyl group, wherein the terms "aryl" and "alkyl" are defined herein, and wherein the aryl group is attached to the alkyl group, which in turn is attached to the parent molecule. An exemplary arylalkyl group includes benzyl.

[0028] The term "aryloxy" refers to a "substituted hydroxyl" of the formula (-OR¹), wherein R¹ is an optionally substituted aryl group, as defined herein, and the oxygen moiety is directly attached to the parent molecule.

[0029] The term "arylamino," refers to a "substituted amino" of the formula (-NR^h ₂), wherein R^h is, independently, a hydrogen or an optionally substituted aryl group, as defined herein, and the nitrogen moiety is directly attached to the parent molecule. [0030] The term "arylthioxy" refers to a "substituted thiol" of the formula (-SR^r), wherein R^r is an optionally substituted aryl group, as defined herein, and the sulfur moiety is directly attached to the parent molecule.

[0031] The term "azido," as used herein, refers to a group of the formula (-N₃).

[0032] The term "cyano," as used herein, refers to a group of the formula (-CN).

[0033] The terms "halo" and "halogen" as used herein refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), and iodine (iodo, -I). [0034] The term "heteroaliphatic," as used herein, refers to an aliphatic moiety, as defined herein, which includes both saturated and unsaturated, nonaromatic, straight chain (i.e., unbranched), branched, acyclic, cyclic (i.e., heterocyclic), or polycyclic hydrocarbons, which are optionally substituted with one or more functional groups, and that contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in place of carbon atoms. In certain embodiments, heteroaliphatic moieties are substituted by independent replacement of one or more of the hydrogen atoms thereon with one or more substituents. As will be appreciated by one of ordinary skill in the art, "heteroaliphatic" is intended herein to include, but is not limited to, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocycloalkyl, heterocycloalkenyl, and heterocycloalkynyl moieties. Thus, the term "heteroaliphatic" includes the terms "heteroalkyl," "heteroalkenyl", "heteroalkynyl", and the like. Furthermore, as used herein, the terms "heteroalkyl", "heteroalkenyl", "heteroalkynyl", and the like encompass both substituted and unsubstituted groups. In certain embodiments, as used herein, "heteroaliphatic" is used to indicate those heteroaliphatic groups (cyclic, acyclic, substituted, unsubstituted, branched or unbranched) having 1—20 carbon atoms. Heteroaliphatic group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, sulfinyl, sulfonyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted). [0035] The term "heteroalkyl," as used herein, refers to an alkyl moiety, as defined herein, which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in place of carbon atoms.

[0036] The term "heteroalkenyl," as used herein, refers to an alkenyl moiety, as defined herein, which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in place of carbon atoms.

[0037] The term "heteroalkynyl," as used herein, refers to an alkynyl moiety, as defined herein, which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in place of carbon atoms. [0038] The term "heteroalkylamino" refers to a "substituted amino" of the formula (-

NR^h ₂), wherein R^h is, independently, a hydrogen or an optionally substituted heteroalkyl group, as defined herein, and the nitrogen moiety is directly attached to the parent molecule. [0039] The term "heteroalkyloxy" refers to a "substituted hydroxyl" of the formula (—

OR¹), wherein R¹ is an optionally substituted heteroalkyl group, as defined herein, and the oxygen moiety is directly attached to the parent molecule.

[0040] The term "heteroalkylthioxy" refers to a "substituted thiol" of the formula (-

SR¹), wherein R^r is an optionally substituted heteroalkyl group, as defined herein, and the sulfur moiety is directly attached to the parent molecule.

[0041] The term "heterocyclic," "heterocycles," or "heterocyclyl," as used herein, refers to a cyclic heteroaliphatic group. A heterocyclic group refers to a non-aromatic, partially unsaturated or fully saturated, 3- to 10-membered ring system, which includes single rings of 3 to 8 atoms in size, and bi- and tri-cyclic ring systems which may include aromatic five- or six— membered aryl or heteroaryl groups fused to a non-aromatic ring. These heterocyclic rings include those having from one to three heteroatoms independently selected from oxygen, sulfur, and nitrogen, in which the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. In certain embodiments, the term heterocylic refers to a non-aromatic 5-, 6—, or 7-membered ring or polycyclic group wherein at least one ring atom is a heteroatom selected from O, S, and N (wherein the nitrogen and sulfur heteroatoms may be optionally oxidized), and the remaining ring atoms are carbon, the radical being joined to the rest of the molecule via any of the ring atoms. Heterocycyl groups include, but are not limited to, a bi- or tri-cyclic group, comprising fused five, six, or seven-membered rings having between one and three heteroatoms independently selected from the oxygen, sulfur, and nitrogen, wherein (i) each 5-membered ring has 0 to 2 double bonds, each 6-membered ring has 0 to 2 double bonds, and each 7-membered ring has 0 to 3 double bonds, (ii) the nitrogen and sulfur heteroatoms may be optionally oxidized, (iii) the nitrogen heteroatom may optionally be quaternized, and (iv) any of the above heterocyclic rings may be fused to an aryl or heteroaryl ring. Exemplary heterocycles include azacyclopropanyl, azacyclobutanyl, 1,3-diazatidinyl, piperidinyl, piperazinyl, azocanyl, thiaranyl, thietanyl, tetrahydrothiophenyl, dithiolanyl, thiacyclohexanyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropuranyl, dioxanyl, oxathiolanyl, morpholinyl, thioxanyl, tetrahydronaphthyl, and the like, which may bear one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, sulfinyl, sulfonyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted). [0042] The term "heteroaryl," as used herein, refer to stable aromatic mono- or polycyclic ring system having 3-20 ring atoms, of which one ring atom is selected from S. O, and N; zero, one, or two ring atoms are additional heteroatoms independently selected from S, O, and N; and the remaining ring atoms are carbon, the radical being joined to the rest of the molecule via any of the ring atoms. Exemplary heteroaryls include, but are not limited to pyrrolyl, pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, pyyrolizinyl, indolyl, quinolinyl, isoquinolinyl, benzoimidazolyl, indazolyl, quinolinyl, isoquinolinyl, quinolizinyl, cinnolinyl, quinazolynyl, phthalazinyl, naphthridinyl, quinoxalinyl, thiophenyl, thianaphthenyl, furanyl, benzofuranyl, benzothiazolyl, thiazolynyl, isothiazolyl, thiadiazolynyl, oxazolyl, isoxazolyl, oxadiaziolyl, oxadiaziolyl, and the like, which may bear one or more substituents. Heteroaryl substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, sulfinyl, sulfonyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted).

[0043] The term "heteroarylene," as used herein, refers to a biradical derived from an heteroaryl group, as defined herein, by removal of two hydrogen atoms. Heteroarylene groups may be substituted or unsubstituted. Additionally, heteroarylene groups may be incorporated as a linker group into an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, or heteroalkynylene group, as defined herein. Heteroarylene group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, and the like, each of which may or may not be further substituted).

[0044] The term "heteroarylamino" refers to a "substituted amino" of the (-NR^h ₂), wherein R^h is, independently, a hydrogen or an optionally substituted heteroaryl group, as defined herein, and the nitrogen moiety is directly attached to the parent molecule.

[0045] The term "heteroaryloxy" refers to a "substituted hydroxyl" of the formula (—

OR¹), wherein R' is an optionally substituted heteroaryl group, as defined herein, and the oxygen moiety is directly attached to the parent molecule.

[0046] The term "heteroarylthioxy" refers to a "substituted thiol" of the formula (-

SR¹), wherein R^r is an optionally substituted heteroaryl group, as defined herein, and the sulfur moiety is directly attached to the parent molecule.

[0047] The term "hydroxy," or "hydroxyl," as used herein, refers to a group of the formula (-OH). A "substituted hydroxyl" refers to a group of the formula (-OR¹), wherein R¹ can be any substitutent which results in a stable moiety (e.g., a suitable hydroxyl protecting group; aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, nitro, alkylaryl, arylalkyl, and the like, each of which may or may not be further substituted).

[0048] The term "imino," as used herein, refers to a group of the formula (=NR^r), wherein R^r corresponds to hydrogen or any substitutent as described herein, that results in the formation of a stable moiety (for example, a suitable amino protecting group; aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, amino, hydroxyl, alkylaryl, arylalkyl, and the like, each of which may or may not be further substituted). In certain embodiments, imino refers to =NH wherein R^r is hydrogen.

[0049] The term "isocyano," as used herein, refers to a group of the formula (-NC).

[0050] The term "nitro," as used herein, refers to a group of the formula (-NO₂).

[0051] The term "oxo," as used herein, refers to a group of the formula (=0).

[0052] The term "stable moiety," as used herein, preferably refers to a moiety which possess stability sufficient to allow manufacture, and which maintains its integrity for a sufficient period of time to be useful for the purposes detailed herein.

[0053] A "suitable amino— protecting group," as used herein, is well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W.

Greene and P. G. M. Wuts, 3^rd edition, John Wiley & Sons, 1999, the entirety of which is incorporated herein by reference. Suitable amino-protecting groups include methyl carbamate, ethyl carbamante, 9-fluorenylmethyl carbamate (Fmoc), 9-(2- sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t- butyl-[9-(l 0, 10-dioxo-10, 10, 10, 10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2- trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1 -(I -adamanty I)-I- methylethyl carbamate (Adpoc), l,l-dimethyl-2-haloethyl carbamate, l,l-dimethyl-2,2- dibromoethyl carbamate (DB-t-BOC), l,l-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1 -methyl- l-(4-bipheny Iy l)ethyl carbamate (Bpoc), l-(3,5-di-t-butylphenyl)-l- methylethyl carbamate (t-Bumeoc), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N₁N- dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, iV-hydroxypiperidinyl carbamate, alkyldithio carbamate, benzyl carbamate (Cbz), /7-methoxybenzyl carbamate (Moz), />-nitobenzyl carbamate, /?-bromobenzyl carbamate, p- chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2— (1,3— dithianyl)]methyl carbamate (Dmoc), 4— methylthiophenyl carbamate (Mtpc), 2,4— dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2- triphenylphosphonioisopropyl carbamate (Ppoc), l,l-dimethyl-2-cyanoethyl carbamate, m- chloro-/?-acyloxybenzyl carbamate, /?-(dihydroxyboryl)benzyl carbamate, 5- benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), /w-nitrophenyl carbamate, 3, 5-dimethoxy benzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl (o-nitrophenyl)methyl carbamate, phenothiazinyl-(10)-carbonyl derivative, jV -/?-toluenesulfbnylaminocarbonyl derivative, iV'-phenylaminothiocarbonyl derivative, t— amyl carbamate, S— benzyl thiocarbamate, /j-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, /?-decyloxybenzyl carbamate, 2,2- dimethoxycarbonylvinyl carbamate, o-(jV₎Λ/-dimethylcarboxamido)benzyl carbamate, 1,1- dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2— furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate, isobutyl carbamate, isonicotinyl carbamate, p—ip - methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methy 1-1— cyclopropylmethyl carbamate, 1-methyl-l— (3,5- dimethoxyphenyl)ethyl carbamate, l-methyl-l-(/?-phenylazophenyl)ethyl carbamate, 1— methyl- 1-phenylethyl carbamate, l-methyl-l-(4-pyridyl)ethyl carbamate, phenyl carbamate, ;?-(phenylazo)benzyl carbamate, 2,4,6-tri-/-butylphenyl carbamate, 4— (trimethylammonium)benzyl carbamate, 2,4,6-trimethylbenzyl carbamate, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3— phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide, /7-phenylbenzamide, o-nitophenylacetamide, o- nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxycarbonylamino)acetamide, 3- (p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o- nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4- chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N— acetylmethionine derivative, o-nitrobenzamide, ø-(benzoyloxymethyl)benzamide, 4,5-diphenyl-3-oxazolin- 2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N—2,5— dimethylpyrrole, 7V-l,l,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5- substituted l,3-<hmethyl-l,3,5-triazacyclohexan-2-one, 5-substituted l,3-dibenzyl-l,3,5- triazacyclohexan-2-one, 1 -substituted 3,5-dinitro-4-pyridone, iV-methylamine, JV- allylamine, /V-[2-(trimethylsilyl)ethoxy]methylamine (SEM), iV-3-acetoxypropylamine, N- (1-isopropyl— 4-nitro-2-oxo— 3-pyroolin-3-yl)amine, quaternary ammonium salts, N— benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, TV- triphenylmethylamine (Tr), 7V-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9- phenylfluorenylamine (PhF), /V-2,7-dichloro-9-fluorenylmethyleneamine, N- ferrocenylmethylamino (Fcm), iV-2-picolylamino N'-oxide, N-1, 1- dimethylthiomethyleneamine, N— benzylideneamine, N-/>-methoxybenzylideneamine, N- diphenylmethyleneamine, N-[(2-pyridy l)mesityl]methyleneamine, N-(N ', N - dimethylaminomethylene)amine, NN'-isopropylidenediamine, N- p-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro— 2— hydroxyphenyl)phenylmethyleneamine, N-cyclohexy lideneamine, N-(5 , 5-dimethyl-3-oxo- l-cyclohexenyl)amine, N-borane derivative, N-diphenylborinic acid derivative, N- [phenyl(pentacarbonylchromium- or tungsten)carbonyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Νps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, 3-nitropyridinesulfenamide (Npys), />-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6- trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3 ,5,6-tetramethyl— 4-methoxy benzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-^- methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9- anthracenesulfonamide, 4-(4' ,8 '-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide. [0054] A "suitable carboxylic acid protecting group," or "protected carboxylic acid," as used herein, are well known in the art and include those described in detail in Greene (1999). Examples of suitably protected carboxylic acids further include, but are not limited to, silyl-, alkyl-, alkenyl-, aryl-, and arylalkyl-protected carboxylic acids. Examples of suitable silyl groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t- butyldiphenylsilyl, triisopropylsilyl, and the like. Examples of suitable alkyl groups include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, tetrahydropyran-2- yl. Examples of suitable alkenyl groups include allyl. Examples of suitable aryl groups include optionally substituted phenyl, biphenyl, or naphthyl. Examples of suitable arylalkyl groups include optionally substituted benzyl (e.g., p-methoxybenzyl (MPM), 3,4- dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p- cyanobenzyl), and 2- and 4-picolyl.

[0055] A "suitable hydroxyl protecting group," as used herein, is well known in the art and include those described in detail in Greene (1999). Suitable hydroxyl protecting groups include methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t— butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), /?-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2- methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2- (trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3- bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4— methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4— methoxytetrahydrothiopyranyl S,S-dioxide, l-[(2-chloro— 4-methyl)phenyl]— 4- methoxypiperidin-4-yl (CTMP), 1 ,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxy ethyl, l-(2-chloroethoxy)ethyl, 1-methyl-l-methoxyethyl, 1-methyl-l-benzyloxyethyl, 1- methyl-l-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2- (phenylselenyl)ethyl, t-butyl, allyl, />-chlorophenyl, /?-methoxyphenyl, 2,4-dinitrophenyl, benzyl, />-methoxybenzyl, 3,4-dimethoxybenzyl, o— nitrobenzyl, p— nitrobenzyl, p— halobenzyl, 2,6-dichlorobenzyl, /?-cyanobenzyl, /?-phenylbenzyl, 2-picolyl, 4-picolyl, 3- methyl-2-picolyl N-oxido, diphenylmethyl, p,p '-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, />-methoxyphenyldiphenylmethyl, di(p- methoxyphenyl)phenylmethyl, tri(/7-methoxyphenyl)methyl, 4— (4'— bromophenacyloxyphenyl)diphenylmethyl, 4,4',4' '— tris(4,5- dichlorophthalimidophenyl)methyl, 4,4',4"-tris(levulinoyloxyphenyl)methyl, 4,4',4"- tris(benzoyloxyphenyl)methyl, 3-(imidazol-l-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1- bis(4-methoxyphenyl)-l '-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl- 10-oxo)anthryl, 1 ,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t- butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri— /?— xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), /-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, /7-chlorophenoxyacetate, 3- phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate, adamantoate, crotonate, 4— methoxycrotonate, benzoate, p— phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9— fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl />-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-l- napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4- methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2- (methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-

(methylthiomethoxymethyl)benzoate, 2,6-dichloro— 4-methylphenoxyacetate, 2,6-dichloro- 4-( 1,1,3 ,3-tetramethylbutyl)phenoxyacetate, 2,4-bis( 1 , 1 -dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o- (methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkyl NJI₁N ',N'- tetramethylphosphorodiamidate, alkyl 7V-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). For protecting 1,2— or 1,3-diols, the protecting groups include methylene acetal, ethylidene acetal, l-/-butylethylidene ketal, 1-phenylethylidene ketal, (4- methoxyphenyl)ethylidene acetal, 2,2,2-trichloroethylidene acetal, acetonide, cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal, p- methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal, 3, 4-dimethoxy benzylidene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene ortho ester, 1-methoxyethylidene ortho ester, 1-ethoxyethylidine ortho ester, 1 ,2-dimethoxyethylidene ortho ester, α-methoxybenzylidene ortho ester, l-(N,N— dimethylamino)ethylidene derivative, α-(iV,N'-dimethylamino)benzylidene derivative, 2- oxacyclopentylidene ortho ester, di-t-butylsilylene group (DTBS), 1,3 — (1,1,3,3— tetraisopropyldisiloxanylidene) derivative (TIPDS), tetra-/-butoxydisiloxane-l,3-diylidene derivative (TBDS), cyclic carbonates, cyclic boronates, ethyl boronate, and phenyl boronate. [0056] A "suitable thiol protecting group," as used herein, are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3^rd edition, John Wiley & Sons, 1999, the entirety of which is incorporated herein by reference. Examples of suitably protected thiol groups further include, but are not limited to, thioesters, carbonates, sulfonates allyl thioethers, thioethers, silyl thioethers, alkyl thioethers, arylalkyl thioethers, and alkyloxyalkyl thioethers. Examples of suitable ester groups include formates, acetates, proprionates, pentanoates, crotonates, and benzoates. Specific examples of suitable ester groups include formate, benzoyl formate, chloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p— chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate, 4,4-

(ethylenedithio)pentanoate, pivaloate (trimethylacetate), crotonate, 4-methoxy-crotonate, benzoate, p-benylbenzoate, 2,4,6-trimethylbenzoate. Examples of suitable carbonates include 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2- (phenylsulfonyl)ethyl, vinyl, allyl, and p-nitrobenzyl carbonate. Examples of suitable silyl groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl ether, and other trialkylsilyl ethers. Examples of suitable alkyl groups include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, and allyl ether, or derivatives thereof. Examples of suitable arylalkyl groups include benzyl, p- methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2- and 4-picolyl ethers.

[0057] The term "thio," or "thiol," as used herein, refers to a group of the formula (-

SH). A "substituted thiol" refers to a group of the formula (-SR^r), wherein R^r can be any substituent that results in the formation of a stable moiety (e.g., a suitable thiol protecting group; aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, sulfmyl, sulfonyl, cyano, nitro, alkylaryl, arylalkyl, and the like, each of which may or may not be further substituted).

[0058] The term "thiooxo," as used herein, refers to a group of the formula (=S).

[0059] As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et ah, describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N⁺(C i_₄alky I)₄ salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate, and aryl sulfonate.

[0060] The following definitions are more general terms used throughout the present application:

[0061] The terms "administer," "administering," or "administration," as used herein refers to implanting, absorbing, ingesting, injecting, or inhaling the inventive compound.

[0062] The term "chemosensitizer," as used herein, refers to a drug that makes tumor cells more sensitive to the effects of chemotherapy.

[0063] The terms "effective amount" and "therapeutically effective amount," as used herein, refer to the amount or concentration of an inventive compound, that, when administered to a subject, is effective to at least partially treat a condition from which the subject is suffering (e.g., a neurodegenerative disease).

[0064] The term "gibberellin" refers to any of the compounds of formula I as described herein. In certain embodiments, gibberellins are diterpenoids with the pentacyclic or tetracyclic scaffold of formula I. In certain embodiments, a gibberellin is a natural product phytohormone found in plants. Gibberellins may be obtained from natural sources, from commercial sources, by fermentation, by semi-synthesis, or by total synthesis. Exemplary compounds include gibberellic acid, gibberellin A9, gibberellin A9 methyl ester, gibberellin

A4, gibberellenic acid, allogibberic acid, and gibberic acid.

[0065] The term "NF-κB," as used herein, refers to the nuclear factor-kappaB protein complex, which acts as a transcription factor. NF-κB is found in almost all animal cell types and is involved in cellular responses to stimuli such as stress, cytokines, free radicals, ultraviolet irradiation, oxidized LDL, and bacterial or viral antigens. The term "NF-κB" may also refer to variants of wild type NF-κB, for example, truncated forms, homologs, mutant forms, and modified forms. Preferably, the variant retains NF-κB's ability to induce transcription of particular genes.

[0066] The term "subject," as used herein, refers to any animal. In certain embodiments, the subject is a mammal. In certain embodiments, the term "subject", as used herein, refers to a human (e.g., a man, a woman, or a child).

[0067] The terms "treat" or "treating," as used herein, refers to partially or completely alleviating, inhibiting, ameliorating, and/or relieving the disease or condition from which the subject is suffering. Brief Description of the Drawings

[0068] Figure 1. Image of a small molecule microarray (SMM) containing 20,800 small molecules. Structures of representative bioactive compounds (outer ring) and scaffolds for diversity-oriented synthesis (inner ring) contained within the SMM are shown. [0069] Figure 2. Signal transduction pathway for NF-κB activation. Proinflammatory signals such as IL-I, LPS, or TNF-α bind their corresponding receptor and trigger serine kinases that phosphorylate inhibitor of KB (IKB) kinase and subsequent phosphorylation of IKB and proteasomal degradation. Upon release, p65 binds to the p50 proteolytic cleavage product of p 105 in the cytoplasm. The active complex is translocated into the nucleus via interactions between p65 and importins. In the nucleus, the complex interacts with other transcription factors and co-regulators to participate in transcriptional activation. NF-κB is a highly conserved transcription factor protein complex found in almost all animal cell types. NF-κB is involved in cellular responses to various types of stress including cytokines, free radicals, UV light, bacterial antigens, and viral antigens. It has also been found to regulated several genes that control cell proliferation and survival. Aberrant NF-κB activity is associated with various cancers, inflammatory diseases, autoimmune diseases, septic shock, and viral infection. Over 700 compounds are known to target various proteins in the NF-κB pathways (e.g., kinase inhibitors, proteasome inhibitors).

[0070] Figure 3. (A) Gibberellic acid (GA₃) was discovered to be a binder of p50 using small molecule microarrays. It was confirmed as a binder by surface plasmon resonance (K_D - 226 nM). Gibberellic acid is a natural product phytohormone found in plants. It was originally isolated from Gibberella fujikuroi as the causative agent of the baka- nae ("foolishly overgrown seedling") disease. Gibberellic acid is the most widely distributed member of the gibberelilin family and has been found to promote cell growth and elongation. It has been shown to be involved in plant stress response and binds to the plant nuclear receptor GIDl. Gibberellic acid is widely used to control fruit and vegetable size, promote the growth of sugarcane, and assist in the malting of barley. Gibberellic acid is generally considered safe in humans (oral rat LD₅₀ = 6,300 mg/kg). (B) Surface plasmon resonance sensogram for binding of gibberellic acid to immobilized p50. The compound was injected at eight concentrations (2:1 dilutions, 39 nM to 5 uM) and the Kd was determined to be -400 nM using both kinetic and steady-state models.

[0071] Figure 4. Connections between the p50 ligand and other small molecules using gene expression profiling in Connectivity Map. (Top) Rank-order of top twenty compounds with connectivity to the p50 ligand are shown for the PC3 instance along with name, number of instances, enrichment scores, permutation p-values and functional annotation where available. (Bottom) Bar views for several connected compounds constructed from 6,100 horizontal lines, each representing an individual treatment instance ordered by their corresponding connectivity scores with the p50 binder signature. Instances shown in black correspond to the compound shown, and colors applied to the other instances reflect the sign of their scores (green = positive, gray = null, red = negative).

[0072] Figure 5. Tetracyclic diterpene inhibitors of the NF-κB pathway. Chen et al. ,

Bioorg. Med. Chem. Lett. 19:5496-99, 2009; which is incorporated herein by reference.

[0073] Figure 6. Gibberellins block nuclear translocation of p50 in HEK293 cells.

[0074] Figure 7. p65 immunofluorescent assays in HUVECs with TNFα

[0075] Figure 8. p65 immunofluorescent assays in HUVECs with allogibberic acid

(AGA) and TNFα.

[0076] Figure 9. p65 immunofluorescent assays in HUVECS with LPS alone and gibberellic acid (GA₃) plus LPS.

[0077] Figure 10. Images of HUVEC cells acquired using the ImageXpress 5000

ArrayScan HCS reader. Cell were exposed to compound (n=4), co-administered with 20 ng/mL of TNFα, and incubated at 37 ⁰C for 20 minutes. A. Cells were exposed to vehicle

0.1% DMSO as control. B. Cells were induced with TNFα in 0.1% DMSO. C. Cells were exposed to 10 μM GA3 in 0.1% DMSO with TNFα. D. Cells were exposed to 10 μM AGA in 0.1% DMSO with TNFα. E. Cells were exposed to 0.2 μM of the proteasome inhibitor clasto-lactacystin β-lactone, a known inhibitor of I-κBα degradation and subsequent NF-κB translocation, as a positive control, in 0.1% DMSO with TNFα. Note the accumulation of p65 in the nucleus when cells are stimulated with TNFα. This accumulation is inhibited in the presence of the two gibberellins as well as the positive control. This behaviour is also observed for p50.

[0078] Figure 11. Gibberellins inhibit transcription in response to TNFα stimulation in a reporter line.

[0079] Figure 12. Gibberellin stability.

[0080] Figure 13A-13E. Connected compounds to the p50 binder gibberellic acid as identified using Connectivity Map. Detailed Description of Certain Embodiments of the Invention

[0081] The present invention provides methods for modulating the NF-κB pathway using gibberellins {e.g., gibberellic acid, see Figure 3A). Modulation of the NF-κB pathway using a gibberellin may be done in vivo (e.g., in a subject) or in vitro (e.g., in cell culture). Gibberellic acid has been identified through screening as a direct binder of the p50 subunit of NF-κB, and p50 binding has been confirmed by surface plasmon resonance (Figure 3B). Gibberellic acid and other gibberellins (see the Examples below) have been found to inhibit TNFα-stimulated transcription.

Gibberellins

[0082] Gibberellins are a series of naturally occurring compounds, which are known as plant growth regulators with wide application in the plant kingdom (MacMillan, et al, Adv. Chem. Ser (1961) 28: 18-24, incorporated herein by reference). They have also been isolated from metabolites of some microorganisms, such as Gibber ella fujikuroi (Cross, et al, J. Chem. Soc. (1954), 4670; Brian, et al, U.S. Patent 2,842,051; Calam et al, U.S. Patent 2,950,288; Birch et al, U.S. Patent 2,977,285, each of which is incorporated herein by reference). Gibberellins, especially gibberellic acid (gibberellin A3), and its mixture with gibberellin A4, and/or gibberellin A7, which are commercially available, have been extensively applied in agriculture to increase the growth of some fruits (e.g., strawberries and grapes) and vegetables (e.g., tomatoes, cabbages and cauliflowers), also as food additive in the malting of barley. Other known gibberellins include, but are not limited to, gibberellin A9, gibberellin A9 methyl ester, gibberellin A4, allogibberic acid (AGA), gibberelenic acid, and gibberic acid.

[0083] Gibberellins as the term is used herein are generally of formula I:

wherein R is hydrogen or a group -OR²⁰, where R²⁰ is hydrogen, a glycoside, C_1-6 alkyl group, or R¹ together with R² or R¹⁰ forms a bond (Ci-C₂ or Ci-Cio double bond, respectively);

R³ is hydrogen, =0, or —OR²², where R²² is hydrogen or a glycoside; or R³ together with R² forms a bond (C₂-C₃ double bond); or R³ together with R² and the intervening atoms forms an epoxide ring;

R⁴ is -OH, or -OR²³, where R²³ is unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, arylalkyl, amidine, -NR²⁴R²⁵ or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur; R ⁴ and R ⁵ may or may not be the same, are hydrogen, or Ci_-20 alkyl, allyl, aryl, arylalkyl or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; or R⁴ together with R²¹ or R²⁸ forms a lactone;

R⁵ is hydrogen, a glycoside, unsubstituted or substituted Cj_-20 alkyl, allyl, aryl, or arylalkyl;

R⁸ is hydrogen, hydroxyl, mercaptan, or halogen, amino, azido, -NR²⁴R²⁵, unsubstituted or substituted Cj_-20 alkyl, allyl, aryl, or arylalkyl, or -OR²⁷, where R²⁷ is hydrogen or a glycoside;

R⁹ is hydrogen or hydroxyl; or R⁹ together with R¹⁵ forms a bond (C₉-Ci₅ double bond); or R⁹ together with R¹⁵ and the intervening atoms forms an epoxide ring;

R¹ ' is hydrogen or hydroxyl or is absent;

R¹² is methyl, -CH₂OH, -CO₂H, or a glycoside ester of said -CO₂H; R¹³ is methylene, or a divalent heteroatom, or -NR²⁹, where R²⁹ is -NHR³⁰ or -OR³⁰ where R is hydrogen, or Ci_-20 alkyl, aryl, alkylaryl; and a double bond is present between C_J6 and R¹³ when R¹¹ is absent; or R¹³ is hydrogen, hydroxyl, methyl, -CHO, -CH₂X, where X is halogen, -CH₂NR²⁹, where R²⁹ is -NR³⁰ or -OR³⁰, where R³⁰ is hydrogen, or C_1-20 alkyl, aryl, or alkylaryl when R¹¹ is hydrogen or hydroxyl;

R¹⁴ is hydrogen or hydroxyl;

R¹⁵ is hydrogen, or R¹⁵ together with R⁹ forms a bond (C₉-Ci₅ double bond); or R is together with R⁹ and the intervening atoms forms an epoxide ring; or a pharmaceutically acceptable salt thereof.

[0084] A very similar genus of gibberellins can be found in U.S. Patent 7,435,756; U.S.

Publ. Appl. 2004/0162248; U.S. Publ. Appl. 2005/0215496; and U.S. Publ. Appl. 2007/0197456; each of which is incorporated herein by reference.

[0085] In certain embodiments, the stereochemistry of the gibberellin is as shown in the formula:

[0086] In certain embodiments, the present invention utilizes gibberellic acid

(gibberellin A3) of formula:

Gibberellic acid has been found to bind the p50 subunit of NF-κB with a Kj of approximately

216 nM.

[0087] In certain embodiments, the present invention utilizes gibberellin A4 of formula:

[0088] In certain embodiments, the present invention utilizes gibberellin A9 methyl ester of formula:

[0089] In certain embodiments, the present invention utilizes gibberellin A7 of formula:

[0090] In certain embodiments, the present invention utilizes allogibberic acid (AGA) of formula:

[0091] In certain embodiments, the present invention provides derivatives of allogibberic acid of formula:

wherein

n is an integer between 2 and 6, inclusive; m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, m is 1. In certain embodiments, m is 2. In certain embodiments, m is 3. Such compounds may be prepared by conjugation of a polyethylene glycol chain to an activated ester of AGA.

[0092] In certain embodiments, the present invention provides an ethylene glycol derivative of allogibberic acid (AGA) of formula:

This derivative may be used in the present invention to modulate the NF-κB pathway. [0093] Gibberellin A3, gibberellin A4, and gibberellin A7 can be obtained by fermentation of microorganisms such as Gibber ella fujikuroi. The crude compound(s) can be isolated and purified to afford a high purity crystalline product. Other gibberellin derivatives can be obtained by either semi-synthesis from gibberellic acid or other gibberellins, or total synthesis, each of which has been well-documented (Corey et al, J. Am. Chem. Soc. (1978) 100: 8031-8034; 8034-8036); Furber et al, J. Org. Chem. (1990) 55: 4860-4870; Mander et al, J. Chem. Soc, Perkin Trans 1 (2000) 2893-2894; Pour et al, Tetrahedron (1998) 54: 13833-13850; Liu, et al, Tetrahedron (1998) 54: 11637-11650; Pour et al, Pure Appl. Chem. (1998) 70: 351-354; Pour et al, Tetrahedron Lett. (1998) 39:1991-1994; Pour et al, Aus. J Chem. (1997) 50:289-299; King et al, J. Am. Chem. Soc. (1997) 119: 3828-3829; Mander et al, Tetrahedron (1997) 53: 2137-2162, each of which is incorporated herein by reference). Furthermore, the extraction and isolation of various gibberellins from different plants, shoots, fruits, and seeds have also been widely published (Pearce et al, Phytochemistry (2002) 59: 679-687; Nakayama et al, Phytochemistry (2001) 57:749-758 (2001); Nakayama et al, Phytochemistry (1998) 48:587-593; Blake et al, Phytochemistry (2000) 55: 887-890; Blake et al, Phytochemistry (2000) 53:519-528; Koshioka et al, J. of the Japanese Society for Horticultural Science (1999) 68:1158-1160; Koshioka et al, J. of the Japanese Society for Horticultural Science (1998) 67:866-871; Mander et al, Phytochemistry (1998) 49: 2195- 2206 (1998); Mander et al, Phytochemistry (1998) 49:1509-1515; Wynne et al, Phytochemistry (1998) 49:1837-1840, each of which is incorporated herein by reference). Gibberellins and methods of preparing gibberellins have been disclosed in the patent literature as well (see, e.g., U.S. Patent No. 7,195,917; 6,723,897; 5,866,779; 5,773,288, 5,767,375; 5,767,042; 5,612,191; 5,580,857; 5,562,831; 4,578,483; 4,349,481; 4,243,594; 4,156,684; 4,154,596; U.S. Patent Publications 2008/0085543; 2007/0174931; 2006/0253933; 2004/0121321 ; 2002/0053095; each of which is incorporated herein by reference.

[0094] Gibberellins have previously been used in the treatment of herpes simplex, prostatitis, psoriasis, tumor, ulcer and wound healing, and diabetes (Parkinson, U.S. Patent No. 4,424,232; French Patent No. 2597339; Davis, U.S. Pat. No. 5,487,899; Oden, U.S. Patent No. 5,580,857; Wu, Australian Patent No. 695054; Wu, U.S. Pat. No. 6,121,317; Jenkins, U.S. Patent No. 7,435,756; U.S. Patent Publication 2004/0162248; U.S. Patent Publication 2005/0215496; U.S. Patent Publication 2007/0197456, each of which is incorporated herein by reference).

Small-molecule microarrays

[0095] Small-molecule microarrays (SMMs) have proven to be a general, robust, and scalable screening platform for discovering protein-small molecule interactions that lead to modulators of protein function (Vegas et al, Chem. Soc. Rev. (2008) 37:1385-1394). SMMs are manufactured by robotically arraying stock solutions of compounds onto functionalized substrates such as glass microscope slides. The arrays are then incubated with proteins of interest and putative protein-small molecule interactions are typically detected using fluorescently labeled antibodies and a standard fluorescence slide scanner. The high- throughput and miniaturized nature of the microarray-based binding assay allows for screening of large panels of proteins against tens of thousands of compounds in a relatively short time frame (Bradner et al, Nature Protocols (2006) 1 : 2344-2352). Multiple approaches to manufacturing SMMs have been developed, including fluorous-based non- covalent attachment for homogeneous display (Vegas et al, Angew. Chem. Int. Ed. (2007) 46: 7960-7964) and isocyanate-based covalent attachment for heterogeneous display (Bradner et al, Chem. Biol (2006) 13:493-504; Bradner et al, PCT Publ. WO 2008/013569, incorporated herein by reference). The isocyanate approach allows for SMMs that contain compounds not intentionally synthesized or modified for immobilization onto a solid substrate, allowing for arrays containing bioactive small molecules, including FDA-approved drugs, synthetic drug-like compounds, natural products, and products of diversity-oriented synthesis on a single slide {Figure 1) (Bradner et al, Chem. Biol. (2006) 13:493-504; incorporated herein by reference).

[0096] SMMs have been used by several labs to identify functional small molecules that target proteases (Urbina et al, Chembiochem (2006) 7: 1790-1797; Uttamchandani et al, J. Am. Chem. Soc. (2007) 129: 13110-13117; each of which is incorporated herein by reference), kinases (Kwon et al, ACS Chem Biol (2007) 2: 419-425; Miao et al, J. Comb. Chem. (2007) 9: 245-253; each of which is incorporated herein by reference), and histone deacetylases (Vegas et al, Angew. Chem. Int. Ed. (2007) 46:7960-7964; incorporated herein by reference). SMMs were also used to identify a ligand to the yeast protein Hap3p, a subunit of the Hap2/3/4/5p (HAP) transcription factor complex involved in aerobic respiration and the nutrient-response signaling network (Koehler et al, J. Am. Chem. Soc. (2003) 125:8420-8421; Hahn et al, Science (1988) 240:317-321; Dang et al, J. Bacteriol (1996) 178:1842-1849; each of which is incorporated herein by reference).

Identification of Modulators ofNF-κB

[0097] The NF-κB transcription factor family plays a key role in inflammation and the immune response. Active NF-κB has been found in the nucleus of several different hematopoietic and solid tumor cell types (Darnell, Nature Rev. Cancer (2002) 2: 740-749; incorporated herein by reference). The NF-κB protein resides in the cytoplasm until extracellular signals trigger activation and translocation into the nucleus as outlined in Figure 2. Both NF-κB subunits of the p65-p50 heterodimer were screened against 20,000 small molecules. After filtering promiscuous and nonspecific binders to the other 98 transcription factors that were screened, 26 compounds scored as binders to p50 (e.g., gibberellic acid), 6 compounds bound to p65, and several compounds bound to both proteins. The SMM positives may be verified in secondary binding assays involving surface plasmon resonance (SPR). The SPR assay involves affinity capture of the protein target onto a carboxymethyldextran sensor chip via an epitope tag and an immobilized antibody. The test compounds are then injected over the surface and binding constants are determined using both kinetic and steady state equilibrium approaches. SPR may also be used to evaluate the ability of the compounds to prevent or disrupt the p50-p65 interaction. For example, small molecules that bind to p50 (e.g., gibberellic acid) may be pre-incubated with the target, and the complexes can be injected over a p65 sensor chip surface. Compounds that disrupt heterodimer formation will lead to a reduced SPR response or altered binding kinetics as compared to a DMSO control. Compounds may also be evaluated for their ability to disrupt preformed p50-p65 complex. Verified p50 and p65 ligands may be evaluated alongside compounds that modulate various stages of the pathway for their ability to modulate NF-κB activity using a luciferase reporter gene assay and EMSA assays using established protocols (Castrillo et al, J. Biol. Chem. (2001) 19: 15854-15860; Witherow, et al, Proc. Natl. Acad. ScL USA (2004) 101 : 8603-8607). The compounds may also be subjected to an image-based nuclear translocation assay to identify compounds that decrease or increase entry of the heterodimer into the nucleus. The effect of compounds on proteasome-mediated degradation of IκBα may also be evaluated. Compounds may also be evaluated for their ability to affect cell cycle progression and apoptosis using flow cytometry. Selected compounds may be studied using gene expression profiling. Active ligands may be subjected to detailed mechanistic studies to narrow down which specific biomolecular interaction(s) are perturbed. For example, compounds that prevent nuclear translocation may disrupt a heterodimer- importin interaction or may prevent formation of the heterodimer itself. Compounds that do not alter translocation but prevent transcription may function by disrupting interactions with a specific transcriptional partner such as a co-activator or parts of the general transcriptional machinery. Once hypotheses about specific interactions are developed, biophysical assays tools such as SPR, fluorescence polarization, or fluorescence resonance energy transfer (FRET) may be used to evaluate the effect of specific compounds on protein-protein or protein-DNA interactions. [0098] The known plant hormone gibberellic acid (GA, gibberellin A3), which stimulates germination and influences plant cellular expansion, was identified as a ligand for NF-κB. Gibberellic acid has not been previously shown to bind to NF-κB. The hormone is of very low toxicity to animals and is used in the grape and cherry industries to regulate fruit size. In plants, the hormone activates transcription when it binds to its receptor and subsequently leads to SCF E3 ubiquitin ligase-mediated degradation of a set of transcriptional repressor proteins (Ueguchi-Tanaka et al, Nature (2005) 437: 693-698; incorporated herein by reference). Gibberellic acid was identified as a selective ligand to p50 using SMMs. Gibberellic acid binds to p50 with a KQ of -400 nM as judged by kinetic and equilibrium analysis of SPR data {Figure 3B). A survey of publicly available data in Chembank shows that gibberellic acid inhibits SIRTl deacetylase activity and inhibits E3 ubiquitin ligase activity of the mammalian apoptosis inhibitor cIAPl . SIRTl, a member of the sirtuin family that regulates transcriptional silencing and cell survival, is known to bind the p65 subunit of NF-κB and inhibits transcription by deacetylating p65 at K310 (Yeung et al, EMBO J. (2004) 23: 2369-2380). In a recent report, HIV-I Tat was shown to bind the deacetylase domain of SIRT and inhibit SIRTl -mediated deacetylation of p65, resulting in increased NF- KB acetylation levels and increased transactivation (K won et al, Cell Host Microb. (2008) 3: 158-167; incorporated herein by reference). Without wishing to be bound to a particular theory, one possible mode of interaction for gibberellic acid may involve binding p50-p65 and preventing SIRTl deacetylation of p65. cIAPl is a member of the tumor-necrosis factor receptor (TNFR)-associated signaling complex and is required for proper polyubiquitination of Rip 1 and NF-κB activation upon TNF-a treatment (Mahoney et al, Proc. Natl. Acad. ScL USA (2008) 105: 11778-11783; incorporated herein by reference). The binding of GA to p50 leads to decreased cIAPl activity, which may be related to altered processing of pi 05 or levels of NIKl (Vince et al, Cell (2007) 131 : 682-693; incorporated herein by reference). [0099] Several gene expression profiles for the p50 ligand gibberellic acid and many known NF-κB pathway modulators may be found in the most recent version of Connectivity Map (CMAP) (Lamb et al, Science (2006) 313: 1929-1935; incorporated herein by reference). The current version of CMAP contains more than 6,100 profiling instances including 1,309 discrete small molecules from a variety of sources (e.g., FDA-approved drugs, known bioactives, and products of diversity-oriented synthesis). Each treatment instance gives rise to a rank-ordered list of -22,000 genes. Profiles were generated for gibberellic acid at 12 μM doses in the MCF7 breast cancer epithelial cell line, the PC3 prostate cancer epithelial cell line, and leukemia HL60 cell line. Query signatures for gibberellic acid in all three cell lines were used to probe CMAP for highly correlated or anti- correlated gene expression signatures for the other 1,308 compounds. For example, the query signature for the PC3 instance consisted of 524 genes (290 up- and 234 down-regulated). High connectivity scores were observed for several known modulators of NF-κB and antiinflammatory agents. The top 20-ranked compounds, both correlated and anti-correlated, are listed in a table in Figure 4 along with several bar views constructed for representative compounds. The top compound, camptothecin, is an anti-cancer agent that acts by inhibiting DNA topoisomerase I activity and introducing DNA double-strand breaks during DNA replication that lead to S-phase cytotoxicity (Liew et al, Curr. Pharm. Des. (2008) 14:1078- 1097; incorporated herein by reference). Previous studies have shown that DNA damage by camptothecin translates into activation of cytoplasmic signaling that results in release of NF- KB from IκKα and activation of NF-κB (Huang et al, J. Biol. Chem. (2000) 13: 9501-9509; incorporated herein by reference). The PC3 signature also recovered a reactive oxygen species-initiating teratogen known as etifenin (also known as phenytoin) that increases NF- KB activity in various tissue types (Kennedy et al, MoI. Pharmacol. (2004) 66: 404-412; incorporated herein by reference). Examples of additional correlated compounds include azacitidine, a DNA methyltransferase inhibitor that inhibits NF-κB translocation (Fabre et al., Cell Cycle (2008) 7:2139-2145; incorporated herein by reference), and the anti-inflammatory agents phenacetin and acetylsalicylic acid. Examples of anti-correlated compounds in the list include chenodeoxycholic acid, which induces cIAPl expression and NF-κB transcriptional activity (Hirano et al, J. Gastroenterol Hepatol. (2006) 21 :1807-1813; incorporated herein by reference), and the anti-inflammatory protease inhibitor tranexamic acid. Similar queries were performed for HL60 and MCF7 instances involving the p50 binder. In all instances, several known pathway modulators and anti-inflammatory compounds were among the top connected compounds to the p50 binder (see Figure 13). Current CMAP analyses involve reverse queries where a selected known pathway inhibitor is used to generate the query and we look for the rank corresponding to the p50 ligand to help generate mechanistic hypotheses related to the p50 compound. For example, across all signatures it is noted that many compounds annotated as NF-κB activators correlated positively with gibberellic acid. The CMAP signatures are supporting evidence that gibberellic acid perturbs NF-κB function. Uses ofGiberellins as Modulators ofNF-kB

[00100] NF-κB contributes to progression of cancers by serving both as a positive regulator of cell growth and as a negative regulator of apoptosis (Rayet et al. , Oncogene (1999) 18: 6938-6947; Mayo et al, Biochim. Biophys. Acta (2000) 1470: M55-M62; each of which is incorporated herein by reference). NF-κB stimulates expression of cell cycle- specific proteins c-Myc and cyclin Dl (Romashkova et al, Nature (1999) 401 :86-90; Hinz et al, MoI Cell Biol. (1999) 19:2690-2698; each of which is incorporated herein by reference). The constitutive expression of these proteins results in sustained cell proliferation. Continued expression of c-Myc ultimately leads to apoptosis. NF-κB can block c-Myc's apoptosis effects, thereby stimulating proliferation without cytotoxicity. NF-κB also inhibits the ability of tumor necrosis factor (TNF) to induce cell death as well as protect cells from the effects of ionizing radiation and chemo therapeutic drugs (Barkett et al, Oncogene (1999) 18:6910- 6924; incorporated herein by reference). Thus, NF-κB promotes both hyperplasia and resistance to oncological treatments, which are hallmarks of many cancers. [00101] Many clinically used chemotherapeutic agents (including the vinca alkaloids, vincristine and vinblastine, camptothecin and many others) have recently been shown to activate NF-κB resulting in a retardation of their cytotoxicity. This form of resistance is commonly referred to as NF-κB -mediated chemoresistance. Inhibition of NF-κB has shown to increase the sensitivity to chemotherapeutic agents of tumor cells and solid tumors (Wang, et al, Science (1996) 274:784-787; incorporated herein by reference). Inhibition of NF-κB activation has been linked to the chemopreventive properties of several anti-cancer compounds {e.g., selenium) (Gasparian et al, MoI Cancer Ther (2002) 1 : 1079-1087; incorporated herein by reference). Down-regulation of NF-κB activity is considered a very attractive strategy for developing new cancer treatments. There is currently intense interest in elucidating the details of the signaling pathway (especially the mechanism of NF-κB activation) in order to identify suitable molecular targets for therapeutic intervention. [00102] In the case of inflammation, NF-κB plays important roles in both the initiation and maintenance of the inflammatory response (Makarov, MoI Med Today (2000) 6:441-448; incorporated herein by reference). Activated T cells, such as activated CD4+ T helper cells, trigger inflammation. The T helper cell population can differentiate further to two subset populations that have opposite effects on the inflammatory response. The ThI subset is considered proinflammatory, as these cells mediate cellular immunity and activate macrophages. The Th2 subset is considered anti-inflammatory, as these cells mediate humoral immunity and down-regulate macrophage activation. The subsets are distinguishable by the different types of cytokine profiles that they express upon differentiation. NF-kB stimulates production of cytokine profiles characteristic of the ThI subset type, leading to a proinflammatory response. Conversely, suppression of NF-κB activation leads to production of cytokine profiles characteristic of the Th2 subset type that mediates an anti-inflammatory response. Once activated, these inflammatory cytokines and growth factors can act through autocrine loops to maintain NF-κB activation in non-immune cells within the lesion (Makarov, MoI Med Today (2000) 6:441-448; incorporated herein by reference). For example, NF-κB regulates the expression of cytokines interleukin 1-beta (IL- lβ (Baldwin, Annu. Rev. Immunol. (1996) 14: 649-683; incorporated herein by reference) and tumor necrosis factor alpha (TNF-α), which are considered essential mediators of the inflammatory response. Conversely, these gene products positively activate NF-κB expression that leads to persistence of the inflammatory state. For example, TNF products have been implicated in promoting inflammation in several gastrointestinal clinical disorders that include: alcoholic liver disease, non-alcoholic steatohepatitis, prancreatitis (including chronic, acute and alcohol-induced), and inflammatory bowel disorders, such as ulcerative colitis and Crohn's Disease.

[00103] Continued NF-κB activation also promotes tissue remodeling in the inflammatory lesions (Makarov, MoI Med Today (2000) 6: 441-448; incorporated herein by reference). Several NF-κB-responsive genes have been implicated in this regard and include growth factors that are important to neovascularization {e.g., VEGF), matrix proteinases (including metalloproteases), cyclooxygenase, nitric oxide synthase, and enzymes that are involved in the synthesis of proinflammatory prostaglandins, nitric oxide, and nitric oxide metabolites. Such tissue remodeling is often accompanied by breakdown of healthy cells as well as by hyperplasia, both of which are often observed in rheumatoid arthritis and other inflammatory diseases.

[00104] Alzheimer's disease represents another example of a condition that displays an inflammatory component in its pathogenesis. Recent studies indicate that abnormal regulation of the NF-κB pathway may be central to the pathogenesis of Alzheimer's disease. NF-κB activation correlates with the initiation of neuritic plaques and neuronal apoptosis during the early phases of the disease. For example, NF-κB immunoreactivity is found predominantly in and around early neuritic plaque types, whereas mature plaque types display reduced NF-κB activity (Kaltschmidt, et al, Proc. Natl. Acad. ScL USA (1999) 96: 9409-9414; incorporated herein by reference). [00105] Modulation of NF-κB may also be important in treating immunodeficiency. In the case of the human immunodeficiency virus (HIV), infection results in NF-κB activation, which results in regular viral persistence (Rabson, et al, Adv Pharmacol, 48, 161-207 (2000); incorporated herein by reference). HIV-I replication is regulated through a variety of viral proteins as well as cellular transcription factors, in particular NF-κB, that interact with the viral long terminal repeat (Asin, et al, J Virol (1999) 73: 3893-3903; incorporated herein by reference). HIV-I is able to enter a latent state in which the integrated provirus remains transcriptionally silent. The ability to continue to infect cells latently aids the virus to establish persistent infections and avoid the host immune system. The latent virus can establish large reservoirs of genetic variants in T-cells residing in lymphoid tissue. In addition, a recent study implicates NF-κB with the reactivation of latent HIV in T-cells in patents undergoing antiviral therapy (Finzi, et al. Science, 278, 1295-1300 (1997); Baba et al, EP 0931544 A2; Callahan, et al, WO 02/30423; each of which is incorporated herein by reference).

[00106] Invasive infection with Gram-positive or Gram-negative bacteria can result in septic shock and death. Invasion of the blood stream by bacteria causes sepsis syndrome in humans as a result of an endotoxin, lipopoly saccharide (LPS) (Bohrer, J. Clin. Invest. (1997) 972-985; incorporated herein by reference), that triggers an overwhelming inflammation response in the host. The mechanism by which LPS causes septic shock is through the activation of the transcription factor NF-κB. Activation of NF-κB by LPS initiates the massive release of cytokines resulting in a potentially fatal septic shock. Septic shock, caused by an exaggerated host response to these endotoxins often leads to multiple organ dysfunction, multiple organ failure, and remains the leading cause of death in trauma patients. Inhibition of NF-κB activation would, therefore, be therapeutically useful in the treatment of septic shock and other bacterial infections. Inhibition of NF-κB activation may also be therapeutically useful in the treatment of certain viral diseases.

[00107] The invention provides methods of treating a disease using giberellins of formula I. The inventive method involves the administration of a therapeutically effective amount of a compound of formula I to a subject (including, but not limited to a human or other animal) in need of it.

[00108] Certain compounds of formula I modulate the NF-κB pathway. Thus, in certain embodiments, the present invention provides a method for treating a NF-κB-mediated disorder comprising the step of administering to a patient in need thereof a compound of formula I or pharmaceutically acceptable composition thereof. [00109] As used herein, the term "NF-κB-mediated" disorders means any disease or other deleterious condition in which NF-κB is known to play a role. Accordingly, another embodiment of the present invention relates to treating or lessening the severity of one or more diseases in which the NF-κB pathway is known to play a role including, but not limited to, cancers or other proliferative diseases, inflammatory disorders, immunodeficiencies, gastrointestinal disorders, neurodegenerative disorders, autoimmune diseases, and sepsis. In another embodiment, compounds of formula I may also act as chemosensitizers. [00110] According to the methods of the current invention, compounds of formula I and pharmaceutical compositions thereof may be used in treating or preventing any disease or condition including, but not limited to, asthma, arthritis, inflammatory diseases (e.g., Crohn's disease, rheumatoid arthritis, psoriasis), proliferative diseases (e.g., cancer, benign neoplasms, diabetic retinopathy), cardiovascular diseases, neurodegenerative diseases, neurodegenerative disorders (e.g., Huntington's disease, Alzheimer's disease), and autoimmune diseases (e.g., rheumatoid arthritis, lupus). The inventive compounds and pharmaceutical compositions may be administered to animals, preferably mammals (e.g., domesticated animals, cats, dogs, mice, rats), and more preferably humans. Any method of administration may be used to deliver the inventive compound or pharmaceutical composition thereof to the animal. In certain embodiments, the compound or pharmaceutical composition thereof is administered orally. In other embodiments, the compound or pharmaceutical composition thereof is administered parenterally.

[00111] The methods of the invention can be particularly used for the treatment of cancer. Examples of cancers that may be treated with compounds of formula I according to the present invention include malignancies of the breast, bladder, blood, bone, bone marrow, brain, central and peripheral nervous system, colon, connective tissue, endocrine glands (e.g., thyroid and adrenal cortex), esophagus, endometrium, eye, germ cells, head and neck, kidney, liver, lung, larynx and hypopharynx, muscle, ovary, pancreas, prostate, rectum, retina, small intestine, soft tissue, testis, stomach, skin, ureter, vagina, and vulva. Inherited cancers exemplified by retinoblastoma and Wilms tumor are included. In addition, cancers include primary tumors in said organs and corresponding secondary tumors in distant organs ("metastases"). Hematological malignancies are exemplified by aggressive and indolent forms of leukemia and lymphoma, namely non-Hodgkin's disease, chronic and acute myeloid leukemia (CML/ AML), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), Hodgkins disease, multiple myeloma, and T-cell lymphoma (e.g., peripheral T cell lymphoma, cutaneous T cell lymphoma). Also included are myelodysplastic syndrome, plasma cell neoplasia, paraneoplastic syndromes, cancers of unknown primary site as well as AIDS-related malignancies.

[00112] In certain embodiments, the current invention provides a method for the treatment of benign neoplasia. Examples of benign neoplasia treated with compounds according to the present invention include, but are not limited to, benign soft tissue tumors, bone tumors, brain and spinal tumors, eyelid and orbital tumors, granuloma, lipoma, meningioma, multiple endocrine neoplasia, nasal polyps, pituitary tumors, prolactinoma, pseudotumor cerebri, seborrheic keratoses, stomach polyps, thyroid nodules, cystic neoplasms of the pancreas, hemangiomas, vocal cord nodules, polyps, and cysts, Castleman disease, chronic pilonidal disease, dermatofibroma, pilar cyst, pyogenic granuloma, and juvenile polyposis syndrome.

[00113] In certain embodiments, the present invention provides methods for treating or lessening the severity of autoimmune diseases including, but not limited to, inflammatory bowel disease, arthritis, systemic lupus erythematosus, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Ord's thyroiditis, Graves' disease, Sjogren's syndrome, multiple sclerosis, Guillain-Barre syndrome, acute disseminated encephalomyelitis, Addison's disease, opsoclonus-myoclonus syndrome, ankylosing spondylosis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, celiac disease, Goodpasture's syndrome, idiopathic thrombocytopenic purpura, optic neuritis, scleroderma, primary biliary cirrhosis, Reiter's syndrome, Takayasu's arteritis, temporal arteritis, warm autoimmune hemolytic anemia, Wegener's granulomatosis, psoriasis, alopecia universalis, Behcet's disease, chronic fatigue, dysautonomia, endometriosis, interstitial cystitis, neuromyotonia, scleroderma, or vulvodynia.

[00114] In some embodiments, the present invention provides a method for treating or lessening the severity of an inflammatory disease including, but not limited to, asthma, appendicitis, Behcet's disease, Blau syndrome, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic recurrent multifocal osteomyelitis (CRMO), colitis, conjunctivitis, cryopyrin associated periodic syndrome (CAPS), cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, familial cold-induced autoinflammatory syndrome, familial Mediterranean fever (FMF), fasciitis, fibrositis, gastritis, gastroenteritis, hepatitis, hidradenitis suppurativa, laryngitis, mastitis, meningitis, mevalonate kinase deficiency (MKD), Muckle-Well syndrome, myelitis myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis, otitis, pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis, pleuritis, phlebitis, pneumonitis, pneumonia, proctitis, prostatitis, pyelonephritis, pyoderma gangrenosum and acne syndrome (PAPA), pyogenic sterile arthritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, systemic juvenile rheumatoid arthritis, tendonitis, TNF receptor associated periodic syndrome (TRAPS), tonsillitis, uveitis, vaginitis, vasculitis, or vulvitis.

[00115] In certain embodiments, the present invention provides methods for treating or lessening the severity of arthropathies and osteopathological diseases including, but not limited to, rheumatoid arthritis, osteoarthrtis, gout, polyarthritis, and psoriatic arthritis. [00116] In certain embodiments, the present invention provides methods for treating or lessening the severity of hyperproliferative diseases including, but not limited to, psoriasis or smooth muscle cell proliferation including vascular proliferative disorders, atherosclerosis, and restenosis. In certain embodiments, the present invention provides methods for treating or lessening the severity of endometriosis, uterine fibroids, endometrial hyperplasia, and benign prostatic hyperplasia.

[00117] In certain embodiments, the present invention provides methods for treating inflammatory diseases including, but not limited to, allergies, ulcerative colitis, inflammatory bowel disease, Crohns disease, allergic rhinitis, allergic dermatitis, cystic fibrosis, chronic obstructive bronchitis, and asthma.

[00118] In certain embodiments, the present invention provides methods for treating or lessening the severity of neuropathological disorders and/or protein aggregation disorders including, but not limited to, Parkinson's disease, Alzheimer's disease, or polyglutamine related disorders including, but not limited to, Huntington's disease, Spinocerebellar ataxia 1 (SCA 1), Machado-Joseph disease (MJD)/Spinocerebella ataxia 3 (SCA 3), Kennedy disease/Spinal and bulbar muscular atrophy (SBMA), Dentatorubral pallidolusyian atrophy (DRPLA), fronto-temporal dementia, Lewy body disease, Pick's disease, multiple system atrophy, amyotrophic lateral sclerosis, and progressive supranuclear palsy (PSP). [00119] The exact amount of compound required to treat the indicated disease or disorder will vary from subject to subject, depending on the species, age, and general condition of the subject, the disease or disorder being treated, the particular compound, its mode of administration, its mode of activity, and the like. The compounds of formula I are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compound of formula I and compositions thereof will be decided by a physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific protein employed; the specific composition employed; the age, body weight, general health, sex, and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors known in the medical arts.

[00120] Furthermore, after formulation with an appropriate pharmaceutically acceptable carrier in a desired dosage, the pharmaceutical compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), bucally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated. In certain embodiments, the proteins of the invention may be administered orally or parenterally at dosage levels sufficient to deliver from about 0.001 mg/kg to about 100 mg/kg, from about 0.01 mg/kg to about 50 mg/kg, preferably from about 0.1 mg/kg to about 40 mg/kg, preferably from about 0.5 mg/kg to about 30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, and more preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect. The desired dosage may be delivered three times a day, two times a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the desired dosage may be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations).

[00121] Liquid dosage forms for oral and parenteral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifϊers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In certain embodiments for parenteral administration, the compounds of the invention are mixed with solubilizing agents such Cremophor (polyethoxylated castor oil), alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and combinations thereof.

[00122] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U. S. P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of iηjectables.

[00123] The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[00124] In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as poly(lactide-co- glycolide). Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.

[00125] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[00126] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[00127] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.

[00128] The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active protein may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes.

[00129] Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this invention. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel. [00130] It will also be appreciated that the compounds and pharmaceutical compositions of the present invention can be employed in combination therapies, that is, the compounds and pharmaceutical compositions can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures. The particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. It will also be appreciated that the therapies employed may achieve a desired effect for the same disorder (for example, an inventive compound may be administered concurrently with another anticancer agent), or they may achieve different effects (e.g., control of any adverse effects).

[00131] Depending upon the particular condition or disease to be treated, additional therapeutic agents that are normally administered to treat that disease or condition may also be present in the compositions of this invention or may be used in combination with compounds of formula I.

[00132] According to the methods of the current invention, a compound of formula I may also be used to advantage in combination with other antiproliferative compounds. Such antiproliferative compounds include, but are not limited to aromatase inhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active compounds; alkylating compounds; histone deacetylase inhibitors; compounds which induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; kinase inhibitors; phosphatase inhibitors; farnesyl transferase inhibitors; antineoplastic antimetabolites; platin compounds; compounds targeting/decreasing a protein or lipid kinase activity and further anti-angiogenic compounds; compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase; gonadorelin agonists; anti-androgens; methionine aminopeptidase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematologic malignancies; compounds which target, decrease or inhibit the activity of Flt-3; Hsp90 inhibitors; temozolomide (Temodal^®); kinesin spindle protein inhibitors; MEK inhibitors; and leucovorin.

[00133] The term "aromatase inhibitor" as used herein relates to a compound that prevents the formation of estradiol, a female hormone, by interfering with an aromatase enzyme. Aromatase inhibitors are used as a type of hormone therapy for post-menopausal women who have hormone-dependent breast cancer. The term "aromatase inhibitor" includes, but is not limited to, steroids, particularly atamestane, exemestane and formestane and, in particular, non-steroidal agents, particularly aminoglutethimide, roglethimide, pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole, fadrozole, anastrozole, and letrozole. Exemestane is marketed under the trade name Aromasin™. Formestane is marketed under the trade name Lentaron™. Fadrozole is marketed under the trade name Afema™. Anastrozole is marketed under the trade name Arimidex™. Letrozole is marketed under the trade names Femara™ or Femar™. Aminoglutethimide is marketed under the trade name Orimeten™. A combination of the invention comprising a compound of formula I and a chemotherapeutic agent which is an aromatase inhibitor is particularly useful for the treatment of hormone receptor positive tumors, such as breast tumors.

[00134] The term "antiestrogen" as used herein relates to a compound which antagonizes the effect of estrogens at the estrogen receptor level. The term "antiestrogen" includes, but is not limited to tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride. Tamoxifen is marketed under the trade name Nolvadex™. Raloxifene hydrochloride is marketed under the trade name Evista™. Fulvestrant can be administered under the trade name Faslodex™. A combination of the invention comprising a chemotherapeutic agent which is an antiestrogen is particularly useful for the treatment of estrogen receptor positive tumors, such as breast tumors. [00135] The term "anti-androgen" as used herein relates to any substance which is capable of inhibiting the biological effects of androgenic hormones and includes, but is not limited to, bicalutamide (Casodex™). The term "gonadorelin agonist" as used herein includes, but is not limited to abarelix, goserelin, and goserelin acetate. Goserelin can be administered under the trade name Zoladex™.

[00136] The term "topoisomerase I inhibitor" as used herein includes, but is not limited to, topotecan, gimatecan, irinotecan, camptothecian and its analogues, 9-nitrocamptothecin, and the macromolecular camptothecin conjugate PNU-166148. Irinotecan can be administered, e.g., in the form as it is marketed under the trademark Camptosar™. Topotecan is marketed under the trade name Hycamptin™.

[00137] The term "topoisomerase II inhibitor" as used herein includes, but is not limited to, anthracyclines such as doxorubicin (including liposomal formulation, such as Caelyx™), daunorubicin, epirubicin, idarubicin, and nemorubicin; anthraquinones such as mitoxantrone and losoxantrone; and podophillotoxines such as etoposide and teniposide. Etoposide is marketed under the trade name Etopophos™. Teniposide is marketed under the trade name VM 26-Bristol Doxorubicin is marketed under the trade name Acriblastin ™ or Adriamycin™. Epirubicin is marketed under the trade name Farmorubicin™. Idarubicin is marketed under the trade name Zavedos™. Mitoxantrone is marketed under the trade name Novantron.

[00138] The term "microtubule active agent" relates to microtubule stabilizing, microtubule destabilizing compounds, and microtublin polymerization inhibitors including, but not limited to, taxanes, such as paclitaxel and docetaxel; vinca alkaloids, such as vinblastine or vinblastine sulfate, vincristine, vincristine sulfate, and vinorelbine; discodermolides; cochicine; and epothilones and derivatives thereof. Paclitaxel is marketed under the trade name Taxol™. Docetaxel is marketed under the trade name Taxotere™. Vinblastine sulfate is marketed under the trade name Vinblastin R.P™. Vincristine sulfate is marketed under the trade name Farmistin™.

[00139] The term "alkylating agent" as used herein includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan, or nitrosourea (BCNU or Gliadel). Cyclophosphamide is marketed under the trade name Cyclostin™. Ifosfamide is marketed under the trade name Holoxan™.

[00140] The term "histone deacetylase inhibitors" or "HDAC inhibitors" relates to compounds which target, decrease or inhibit activity of histone deacetylase (HDAC), such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA). This includes, but is not limited to, suberoylanilide hydroxamic acid (SAHA), sodium butyrate, MS275, FK228 (formerly FR901228), Trichostatin A, and compounds disclosed in US 6,552,065 including, but not limited to, N-hydroxy-3-[4-[[[2-(2-methyl-lH-indol-3-yl)-ethyl]- amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof and N-hydroxy-3-[4-[(2-hydroxyethyl){2-(lH-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2- propenamide, or a pharmaceutically acceptable salt thereof, particularly the lactate salt. [00141] The term "antineoplastic antimetabolite" includes, but is not limited to, 5- fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds, such as 5- azacytidine and decitabine, methotrexate and edatrexate, and folic acid antagonists such as pemetrexed. Capecitabine is marketed under the trade name Xeloda™. Gemcitabine is marketed under the trade name Gemzar™.

[00142] The term "platin compound" as used herein includes, but is not limited to, carboplatin, cis-platin, cisplatinum and oxaliplatin. Carboplatin can be administered, e.g., in the form as it is marketed under the trademark Carboplat™. Oxaliplatin can be administered, e.g., in the form as it is marketed under the trademark Eloxatin™.

[00143] The term "compounds targeting/decreasing a protein or lipid kinase activity; or a protein or lipid phosphatase activity; or further anti-angiogenic compounds" as used herein includes, but is not limited to, protein tyrosine kinase and/or serine and/or threonine kinase inhibitors or lipid kinase inhibitors, such as a) compounds targeting, decreasing or inhibiting the activity of the platelet-derived growth factor-receptors (PDGFR), such as compounds which target, decrease or inhibit the activity of PDGFR, especially compounds which inhibit the PDGF receptor, such as an N-phenyl-2-pyrimidine-amine derivative, such as imatinib, SUlOl, SU6668 and GFB-11 1; b) compounds targeting, decreasing or inhibiting the activity of the fibroblast growth factor-receptors (FGFR); c) compounds targeting, decreasing or inhibiting the activity of the insulin-like growth factor receptor I (IGF-IR), such as compounds which target, decrease or inhibit the activity of IGF-IR, especially compounds which inhibit the kinase activity of IGF-I receptor, or antibodies that target the extracellular domain of IGF-I receptor or its growth factors; d) compounds targeting, decreasing or inhibiting the activity of the Trk receptor tyrosine kinase family, or ephrin B4 inhibitors; e) compounds targeting, decreasing or inhibiting the activity of the AxI receptor tyrosine kinase family; f) compounds targeting, decreasing or inhibiting the activity of the Ret receptor tyrosine kinase; g) compounds targeting, decreasing or inhibiting the activity of the Kit/SCFR receptor tyrosine kinase, such as imatinib; h) compounds targeting, decreasing or inhibiting the activity of the C-kit receptor tyrosine kinases, which are part of the PDGFR family, such as compounds which target, decrease or inhibit the activity of the c-Kit receptor tyrosine kinase family, especially compounds which inhibit the c-Kit receptor, such as imatinib; i) compounds targeting, decreasing or inhibiting the activity of members of the c-Abl family, their gene-fusion products (e.g. BCR-AbI kinase) and mutants, such as compounds which target decrease or inhibit the activity of c-Abl family members and their gene fusion products, such as an N-phenyl-2-pyrimidine-amine derivative, such as imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410; PD173955 from ParkeDavis; or dasatinib (BMS-354825); j) compounds targeting, decreasing or inhibiting the activity of members of the protein kinase C (PKC) and Raf family of serine/threonine kinases, members of the MEK, SRC, JAK, FAK, PDKl, PKB/Akt, and Ras/MAPK family members, and/or members of the cyclin-dependent kinase family (CDK) including staurosporine derivatives, such as midostaurin; examples of further compounds include UCN-Ol, safingol, BAY 43-9006, Bryostatin 1, Perifosine; llmofosine; RO 318220 and RO 320432; GO 6976; lsis 3521; LY333531/LY379196; isochinoline compounds; FTIs; PD184352 or QAN697 (a P13K inhibitor) or AT7519 (CDK inhibitor), ARRY142886 from Array BioPharma, AZD6244 from AstraZeneca, PDl 81461 from Pfizer; k) compounds targeting, decreasing or inhibiting the activity of protein-tyrosine kinase inhibitors, such as compounds which target, decrease or inhibit the activity of protein-tyrosine kinase inhibitors include imatinib mesylate (Gleevec™) or tyrphostin such as Tyrphostin A23/RG-50810; AG 99; Tyrphostin AG 213; Tyrphostin AG 1748; Tyrphostin AG 490; Tyrphostin B44; Tyrphostin B44 (+) enantiomer; Tyrphostin AG 555; AG 494; Tyrphostin AG 556, AG957 and adaphostin (4-{[(2,5- dihydroxyphenyl)methyl]amino} -benzoic acid adamantyl ester; NSC 680410, adaphostin); 1) compounds targeting, decreasing or inhibiting the activity of the epidermal growth factor family of receptor tyrosine kinases (EGFRl ErbB2, ErbB3, ErbB4 as homo- or heterodimers) and their mutants, such as compounds which target, decrease or inhibit the activity of the epidermal growth factor receptor family are especially compounds, proteins or antibodies which inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptor, ErbB2, ErbB3 and ErbB4 or bind to EGF or EGF related ligands, CP 358774, ZD 1839, ZM 105180; trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, Cl- 1033, EKB-569, GW-2016, El.1, E2.4, E2.5, E6.2, E6.4, E2.l l, E6.3 or E7.6.3, and 7H- pyrrolo-[2,3-d]pyrimidine derivatives; and m) compounds targeting, decreasing or inhibiting the activity of the c-Met receptor, such as compounds which target, decrease or inhibit the activity of c-Met, especially compounds which inhibit the kinase activity of c-Met receptor, or antibodies that target the extracellular domain of c-Met or bind to HGF. [00144] Further anti-angiogenic compounds include compounds having another mechanism for their activity, e.g., unrelated to protein or lipid kinase inhibition e.g. thalidomide (Thalomid™) and TNP-470.

[00145] Compounds which target, decrease, or inhibit the activity of a protein or lipid phosphatase are inhibitors of phosphatase 1 , inhibitors of phosphatase 2 A, or inhibitors of

CDC25, such as okadaic acid or derivatives thereof.

[00146] Compounds which induce cell differentiation processes include, but are not limited to, retinoic acid, α- γ- or δ- tocopherol, or α- γ- or δ-tocotrienol.

[00147] The term cyclooxygenase inhibitor as used herein includes, but is not limited to,

Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acid and derivatives, such as celecoxib (Celebrex™), rofecoxib (Vioxx™), etoricoxib, valdecoxib or a 5 -alky 1-2- arylaminophenylacetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenyl acetic acid, lumiracoxib.

[00148] The term "bisphosphonates" as used herein includes, but is not limited to, etridonic, clodronic, tiludronic, pamidronic, alendronic, ibandronic, risedronic and zoledronic acid. Etridonic acid is marketed under the trade name Didronel™. Clodronic acid is marketed under the trade name Bonefos™. Tiludronic acid is marketed under the trade name

Skelid™. Pamidronic acid is marketed under the trade name Aredia™. Alendronic acid is marketed under the trade name Fosamax™. Ibandronic acid is marketed under the trade name

Bondranat™. Risedronic acid is marketed under the trade name Actonel™. Zoledronic acid is marketed under the trade name Zometa™. The term "mTOR inhibitors" relates to compounds which inhibit the mammalian target of rapamycin (mTOR) and which possess antiproliferative activity such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779 and ABT578.

[00149] The term "heparanase inhibitor" as used herein refers to compounds which target, decrease or inhibit heparan sulfate degradation. The term "heparanase inhibitor" includes, but is not limited to, PI-88.

[00150] The term "biological response modifier" as used herein refers to an agent used in biological response modifier therapy. "Biological response modifier therapy" or "BRM therapy" as used herein refers to treatment to boost or restore the ability of the immune system to fight cancer, infections, and other diseases. BRM therapy may also be used to lessen certain side effects that may be caused by some cancer treatments. Agents used in

BRM therapy include monoclonal antibodies, growth factors, and vaccines. These agents may also have a direct antitumor effect. BRM therapy is also sometimes called immunotherapy, biological therapy, or biotherapy.

[00151] The term "inhibitor of Ras oncogenic isoforms", such as H-Ras, K-Ras, or N-

Ras, as used herein refers to compounds which target, decrease, or inhibit the oncogenic activity of Ras; for example, a "farnesyl transferase inhibitor" such as L-744832, DK8G557, or Rl 15777 (Zarnestra™). The term "telomerase inhibitor" as used herein refers to compounds which target, decrease, or inhibit the activity of telomerase. Compounds which target, decrease, or inhibit the activity of telomerase are especially compounds which bind to the telomerase receptor, such as telomestatin.

[00152] The term "methionine aminopeptidase inhibitor" as used herein refers to compounds which target, decrease, or inhibit the activity of methionine aminopeptidase.

Compounds which target, decrease, or inhibit the activity of methionine aminopeptidase include, but are not limited to, bengamide or a derivative thereof.

[00153] The term "proteasome inhibitor" as used herein refers to compounds which target, decrease or inhibit the activity of the proteasome. Compounds which target, decrease or inhibit the activity of the proteasome include, but are not limited to, Bortezomib

(Velcade™) and MLN 341.

[00154] The term "matrix metalloproteinase inhibitor" or ("MMP" inhibitor) as used herein includes, but is not limited to, collagen peptidomimetic and nonpeptidomimetic inhibitors; tetracycline derivatives, e.g., hydroxamate peptidomimetic inhibitor batimastat and its orally bioavailable analogue marimastat (BB-2516); prinomastat (AG3340); metastat

(NSC 683551); BMS-279251 ; BAY 12-9566; TAA211 ; MMI270B; or AAJ996.

[00155] The term "compounds used in the treatment of hematologic malignancies" as used herein includes, but is not limited to, FMS-like tyrosine kinase inhibitors, which are compounds targeting, decreasing or inhibiting the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, 1 -β-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors, which are compounds which target, decrease or inhibit anaplastic lymphoma kinase.

[00156] Compounds which target, decrease or inhibit the activity of FMS-like tyrosine kinase receptors (FK-3R) are especially compounds, proteins or antibodies which inhibit members of the Flt-3R receptor kinase family, such as PKC412, midostaurin, a staurosporine derivative, SUl 1248 and MLN518.

[00157] The term "HSP90 inhibitors" as used herein includes, but is not limited to, compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90; degrading, targeting, decreasing or inhibiting the HSP90 client proteins via the ubiquitin proteosome pathway. Compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90 are especially compounds, proteins or antibodies which inhibit the ATPase activity of HSP90, such as 17-allylamino, 17-demethoxygeldanamycin (17AAG), a geldanamycin derivative; other geldanamycin related compounds; radicicol and HDAC inhibitors.

[00158] The term "antiproliferative antibodies" as used herein includes, but is not limited to, trastuzumab (Herceptin™), Trastuzumab-DMl, erbitux, bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40), and 2C4 Antibody. By antibodies is meant intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least 2 antibodies, humanized antibodies, and antibodies fragments so long as they exhibit the desired biological activity.

[00159] The term "antileukemic compounds" includes, for example, Ara-C, a pyrimidine analog, which is the 2'-alpha-hydroxy ribose (arabinoside) derivative of deoxycytidine. Also included is the purine analog of hypoxanthine, 6-mercaptopurine (6-

MP) and fludarabine phosphate.

[00160] Somatostatin receptor antagonists as used herein refer to compounds which target, treat or inhibit the somatostatin receptor such as octreotide, and SOM230.

[00161] The term "EDG binders" as used herein refers to a class of immunosuppressants that modulates lymphocyte recirculation, such as FTY720.

[00162] The term "ribonucleotide reductase inhibitors" refers to pyrimidine or purine nucleoside analogs including, but not limited to, fludarabine and/or cytosine arabinoside (ara-

C), 6-thioguanine, 5-fluorouracil, cladribine, 6-mercaptopurine, and/or pentostatin. In certain embodiments, ribonucleotide reductase inhibitors are hydroxyurea or 2-hydroxy-lH- isoindole-1, 3-dione derivatives.

[00163] Also included are in particular those compounds, proteins or monoclonal antibodies of VEGF such as l-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, 1 -(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate; Angiostatin™; Endostatin™; anthranilic acid amides; ZD4190; ZD6474; SU5416;

SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies, such as rhuMAb and RHUFab, VEGF aptamer such as Macugon; FLT-4 inhibitors, FLT-3 inhibitors,

VEGFR-2 IgGI antibody, Angiozyme (RPI 4610) and Bevacizumab (Avastin™).

[00164] Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, antisense oligonucleotides or oligonucleotide derivatives; shRNA, microRNA, or siRNA; or miscellaneous compounds or compounds with other or unknown mechanism of action.

[00165] Tumor cell damaging approaches refer to approaches such as ionizing radiation. The term "ionizing radiation" referred to above and hereinafter means ionizing radiation that occurs as either electromagnetic rays (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation is provided in, but not limited to, radiation therapy and is known in the art. See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al, Eds., 4th Edition, Vol. 1 , pp. 248-275 (1993).

[00166] Photodynamic therapy as used herein refers to therapy which uses certain chemicals known as photosensitizing compounds to treat or prevent cancers. Examples of photodynamic therapy include treatment with compounds, such as Visudyne™ and porfimer sodium.

[00167] According to the methods of the invention, compounds of formula I are also useful as co-therapeutic agents for use in combination with other therapeutic agents such as anti-inflammatory agents, bronchodilators, or antihistamines, particularly in the treatment of inflammatory airways diseases such as those mentioned hereinbefore, for example as potentiators of therapeutic activity of such drugs or as a means of reducing required dosaging or potential side effects of such drugs. A compound of formula I may be mixed with the other drug substance in a fixed pharmaceutical composition, or it may be administered separately, before, simultaneously with, or after the other drug substance. Accordingly the invention includes a combination of a compound of formula I as hereinbefore described with an anti-inflammatory, bronchodilatory, antihistamine or anti-tussive drug substance, said compound of the invention and said drug substance being in the same or different pharmaceutical composition.

[00168] Suitable anti-inflammatory agents include steroids, in particular glucocorticosteroids such as budesonide, beclamethasone dipropionate, fluticasone propionate, ciclesonide, or mometasone furoate; non-steroidal glucocorticoid receptor agonists; LTB4 antagonists, such LY2931 11, CGS025019C, CP-195543, SC-53228, BIIL 284, ONO 4057, and SB 209247; LTD4 antagonists such as montelukast and zafirlukast; PDE4 inhibitors such cilomilast (Ariflo® GlaxoSmithKline), Roflumilast (Byk Gulden), V- 11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering- Plough), Arofylline (Almirall Prodesfarma), PDl 89659 / PD168787 (Parke-Davis), AWD-12- 281 (Asta Medica), CDC-801 (Celgene), SeICID(TM) CC-10004 (Celgene), VM554/UM565 (Vernalis), T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo); A2a agonists; A2b antagonists; and beta-2 adrenergic receptor agonists such as albuterol (salbutamol), metaproterenol, terbutaline, salmeterol fenoterol, procaterol, formoterol, and pharmaceutically acceptable salts thereof. Suitable bronchodilatory drugs include anticholinergic and antimuscarinic compounds, in particular ipratropium bromide, oxitropium bromide, tiotropium salts, CHF 4226 (Chiesi), and glycopyrrolate.

[00169] Suitable antihistamines include cetirizine hydrochloride, acetaminophen, clemastine fumarate, promethazine, loratidine, desloratidine, diphenhydramine and fexofenadine hydrochloride, activastine, astemizole, azelastine, ebastine, epinastine, mizolastine and tefenadine.

[00170] Other useful combinations of compounds of the invention with antiinflammatory drugs are those with antagonists of chemokine receptors, e.g., CCR-I , CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9 and CCRlO, CXCRl , CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5 antagonists such as Schering-Plough antagonists SC-351125, SCH- 55700 and SCH-D, and Takeda antagonists such as N-[[4- [[[6,7-dihydro-2-(4-methylphenyl)-5H-benzo-cyclohepten-8-yl]carbonyl]amino]phenyl]- methyl]tetrahydro-N,N-dimethyl-2H-pyran-4- aminium chloride (TAK-770). [00171] The structure of the active compounds identified by code numbers, generic, or trade names may be taken from The Merck Index or from databases, e.g., Patents International (e.g., IMS World Publications).

[00172] The above-mentioned compounds, which can be used in combination with a compound of the current invention, can be prepared and administered as described in the art, such as in the documents cited above.

[00173] A compound of the current invention may also be used to advantage in combination with known therapeutic processes, for example, the administration of hormones or especially radiation.

[00174] Those additional agents may be administered separately from a gibberellin- containing composition, as part of a multiple dosage regimen. Alternatively, those agents may be part of a single dosage form, mixed together with a compound of this invention in a single composition. If administered as part of a multiple dosage regime, the two active agents may be submitted simultaneously, sequentially, or within a period of time from one another normally within twenty-four hours from one another.

[00175] As used herein, the term "combination," "combined," and related terms refers to the simultaneous or sequential administration of therapeutic agents in accordance with this invention. For example, a compound of the present invention may be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present invention provides a single unit dosage form comprising a compound of the current invention, an additional therapeutic agent, and a pharmaceutically acceptable excipient, carrier, adjuvant, or vehicle. [00176] In still another aspect, the present invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention, and in certain embodiments, includes an additional approved therapeutic agent for use as a combination therapy. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceutical products, which notice reflects approval by the agency of manufacture, use or sale for human administration.

Examples

[00177] We have developed a general approach to small-molecule probe discovery for transcription factors by coupling a high-throughput binding assay involving small-molecule microarrays (SMMs) with functional assays involving transcriptional and cellular readouts (Koehler et al, J. Am. Chem. Soc. 125, 8420-8241 (2003); Vegas et al, Chem. Soc. Rev. 37, 1385-1394 (2008); each of which is incorporated herein by reference). We applied this discovery approach to identify ligands for the transcription factor NF-κB. Aberrant NF-κB activity is associated with various cancers as well as inflammatory and autoimmune diseases. Although more than 700 compounds are known to affect the NF-κB signaling pathway by targeting various signaling enzymes, the proteasome, or through modification of DNA, none of the compounds are known to directly target the transcription factor itself. [00178] Small-molecule microarrays (SMMs) were used to identify gibberellic acid (GA₃) (Figure 3A) as a binder of the p50 subunit of NF-κB. p50 was screened as a purified His-tag fusion protein and binding to the array was detected through an Alexa647-labeled antibody against the His-tag. Gibberellic acid was verified as a binder of p50 using surface plasmon resonance (Biacore) having a K_D of approximately 226 nM (Figure 3B). Gibberelilic acid is produced industrially by stirred fermentation of Gibber ella fujikuroi and has been used widely in the agricultural industry to stimulate growth of sugarcane as well as a variety of fruits and vegetables. Gibberellic acid has also been evaluated for its ability to affect TNF-alpha stimulated transcription using a commercially available stable reporter cell line (GloResponse™ NF-kB-RE-luc2P HEK293, Promega) alongside control inhibitors such as parthenolide. GA₃ inhibits TNF-alpha stimulated transcription in reporter gene assays at submicromolar doses. GA3, GA4, and AGA appear to prevent translocation of p50 and p65 into the nucleus at 10 μM {Figures 6-10). Commercially available analogs of gibberellic acid, GA₄ and GA₉ methyl ester {Figure 5), also inhibit transcription by blocking translocation at 10 μM doses {Figure 11). Surveying public sites such as ChemBank, Pubchem, and Connectivity Map has also helped us identify supporting information to suggest a connection between gibberellins and NF-κB signaling {Figure 13). GA₃ scored as an assay positive in several cellular assays with connections to NF-κB including an IKB signaling assay, a cIAPl-TNFR assay, and two assays involving TNF-a mediated expression of either VCAM or E-selectin. GA₃ was profiled in three cell lines (HL-60, PC3, MCF7) using the Connectivity Map. We created a variety of signatures and consistently the most connected compounds to GA₃, across the three lines, were known NF-κB pathway modulators or anti-inflammatory agents {Figure 13).

[00179] Initial studies based on a commercial sample of GA₃ provided a dissociation constant of 216 nM for p50 using surface plasmon resonance (SPR). Upon purification, we observed a decrease in affinity to 2,300 nM for p50 and 13,000 nM for p65. Characterization of the commercial sample revealed the presence of allogibberic acid (AGA). AGA was also evaluated using SPR and in reporter gene and translocation assays. AGA also binds to p50 selectively over p65 with affinities of 76 nM and 6,340 nM, respectively. AGA also inhibits in the reporter assay < 1 μM and the translocation assay (a dose of 10 μM is effective and preliminary data suggests the EC50 will be < 1 μM). The compound does not appear to directly block binding of the transcription factor to DNA in vitro, but cellular TF-ELISA data involving nuclear fractions from treated cells show decreased binding of NF-κB to immobilized consensus sequence, consistent with the hypothesis that the compound disrupts NF-κB function in the cytoplasm.

[00180] While the gibberellins affect transcription in HEK293 cells and translocation in multiple cell types, the compounds do not appear to have an effect on viability of multiple cell lines in the absence of TNF-alpha stimulation.

[00181] Structures for five of the gibberellins evaluated to date, including GA₃ and AGA, are shown in Figure 12. Both LCMS and NMR methods have been used to evaluate samples of 1-4 that had been stored in both dry powder form and DMSO stock solutions. Consistent with the vendor's claim, the sample purities for 1, 3, and 4 are 80-95% of the major gibberellin with minor products corresponding to other gibberellins or common byproducts described below. It is important to note that LCMS indicates that our GA₉ME (2) sample contains a mixture of both the methyl ester and the carboxylic acid. Previous studies have shown that the A ring of GA₃ is reactive under several conditions and structures for selected byproducts are also shown. In fact, AGA (4) is prepared by treating GA₃ (1) with dilute mineral acid or boiling water, which leads to decarboxylase elimination of water and CO₂, aromatization of the A ring, and inverted stereochemistry at C9. This epimerization, which occurs via protonation of a C9-C10 unsaturated derivative, leads to a thermodynamically favored trans B-C ring fusion. When exposed to cold dilute aqueous alkali, the C4-C10 lactone bridge of GA₃ experiences an allylic transposition to give compound 5. Upon standing in distilled water at pH 7, GA₃ (1) slowly undergoes a self- induced acid-catalyzed trans elimination of the lactone bridge, yielding gibberellenic acid (6). Gibberellins containing saturated A rings, such as GA₄ (3), undergo facile epimerization of the C3 hydroxyl upon treatment with base; retroaldol cleavage of the C3-C4 bond leads to an aldehyde, which recyclizes to the more stable, equatorial alcohol (e.g., GA₄->7). Finally, it is worth noting the notoriously sensitive allylic tertiary alcohol of the C and D rings. Exposing this system to conditions that develop significant electron deficiency at C16 initiates a rearrangement of the C12-C13 bond, resulting in formation of a new bicyclo[3.2.1]octane with inverted configuration of the D ring (8). An ethylene glycol derivative of AGA has been preapred and has higher potency than AGA in the translocation assay (EC50 < 1 μM), presumably due to increased water solubility or cellular permeability.

* Note that this compound batch contains the COOH as an impurity

Claims

ClaimsWhat is claimed is:

1. A method of modulating the NF-κB pathway in a cell comprising contacting NF-κB with a compound of formula I:

wherein

R¹ is hydrogen or a group -OR²⁰, where R²⁰ is hydrogen, a glycoside, Cj_-6 alkyl group, or R¹ together with R² or R¹⁰ forms a bond (Ci-C₂ or Ci-Ci₀ double bond, respectively);

R² is hydrogen or a group -OR²¹, where R²¹ is hydrogen, a glycoside; or R² together with R⁴ forms a lactone; or R² together with R¹ or R³ forms a bond (Cj-C₂ or C₂-C₃ double bond, respectively); or R² together with R¹ or R³ and the intervening atoms forms an epoxide ring;

R³ is hydrogen, =0, or -OR²², where R²² is hydrogen or a glycoside; or R³ together with R forms a bond (C₂-C₃ double bond); or R together with R and the intervening atoms forms an epoxide ring;

R⁵ is hydrogen, a glycoside, unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, or arylalkyl; R⁶ is hydrogen or -OH; or R⁶ together with R⁷ forms a bond (Cn-Ci₂ double bond); or R⁶ together with R⁷ and the intervening atoms forms an epoxide ring;

R⁸ is hydrogen, hydroxyl, mercaptan, or halogen, amino, azido, -NR²⁴R²⁵, unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, or arylalkyl, or -OR , where R is hydrogen or a glycoside;

R¹¹ is hydrogen or hydroxyl or is absent;

R¹² is methyl, -CH₂OH, -CO₂H, or a glycoside ester of said -CO₂H;

R¹³ is methylene, or a divalent heteroatom, or -NR²⁹, where R²⁹ is -NHR³⁰ or -OR³⁰ where R³⁰ is hydrogen, or Ci_-20 alkyl, aryl, alkylaryl; and a double bond is present between Ci₆ and R¹³ when R¹¹ is absent; or R¹³ is hydrogen, hydroxyl, methyl, -CHO, -CH₂X, where X is halogen, -CH₂NR²⁹, where R²⁹ is -NR³⁰ or -OR³⁰, where R³⁰ is hydrogen, or Ci_-20 alkyl, aryl, or alkylaryl when R¹ ' is hydrogen or hydroxyl;

R¹⁴ is hydrogen or hydroxyl;

R¹⁵ is hydrogen, or R¹⁵ together with R⁹ forms a bond (C₉-Ci₅ double bond); or R¹⁵ together with R⁹ and the intervening atoms forms an epoxide ring; or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the compound of formula I is gibberellic acid of formula:

3. The method of claim 1 , wherein the compound of formula I is of formula:

4. The compound of claim 1, wherein the compound of formula I is of formula:

5. The compound of claim 1, wherein the compound of formula I is of formula:

6. The compound of claim 1, wherein the compound of formula I is of formula:

7. The method of claim 1, wherein the NF-κB is in an organism.

8. The method of claim 1 , wherin the NF-κB is in a mammal.

9. The method of claim 1 , wherin the NF-κB is in a human.

10. The method of claim 1, wherin the NF-κB is in a cell.

11. The method of claim 1 , wherein the compound binds the p50 subunit of NFKB.

12. The method of claim 1, wherein the compound binds the p65 subunit of NFKB.

13. The method of claim 1, wherein the compounds stimulates the Th2 response.

14. A method of treating a subject having an NFκB-mediated disorder comprising administering to a subject a therapeutically effective amount of a compound of formula I:

wherein

R¹ is hydrogen or a group -OR²⁰, where R²⁰ is hydrogen, a glycoside, Ci_-6 alkyl group, or R¹ together with R² or R¹⁰ forms a bond (Ci-C₂ or Ci-Ci₀ double bond, respectively);

R² is hydrogen or a group -OR²¹, where R²¹ is hydrogen, a glycoside; or R² together with R⁴ forms a lactone; or R² together with R¹ or R³ forms a bond (Ci-C₂ or C₂-C₃ double bond, respectively); or R together with R or R and the intervening atoms forms an epoxide ring; R³ is hydrogen, =0, or -OR²², where R²² is hydrogen or a glycoside; or R³ together with R² forms a bond (C₂-C₃ double bond); or R³ together with R² and the intervening atoms forms an epoxide ring;

R⁵ is hydrogen, a glycoside, unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, or arylalkyl;

R⁸ is hydrogen, hydroxyl, mercaptan, or halogen, amino, azido, -NR²⁴R²⁵, unsubstituted or substituted C_{I -20} alkyl, allyl, aryl, or arylalkyl, or -OR²⁷, where R²⁷ is hydrogen or a glycoside;

R⁹ is hydrogen or hydroxyl; or R⁹ together with R¹⁵ forms a bond (C₉-C1₅ double bond); or R⁹ together with R¹⁵ and the intervening atoms forms an epoxide ring;

R¹⁰ is hydrogen, methyl, -CHO, -CO₂H, or a glycoside ester of said -CO₂H, -CH₂OR²* or -OR²⁸, where R²⁸ is hydrogen or together with R⁴ forms a lactone; or R¹⁰ together with R¹ forms a bond (Ci-Ci₀ double bond); or R¹⁰ together with R¹ and the intervening atoms forms an epoxide ring;

R¹ ' is hydrogen or hydroxyl or is absent;

R¹² is methyl, -CH₂OH, -CO₂H, or a glycoside ester of said -CO₂H;

R¹³ is methylene, or a divalent heteroatom, or -NR²⁹, where R²⁹ is -NHR³⁰ or -OR³⁰ where R³⁰ is hydrogen, or Ci_-20 alkyl, aryl, alkylaryl; and a double bond is present between Ci₆ and R¹³ when R^{1 1} is absent; or R¹³ is hydrogen, hydroxyl, methyl, -CHO, -CH₂X, where X is halogen, -CH₂NR²⁹, where R²⁹ is -NR³⁰ or -OR³⁰, where R³⁰ is hydrogen, or C_1-20 alkyl, aryl, or alkylaryl when R¹ ' is hydrogen or hydroxyl;

R¹⁴ is hydrogen or hydroxyl; R¹⁵ is hydrogen, or R¹⁵ together with R⁹ forms a bond (C₉-Ci₅ double bond); or R¹⁵ togethe rr wwiitthh RR a anndd tthhee iinntteerrvveenniinngg aattoommss ffoorrmmss aann < epoxide ring; or a pharmaceutically acceptable salt thereof.

15. The method of claim 14, wherein the compound of formula I is gibberellic acid of formula:

16. The method of claim 14, wherein the compound of formula I is of formula:

17. The compound of claim 14, wherein the compound of formula I is of formula:

18. The compound of claim 14, wherein the compound of formula I is of formula:

19. The compound of claim 14, wherein the compound of formula I is of formula:

20. The method of claim 14, wherein the disorder is a proliferative disorder.

21. The method of claim 14, wherein the disorder is an inflammatory disorder.

22. The method of claim 14, wherein the disorder is an autoimmune disorder.

23. The method of claim 14, wherein the disorder is an immunodeficiency disorder.

24. The method of claim 14, wherein the disorder is a neurodegenerative disorder.

25. The method of claim 14, wherein the disorder is a gastrointestinal disorder.

26. The method of claim 14, wherein the disorder is sepsis.

27. The method of claim 14, wherein the disorder is an infectious disease.

28. A method of treating a drug-resistant cancer comprising administering to a subject a therapeutically effective amount of a chemotherapeutic agent; and a therapeutically effective amount of a compound of formula I:

wherein

R is hydrogen, =0, or -OR ,22 , where R ,22 is hydrogen or a glycoside; or R together with R² forms a bond (C₂-C₃ double bond); or R³ together with R² and the intervening atoms forms an epoxide ring;

R⁴ is —OH, or -OR²³, where R²³ is unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, arylalkyl, amidine, -NR²⁴R²⁵ or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur; R²⁴ and R²⁵ may or may not be the same, are hydrogen, or Ci_-20 alkyl, allyl, aryl, arylalkyl or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; or R⁴ together with R²¹ or R²⁸ forms a lactone;

R is hydrogen or -OH; or R⁶ together with R⁷ forms a bond (Ci i-Ci₂ double bond); or R⁶ together with R⁷ and the intervening atoms forms an epoxide ring;

R⁷ is hydrogen, =0, or -OR²⁶, where R²⁶ is hydrogen or a glycoside; or R⁷ together with R⁶ forms a bond (Cn-Ci₂ double bond); or R⁷ together with R⁶ and the intervening atoms forms an epoxide ring; R⁸ is hydrogen, hydroxyl, mercaptan, or halogen, amino, azido, -NR²⁴R²⁵, unsubstituted or substituted C_1-2O alkyl, allyl, aryl, or arylalkyl, or -OR²⁷, where R²⁷ is hydrogen or a glycoside;

R⁹ is hydrogen or hydroxyl; or R⁹ together with R¹⁵ forms a bond (C₉-C_J5 double bond); or R together with R¹⁵ and the intervening atoms forms an epoxide ring;

R¹ ' is hydrogen or hydroxyl or is absent;

R¹² is methyl, -CH₂OH₅ -CO₂H, or a glycoside ester of said -CO₂H;

R¹³ is methylene, or a divalent heteroatom, or -NR²⁹, where R²⁹ is -NHR³⁰ or -OR³⁰ where R is hydrogen, or Cj_-20 alkyl, aryl, alkylaryl; and a double bond is present between Ci₆ and R¹³ when R¹¹ is absent; or R¹³ is hydrogen, hydroxyl, methyl, -CHO, -CH₂X, where X is halogen, -CH₂NR²⁹, where R²⁹ is -NR³⁰ or -OR³⁰, where R³⁰ is hydrogen, or Ci_-20 alkyl, aryl, or alkylaryl when R¹¹ is hydrogen or hydroxyl;

R¹⁴ is hydrogen or hydroxyl;

29. The method of claim 28, wherein the compound of formula I is gibberellic acid of formula:

30. The method of claim 28, wherein the compound of formula I is of formula:

31. The compound of claim 28, wherein the compound of formula I is of formula:

32. The compound of claim 28, wherein the compound of formula I is of formula:

33. The compound of claim 28, wherein the compound of formula I is of formula:

34. A method of screening for a modulator of the NF-KB pathway comprising: providing a small molecule microarray; and contacting the small molecule microarray with NF-κB or a subunit thereof.

35. The method of claim 34, wherein the NF-κB subunit is p50.

36. The method of claim 34, wherein the NF-κB subunit is p65.

37. The method of claim 34, wherein the step of contacting utilizes NF-κB.

38. A method for identifying binders of NF-κB comprising: providing the p50 subunit of NF-κB preincubated with gibberellic acid; providing a test compound; and identifying binding of the test compound by release of gibberellic acid.

39. The method of claim 38, further comprising providing a small molecule microarray on which the test compound is attached.

40. A method for identifying binders of a transcription factor comprising: providing a transcription factor or a subunit thereof preincubated with a known binder of transcription factor or a subunit thereof; providing a test compound; and identifying binding of the test compound by release of the known binder.

41. The method of claim 40, further comprising providing a small molecule microarray on which the test compound is attached.

42. A method for inhibiting expression, in a eukaryotic cell, of a gene whose transcription is regulated by NF-κB comprising administering to said cell a compound of formula I:

wherein

R¹ is hydrogen or a group -OR²⁰, where R²⁰ is hydrogen, a glycoside, Cj_-6 alkyl group, or R¹ together with R² or R¹⁰ forms a bond (CpC₂ or Ci-Qo double bond, respectively);

R³ is hydrogen, =0, or -OR²², where R²² is hydrogen or a glycoside; or R³ together with R² forms a bond (C₂-C₃ double bond); or R³ together with R² and the intervening atoms forms an epoxide ring;

R⁴ is -OH, or -OR²³, where R²³ is unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, arylalkyl, amidine, -NR²⁴R²⁵ or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur; R²⁴ and R²⁵ may or may not be the same, are hydrogen, or Ci_-20 alkyl, allyl, aryl, arylalkyl or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; or R⁴ together with R Or R forms a lactone;

R⁷ is hydrogen, =0, or -OR²⁶, where R²⁶ is hydrogen or a glycoside; or R⁷ together with R forms a bond (Cn-Ci₂ double bond); or R⁷ together with R⁶ and the intervening atoms forms an epoxide ring; R⁸ is hydrogen, hydroxyl, mercaptan, or halogen, amino, azido, -NR²⁴R²⁵, unsubstituted or substituted C₁.₂₀ alkyl, allyl, aryl, or arylalkyl, or -OR²⁷, where R²⁷ is hydrogen or a glycoside;

R¹⁰ is hydrogen, methyl, -CHO, -CO₂H, or a glycoside ester of said -CO₂H, -CH₂OR²⁸ or -OR²⁸, where R²⁸ is hydrogen or together with R⁴ forms a lactone; or R¹⁰ together with R¹ forms a bond (Ci-Ci₀ double bond); or R¹ together with R¹ and the intervening atoms forms an epoxide ring;

R¹¹ is hydrogen or hydroxyl or is absent;

R¹² is methyl, -CH₂OH, -CO₂H, or a glycoside ester of said -CO₂H;

R¹³ is methylene, or a divalent heteroatom, or -NR²⁹, where R²⁹ is -NHR³⁰ or -OR³⁰ where R³⁰ is hydrogen, or Ci_-20 alkyl, aryl, alkylaryl; and a double bond is present between Ci₆ and R¹³ when R¹¹ is absent; or R¹³ is hydrogen, hydroxyl, methyl, -CHO, -CH₂X, where X is halogen, -CH₂NR²⁹, where R²⁹ is -NR³⁰ or -OR³⁰, where R³⁰ is hydrogen, or Ci_-20 alkyl, aryl, or alkylaryl when R¹¹ is hydrogen or hydroxyl;

R¹⁴ is hydrogen or hydroxyl;

43. A method for reducing the level of expression of a viral gene whose transcription is regulated by NF-κB in a eukaryotic cell comprising administering to the cell a compound of formula I:

wherein R¹ is hydrogen or a group -OR²⁰, where R²⁰ is hydrogen, a glycoside, Ci_-6 alkyl group, or R¹ together with R² or R¹⁰ forms a bond (Ci-C₂ or Ci-Ci₀ double bond, respectively);

R is hydrogen, =0, or -OR , where R is hydrogen or a glycoside; or R together with R² forms a bond (C₂-C₃ double bond); or R³ together with R² and the intervening atoms forms an epoxide ring;

R⁴ is -OH, or -OR²³, where R²³ is unsubstituted or substituted Ci_-20 alkyl, allyl, aryl, arylalkyl, amidine, -NR²⁴R²⁵ or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur; R ⁴ and R may or may not be the same, are hydrogen, or Ci_-20 alkyl, allyl, aryl, arylalkyl or an unsaturated or saturated ring containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur; or R⁴ together with R²¹ or R²⁸ forms a lactone;

R⁹ is hydrogen or hydroxyl; or R⁹ together with R¹⁵ forms a bond (C₉-Ci₅ double bond); or R together with R¹⁵ and the intervening atoms forms an epoxide ring;

R¹⁰ is hydrogen, methyl, -CHO, -CO₂H, or a glycoside ester of said -CO₂H, -CH₂OR²⁸ or -OR , where R²⁸ is hydrogen or together with R⁴ forms a lactone; or R¹⁰ together with R¹ forms a bond (Ci-Ci₀ double bond); or R¹⁰ together with R¹ and the intervening atoms forms an epoxide ring;

R¹ ' is hydrogen or hydroxyl or is absent;

R¹² is methyl, -CH₂OH, -CO₂H, or a glycoside ester of said -CO₂H; R¹³ is methylene, or a divalent heteroatom, or -NR²⁹, where R²⁹ is -NHR³⁰ or -OR³⁰ where R³⁰ is hydrogen, or Ci_-20 alkyl, aryl, alkylaryl; and a double bond is present between C₎₆ and R¹³ when R¹¹ is absent; or R¹³ is hydrogen, hydroxyl, methyl, -CHO, -CH₂X, where X is halogen, -CH₂NR²⁹, where R²⁹ is -NR³⁰ or -OR³⁰, where R³⁰ is hydrogen, or C_-20 alkyl, aryl, or alkylaryl when R¹ ' is hydrogen or hydroxyl;

R¹⁴ is hydrogen or hydroxyl;

44. A compound of formula:

wherein

n is an integer between 2 and 6, inclusive; m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof.

45. The compound of claim 44 of formula: