WO2016025640A1

WO2016025640A1 - Heteroaryl compounds and uses thereof

Info

Publication number: WO2016025640A1
Application number: PCT/US2015/044918
Authority: WO
Inventors: Upendra Purush DAHAL; Matthew T. LABENSKI; Lixin Qiao; Sekhar S. Surapaneni
Original assignee: Celgene Avilomics Research, Inc.
Priority date: 2014-08-13
Filing date: 2015-08-12
Publication date: 2016-02-18

Abstract

The present invention provides compounds, pharmaceutically acceptable compositions thereof, and methods of using the same, for use in inhibiting one or both of ERK1 and ERK2.

Description

HETEROARYL COMPOUNDS AND USES THEREOF

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority to U.S. provisional application number 62/037,077, filed August 13, 2014, and U.S. provisional application number 62/069,750 filed October 28, 2014, the entirety of each of which is hereby incorporated by reference.

FIELD OF THE INVENTION

[0002] The present invention provides various forms and compositions useful as inhibitors of ER kinases, for example one or both of ERKl and ERK2 kinases.

BACKGROUND OF THE INVENTION

[0003] The search for new therapeutic agents has been greatly aided in recent years by a better understanding of the structure of enzymes and other biomolecules associated with diseases. One important class of enzymes that has been the subject of extensive study is protein kinases.

[0004] Protein kinases constitute a large family of structurally related enzymes that are responsible for the control of a variety of signal transduction processes within the cell. Protein kinases are thought to have evolved from a common ancestral gene due to the conservation of their structure and catalytic function. Almost all kinases contain a similar 250-300 amino acid catalytic domain. The kinases may be categorized into families by the substrates they phosphorylate (e.g., protein-tyrosine, protein-serine/threonine, lipids, etc.).

[0005] The processes involved in tumor growth, progression, and metastasis are mediated by signaling pathways that are activated in cancer cells. The ERK pathway plays a central role in regulating mammalian cell growth by relaying extracellular signals from ligand-bound cell surface tyrosine kinase receptors such as erbB family, PDGF, FGF, and VEGF receptor tyrosine kinase. Activation of the ERK pathway is via a cascade of phosphorylation events that begins with activation of Ras. Activation of Ras leads to the recruitment and activation of Raf, a serine- threonine kinase. Activated Raf then phosphorylates and activates MEKl/2, which then phosphorylates and activates one or both of ERKl and ERK2. When activated, one or both of ERKl and ERK2 phosphorylates several downstream targets involved in a multitude of cellular events including cytoskeletal changes and transcriptional activation. The ERK/MAPK pathway is one of the most important for cell proliferation, and it is believed that the ERK MAPK pathway is frequently activated in many tumors. Ras genes, which are upstream of one or both of ERKl and ERK2, are mutated in several cancers including colorectal, melanoma, breast and pancreatic tumors. The high Ras activity is accompanied by elevated ERK activity in many human tumors. In addition, mutations of BRAF, a serine-threonine kinase of the Raf family, are associated with increased kinase activity. Mutations in BRAF have been identified in melanomas (60%), thyroid cancers (greater than 40%) and colorectal cancers.

[0006] Many diseases are associated with abnormal cellular responses triggered by protein kinase-mediated events as described above. Accordingly, there remains a need to find protein kinase inhibitors useful as therapeutic agents.

SUMMARY OF THE INVENTION

[0007] It has now been found that compounds of the present invention, and compositions thereof, are useful as inhibitors of one or both of ERKl and ERK2 and exhibit desirable characteristics for the same.

[0008] In some embodiments, such compounds have general Formula I:

I

or a pharmaceutically acceptable salt thereof, wherein R¹, R⁴, R^w, R^x, R^y, R^z, R^z , R^z , X, X', X", m, n, p, q, and r are as defined herein.

[0009] In some embodiments, such compounds have general Formula II:

or a pharmaceutically acceptable salt thereof, wherein R¹, R⁴, R⁶, R⁷, R^w, R^x, R^y, R^z, R^z , R^z , X, X', X", m, n, p, q, and r are as defined herein.

[0010] Compounds of the present invention, and pharmaceutically acceptable compositions thereof, are useful for treating a variety of diseases, disorders or conditions, associated with abnormal cellular responses triggered by certain protein kinase-mediated events. Such diseases, disorders, or conditions include those described herein.

[0011] Compounds provided by this invention are also useful for the study of certain kinases in biological and pathological phenomena; the study of intracellular signal transduction pathways mediated by such kinases; and the comparative evaluation of new kinase inhibitors. Additional compounds and methods can be found in PCT patent application serial number PCT/US14/15256, filed February 7, 2014 and published as WO 2014/124230 on August 14, 2014 ("the '230 publication," the entirety of which is hereby incorporated herein by reference). The '230 publication describes certain ER inhibitor compounds which covalently and irreversibly inhibit activity of one or both of ERKl and ERK2 kinases.

DETAILED DESCRIPTION OF THE INVENTION

1. General Description of Certain Aspects of the Invention

[0012] In certain embodiments, the present invention provides inhibitors of one or both of ERKl and ERK2 and conjugates thereof. In some embodiments, such compounds include those of the formulae described herein, or a pharmaceutically acceptable salt thereof, wherein each variable is as defined and described herein. [0013] In some embodiments such compounds have the structure of Formula I:

I

or a pharmaceutically acceptable salt thereof, wherein:

X, X', and X" are each independently O;

p, q, and r are each independently 0 or 1;

R¹ is -OR';

R' is -H, -CH₃, -S0₃H or -Glu;

each Glu is a glucuronyl moiety;

R² and R³ are each independently -H, -OH, -OS0₃H, -OGlu, -SR⁵, or:

R² and R³ are taken together to form a double bond; or:

R² and R³ are taken together with their intervening atoms to form an epoxide moiety; R^2' is -H, or:

R² and R^2' are taken together to form =0;

R³ and R³ are each -H, or:

R^3' and R^3" are taken together to form =0;

R is -H, -OH, -OS0₃H or -OGlu;

each R⁵ is independently selected from:

II III IV

R^x and R^y are each independently -OH, -OSO₃H, or -OGlu;

R^z, R^z' and R^z" are each independently -H, -CH₃, -OH, -OSO3H, or -OGlu; or

R^z" and R³ are taken together to form -0-;

m is 0, 1, 2, or 3;

n is 0, 1, or 2;

wherein the compound is other than:

[0014] In some embodiments, when R² and R³ are taken together to form a double bond, at least one of the following is true:

(a) at least one of R¹, R⁴, R^x, R^y, R^z, R^z' and R^z" is -OH, -OS0₃H or -OGlu; or (b) one of p, q, or r is 1.

[0015] In some embodiments such compounds have the structure of Formula II:

or a pharmaceutically acceptable salt thereof, wherein:

X, X', and X" are each independently O;

p, q, and r are each independently 0 or 1;

R¹ is -OR';

R' is -H, -CH₃, -S0₃H or -Glu;

each Glu is a glucuronyl moiety;

R² and R³ are each independently -H, -OH, -OS0₃H, -OGlu, -SR⁵, or:

R² and R³ are taken together to form a double bond; or:

R² and R³ are taken together with their intervening atoms to form an epoxide moiety; R²' is -H, or:

R² and R²' are taken together to form =0;

R³ and R³ are each -H, or:

R³' and R³" are taken together to form =0;

R⁴ is -H, -OH, -OS0₃H or -OGlu;

each R⁵ is independently selected from:

i ii Hi iv

R⁶ is -CH₃, -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu;

R⁷ is -CH₃, -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu;

R^x and R^y are each independently -OH, -OS0₃H, or -OGlu;

R^z, R^z' and R^z" are each independently -H, -CH₃, -OH, -OS0₃H, or -OGlu; or

R^z" and R³ are taken together to form -0-;

m is 0, 1, 2, or 3;

n is 0, 1, or 2;

wherein the compound is other than:

[0016] In some embodiments, when R² and R³ are taken together to form a double bond, at least one of the following is true:

(a) at least one of R¹, R⁴, R^x, R^y, R^z, R^z' and R^z" is -OH, -OSO3H or -OGlu; or

(b) at least one of R⁶ and R⁷ is -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu; or

(c) at least one of p, q, or r is 1.

2. Compounds and Definitions

[0017] Compounds of this invention include those described generally above, and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. 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. Additionally, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5^th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

[0018] The term "aliphatic" or "aliphatic group", as used herein, means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as "carbocycle," "carbocyclic", "cycloaliphatic" or "cycloalkyl"), that has a single point of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms. In some embodiments, "carbocyclic" (or "cycloaliphatic" or "carbocycle" or "cycloalkyl") refers to a monocyclic C3-C8 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0019] As used herein, "aralkyl" refers to a straight or branched aliphatic group in which one of the hydrogen atoms of the aliphatic is replaced by an aryl group, wherein the aliphatic group has from 1 to 10 carbon atoms. Substituted aralkyl groups may be substituted at the aliphatic, the aryl, or both the aliphatic and the aryl portions of the group. Representative aralkyl groups include but are not limited to benzyl and phenethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-ethyl-indanyl.

[0020] As used herein, an "amino acid fragment" The term "amino acid fragment" refers to a portion of an amino acid, such as by way of example only, the 20 common, genetically-encoded amino acids (i.e., alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine), or a dipeptide, tripeptide or other polypeptide comprising a combination of the 20 common amino acids or a non-natural amino acid. In some embodiments, the amino acid fragment is attached to a compound of Formula I or II via the side chain of the amino acid. In one embodiment, the amino acid fragment is a cysteine fragment wherein the remaining portion of a compound of Formula I or II is bound via a sulfur bond. In another embodiment, the amino acid fragment is a glutathione fragment wherein the remaining portion of a compound of Formula I or II is bound via a sulfur bond of the glutathione fragment. In another embodiment, the amino acid fragments are derived from beta-amino acids. In further embodiments, the amino acid fragments are derived from portions of polypeptides or proteins. In yet further embodiments, the amino acid fragment is attached to a compound of Formula I or II via the N-terminal or the acyl -terminal of the amino acid.

[0021] As used herein, the term "bridged bicyclic" refers to any bicyclic ring system, i.e. carbocyclic or heterocyclic, saturated or partially unsaturated, having at least one bridge. As defined by IUPAC, a "bridge" is an unbranched chain of atoms or an atom or a valence bond connecting two bridgeheads, where a "bridgehead" is any skeletal atom of the ring system which is bonded to three or more skeletal atoms (excluding hydrogen). In some embodiments, a bridged bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groups are well known in the art and include those groups set forth below where each group is attached to the rest of the molecule at any substitutable carbon or nitrogen atom. Unless otherwise specified, a bridged bicyclic group is optionally substituted with one or more substituents as set forth for aliphatic groups. Additionally or alternatively, any substitutable nitrogen of a bridged bicyclic group is optionally substituted. Exemplary bridged bicyclics include:

[0022] The term "lower alkyl" refers to a Ci_₄ straight or branched alkyl group. Exemplary lower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert-butyl.

[0023] The term "lower haloalkyl" refers to a Ci_₄ straight or branched alkyl group that is substituted with one or more halogen atoms.

[0024] The term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), ΝΗ (as in pyrrolidinyl) or NR⁺ (as in N-substituted pyrrolidinyl)).

[0025] The term "unsaturated," as used herein, means that a moiety has one or more units of unsaturation. [0026] As used herein, the term "bivalent Ci_g (or Ci_₆) saturated or unsaturated, straight or branched, hydrocarbon chain", refers to bivalent alkylene, alkenylene, and alkynylene chains that are straight or branched as defined herein.

[0027] The term "alkylene" refers to a bivalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., -(CH₂)_n-, wherein n is a positive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.

[0028] The term "alkenylene" refers to a bivalent alkenyl group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.

[0029] As used herein, the term "cyclopropylenyl" refers to a bivalent cyclopropyl group of the following structure:

[0030] The term "cycloalkylalkyl" refers to a bivalent straight or branched aliphatic group that is substituted with a cycloalkyl group, wherein the aliphatic group has from 1 to 10 carbon atoms. Examples of a cycloalkylalkyl group include -(CH₂)_x-cyclopentyl, -(CH₂)_x-cyclohexyl, - (CH₂)_x-cycloheptyl, etc., wherein x is 1-10. In some embodiments, the aliphatic chain of a cycloalkylalkyl group may be straight or branched, for example, -(CH₂)_yCH(CH₃)(CH₂)_y- cyclopentyl, -(CH₂)_yCH(CH₃)(CH₂)_y-cyclohexyl, -(CH₂)_yCH(CH₃)(CH₂)_y-cycloheptyl, etc., wherein y is 0-10. In some embodiments, the cycloalkyl ring of a cycloalkylalkyl group is saturated or partially unsaturated. Substituted cycloalkylalkyl groups may be substituted at the aliphatic, the cycloalkyl, or both the aliphatic and the cycloalkyl portions of the group.

[0031] The term "halogen" means F, CI, Br, or I.

[0032] The term "aryl" used alone or as part of a larger moiety as in "aralkyl," "aralkoxy," or "aryloxyalkyl," refers to monocyclic or bicyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments of the present invention, "aryl" refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term "aryl," as it is used herein, is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, 1,3- dihydro-2H-benzo[d]imidazole-2-one, and the like.

[0033] The terms "heteroaryl" and "heteroar-," used alone or as part of a larger moiety, e.g., "heteroaralkyl," or "heteroaralkoxy," refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 π electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quatemized form of a basic nitrogen. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms "heteroaryl" and "heteroar-", as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring. Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin- 3(4H)-one. A heteroaryl group may be mono- or bicyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which terms include rings that are optionally substituted.

[0034] The term "heteroaralkyl" refers to an straight or branched aliphatic group substituted by a heteroaryl, wherein the aliphatic group has from 1 to 10 carbon atoms and wherein the aliphatic and heteroaryl portions independently are optionally substituted.

[0035] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic radical," and "heterocyclic ring" are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7-10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4- dihydro-2H-pyrrolyl), ΝΗ (as in pyrrolidinyl), or ⁺NR (as in N-substituted pyrrolidinyl).

[0036] A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical," are used interchangeably herein, and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl, where the radical or point of attachment is on the heterocyclyl ring. A heterocyclyl group may be mono- or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0037] As used herein, the term "heterocyclylalkyl" refers to a bivalent straight or branched aliphatic group that is substituted with a heterocyclyl moiety, wherein the aliphatic group has from 1 to 10 carbon atoms. Examples of a heterocyclylalkyl group include, without limitation, - (CH₂)_x-piperidinyl, -(CH₂)_x-piperazinyl, -(CH₂)_x-pyrrolidinyl, -(CH₂)_x-morpholinyl, -(CH₂)_X- pyrrolidinyl, -(CH₂)_x-tetrahydrofuranyl, -(CH₂)_x-tetrahydropyranyl, etc., wherein x is 1-10. In some embodiments, the aliphatic chain of a heterocyclylalkyl group may be straight or branched, for example, -(CH₂)_yCH(CH₃)(CH₂)_y-piperidinyl, -(CH₂)_yCH(CH₃)(CH₂)_y-tetrahydrofuranyl, - (CH₂)_yCH(CH₃)(CH₂)_y-tetrahydropyranyl, -(CH₂)_yCH(CH₃)(CH₂)_y-morpholinyl, etc., wherein y is 0-10. In some embodiments, the heterocyclyl ring of a heterocyclylalkyl group is saturated or partially unsaturated. Substituted heterocyclylalkyl groups may be substituted at the aliphatic, the heterocyclyl, or both the aliphatic and the heterocyclyl portions of the group.

[0038] An "alkoxy" group is -O-(aliphatic), wherein aliphatic is defined above.

[0039] An "alkoxyalkyl" group is -(aliphatic)-O-(aliphatic), wherein aliphatic is defined above.

[0040] An "amino" group is a radical of the formula -NH₂. [0041] An "alkylamino" group is a radical of the formula: -NH-aliphatic or -N(aliphatic)₂, wherein each aliphatic is independently as defined above.

[0042] A "carboxy" group is a radical of the formula -C(0)OH.

[0043] An "aminocarbonyl" group is a radical of the formula -C(0)N(R )₂, -C(0)NH(R ) or -C(0)NH₂, wherein each R is independently a substituted or unsubstituted aliphatic, cycloalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroaralkyl group as defined herein.

[0044] An "acylamino" group is a radical of the formula: -NHC(0)(R ) or -N(alkyl)C(0)( R ), wherein each alkyl and R are independently as defined above.

[0045] An "alkylsulfonylamino" group is a radical of the formula: -NHS0₂(R ) or

-N(alkyl)S0₂(R ), wherein each aliphatic and R are defined above.

[0046] As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as herein defined.

[0047] As described herein, compounds of the invention may contain "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally" or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and 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. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0048] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently halogen; -(CH₂)o-₄R°; -(CH₂)o^OR°; -O(CH₂)₀_₄R^o, -O- (CH₂)_{0 4}C(0)OR°; -(CH₂)_{0 4}CH(OR°)₂; -(CH₂)₀^SR°; -(CH₂)₀^Ph, which may be substituted with R°;

which may be substituted with R°; -CH=CHPh, which may be substituted with R°; -(CH₂)o-₄0(CH₂)₀-i-pyridyl which may be substituted with R°; -N0₂; -CN; -N₃; -(CH₂)o ₄N(R°)₂; -(CH₂)_{0 4}N(R°)C(0)R°; -N(R°)C(S)R°; -(CH₂)_{0 4}N(R°)C(0)NR°₂; -N(R°)C(S)NR°₂; -(CH₂)_{0 4}N(R°)C(0)OR°; -N(R°)N(R°)C(0)R°; -N(R°)N(R°)C(0)NR°₂; -N(R°)N(R°)C(0)OR⁰; -(CH₂)_{0 4}C(0)R°; -C(S)R°; -(CH₂)_{0 4}C(0)OR°; -(CH₂)_{0 4}C(0)SR°; -(CH₂)o^C(0)OSiR°₃; -(CH₂)_{0 4}OC(0)R°; -OC(O)(CH₂)_{0 4}SR- SC(S)SR°; -(CH₂)_{0 4}SC(0)R°; -(CH₂)o ₄C(0)NR°₂; -C(S)NR°₂; -C(S)SR°; -SC(S)SR°, -(CH₂)_{0 4}OC(0)NR°₂; -C(0)N(OR°)R°; -C(0)C(0)R°; -C(0)CH₂C(0)R°; -C(NOR°)R°; -(CH₂)₀^SSR°; -(CH₂)₀ ₄S(0)₂R°; -(CH₂)o ₄S(0)₂OR°; -(CH₂)_{0 4}OS(0)₂R°; -S(0)₂NR°₂; -(CH₂)_{0 4}S(0)R°; -N(R°)S(0)₂NR°₂; -N(R°)S(0)₂R°; -N(OR°)R°; -C(NH)NR°₂; -P(0)₂R°; -P(0)R°₂; -OP(0)R°₂; -OP(0)(OR°)₂; SiR°₃; -(Ci_₄ straight or branched alkylene)0-N(R°)₂; or

straight or branched alkylene)C(0)0-N(R°)₂, wherein each R° may be substituted as defined below and is independently hydrogen, Ci_₆ aliphatic, -CH₂Ph, -O(CH₂)₀ iPh, -CH₂-(5-6 membered heteroaryl ring), or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0049] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms), are independently halogen, -(CH₂)_{0 2}R*, -(haloR*), -(CH₂)_{0 2}OH, -(CH₂)_{0 2}OR*, -(CH₂)_{0 2}CH(OR*)₂; -O(haloR'), -CN, -N₃, -(CH₂)_{0 2}C(0)R*, -(CH₂)_{0 2}C(0)OH, -(CH₂)_{0 2}C(0)OR*, -(CH₂)_{0 2}SR*, -(CH₂)o ₂SH, -(CH₂)o ₂NH₂, -(CH₂)_{0 2}NHR*, -(CH₂)_{0 2}NR*₂, -N0₂, -SiR*₃, -OSiR*₃, -C(0)SR* -(Ci_₄ straight or branched alkylene)C(0)OR*, or -SSR* wherein each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently selected from C^ aliphatic, -CH₂Ph, -O(CH₂)₀_iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =0 and =S.

[0050] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: =0 ("oxo"), =S, = NR^* ₂, =NNHC(0)R^*, =NNHC(0)OR^*, =NNHS(0)₂R^*, =NR^*, =NOR^*, -0(C(R^* ₂))_{2 3}0- or -S(C(R^* ₂))₂ _₃S- wherein each independent occurrence of R is selected from hydrogen, Ci_₆ aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an "optionally substituted" group include: -0(CR ₂)₂_₃0-, wherein each independent occurrence of R is selected from hydrogen, Ci_6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0051] Suitable substituents on the aliphatic group of R^* include halogen, -R*, -(haloR*), -OH, -OR*, -O(haloR'), -CN, -C(0)OH, -C(0)OR*, -NH₂, -NHR*, -NR*₂, or -N0₂, wherein each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently Ci_₄ aliphatic, -CH₂Ph, -O(CH₂)₀-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0052] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R^†, -NR^† ₂, -C(0)R^†, -C(0)OR^†, -C(0)C(0)R^†, -C(0)CH₂C(0)R^†, -S(0)₂R^†, -S(0)₂NR^† ₂, -C(S)NR^† ₂, -C(NH)NR^† ₂, or -N(R^†)S(0)₂R^†; wherein each R^† is independently hydrogen, Ci_₆ aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R^, taken together with their intervening atom(s) form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0053] Suitable substituents on the aliphatic group of R^† are independently halogen, -R*, -(haloR*), -OH, -OR*, -O(haloR'), -CN, -C(0)OH, -C(0)OR*, -NH₂, -NHR*, -NR*₂, or -N0₂, wherein each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently Ci_₄ aliphatic, -CH₂Ph, -O(CH₂)₀ iPh, or a 5-6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. [0054] 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, S. M. Berge et al., 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, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like.

[0055] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and

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, lower alkyl sulfonate and aryl sulfonate.

[0056] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention.

[0057] Unless otherwise stated, all tautomeric forms of compounds of the invention are within the scope of the invention. By way of non-limiting example, in some embodiments

wherein a compound is depicted as containing a ring of the following structure:

, the

present invention also contemplates the corresponding tautomer:

[0058] Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present invention. In certain embodiments, a warhead moiety, R¹, of a provided compound comprises one or more deuterium atoms.

[0059] As used herein, the term "irreversible" or "irreversible inhibitor" refers to an inhibitor (i.e. a compound) that is able to be covalently bonded to a target protein kinase in a substantially non-reversible manner. That is, whereas a reversible inhibitor is able to bind to (but is generally unable to form a covalent bond) the target protein kinase, and therefore can become dissociated from the target protein kinase, an irreversible inhibitor will remain substantially bound to the target protein kinase once covalent bond formation has occurred. Irreversible inhibitors usually display time dependency, whereby the degree of inhibition increases with the time with which the inhibitor is in contact with the enzyme. Methods for identifying if a compound is acting as an irreversible inhibitor are known to one of ordinary skill in the art. Such methods include, but are not limited to, enzyme kinetic analysis of the inhibition profile of the compound with the protein kinase target, the use of mass spectrometry of the protein drug target modified in the presence of the inhibitor compound, discontinuous exposure, also known as "washout," experiments, and the use of labeling, such as radiolabeled inhibitor, to show covalent modification of the enzyme, as well as other methods known to one of skill in the art.

Description of Exemplary Compounds

According to one as ect, the present invention provides a compound of Formula I

I

or a pharmaceutically acceptable salt thereof, wherein:

X, X', and X" are each independently O;

p, q, and r are each independently 0 or 1;

R¹ is -OR';

R' is -H, -CH₃, -S0₃H or -Glu;

each Glu is a glucuronyl moiety;

R² and R³ are each independently -H, -OH, -OS0₃H, -OGlu, -SR⁵, or:

R² and R³ are taken together to form a double bond; or:

R² and R^2' are taken together to form =0;

R³ and R³ are each -H, or: R^3' and R^3" are taken together to form =0;

R⁴ is -H, -OH, -OSO₃H or -OGlu;

eac ⁵ is independently selected from:

11 III IV

R^x and R^y are each independently -OH, -OSO₃H, or -OGlu;

R^z, R^z' and R^z" are each independently -H, -CH₃, -OH, -OSO3H, or -OGlu; or

R^z" and R³ are taken together to form -0-;

m is 0, 1, 2, or 3;

n is 0, 1, or 2;

wherein the compound is other than:

[0061] In some embodiments, when R² and R³ are taken together to form a double bond, at least one of the following is true:

(b) one of p, q, or r is 1.

[0062] In some embodiments, the present invention provides a compound of Formula II:

or a pharmaceutically acceptable salt thereof, wherein:

X, X', and X" are each independently O;

p, q, and r are each independently 0 or 1;

R¹ is -OR';

R' is -H, -CH₃, -S0₃H or -Glu;

each Glu is a glucuronyl moiety;

R² and R³ are each independently -H, -OH, -OS0₃H, -OGlu, -SR⁵, or:

R² and R³ are taken together to form a double bond; or:

R² and R^2' are taken together to form =0;

R³ and R³ are each -H, or:

R^3' and R^3" are taken together to form =0; R⁴ is -H, -OH, -OSO₃H or -OGlu;

eac ⁵ is independently selected from:

i ii Hi iv

R⁶ is -CH₃, -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu;

R⁷ is -CH₃, -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu;

R^x and R^y are each independently -OH, -OS0₃H, or -OGlu;

R^z" and R³ are taken together to form -0-;

m is 0, 1, 2, or 3;

n is 0, 1, or 2;

wherein the compound is other than:

[0063] In some embodiments, when R² and R³ are taken together to form a double bond, at least one of the following is true:

(b) at least one of R⁶ and R⁷ is -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu; or

(c) at least one of p, q, or r is 1.

[0064] As used herein, the term "glucuronyl moiety" refers to a group having the structure:

wherein the wavy line depicted designates the point of attachment to a compound of Formula I or II.

[0065] In some embodiments, an -OH group on a compound of Formula I or II is glucuronidated to a -OGlu group. In some embodiments, an -OH group of Formula I or II is sulfated to a -OSO₃H group.

[0066] As defined generally above, R^w is H or

. In some embodiments, R^w is

H. In some embodiments, R^w is

, wherein each of R², R² , R³, R³ , and R³ are as defined above and herein.

[0067] As defined generally above, R¹ is -OR'. In some embodiments, R' is -H, -CH₃, - S0₃H or -Glu. In some embodiments, R' is -H. In some embodiments, R' is selected from - CH₃, -S0₃H and -Glu. In some embodiments, R' is -CH₃. In some embodiments, R' is -S0₃H. In some embodiments, R' is -Glu. Accordingly, in some embodiments, R¹ is selected from -OH, -OCH₃, -OS0₃H, -OGlu.

[0068] In some embodiments, R¹ is -OH. In some embodiments, R¹ is -OCH₃. In some embodiments, R¹ is -OS0₃H. In some embodiments, R¹ is -OGlu.

[0069] As defined generally above, R² is selected from -H, -OH, -OS0₃H -OGlu, and -SR⁵. In some embodiments, R² is -H. In some embodiments, R² is selected from -OH, -OS0₃H and - OGlu. In some embodiments, R² is -OH. In some embodiments, R² is -OSO3H. In some embodiments, R² is -OGlu. In some embodiments, R² is -SR⁵.

[0070] As defined generally above, R³ is selected from -H, -OH, -OSO3H, -OGlu and -SR⁵. In some embodiments, R³ is -H. In some embodiments, R³ is selected from -OH, -OSO3H and - OGlu. In some embodiments, R³ is -OH. In some embodiments, R³ is -OSO3H. In some embodiments, R³ is -OGlu.

[0071] In some embodiments, R² is -SR⁵. In some embodiments, R³ is -SR⁵. In some such

embodiments,

Such moiety is generally referred to glutathione fragment.

[0072] In some embodiments, R³ is -SR⁵, wherein R⁵ is

In

some embodiments,

Such moiety is generally referred to as a cysteine-glycine fragment.

[0073] In some embodiments, R³ is -SR⁵, wherein R⁵ is

Such moiety is generally referred to as a cysteine fragment.

[0074] In some embodiments, R³ is -SR⁵, wherein R⁵ is

. Such moiety is generally referred to as an N-acetyl cysteine fragment.

[0075] In some embodiments, R² is H and R³ is -SR⁵. In some such embodiments, R⁵ is

² is H and R³ is -SR⁵, wherein R⁵ is

some embodiments, R is H and R is -SR , wherein R is

[0076] In some embodiments, R² and R³ are the same. In some embodiments, R² and R³ are each -OH. In some embodiments, R² and R³ are each -SO3H. In some embodiments, R² and R³ are each -OGlu.

[0077] In some embodiments, R² and R³ are different. In some embodiments, one of R² and

R³ is -H and the other is -OH. In some embodiments, one of R² and R³ is -H and the other is

OSO3H. In some embodiments, one of R² and R³ is -H and the other is -OGlu. In some embodiments, one of R² and R³ is -OH and the other is -OGlu. In some embodiments, one of R² and R³ is -OH and the other is -OSO3H.

[0078] In some embodiments, R² is -OH and R³ is -OGlu. In some embodiments, R² is - OGlu and R³ is -OH. In some embodiments, R² is -OH and R³ is -OSO3H. In some embodiments, R² is -OSO₃H and R³ is -OH. In some embodiments, each of R² and R³ is - OSO3H. In some embodiments, each of R² and R³ is -OGlu.

[0079] In some embodiments, R² and R³ are taken together to form a double bond.

[0080] In some embodiments, R² and R³ are taken together with their intervening atoms to form an epoxide moiety.

[0081] In some embodiments, R² is -H. In some embodiments, R² and R² are each -H. In some embodiments, R² is -OH and R² is -H. In some embodiments, R² is -OSO3H and R² is - H. In some embodiments, R² is -OGlu and R² is -H.

[0082] In some embodiments, R³ and R³ are each -H. In some embodiments, R³ is -OH

3 ' 3 " 3 3 ' 3 "

and R and R are each -H. In some embodiments, R is -OSO3H and R and R are each -H.

3 3 ' 3 " 3 3'

In some embodiments, R is -OGlu and R and R are each -H. In some embodiments, R , R and R³ are each -H. [0083] In some embodiments, R² and R² are taken together to form =0. In some

2 2' 3 3' 3"

embodiments, R and R are taken together to form =0 and R , R and R are each hydrogen. In some embodiments, R² and R² are taken together to form =0 and R³ is -OH.

[0084] In some embodiments, R³ and R³ are taken together to form =0. In some embodiments, R³ and R³ are taken together to form =0 and R³ is -H. In some embodiments, R³ and R³ are taken together to form =0 and R³ is -OH.

[0085] As defined generally above, R⁴ is selected from -H, -OH, -OSO3H and -OGlu. In some embodiments, R⁴ is -H. In some embodiments, R⁴ is selected from -OH, -OSO3H and - OGlu. In some embodiments, R⁴ is selected from -OH, -OSO3H and OGlu. In some embodiments, R⁴ is -OH. In some embodiments, R⁴ is -OSO₃H. In some embodiments, R⁴ is - OGlu.

[0086] As defined generally above, R⁶ is selected from -CH₃, -CH₂OH, -CH₂OS0₃H and - CH₂OGlu. In some embodiments, R⁶ is -CH₃. In some embodiments, R⁶ is selected from - CH₂OH, -CH₂OS0₃H and -CH₂OGlu. In some embodiments, R⁶ is -CH₂OH. In some embodiments, R⁶ is -CH₂OS0₃H. In some embodiments, R⁶ is -CH₂OGlu.

[0087] As defined generally above, R⁷ is selected from -CH₃, -CH₂OH, -CH₂OS0₃H and - CH₂OGlu. In some embodiments, R⁷ is -CH₃. In some embodiments, R⁷ is selected from - CH₂OH, -CH₂OS0₃H and -CH₂OGlu. In some embodiments, R⁷ is -CH₂OH. In some embodiments, R⁷ is -CH₂OS0₃H. In some embodiments, R⁷ is -CH₂OGlu.

[0088] As defined generally above, p, q, and r are each independently selected from 0 and 1. In some embodiments, one of p, q, and r is 1. In some embodiments, p is 1. In some embodiments, p is 0. In some embodiments, q is 1. In some embodiments, q is 0. In some embodiments, r is 1. In some embodiments, r is 0. In some embodiments, both p and q are 0. In some embodiments, both q and r are 0. In some embodiments, both p and r are 0. In some embodiments, each of p, q, and r are 0.

[0089] As defined generally above, m is selected from 0, 1, 2, and 3. In some embodiments, m is 1, 2, or 3. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3.

[0090] As defined generally above, n is selected from 0, 1, and 2. In some embodiments, n is 1 or 2. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. [0091] As defined generally above, R^x is selected from -H, -OH, -OSO₃H and -OGlu. In some embodiments, R^x is -H. In some embodiments, R^x is selected from -OH, -OSO₃H and - OGlu. In some embodiments, R^x is selected from -OH, -OSO₃H and -OGlu. In some embodiments, R^x is -OH. In some embodiments, R^x is -OSO₃H. In some embodiments, R^x is - OGlu.

[0092] As defined generally above, R^y is selected from -H, -OH, -OSO₃H and -OGlu. In some embodiments, R^y is -H. In some embodiments, R^y is selected from -OH, -OSO₃sHi and - OGlu. In some embodiments, R^y is selected from -OH, J

-OSO₃SHi and -OGlu. In some embodiments, R^y is -OH. In some embodiments, R^y is -OSO₃H. In some embodiments, R^y is -

OGlu.

[0093] As defined generally above, R^z, R^z and R^z are each independently selected from -H, -CH₃, -OH, -OSO₃H and -OGlu. In some embodiments, R^z, R^z and R^z are each independently selected from -CH₃, -OH, -OSO₃H and -OGlu. In some embodiments, at least one of R^z, R^z and R^z is -OH. In some embodiments, one of R^z, R^z and R^z is -CH₃. In some embodiments, R^z is -H. In some embodiments, R^z is selected from -CH₃, -OH, -OSO₃H and -OGlu. In some embodiments, R^z is -OH. In some embodiments, R^z is -CH₃. In some embodiments, R^z is - OSO₃H. In some embodiments, R^z is -OGlu. In some embodiments, R^z is -H. In some embodiments, R^z is selected from -CH₃, -OH, -OSO₃H and -OGlu. In some embodiments, R^z is -OH. In some embodiments, R^z is -CH₃. In some embodiments, R^z is -OSO₃H. In some embodiments, R^z is -OGlu. In some embodiments, R^z is -H. In some embodiments, R^z is selected from -CH₃, -OH, -OSO₃H and -OGlu. In some embodiments, R^z is -OH. In some embodiments, R^z is -CH₃. In some embodiments, R^z is -OSO₃H. In some embodiments, R^z is -OGlu.

[0094] In some embodiments, R^z and R³ are taken together to form -0-.

R² R^2'

[0095] As described above, in some embodiments, R^w is R^{3 0} , wherein each of R¹, R², R² , R³, R^3', R^3", R⁴, R^z, R^z'. R^z", R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein. In some such embodiments, at least one of R² and R³ is -SR⁵ and at least one of the following is true:

(a) at least one of R¹, R⁴, R^x, R^y, R^z, R^z' and R^z" is -OH, -OSO₃H or -OGlu; or (b) at least one of R⁶ and R⁷, when present, is -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu; or

(c) at least one of p, q, or r is 1.

embodiments, R³ is -SR⁵. In some such embodiments, R⁵ is

[0097] In some embodiments, a compound of formula I or II is of any of the structures described above and herein, wherein the structure is prone to further oxidation. For instance, in some embodiments, a compound is of formula I or II, wherein m is 1 or 2 and each R^x is -OH, such that the compound comprises a phenol or hydroquinone moiety. One of skill in the art will appreciate that such a phenol or hydroquinone moiety may undergo further oxidation to afford the corresponding quinone configuration. One of skill in the art will further appreciate that heteroaryl rings such as those of compounds of formula I or II may undergo similar such reactions, and that such further oxidized compounds are accordingly contemplated herein.

[0098] In some embodiments, a compound is of formula I or II wherein R^w is H.

[0099] n some embodiments, a compound is of formula I or

[00100] In some embodiments, a compound is of formula I or II wherein R¹ is - -OH.

[00101] In some embodiments, a compound is of formula I or II wherein R¹ is - -OCH₃.

[00102] In some embodiments, a compound is of formula I or II wherein R¹ is - -OS0₃H.

[00103] In some embodiments, a compound is of formula I or II wherein R¹ is - -OGlu.

[00104] In some embodiments, a compound is of formula I or II wherein R⁴ is - -H.

[00105] In some embodiments, a compound is of formula I or II wherein R⁴ is - -OH.

[00106] In some embodiments, a compound is of formula I or II wherein R⁴ is - -OS0₃H.

[00107] In some embodiments, a compound is of formula I or II wherein R⁴ is - -OGlu.

[00108] In some embodiments, a compound is of formula I or II wherein R^x is - -OH.

[00109] In some embodiments, a compound is of formula I or II wherein R^x is - -OS0₃H.

[00110] In some embodiments, a compound is of formula I or II wherein R^x is - -OGlu.

[00111] In some embodiments, a compound is of formula I or II wherein m is 0

[00112] In some embodiments, a compound is of formula I or II wherein m is 1 [00113] In some embodiments, a compound is of formula I or II wherein m is 2

[00114] In some embodiments, a compound is of formula I or II wherein m is 3

[00115] In some embodiments, a compound is of formula I or II wherein R^y is - OH.

[00116] In some embodiments, a compound is of formula I or II wherein R^y is - OSO₃H.

[00117] In some embodiments, a compound is of formula I or II wherein R^y is - OGlu.

[00118] In some embodiments, a compound is of formula I or II wherein n is 0.

[00119] In some embodiments, a compound is of formula I or II wherein n is 1.

[00120] In some embodiments, a compound is of formula I or II wherein n is 2.

[00121] In some embodiments, a compound is of formula I or II wherein p is 1.

[00122] In some embodiments, a compound is of formula I or II wherein q is 1.

[00123] In some embodiments, a compound is of formula I or II wherein r is 1.

[00124] In some embodiments, a compound is of formula I or II wherein R^z, R^z and R^z are each -H.

[00125] In some embodiments, a compound is of formula I or II wherein one of R^z, R^z or R^z is -CH₃, -OH, -OS0₃H, or -OGlu.

Compounds of Formula I-a

[00126] As described above, in some embodiments, R² and R³ are taken together to form a double bond. Accordingly, in some embodiments, the present invention provides a compound of Formula I-a:

I-a wherein each of R¹, R⁴, R^z, R^z', R^{z "}, R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00127] In some embodiments, a compound is of formula I-a, wherein at least one of R¹, R⁴, R^x, and R^y is independently selected from -OH, -OSO3H and -OGlu. In some embodiments, a compound is of formula I-a, wherein at least one of R¹, R⁴, R^x, and R^y is -OH. In some embodiments, a compound is of formula I-a, wherein at least one of R¹, R⁴, R^x, and R^y is - OSO3H. In some embodiments, a compound is of formula I-a, wherein at least one of R¹, R⁴, R^x, and R^y is -OGlu.

[00128] In some embodiments, a compound is of formula I-a and R¹ is -OH, -OSO3H or - OGlu. In some embodiments, a compound is of formula I-a and R¹ is -OH. In some embodiments, a compound is of formula I-a and R¹ is -OSO3H. In some embodiments, a compound is of formula I-a and R¹ is -OGlu.

[00129] In some embodiments, a compound is of formula I-a and n is 0. In some embodiments, a compound is of formula I-a and n is 1. In some embodiments, a compound is of formula I-a and n is 2. In some embodiments, a compound is of formula I-a and m is 0. In some embodiments, a compound is of formula I-a and m is 1. In some embodiments, a compound is of formula I-a and m is 2. In some embodiments, a compound is of formula I-a and m is 3.

[00130] In certain embodiments, at least one of m or n is 0 and R¹ is OH. In certain embodiments, both m and n are 0 and R¹ is OH.

[00131] In some embodiments, a compound is of formula I-a and at least one of p, q, and r is

1. In some embodiments, a compound is of formula I-a and p is 1. In some embodiments, a compound is of formula I-a and q is 1. In some embodiments, a compound is of formula I-a and r is 1.

[00132] In some embodiments, a compound is of formula I-a and R¹ is -OMe.

[00133] In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe and at least one of p, q, and r is 1. In some embodiments, a compound is of formula I-a, wherein R¹ is - OMe and p is 1. In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe and and q is 1. In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe and r is 1.

[00134] In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe and m is 1,

2, or 3. In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe and m is 1. In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe and m is 2. In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe and m is 3. In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe, m is 1, 2, or 3, and each R^x is -OH. In some embodiments, a compound is of formula I-a, wherein R¹ is -OMe, m is 1, 2, or 3, and each R^x is -OSO3H. In some embodiments, a compound is of formula I-a, wherein R¹ is - OMe, m is 1, 2, or 3, and each R^x is -OGlu.

[00135] In certain embodiments, a compound is of formula I-a, wherein R¹ is -OMe, m is 1, and R^x is -OH. In certain embodiments, a compound is of formula I-a, wherein R¹ is -OMe, m is 1, and R^x is -OSO3H. In certain embodiments, a compound is of formula I-a, wherein R¹ is - OMe, m is 1, and R^x is -OGlu.

[00136] In some embodiments, a compound is of formula I-a, wherein R^z, R^z and R^z are each -H.

[00137] In some embodiments, a compound is of formula I-a, wherein one of R^z, R^z or R^z is -

CH₃, -OH, -OS0₃H, or -OGlu.

[00138]

[00139] In some embodiments, the compound is of formula I-a, wherein R⁴ is hydrogen. In some embodiments the present invention provides a compound of formula I-a of any one of the followin formulae:

I-a(l) I-a(2)

I-a(3) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R^z, R^z , R^z , R^x, R^y, X, X', X", n, p, q, and r is as defined above and described herein.

[00140] In some embodiments the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(4) I-a(5)

I-a(6) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R^z, R^z , R^z , R^y, X, X', X", n, p, q, and r is as defined above and described herein.

[00141] In some embodiments the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(7)

I-a(9) or a pharmaceutically acceptable salt thereof, wherein each of R^z, R^z , R^y, X, X', X", n, p, q, and r is as defined above and described herein.

[00142] In some embodiments the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(12) or a pharmaceutically acceptable salt thereof, wherein each of R^y and n are as defined above and described herein.

[00143] In some embodiments, the present invention provides a compound of formula I-a having any one of the structures depicted below in Table 1 :

Table 1

1-1



-9 -2-a

I-5-a I-8-a

harmaceutically acceptable salt thereof.

[00144] In some embodiments the present invention provides a compound of formula I-a an one of the following formulae:

I-a(13) I-a(14)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R^z, R^z , R^z , R^x, R^y, X, X', X", m, p, q, and r is as defined above and described herein.

[00145] In some embodiments the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(15) I-a(16) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R^z, R^z , R^z , R^x, X, X', X", m, p, q, and r is as defined above and described herein.

[00146] In some embodiments the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(17) I-a(18) or a pharmaceutically acceptable salt thereof, wherein each of R^z, R^z , R^x, X, X', X", m, p, q, and r is as defined above and described herein. [00147] In some embodiments the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(19) I-a(20) or a pharmaceutically acceptable salt thereof, wherein each of R^x and m are as defined above and described herein.

[00148] In some embodiments, the present invention provides a compound of formula I-a having any one of the structures depicted below in Table 2:

Tabl

1-10 1-11

or a pharmaceutically acceptable salt thereof.

[00149] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(21) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R^x, R^y, X, X', X", n, m, p, q, and r is as defined above and described herein.

[00150] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(22)

or a pharmaceutically acceptable salt thereof, wherein each of R^x, R^y, n, and m is as defined above and described herein.

[00151] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(23)

or a pharmaceutically acceptable salt thereof, wherein R¹ is as defined above and described herein.

[00152] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(24)

or a pharmaceutically acceptable salt thereof, wherein each of R⁴, R^z, R^z , R^z , R^x, R^y, X, X', X", n, m, p, q, and r is as defined above and described herein.

[00153] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(25)

[00154] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(26)

[00155] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(27)

or a pharmaceutically acceptable salt thereof, wherein each of R^z, R^z , R^x, R^y, X, X', X", n, m, p, q, and r is as defined above and described herein.

[00156] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(28)

[00157] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(29)

[00158] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(30)

[00159] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(31)

[00160] In some embodiments, the present invention provides a compound of formula I-a of the following formula:

I-a(32)

[00161] In some embodiments, the present invention provides a compound of formula I-a having any one of the structures depicted below in Table 3 :

Table

1-16 1-17 or a pharmaceutically acceptable salt thereof.

[00162] In some embodiments, the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(35) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R^z, R^z , R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00163] In some embodiments, the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(37)

I-a(38) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R^x, R^y, m, and n is as defined above and described herein.

[00164] In some embodiments, the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(40)

I-a(41) or a pharmaceutically acceptable salt thereof, wherein each of R^x, R^y, m, and n is as defined above and described herein.

[00165] In some embodiments, the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(43)

I-a(44) or a pharmaceutically acceptable salt thereof, wherein each of R^y and n is as defined above and described herein.

[00166] In some embodiments, the present invention provides a compound of formula I-a of any one of the following formulae:

I-a(46)

I-a(47) or a pharmaceutically acceptable salt thereof, wherein each of R^x and m is as defined above and described herein.

[00167] In some embodiments, the present invention provides a compound of formula I-a having any one of the structures depicted below in Table 4:

Table 4

1-20

or a pharmaceutically acceptable salt thereof.

Compounds of Formula I-b

[00168] As described above, in some embodiments, R² and R³ do not form a double bond. Accordingly, in some embodiments, the present invention provides a compound of Formula I-b:

I-b

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R² , R³ , R³ , R⁴, R^z, R^z . R^z ,

R^x, R^y, X, X', X", m, n, p, q, and r are as defined herein, and wherein:

R² and R³ are each independently -H, -OH, -OSO₃H, -OGlu, or -SR⁵, or:

R² and R³ are taken together with their intervening atoms to form an epoxide moiety.

[00169] In some embodiments, the present invention provides a compound of Formula I-b wherein R¹ is -OMe.

[00170] In some embodiments, the present invention provides a compound of Formula I-b wherein R¹ is selected from -OH, -OSO₃H and -OGlu.

[00171] In some embodiments, the present invention provides a compound of Formula I-b wherein m is 1.

[00172] In some embodiments, the present invention provides a compound of Formula I-b wherein n is 1.

[00173] In some embodiments, the present invention provides a compound of Formula I-b wherein p is 1.

[00174] In some embodiments, the present invention provides a compound of Formula I-b wherein q is 1.

[00175] In some embodiments, the present invention provides a compound of Formula I-b wherein r is 1.

[00176] In some embodiments, the present invention provides a compound of Formula I-b wherein R^z, R^z and R^z are each -H. [00177] In some embodiments, the present invention provides a compound of Formula I-b wherein one of R^z, R^z' or R^{z "} is -CH₃, -OH, -OS0₃H, or -OGlu.

[00178] In some embodiments, the present invention provides a compound of Formula I-b wherein R² is -H.

[00179] In some embodiments, the present invention provides a compound of Formula I-b wherein R² is -OH.

[00180] In some embodiments, the present invention provides a compound of Formula I-b wherein R² is -OS0₃H.

[00181] In some embodiments, the present invention provides a compound of Formula I-b wherein R² is -OGlu.

[00182] In some embodiments, the present invention provides a compound of Formula I-b wherein R³ is -H.

[00183] In some embodiments, the present invention provides a compound of Formula I-b wherein R³ is -OH.

[00184] In some embodiments, the present invention provides a compound of Formula I-b wherein R³ is -OS0₃H.

[00185] In some embodiments, the present invention provides a compound of Formula I-b wherein R³ is -OGlu.

[00186] In some embodiments, the present invention provides a compound of Formula I-b wherein R² and R³ are the same.

[00187] In some embodiments, the present invention provides a compound of Formula I-b wherein R² and R³ are -OH.

[00188] In some embodiments, the present invention provides a compound of Formula I-b wherein R² and R³ are different. In some such embodiments, the present invention provides a compound of Formula I-b wherein R² is H. In some such embodiments, the present invention provides a compound of Formula I-b wherein R² is H and R³ is -SR⁵. In some such

embodiments, R⁵ such embodiments, R⁵ is

. In some such embodiments, R⁵ is [00189] In some embodiments, the present invention provides a compound of Formula I-b wherein R⁴ is H. In some embodiments, the present invention provides a compound of Formula

I-b wherein at least one of R 2 , R 2' , R 3 , R 3' , and R 3" is not hydrogen.

[00190] In some embodiments, a compound is of formula I-b and R² and R³ are taken together with their intervening atoms to form an epoxide moiety.

[00191] In some embodiments, R² and R³ are each independently -H, -OH, -OSO₃H, -OGlu, or -SR⁵.

[00192] In some embodiments, a compound is of formula I-b and at least one of R² and R³ is - OH. In some embodiments, a compound is of formula I-b and both of R² and R³ are -OH. In some embodiments, a compound is of formula I-b and at least one of R² and R³ is -OS0₃H. In some embodiments, a compound is of formula I-b and both of R² and R³ are -OSO3H. In some embodiments, a compound is of formula I-b and at least one of R² and R³ is -OGlu. In some embodiments, a compound is of formula I-b and both of R² and R³ are -OGlu. In some embodiments, one of R² or R³ is -OH and the other of R² or R³ is -OS0₃H. In some embodiments, one of R² or R³ is -OH and the other of R² or R³ is -OGlu. In some embodiments, one of R² or R³ is -OGlu and the other of R² or R³ is -OS0₃H.

[00193] In certain embodiments, a compound is of formula I-b, wherein R² is -OH, -OS0₃H, or -OGlu, and the resulting chiral center of the adjacent carbon atom is in the (S) configuration. In certain embodiments, a compound is of formula I-b, wherein R² is -OH, -OS0₃H, or -OGlu, and the resulting chiral center of the adjacent carbon atom is in the (R) configuration. In certain embodiments, a compound is of formula I-b, wherein R² is -OH and the resulting chiral center of the adjacent carbon atom is in the (S) configuration. In certain embodiments, a compound is of formula I-b, wherein R² is -OH and the resulting chiral center of the adjacent carbon atom is in the (R) configuration.

[00194] In some embodiments, a compound is of formula I-b wherein at least one of R² and R³ is -OH and R¹ is -OMe. In some embodiments, a compound is of formula I-b wherein both of R² and R³ are -OH and R¹ is -OMe.

[00195] In some embodiments, a compound is of formula I-b wherein at least one of R² and R³ is -OH and R⁴ is -H. In some embodiments, a compound is of formula I-b wherein both of R² and R³ are -OH and R⁴ is -H. [00196] In some embodiments, a compound is of formula I-b, wherein R³ is -SR⁵. In some embodiments, a compound is of formula I-b, wherein R³ is -SR⁵, and each of R² and R² are H.

ments, a compound is of formula I-b, wherein R³ is -SR⁵ and R⁵ is

embodiments, a compound is of formula I-b, wherein R³

i

s -SR⁵ and R⁵ is n some embodiments, a compound is of formula I-b,

wherein R³ is -SR⁵ and R⁵ is In some embodiments, a compound is of formula I-b,

wherein R³ is -SR⁵ and R⁵ is

[00198] In some embodiments, a compound is of formula I-b, wherein R³ is -SR⁵ and R¹ is - OMe. In some embodiments, a compound is of formula I-b, wherein R³ is -SR⁵ and R⁴ is -H.

[00199] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R², R³, R⁴, R^z, R^z , R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein. [00200] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(2)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R², R³, R^x, R^y, m and n is as defined above and described herein.

[00201] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(3)

or a pharmaceutically acceptable salt thereof, wherein each of R², R³, R^x, and m is as defined above and described herein.

[00202] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(4)

or a pharmaceutically acceptable salt thereof, wherein each of R², R³, R^y, and n is as defined above and described herein.

[00203] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(5)

or a pharmaceutically acceptable salt thereof, wherein each of R² and R³ is as defined above and described herein.

[00204] In some embodiments, the present invention provides a compound of formula I-b of either of the following formulae:

I-b(6) I-b(7) or a pharmaceutically acceptable salt thereof, wherein each of R² and R³ is as defined above and described herein.

[00205] In some embodiments, the present invention provides a compound of formula I-b of either of th following formulae:

I-b(8) I-b(9)

or a pharmaceutically acceptable salt thereof, wherein R³ is as defined above and described herein.

[00206] In some embodiments, the present invention provides a compound of formula I-b having any one of the structures depicted below in Table 5 :

Table 5

60

61 or a pharmaceutically acceptable salt thereof.

[00207] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(10)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R², R⁴, R⁵, R^z, R^z , R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00208] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(ll)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R², R⁴, R⁵, R^x, R^y, m, and n is as defined above and described herein.

[00209] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(12)

or a pharmaceutically acceptable salt thereof, wherein each of R², R⁵, R^y, and n is as defined above and described herein.

[00210] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(13)

or a pharmaceutically acceptable salt thereof, wherein each of R², R⁵, R^x, and m is as defined above and described herein.

[00211] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(14)

or a pharmaceutically acceptable salt thereof, wherein each of R² and R⁵ is as defined above and described herein.

[00212] In some embodiments, the present invention provides a compound of formula I-b of the following formula:

I-b(15)

or a pharmaceutically acceptable salt thereof, wherein R⁵ is as defined above and described herein.

[00213] In some embodiments, the present invention provides a compound of formula I-b having any one of the structures depicted below in Table 6:

65

Compounds of formula I-c

[00214] In some embodiments, the present invention provides a compound of Formula I-c:

I-c

or a pharmaceutically acceptable salt thereof, wherein R¹, R⁴, R^z, R^z , R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00215] In some embodiments, a compound is of formula I-c, wherein R^z is -H.

[00216] In some embodiments, a compound is of formula I-c, wherein R¹ is -OMe.

[00217] In some embodiments, a compound is of formula I-c, wherein R¹ is selected from -

[00218] In some embodiments, a compound is of formula I-c, wherein R⁴ is -H.

[00219] In some embodiments, a compound is of formula I-c, wherein m is 1.

[00220] In some embodiments, a compound is of formula I-c, wherein n is 1.

[00221] In some embodiments, a compound is of formula I-c, wherein p is 1.

[00222] In some embodiments, a compound is of formula I-c, wherein q is 1.

[00223] In some embodiments, a compound is of formula I-c, wherein r is 1.

[00224] In some embodiments, a compound is of formula I-c, wherein R^z, R^z and R^z are each

-H.

[00225] In some embodiments, a compound is of formula I-c, wherein one of R^z, R^z or R^z is - CH₃, -OH, -OS0₃H, or -OGlu.

[00226] In some embodiments, a compound is of formula I-c, wherein R^z is -H and wherein R¹ is -OMe. In some embodiments, a compound is of formula I-c, wherein R^z is -H and wherein R⁴ is -H.

[00227] In some embodiments, the present invention provides a compound of Formula I-c of the following formula:

I-c(l)

or a pharmaceutically acceptable salt thereof, wherein R¹, R⁴, R^z, R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00228] In some embodiments, the present invention provides a compound of Formula I-c of the following formula:

I-c(2)

or a pharmaceutically acceptable salt thereof, wherein R^z, R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00229] In some embodiments, the present invention provides a compound of Formula I-c of the following formula:

I-C(3)

or a pharmaceutically acceptable salt thereof, wherein R^x, R^y, m, and n is as defined above and described herein.

[00230] In some embodiments, the present invention provides a compound of Formula I-c of the following formula:

I-c(4)

or a pharmaceutically acceptable salt thereof, wherein R^y and n is as defined above and described herein.

[00231] In some embodiments, the present invention provides a compound of Formula I-c of the following formula:

I-c(5)

or a pharmaceutically acceptable salt thereof, wherein R^x and m is as defined above and described herein.

[00232] In some embodiments, the present invention provides a compound of formula I-c having the structure depicted below in Table 7:

1-40

or a pharmaceutically acceptable salt thereof.

Compounds of Formula Il-a

[00233] As described above, in some embodiments, R² and R³ are taken together to form a double bond. Accordingly, in some embodiments, the present invention provides a compound of Formula Il-a:

wherein each of R¹, R⁴, R⁶, R⁷, R^z, R^z', R^z", R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00234] In some embodiments, a compound is of formula Il-a, wherein at least one of R¹, R⁴, R^x, and R^y is independently selected from -OH, -OSO₃H and -OGlu. In some embodiments, a compound is of formula Il-a, wherein at least one of R¹, R⁴, R^x, and R^y is -OH. In some embodiments, a compound is of formula Il-a, wherein at least one of R¹, R⁴, R^x, and R^y is - OSO3H. In some embodiments, a compound is of formula Il-a, wherein at least one of R¹, R⁴, R^x, and R^y is -OGlu.

[00235] In some embodiments, a compound is of formula Il-a and R¹ is -OH, -OSO3H or - OGlu. In some embodiments, a compound is of formula Il-a and R¹ is -OH. In some embodiments, a compound is of formula Il-a and R¹ is -OSO3H. In some embodiments, a compound is of formula Il-a and R¹ is -OGlu.

[00236] In some embodiments, a compound is of formula Il-a and n is 0. In some embodiments, a compound is of formula Il-a and n is 1. In some embodiments, a compound is of formula Il-a and n is 2. In some embodiments, a compound is of formula Il-a and m is 0. In some embodiments, a compound is of formula Il-a and m is 1. In some embodiments, a compound is of formula Il-a and m is 2. In some embodiments, a compound is of formula Il-a and m is 3.

[00237] In certain embodiments, at least one of m or n is 0 and R¹ is OH. In certain embodiments, both m and n are 0 and R¹ is OH.

[00238] In some embodiments, a compound is of formula Il-a and at least one of p, q, and r is 1. In some embodiments, a compound is of formula Il-a and p is 1. In some embodiments, a compound is of formula Il-a and q is 1. In some embodiments, a compound is of formula Il-a and r is 1.

[00239] In some embodiments, a compound is of formula Il-a and R¹ is -OMe.

[00240] In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe and at least one of p, q, and r is 1. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe and p is 1. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe and and q is 1. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe and r is 1.

[00241] In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe and m is 1, 2, or 3. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe and m is 1. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe and m is 2. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe and m is 3. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe, m is 1, 2, or 3, and each R^x is -OH. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe, m is 1, 2, or 3, and each R^x is -OSO₃H. In some embodiments, a compound is of formula Il-a, wherein R¹ is -OMe, m is 1, 2, or 3, and each R^x is -OGlu.

[00242] In certain embodiments, a compound is of formula Il-a, wherein R¹ is -OMe, m is 1, and R^x is -OH. In certain embodiments, a compound is of formula Il-a, wherein R¹ is -OMe, m is 1, and R^x is -OSO₃H. In certain embodiments, a compound is of formula Il-a, wherein R¹ is - OMe, m is 1, and R^x is -OGlu.

[00243] In some embodiments, the compound is of formula Il-a, wherein R⁴ is hydrogen.

[00244] In some embodiments, the compound is of formula Il-a, wherein R⁶ is selected from -CH₃, -CH₂OH, -CH₂SO₃H, and -CH₂OGlu. In some embodiments, the compound is of formula Il-a, wherein R⁶ is -CH₃. In some embodiments, the compound is of formula Il-a, wherein R⁶ is selected from -CH₂OH, -CH₂S0₃H, and -CH₂OGlu. In some embodiments, the compound is of formula Il-a, wherein R⁶ is selected from -CH₂OH, -CH₂OSO₃H, and -CH₂OGlu. In some embodiments, the compound is of formula Il-a, wherein R⁶ is -CH₂OH. In some embodiments, the compound is of formula Il-a, wherein R⁶ is -CH₂OSO₃H. In some embodiments, the compound is of formula Il-a, wherein R⁶ is -CH₂OGlu.

[00245] In some embodiments, the compound is of formula Il-a, wherein R⁷ is selected from -CH₃, -CH₂OH, -CH₂SO₃H, and -CH₂OGlu. In some embodiments, the compound is of formula Il-a, wherein R⁷ is -CH₃. In some embodiments, the compound is of formula Il-a, wherein R⁷ is selected from -CH₂OH, -CH₂S0₃H, and -CH₂OGlu. In some embodiments, the compound is of formula Il-a, wherein R⁷ is selected from -CH₂OH, -CH₂OS0₃H, and -CH₂OGluIn some embodiments, the compound is of formula Il-a, wherein R⁷ is -CH₂OH. In some embodiments, the compound is of formula Il-a, wherein R⁷ is -CH₂OSO₃H. In some embodiments, the compound is of formula Il-a, wherein R⁷ is -CH₂OGlu.

[00246] In some embodiments, the compound is of formula Il-a, wherein R^z, R^z and R^z are each -H.

[00247] In some embodiments, the compound is of formula Il-a, wherein one of R^z, R^z or R^z is -CH₃, -OH, -OSO₃H, or -OGlu.

[00248] In some embodiments the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(3) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R⁶, R⁷, R^z, R^z , R^z , R^x, R^y, X, X', X", n, p, q, and r is as defined above and described herein.

[00249] In some embodiments the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(6) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R⁶, R⁷, R^z, R^z , R^z , R^y, X, X', X", n, p, q, and r is as defined above and described herein.

[00250] In some embodiments the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(9) or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^z, R^z , R^y, X, X', X", n, p, q, and r is as defined above and described herein.

[00251] In some embodiments the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(12) or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^y and n are as defined above and described herein.

[00252] In some embodiments, the present invention provides a compound of formula Il-a having any one of the structures depicted below in Table 8:

Table 8

II-5 II-6 [00253] In some embodiments, the present invention provides a compound of formula II, wherein the compound is other than a compound depicted in Table 8.

[00254] In some embodiments the present invention provides a compound of formula Il-a of either of the following formulae:

II-a(13) II-a(14)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R⁶, R⁷, R^z, R^z , R^z , R^x, R^y, X, X', X", m, p, q, and r is as defined above and described herein.

[00255] In some embodiments the present invention provides a compound of formula Il-a of either of the following formulae:

II-a(15) II-a(16) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R⁶, R⁷, R^z, R^z , R^z , R^x, X, X', X", m, p, q, and r is as defined above and described herein.

[00256] In some embodiments the present invention provides a compound of formula Il-a of either of the following formulae:

II-a(17) II-a(18) or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^z, R^z , R^x, X, X', X", m, p, q, and r is as defined above and described herein.

[00257] In some embodiments the present invention provides a compound of formula Il-a of either of the following formulae:

II-a(19) II-a(20) or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^x and m is as defined above and described herein.

[00258] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(21)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁶, R⁷, R^x, R^y, X, X', X", n, p, q, and r is as defined above and described herein.

[00259] In some embodiments, the present invention provides a compound of formula Il-a the following formula:

II-a(22)

or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^x, R^y, n, and m is as defined above and described herein.

[00260] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(23)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁶, and R⁷ is as defined above and described herein.

[00261] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(24)

or a pharmaceutically acceptable salt thereof, wherein each of R⁴, R⁶, R⁷, R^z, R^z , R^z , R^x, R^y, X, X', X", n, m, p, q, and r is as defined above and described herein.

[00262] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(25)

[00263] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(26)

[00264] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(27)

or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^z, R^z , R^x, R^y, X, X', X", n, m, p, q, and r is as defined above and described herein.

[00265] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(28)

[00266] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(29)

[00267] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(30)

[00268] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(31)

[00269] In some embodiments, the present invention provides a compound of formula Il-a of the following formula:

II-a(32)

[00270] In some embodiments, the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(35) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R⁶, R⁷, R^z, R^z , R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00271] In some embodiments, the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(38) or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁶, R⁷, R^x, R^y, m, and n is as defined above and described herein.

[00272] In some embodiments, the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(41) or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^x, R^y, m, and n is as defined above and described herein.

[00273] In some embodiments, the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(44) or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^y and n is as defined above and described herein.

[00274] In some embodiments, the present invention provides a compound of formula Il-a of any one of the following formulae:

II-a(47) or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^x and m is as defined above and described herein.

Compounds of Formula H-b

[00275] As described above, in some embodiments, R² and R³ do not form a double bond. Accordingly, in some embodiments, the present invention provides a compound of Formula Il-b:

Il-b

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R² , R³ , R³ , R⁴, R⁶, R⁷, R^z, R^z .

R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined herein, and wherein:

R² and R³ are each independently -H, -OH, -OSO₃H, -OGlu, or -SR⁵, or:

[00276] In some embodiments, the present invention provides a compound of Formula Il-b wherein at least one of R 2 , R 2' , R 3 , R 3' , and R 3" is not hydrogen.

[00277] In some embodiments, the present invention provides a compound of Formula Il-b wherein R¹ is -OMe.

[00278] In some embodiments, the present invention provides a compound of Formula Il-b wherein R¹ is selected from -OH, -OSO₃H and -OGlu.

[00279] In some embodiments, the present invention provides a compound of Formula Il-b wherein m is 1.

[00280] In some embodiments, the present invention provides a compound of Formula Il-b wherein n is 1.

[00281] In some embodiments, the present invention provides a compound of Formula Il-b wherein p is 1.

[00282] In some embodiments, the present invention provides a compound of Formula Il-b wherein q is 1.

[00283] In some embodiments, the present invention provides a compound of Formula Il-b wherein r is 1.

[00284] In some embodiments, the present invention provides a compound of Formula Il-b wherein R^z, R^z and R^z are each -H.

[00285] In some embodiments, the present invention provides a compound of Formula Il-b wherein one of R^z, R^z' or R^{z "} is -CH₃, -OH, -OS0₃H, or -OGlu.

[00286] In some embodiments, the present invention provides a compound of Formula Il-b wherein R² is -H.

[00287] In some embodiments, the present invention provides a compound of Formula Il-b wherein R² is -OH.

[00288] In some embodiments, the present invention provides a compound of Formula Il-b wherein R² is -OS0₃H.

[00289] In some embodiments, the present invention provides a compound of Formula Il-b wherein R² is -OGlu.

[00290] In some embodiments, the present invention provides a compound of Formula Il-b wherein R³ is -H.

[00291] In some embodiments, the present invention provides a compound of Formula Il-b wherein R³ is -OH.

[00292] In some embodiments, the present invention provides a compound of Formula Il-b wherein R³ is -OS0₃H.

[00293] In some embodiments, the present invention provides a compound of Formula Il-b wherein R³ is -OGlu.

[00294] In some embodiments, the present invention provides a compound of Formula Il-b wherein R² and R³ are the same.

[00295] In some embodiments, the present invention provides a compound of Formula Il-b wherein R² and R³ are different. In some such embodiments, the present invention provides a compound of Formu -b wherein R² is H. In some such embodiments, R³ is -SR⁵. In some

such embodiments, In some such

embodiments, R³ is -

SR⁵ wherein R⁵ is In some such embodiments, R³ is SR⁵ wherein R⁵ is

[00296] In some embodiments, a compound is of formula Il-b and R² and R³ are taken together with their intervening atoms to form an epoxide moiety.

[00297] In some embodiments, R² and R³ are each independently -H, -OH, -OSO3H, -OGlu, or -SR⁵.

[00298] In some embodiments, a compound is of formula Il-b and at least one of R² and R³ is -OH. In some embodiments, a compound is of formula Il-b and both of R² and R³ are -OH. In some embodiments, a compound is of formula Il-b and at least one of R² and R³ is -OSO3H. In some embodiments, a compound is of formula Il-b and both of R² and R³ are -OSO3H. In some embodiments, a compound is of formula Il-b and at least one of R² and R³ is -OGlu. In some embodiments, a compound is of formula Il-b and both of R² and R³ are -OGlu. In some embodiments, one of R² or R³ is -OH and the other of R² or R³ is -OSO3H. In some embodiments, one of R² or R³ is -OH and the other of R² or R³ is -OGlu. In some embodiments, one of R² or R³ is -OGlu and the other of R² or R³ is -OSO₃H.

[00299] In certain embodiments, a compound is of formula Il-b, wherein R² is -OH, -OSO3H, or -OGlu, and the resulting chiral center of the adjacent carbon atom is in the (S) configuration. In certain embodiments, a compound is of formula Il-b, wherein R² is -OH, -OSO₃H, or -OGlu, and the resulting chiral center of the adjacent carbon atom is in the (R) configuration. In certain embodiments, a compound is of formula Il-b, wherein R² is -OH and the resulting chiral center of the adjacent carbon atom is in the (S) configuration. In certain embodiments, a compound is of formula Il-b, wherein R² is -OH and the resulting chiral center of the adjacent carbon atom is in the (R) configuration.

[00300] In some embodiments, a compound is of formula Il-b wherein at least one of R² and R³ is -OH and R¹ is -OMe. In some embodiments, a compound is of formula Il-b wherein both of R² and R³ are -OH and R¹ is -OMe.

[00301] In some embodiments, a compound is of formula Il-b wherein R⁴ is H.

[00302] In some embodiments, a compound is of formula Il-b wherein at least one of R² and

R³ is -OH and R⁴ is -H. In some embodiments, a compound is of formula Il-b wherein both of R² and R³ are -OH and R⁴ is -H. [00303] In some embodiments, a compound is of formula Il-b, wherein R³ is -SR⁵. In some embodiments, a compound is of formula Il-b, wherein R³ is -SR⁵, and each of R² and R² are H.

[00304] In some embodiments, a compound is of formula Il-b, wherein R³ is -SR⁵ and R⁵ is

embodiments, a compound is of formula Il-b, wherein R³

i

s -SR⁵ and R⁵ is some embodiments, a compound is of formula Il-b,

wherein R³ is -SR⁵ and R⁵ is some embodiments, a compound is of formula II-

b, wherein R³ is -SR⁵ and R⁵ i

s

[00305] In some embodiments, a compound is of formula Il-b, wherein R³ is -SR⁵ and R¹ is - OMe. In some embodiments, a compound is of formula Il-b, wherein R³ is -SR⁵ and R⁴ is -H.

[00306] In some embodiments, a compound is of formula Il-b, wherein R⁶ is selected from - CH₂OH, -CH₂OSO₃H, or -CH₂OGlu.

[00307] In some embodiments, a compound is of formula Il-b, wherein R⁷ is selected from - CH₂OH, -CH₂OS0₃H, or -CH₂OGlu.

[00308] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(l)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R², R³, R⁴, R⁶, R⁷, R^z, R^z , R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00309] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(2)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R², R³, R⁶, R⁷, R^x, R^y, m and n is as defined above and described herein.

[00310] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(3)

or a pharmaceutically acceptable salt thereof, wherein each of R², R³, R⁶, R⁷, R^x, and m is as defined above and described herein. [00311] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(4)

or a pharmaceutically acceptable salt thereof, wherein each of R², R³, R⁶, R⁷, R^y, and n is as defined above and described herein.

[00312] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(5)

or a pharmaceutically acceptable salt thereof, wherein each of R², R³, R⁶, and R⁷ is as defined above and described herein.

[00313] In some embodiments, the present invention provides a compound of formula Il-b of either of the following formulae:

[00314] In some embodiments, the present invention provides a compound of formula Il-b of either of th following formulae:

II-b(8) II-b(9)

or a pharmaceutically acceptable salt thereof, wherein each of R³, R⁶, and R⁷ is as defined above and described herein.

[00315] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

Il-b(lO)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R², R⁴, R⁵, R⁶, R⁷, R^z, R^z , R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00316] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

Il-b(ll)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R², R⁴, R⁵, R⁶, R⁷, R^x, R^y, m, and n is as defined above and described herein.

[00317] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(12)

or a pharmaceutically acceptable salt thereof, wherein each of R², R⁵, R⁶, R⁷, R^y, and n is as defined above and described herein.

[00318] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(13)

or a pharmaceutically acceptable salt thereof, wherein each of R², R⁵, R⁶, R⁷, R^x, and m is as defined above and described herein.

[00319] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(14)

or a pharmaceutically acceptable salt thereof, wherein each of R², R⁵, R⁶, and R⁷ is as defined above and described herein.

[00320] In some embodiments, the present invention provides a compound of formula Il-b of the following formula:

II-b(15)

or a pharmaceutically acceptable salt thereof, wherein each of R⁵, R⁶, and R⁷ is as defined above and described herein.

Compounds of formula II-c

[00321] In some embodiments, the present invention provides a compound of Formula II-c:

II- or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R⁶, R⁷ R^z, R^z , R^z , R^x, R^y,

X, X', X", m, n, p, q, and r is as defined above and described herein.

[00322] In some embodiments, a compound is of formula II-c, wherein R^z is -H.

[00323] In some embodiments, a compound is of formula II-c, wherein R¹ is -OMe.

[00324] In some embodiments, a compound is of formula II-c, wherein R¹ is selected from -

OH, -OSO₃H and -OGlu.

[00325] In some embodiments, a compound is of formula II-c, wherein R⁴ is -H.

[00326] In some embodiments, a compound is of formula II-c, wherein R^z is -H and wherein R¹ is -OMe. In some embodiments, a compound is of formula II-c, wherein R^z is -H and wherein R⁴ is -H.

[00327] In some embodiments, a compound is of formula II-c, wherein R⁶ is selected from - CH₂OH, -CH₂OSO₃H, or -CH₂OGlu.

[00328] In some embodiments, a compound is of formula II-c, wherein R⁷ is selected from - CH₂OH, -CH₂OSO₃H, or -CH₂OGlu.

[00329] In some embodiments, a compound is of formula II-c, wherein m is 1.

[00330] In some embodiments, a compound is of formula II-c, wherein n is 1.

[00331] In some embodiments, a compound is of formula II-c, wherein p is 1.

[00332] In some embodiments, a compound is of formula II-c, wherein q is 1.

[00333] In some embodiments, a compound is of formula II-c, wherein r is 1.

[00334] In some embodiments, a compound is of formula II-c, wherein R^z, R^z and R^z are each -H. [00335] In some embodiments, a compound is of formula II-c, wherein one of R^z, R^z or R^z is -CH₃, -OH, -OS0₃H, or -OGlu.

[00336] In some embodiments, the present invention provides a compound of Formula II-c of the following formula:

II-c(l)

or a pharmaceutically acceptable salt thereof, wherein each of R¹, R⁴, R⁶, R⁷, R^z, R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00337] In some embodiments, the present invention provides a compound of Formula II-c of the following formula:

II-c(2)

or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^z, R^z , R^x, R^y, X, X', X", m, n, p, q, and r is as defined above and described herein.

[00338] In some embodiments, the present invention provides a compound of Formula II-c of the following formula:

II-c(3)

or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^x, R^y, m, and n is defined above and described herein.

[00339] In some embodiments, the present invention provides a compound of Formula II-c the following formula:

II-c(4)

or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^y and n is as defined above and described herein.

[00340] In some embodiments, the present invention provides a compound of Formula II-c of the following formula:

II-c(5)

or a pharmaceutically acceptable salt thereof, wherein each of R⁶, R⁷, R^x and m is as defined above and described herein.

[00341] In certain embodiments, the present invention provides any compound described above and herein, or a pharmaceutically acceptable salt thereof.

[00342] In some embodiments, the present invention provides any compound described above and herein in isolated form.

[00343] In some embodiments, the present invention provides a composition comprising a compound of Formula I or Formula II, and a pharmaceutically acceptable adjuvant, carrier, or vehicle.

Uses of Compounds and Pharmaceutically Acceptable Compositions

[00344] As described generally above, compounds of Formula I or II, and pharmaceutically acceptable salts thereof described herein, are inhibitors of one or both of ERKl and ERK2. One of ordinary skill in the art will recognize that ERK is one of the key components in the RAS- RAF-MEK-ERK MAPK pathway and that ERKl and ERK2 are downstream nodes within the MAPK pathway. Without wishing to be bound by theory, because of the downstream location of ERKl and ERKl in the MAPK pathway, an ERK inhibitor can treat disease or disorders in which activation of the MAPK pathway at any level (Ras-Raf-Mek-ERK) is known or suspected to play a role, including one or both of ERKl and ERK2 as well as other nodes in the MAPK pathway upstream from ERK (such as Ras, Raf and Mek). Furthermore, because ERK is a downstream target, ERK inhibitors are believed to be able to overcome, in some instances, drug resistance induced by inhibitors of targets upstream of ERK within the MAPK pathway. For example, small molecule inhibitors of RAF or MEK utilized in the treatment of K-RAS and B- RAF mutant tumors have resulted in such drug resistance. Similarly, drug resistance has been associated with other tumors driven by hyperactivation of the MAPK pathway (such as NF1 mutant tumors). Kinase selectivity was achieved through silencing the selective Cys in a combination of the interactions between the covalent inhibitors of the invention and unique amino acids in the ATP binding pocket. Targeting the selective Cys provides for prolonged pharmacodynamics in silencing ERK activity, as well as potential lower doses in cancer treatment, compared to reversible inhibitors.

[00345] The activity of a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, as an inhibitor of one or both of an ERKl and ERK2 kinase, or a mutant thereof, may be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of downstream phosphorylation, changes in gene expression, subsequent functional markers and consequences, and/or kinase activity of one or both of activated ERKl and ERK2 kinase, or a mutant thereof. Alternate in vitro assays quantitate the ability of the test compound to bind to one or both of ERKl and ERK2. Test compound binding may be measured by radiolabeling the test compound prior to binding, isolating one or both of the compound / ERKl complex and the compound / ERK2 complex, and determining the amount of radiolabel bound. Alternatively, test compound binding may be determined by running a competition experiment where test compounds are incubated with one or both of ERKl and ERK2 kinase bound to known radioligands. Test compound binding may be determined by competition with an ERK covalent probe that is amenable to further functionalization with a detection probe, such as, for example, a fluorophore, biotin conjugate, radiolabel, or any other probe that facilitates its quantification. Detailed conditions for assaying a compound utilized in this invention as an inhibitor of one or both of ERKl and ERK2, or a mutant thereof, are also set forth below and/or in the Examples of the '230 publication.

[00346] The term "measurably inhibit", as used herein means a measurable change in one or both of ERKl and ERK2 protein kinase activity between a sample comprising a provided composition, and one or both of an ERKl and ERK2 protein kinase and an equivalent sample comprising one or both of ERKl and ERK2 protein kinase in the absence of a provided composition. Such measurements of protein kinase activity are known to one of ordinary skill in the art and include those methods set forth herein below and/or in the Examples of the '230 publication.

[00347] As described above, in some embodiments, a compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is an inhibitor of one or both of ER 1 and ERK2 protein kinases, and ERK1 and ERK2 are downstream targets within the MAPK pathway. Without wishing to be bound by any particular theory, such compounds and compositions are particularly useful for treating or lessening the severity of a disease, condition, or disorder in which activation of the MAPK pathway at any level (Ras-Raf-Mek-ERK) is known or suspected to play a role. Such disease, condition, or disorder may be referred to herein as associated with the MAPK pathway or alternatively as associated with one or both of ERK1 and ERK2. Such diseases, conditions, or disorders may also be referred to herein as an "ERK1- or ERK2- mediated disease, condition, or disorder."

[00348] In some embodiments, the present invention provides a method for treating or lessening the severity of a disease, condition, or disorder where activation of the MAPK pathway (at any level in Ras-Raf-Mek-ERK), including one or both of ERKl and ERK2 protein kinases, is implicated in said disease, condition, or disorder, wherein said method comprises administering to a patient in need thereof a compound of the present invention.

[00349] In some embodiments, the present invention relates to a method of inhibiting one or both of ERKl and ERK2 protein kinase activity in a patient comprising the step of administering to said patient a composition comprising a compound of the present invention.

[00350] In other embodiments, the present invention provides a method for treating a disease, condition, or disorder mediated by one or both of ERKl and ERK2 kinase, or a mutant thereof, in a patient in need thereof, comprising the step of administering to said patient a compound of the present invention.

[00351] In certain embodiments, the present invention provides a method for overcoming drug resistance to Raf or MEK inhibitors, comprising the step of administering to a patient an inhibitor compound of one or both of ERKl and ERK2, such as a compound of the present invention. In certain embodiments, the mechanism of drug resistance is through mutation of a target protein or reactivation of the MAPK pathway.

[00352] As used herein, the term "resistance" may refer to changes in a wild-type nucleic acid sequence coding a target protein, and/or to the amino acid sequence of the target protein and/or to the amino acid sequence of another protein, which changes, decreases or abolishes the inhibitory effect of the inhibitor on the target protein. The term "resistance" may also refer to overexpression or silencing of a protein differing from a target protein that can reactivate the MAPK pathway or other survival pathways.

[00353] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein. In some embodiments, treatment is administered after one or more symptoms have developed. In other embodiments, treatment is administered in the absence of symptoms. For example, treatment is administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors). Treatment is also continued after symptoms have resolved, for example to prevent, delay or lessen the severity of their recurrence.

[00354] In some embodiments, the present invention provides a method of treating, stabilizing or lessening the severity or progression of one or more diseases or disorders associated with one or both of ER 1 and ERK2 comprising administering to a patient in need thereof a composition comprising a compound of the present invention.

[00355] General diseases, conditions, or disorders treated by a compound of the present invention include cancer, an autoimmune disorder, a neurodegenerative or neurological disorder, liver disease, a cardiac disorder, schizophrenia, or a bone-related disorder.

[00356] In some embodiments, the present invention relates to a method of treating or lessening the severity of a disease, condition, or disorder selected from cancer, stroke, diabetes, hepatomegaly, cardiovascular disease including cardiomegaly, Alzheimer's disease, cystic fibrosis, viral disease, autoimmune diseases, atherosclerosis, restenosis, psoriasis, allergic disorders including asthma, inflammation, neurological disorders and hormone -related diseases, wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention.

[00357] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention. In some embodiments, the cancer is recurring. In certain embodiments, the cancer is refractory. In some embodiments, the cancer is metastatic. In some embodiments, the cancer is locally advanced. [00358] In certain embodiments, the cancer is a RAF inhibitor-resistant cancer. In some such embodiments, the RAF inhibitor-resistant cancer is a BRAF inhibitor-resistant cancer.

[00359] In certain embodiments, the cancer is a MEK inhibitor-resistant cancer.

[00360] In certain embodiments, the cancer is a MAPK pathway-mediated cancer.

[00361] In some embodiments, the cancer is a BRAF-mutated cancer. In certain embodiments, the BRAF-mutated cancer is a BRAF^V600-mutated cancer, such as BRAF

_BRAFV600K _BRAFV600R _{and BRAF}V600D

[00362] In some embodiments, the cancer is a RAS-mutated cancer. In certain embodiments, the RAS-mutated involves codons 12, 13, or 61. In certain embodiments, the RAS-mutated cancer is a KRAS-mutated cancer, including, but not limited to, KRAS^G12C/D/V, KRAS^G13C/D,or KRAS^Q61L/H/R. In certain embodiments, the RAS-mutated cancer is an NRAS-mutated cancer, including, but not limited to, NRAS^Q61R, NRAS^Q61K, NRAS^Q61L, or NRAS^Q61H. In certain embodiments, the RAS-mutated cancer is an HRAS-mutated cancer, including, but not limited to, HRAS^G12V, HRAS^Q61R, and HRAS^G12S.

[00363] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is selected from multiple myeloma, breast, ovary, cervix, prostate, testis, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach (gastric), skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung, bone, colon, thyroid, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma (including uveal melanoma) sarcoma, bladder carcinoma, liver carcinoma (e.g., hepatocellular carcinoma (HCC)) and biliary passage carcinoma), kidney carcinoma, myeloid disorders, lymphoid disorders, Hodgkin's, hairy cells, buccal cavity and pharynx (oral), lip, tongue, mouth, pharynx, small intestine, colorectal carcinoma, large intestine, rectum, brain and central nervous system, endometrial, multiple myeloma (MM), prostate, AML, and leukemia. In some such embodiments, the cancer is relapsed. In some embodiments, the cancer is refractory. In some embodiments, the cancer is locally advanced. In some embodiments, the cancer is metastatic.

[00364] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is selected from carcinoma, lymphoma, blastoma, sarcoma, and leukemia. In some embodiments, a sarcoma is a soft tissue sarcoma. In some embodiments, a lymphoma is non-Hodgkin's lymphoma. In some embodiments, a lymphoma is large cell immunoblastic lymphoma. In some embodiments, the cancer is selected from adenocarcinoma; adenoma; adrenocortical cancer; bladder cancer; bone cancer; brain cancer; breast cancer; cancer of the buccal cavity; cervical cancer; colon cancer; colorectal cancer; endometrial or uterine carcinoma; epidermoid carcinoma; oesophageal cancer; eye cancer; follicular carcinoma; gallbladder cancer; prostate, AML, multiple myeloma (MM), gastrointestinal cancer, such as, for example, gastrointestinal stromal tumor; cancer of the genitourinary tract; glioblastoma; hairy cell carcinoma; various types of head and neck cancer; hepatic carcinoma; hepatocellular cancer; Hodgkin's disease; keratoacanthoma; kidney cancer; large cell carcinoma; cancer of the large intestine; laryngeal cancer; liver cancer; lung cancer, such as, for example, adenocarcinoma of the lung, anaplastic carcinoma of the lung, papillary lung adenocarcinoma, small-cell lung cancer, squamous carcinoma of the lung, non-small cell lung cancer; melanoma and nonmelanoma skin cancer; lymphoid disorders; myeloproliferative disorders, such as, for example, polycythemia vera, essential thrombocythemia, chronic idiopathic myelofibrosis, myeloid metaplasia with myelofibrosis, chronic myeloid leukemia (CML), chronic myelomonocytic leukemia, chronic eosinophilic leukemia, chronic lymphocytic leukemia (CLL), hypereosinophilic syndrome, systematic mast cell disease, atypical CML, AML, or juvenile myelomonocytic leukemia; plasmacytoma; multiple myeloma; neuroblastoma; ovarian cancer; papillary carcinoma; pancreatic cancer; cancer of the peritoneum; prostate cancer, including benign prostatic hyperplasia; rectal cancer; salivary gland carcinoma; sarcoma; seminoma; squamous cell cancer; small cell carcinoma; cancer of the small intestine; stomach cancer; testicular cancer; thyroid cancer; undifferentiated carcinoma; and vulval cancer. In some such embodiments, the cancer is relapsed. In some embodiments, the cancer is refractory. In some embodiments, the cancer is locally advanced. In some embodiments, the cancer is metastatic.

[00365] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is selected from melanoma, pancreatic cancer, thyroid cancer, colorectal cancer, lung cancer (e.g., non-small cell lung cancer), breast cancer, endometrial cancer, prostate cancer, ovarian cancer, hepatocellular carcinoma (HCC), multiple myeloma (MM), and leukemia. In some embodiments, a leukemia is an acute leukemia. In certain embodiments, a leukemia is acute myeloid leukemia. In certain embodiments, a leukemia is acute lymphoblastic leukemia.

[00366] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is selected from melanoma, colorectal cancer, lung cancer, or pancreatic.

[00367] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is melanoma. In certain embodiments, the melanoma is uveal melanoma. In some embodiments, the melanoma is a melanoma of the skin. In certain embodiments, the melanoma is locally advanced. In some embodiments, the melanoma is metastatic. In some embodiments, the melanoma is recurring. In some embodiments, the melanoma is BRAF^v600-mutated melanoma. In certain embodiments, the melanoma is a RAS-mutated melanoma. In some embodiments, the melanoma is NRAS- mutated melanoma. In certain embodiments, the melanoma is wild type for KRAS, NRAS or BRAF. In certain embodiments, the melanoma is a BRAF inhibitor-resistant (e.g., Vemurfenib- resistant, dabrafenib-resistant, etc.) melanoma. In certain embodiments, the cancer is a VemR (i.e., Vemurfenib-resistant) BRAF -mutated melanoma. In some embodiments, the melanoma is relapsed. In some embodiments, the melanoma is refractory.

[00368] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is colorectal cancer. In certain embodiments, the colorectal cancer is locally advanced. In certain embodiments, the colorectal cancer is metastatic. In certain embodiments, the colorectal cancer is a BRAF- mutated colorectal cancer. In certain embodiments, the colorectal cancer is a BRAF^v600-mutated colorectal cancer. In certain embodiments, the colorectal cancer is a RAS-mutated colorectal cancer. In certain embodiments, the colorectal cancer is a KRAS-mutated colorectal cancer. In certain embodiments, the colorectal cancer is a NRAS-mutated colorectal cancer. In some embodiments, the colorectal cancer is relapsed. In some embodiments, the colorectal cancer is refractory.

[00369] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is pancreatic cancer. In certain embodiments, the pancreatic cancer is locally advanced. In certain embodiments, the pancreatic cancer is metastatic. In certain embodiments, the pancreatic cancer is a pancreatic ductal adenocarcinoma (PDAC). In certain embodiments, the pancreatic cancer is a RAS- mutated pancreatic cancer. In certain embodiments, the pancreatic cancer is a KRAS-mutated pancreatic cancer. In certain embodiments, the pancreatic cancer is KRAS-mutated pancreatic cancer, including, but not limited to, KRAS^G12C/D/V, KRAS^G13C/D,or KRAS^Q61L/H/R. In some embodiments, the pancreatic cancer is relapsed. In some embodiments, the pancreatic cancer is refractory.

[00370] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is a papillary thyroid cancer. In certain embodiments, the papillary thyroid cancer is locally advanced. In some embodiments, the papillary thyroid cancer is metastatic. In some embodiments, the papillary thyroid cancer is recurring. In some embodiments, the papillary thyroid cancer is BRAF- mutated papillary thyroid cancer. In some embodiments, the papillary thyroid cancer is BRAF^v600-mutated papillary thyroid cancer. In some embodiments, the papillary thyroid cancer is relapsed. In some embodiments, the papillary thyroid cancer is refractory. In some embodiments, the papillary thyroid cancer includes undifferentiated or dedifferentiated histology.

[00371] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is lung cancer. In certain embodiments, the lung cancer is non-small cell lung cancer (NSCLC). In certain embodiments, the lung cancer is locally advanced. In certain embodiments, the lung cancer is metastatic. In certain embodiments, the lung cancer is a RAS-mutated lung cancer. In certain embodiments, the lung cancer is KRAS-mutated lung cancer. In certain embodiments, the lung cancer is a KRAS-mutated lung cancer, including, but not limited to, KRAS^W , KRAS ,or KRAS^Q61L/H/R In some embodiments, the lung cancer is relapsed. In some embodiments, the lung cancer is refractory.

[00372] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is a leukemia. In some embodiments, a leukemia is a chronic leukemia. In certain embodiments, a leukemia is chronic myeloid leukemia. In some embodiments, a leukemia is an acute leukemia. In certain embodiments, a leukemia is acute myeloid leukemia (AML). In certain embodiments, a leukemia is acute monocytic leukemia (AMoL, or AML-M5). In certain embodiments, a leukemia is acute lymphoblastic leukemia (ALL). In certain embodiments, a leukemia is acute T cell leukemia. In certain embodiments, a leukemia is myelomonoblastic leukemia. In certain embodiments, a leukemia is human B cell precursor leukemia. In certain embodiments, a leukemia has a Flt3 mutation or rearrangement. In some embodiments, the leukemia is relapsed. In some embodiments, the leukemia is refractory.

[00373] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is a CNS cancer, for instance CNS tumors. In certain embodiments, a CNS tumor is a glioblastoma or glioblastoma multiforme (GBM). In some embodiments, the present invention relates to a method of treating stomach (gastric) and esophageal tumors and cancers.

[00374] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is multiple myeloma (MM). In certain embodiments, the multiple myeloma is locally advanced. In certain embodiments, the multiple myeloma is metastatic. In certain embodiments, the multiple myeloma is a RAS- mutated multiple myeloma. In certain embodiments, the multiple myeloma is KRAS-mutated multiple myeloma. In certain embodiments, the multiple myeloma is a KRAS-mutated multiple myeloma, including, but not limited to, KRAS^G12C/D/V, KRAS^G13C/D, or KRAS^Q61L/H/R. In some embodiments, the multiple myeloma is relapsed. In some embodiments, the multiple myeloma is refractory. [00375] In some embodiments, the present invention relates to a method of treating a cancer, wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is hepatocellular carcinoma (HCC). In certain embodiments, the HCC is locally advanced. In certain embodiments, the HCC is metastatic. In certain embodiments, the HCC is a RAS-mutated HCC. In certain embodiments, the HCC is KRAS-mutated HCC. In certain embodiments, the HCC is a KRAS- mutated HCC, including, but not limited to, KRAS^G12C/D/V, KRAS^G13C/D, or KRAS^Q61L/H/R. In some embodiments, the hepatocellular carcinoma is relapsed. In some embodiments, the hepatocellular carcinoma is refractory.

[00376] In some embodiments, the present invention relates to a method of treating a cancer wherein the method comprises administering to a patient in need thereof a composition comprising a compound of the present invention, wherein the cancer is selected from breast, colorectal, endometrial, hematological, leukemia (e.g., AML), liver, lung, melanoma, ovarian, pancreatic, prostate, or thyroid.

Pharmaceutically Acceptable Compositions

[00377] The compounds and compositions, according to the method of the present invention, are administered using any amount and any route of administration effective for treating or lessening the severity of a disorder provided above. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, and the like. Compounds of the invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression "dosage unit form" as used herein refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific 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 compound 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 well known in the medical arts.

[00378] Pharmaceutically acceptable 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), buccally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated. In certain embodiments, the compounds of the invention are administered orally or parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg and 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.

[00379] Liquid dosage forms for oral 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 emulsifiers 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.

[00380] 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 injectables.

[00381] 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.

[00382] In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound 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 compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide- polyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

[00383] 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.

[00384] 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.

[00385] 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 that 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 polyethylene glycols and the like.

[00386] 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 compound 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 that can be used include polymeric substances and waxes.

[00387] 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.

[00388] According to one embodiment, the invention relates to a method of inhibiting protein kinase activity in a biological sample comprising the step of contacting said biological sample with a compound of this invention, or a composition comprising said compound.

[00389] According to another embodiment, the invention relates to a method of inhibiting one or both of ER 1 and ERK2 kinase, or a mutant thereof, activity in a biological sample comprising the step of contacting said biological sample with a compound of this invention, or a composition comprising said compound. In certain embodiments, the invention relates to a method of irreversibly inhibiting one or both of ERK1 and ERK2 kinase, or a mutant thereof, activity in a biological sample comprising the step of contacting said biological sample with a compound of this invention, or a composition comprising said compound.

[00390] The term "biological sample", as used herein, includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.

[00391] Inhibition of one or both of ER 1 and ERK2, or a mutant thereof, activity in a biological sample is useful for a variety of purposes that are known to one of skill in the art. Examples of such purposes include, but are not limited to, blood transfusion, organ- transplantation, biological specimen storage, and biological assays.

[00392] Another embodiment of the present invention relates to a method of inhibiting protein kinase activity in a patient comprising the step of administering to said patient a compound of the present invention, or a composition comprising said compound.

[00393] According to another embodiment, the invention relates to a method of inhibiting one or both of ER 1 and ERK2 kinase, or a mutant thereof, activity in a patient comprising the step of administering to said patient a compound of the present invention, or a composition comprising said compound. According to certain embodiments, the invention relates to a method of irreversibly inhibiting one or both of ERK1 and ERK2 kinase, or a mutant thereof, activity in a patient comprising the step of administering to said patient a compound of the present invention, or a composition comprising said compound. In certain embodiments, the activity is inhibited irreversibly by covalently modifying Cys 183 of ERKl . In certain embodiments, the activity is inhibited irreversibly by covalently modifying Cys 166 of ERK2. In certain embodiments, the activity is inhibited irreversibly by covalently modifying Cys 183 of ERKl and Cys 166 of ERK2. In other embodiments, the present invention provides a method for treating a disease, disorder, or condition mediated by one or both of ERKl and ERK2 kinase, or a mutant thereof, in a patient in need thereof, comprising the step of administering to said patient a compound according to the present invention or pharmaceutically acceptable composition thereof. Such disorders are described in detail herein.

[00394] All features of each of the aspects of the invention apply to all other aspects mutatis mutandis.

[00395] In order that the invention described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner.

EXEMPLIFICATION

[00396] As depicted in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It will be appreciated that, although the general methods depict the synthesis of certain compounds of the present invention, the following general methods, and other methods known to one of ordinary skill in the art, can be applied to all compounds and subclasses and species of each of these compounds, as described herein.

General Procedures

[00397] Proton Nuclear Magnetic Resonance (¹H NMR) spectra were obtained on a Bruker model AVANCE-III 400 MHz NMR spectrometer. Deuterated DMSO was used as solvent.

[00398] LC-MS was recorded on the Agilent Technology LC-1200 system, and quadrupole MS model-6120.

[00399] Compound numbers utilized in the Examples below correspond to compound numbers set forth in the Tables, supra.

[00400] The following Examples provide representative syntheses of certain of the compounds described herein. Additional synthetic methods useful in the preparation of compounds of formulae I-a, I-b, I-c, Il-a, Il-b, and II-c can be found in PCT patent application serial number PCT/US14/15256, filed February 7, 2014 and published as WO2014/124230 on August 14, 2014 (herein "the '230 publication," the entirety of which is hereby incorporated herein by reference). In instances where a compound of the present invention contains one or more points of oxidation, one of skill in the art will recognize that such a compound can be obtained using synthetic methods described herein and in the '230 publication, in combination with one or more methods of oxidation known in the art. Exemplary such methods and reagents are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5^th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

Example 1. Synthesis of N⁴-(2-amino-4-methylphenyl)-N²-(2-methoxy-5-methylpyridin-4- yl)-5-(trifluoromethyl) pyrimidine-2,4-diamine (Compound P-1)

Compound P-1

[00401] Compound P-1 was prepared following the synthetic approach depicted below in Scheme 1.

Scheme

[00402] Preparation of 2-chloro-N-(4-methyl-2-nitrophenyl)-5-(trifluoromethyl)pyrimidin-4- amine (E-1):

E-1

[00403] To a solution of 4-methyl-2-nitroaniline (1.0 g, 6.57 mmol) in N-methyl-2- pyrrolidone (5.0 mL), 5-trifluoromethyl-2,4-dichloropyrimidine (2.15 g, 9.93 mmol) and sodium hydride (472 mg, 19.71 mmol) were added. The reaction mixture was stirred at 0 °C for 2 h. TLC showed completion of starting material (TLC system: 10% ethyl acetate in hexane, R/ = 0.4). Reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude obtained was purified by silica gel column chromatography to get the 2-chloro-N-(4-methyl-2- nitrophenyl)-5-(trifluoromethyl)pyrimidin-4-amine as yellow solid. (900 mg, 41%)

[00404] Preparation of N²-(2-methoxy-5-methylpyridin-4-yl)-] ⁴-(4-methyl-2-nitrophenyl)-5- ( trifluoromethyl)pyrimidine-2, 4-diamine (E-2) :

E-2

[00405] To a solution of 2-chloro-N-(4-methyl-2-nitrophenyl)-5-(trifluoromethyl)pyrimidin-4- amine (500 mg, 1.5 mmol) in 2-methyl-2-butanol (5.0 mL), 2-methoxy-5-methylpyridin-4-amine (207 mg, 1.5 mmol), sodium carbonate (953 mg, 9.0 mmol), tris(dibenzylideneacetone) dipalladium (0) (164 mg, 0.18 mmol) and davephos (259 mg, 0.66 mmol) were added. The reaction mixture was then degassed with nitrogen for 5 minutes and stirred at 90 °C for 6 h. TLC showed completion of starting material (TLC system: 50% ethyl acetate in hexane, R = 0.4). The reaction mixture was filtered through celite and washed with water. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography to afford N²-(2-methoxy-5- methylpyridin-4-yl)-N⁴-(4-methyl-2-nitrophenyl)-5-(trifluoromethyl) pyrimidine-2,4-diamine as light yellow solid. (200 mg, 30%). [00406] Preparation of rf-(2 -amino-4-methylphenyl)-N -(2-methoxy-5-methylpyridin

(trifluoromethyl)pyrimidine-2,4-diamine (Compound P-1):

P-1

[00407] To a solution of N²-(2-methoxy-5-methylpyridin-4-yl)-N⁴-(4-methyl-2-nitrophenyl)- 5-(trifluoromethyl) pyrimidine-2,4-diamine (150 mg, 0.34 mmol) in 1,4-dioxane: water (3: 1, 5.0 mL), was added zinc (221 mg, 3.4 mmol) and ammonium chloride (181 mg, 3.4 mmol). The reaction mixture was stirred at room temperature for 4 h, at which point TLC showed consumption of starting material (TLC system: 5% methanol in dichloromethane, R/ = 0.5). The reaction mixture was filtered through celite. The filtrate was diluted with water and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography to afford N⁴-(2-amino-4-methylphenyl)-N²-(2-methoxy-5-methylpyridin-4-yl)- 5-(trifluoromethyl) pyrimidine-2,4-diamine as pale yellow solid. (100 mg, 72%). LC-MS: m/z = 405.2 (ES+, M+H); 1H NMR (400 MHz, DMSO-<¾): δ 2.09 (s, 3H), 2.20 (s, 3H), 3.69 (s, 3H), 4.58 (s, 2H), 6.39 (d, J = 7.8 Hz, 1H), 6.59 (s, 1H), 6.86 (d, J = 7.9 Hz, 1H), 7.12 (s, 1H), 7.73 (s, 1H), 8.31 (s, 2H), 8.44 (s, 1H).

Example 2. Synthesis of N-(5-methyl-2-(2-(5-methyl-2-oxo-l,2-dihydropyridin-4-ylamino)- 5-(trifluoromethyl)p rimidin-4-ylamino)phenyl)acrylamide (C ompound P-2)

Compound P-2 Compound P-2 was prepared following the synthetic approach depicted below

1-90 E-3 P-2

[00409] Preparation of 4-((4-((2-amino-4-methylphenyl)amino)-5-(trifluoromethyl)pyrimidin- 2-yl)amino)-5-methylpyridin-2-ol (E-3):

E-3

[00410] To a solution of N-(2-(2-(2-methoxy-5-methylpyridin-4-ylamino)-5-(trifluoromethyl) pyrimidin-4-ylamino)-5-methylphenyl)acrylamide (compound 1-90 in the '230 publication), 200 mg, 0.4 mmol) in tetrahydrofuran (4 mL), was added aq. hydrogen bromide solution (4 mL). The reaction was stirred at 100 °C for 16 h, at which point TLC indicated consumption of starting material (TLC system: 10% methanol in chloroform, R = 0.3). The reaction mixture was diluted with saturated sodium bicarbonate solution and filtered. The resulting solid was diluted in 10% methanol/ chloroform to afford 4-((4-((2-amino-4-methylphenyl)amino)-5- (trifluoromethyl)pyrimidin-2-yl)amino)-5-methylpyridin-2-ol as pale green solid. (60 mg, 35%). LC-MS: m/z = 391.2 (ES+, M+H).

[00411] Preparation of N-(2-((2-((2-hydroxy-5-methylpyridin-4-yl)amino)-5-

( trifluoromethyl)pyrimidin-4-yl)amino)-5-methylphenyl)acrylamide (Compound P-2) :

P-2

[00412] To a solution of 4-((4-((2-amino-4-methylphenyl)amino)-5-

(trifluoromethyl)pyrimidin-2-yl)amino)-5-methylpyridin-2-ol (60 mg, 0.1 mmol) in dimethylacetamide (2.0 mL) was added diisopropylethylamme (29 mg, 0.2 mmol) and acryloyl chloride (15.3 mg, 0.2 mmol). The reaction mixture was stirred at 0 °C for 1 h, at which point TLC showed consumption of starting material (TLC system: 5% methanol in chloroform, R = 0.5). The reaction mixture was diluted with water and extracted with ethylacetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduce pressure to afford N-(2-((2-((2-hydroxy-5 -methylpyridin-4-yl)amino)-5 -(trifluoromethyl)pyrimidin-4-yl)amino)-5 - methylphenyl)acrylamide as off-white solid. (10 mg, 15%). LC-MS: m/z = 445.2 (ES+, M+H); 1H NMR (400 MHz, OMSO-d₆): δ 1.93 (s, 3H), 2.29 (s, 3H), 5.78-5.79 (dd, J = 1.5, 10.0 Hz, 1H), 6.26-6.30 (dd, J = 1.7, 17.0 Hz, 1H), 6.40-6.47 (dd, J = 10.0, 17.0 Hz, 1H), 6.68 (s, 1H), 7.05 (d, J = 5.5 Hz, 2H), 7.12 (d, J = 8.3 Hz, 1H), 7.55 (d, J = 8.2 Hz, 1H), 8.36-8.39 (m, 3H), 10.26 (s, 1H), 10.90 (s, 1H).

Example 3. Synthesis of 4-(4-(2-acrylamido-4-methylphenylamino)-5- (trifluoromethyl)pyrimidin-2-ylamino)-2-methoxy-5-methylpyridine-l-oxide (Compound I-

18)

Compound 1-18 [00413] To a solution of N-(2-(2-(2-methoxy-5-methylpyridin-4-ylamino)-5-(trifluoromethyl) pyrimidin-4-ylamino)-5-methylphenyl)acrylamide (compound 1-90 in the '230 publication, 500 mg, 1.09 mmol) in dichloromethane (20.0 mL) was added meta-chloroperoxybenzoic acid (751 mg, 4.36 mmol) at room temperature. The reaction mixture was stirred for 3 h, at which point TLC indicated consumption of starting material (TLC system: 10% methanol in chloroform, R/ = 0.5). The reaction mixture was diluted with dichloromethane and washed with sodium carbonate solution. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduce pressure. The resulting crude product was purified by preparative HPLC to get 10 mg of

4- (4-(2-acrylamido-4-methylphenylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-2-methoxy-

5- methyl pyridine- 1 -oxide as pale brown solid. LC-MS: m/z 475.2 (ES+, M+H); 1H NMR (400

MHz, DMSO-<¾): δ 2.15 (s, 3H), 2.36 (s, 3H), 3.76 (s, 3H), 5.77-5.80 (dd, J = 1.9, 10.03 Hz, 1H), 6.27-6.31 (dd, J = 1.9, 17.0 Hz, 1H), 6.41-6.47 (dd, J = 10.1, 17.0 Hz, 1H), 7.13-7.16 (m, 3H), 7.52 (d, J = 8.1 Hz, 1H), 7.89 (s, 1H), 8.65 (s, 1H), 8.73 (s, 1H), 9.95 (br s, 1H), 10.24 (s,

1H).

Example 4. (R)-2,3-dihydroxy-N-(2-((2-((2-methoxy-5-methylpyridin-4-yl)amino)-5- (trifluoromethyl) pyrimidin-4-yl)amino)-5-methylphenyl)propanamide (Compound 1-31):

Compound 1-31

[00414] To a mixture of N⁴-(2 -amino-4-methylphenyl)-N²-(2-methoxy-5-methylpyridin-4-yl)- 5-(trifluoromethyl)pyrimidine-2,4-diamine (32 mg, 0.08 mmol), (R)-2,2-dimethyl-l,3-dioxolane- 4-carboxylic acid (21 mg, 0.14 mmol) and 50 mg of DIPEA in 1 mL of DMA, was added HATU (73 mg, 0.19 mmol). After stirring at room temperature overnight, the residue was extracted with ethyl acetate and washed with water, brine. The organic layer was concentrated, then re- diluted with 2 mL of methanol-dichloromethane (v/v 1/1), followed by 0.5 mL of trifluoroacetic acid. The reaction mixture was stirred at room temperature for 1 hr, then concentrated. The residue was subject to prep-HPLC, giving 10 mg of the desired diol (25%). LC-MS: m/z = 519.1 (ES+, M+H).

Example 5. Synthesis of (S)-2-amino-5-(((R)-l-((carboxymethyl)amino)-3-((3-((2-((2-((2- methoxy-5-methylpyridin-4-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)-5- methylphenyl)amino)-3-oxopropyl)thio)-l-oxopropan-2-yl)amino)-5-oxopentanoic acid

(Compound 1-33)

Compound 1-33

[00415] Into a 15 mL centrifuge tube, were added N-(2-(2-(2-methoxy-5-methylpyridin-4- ylamino)-5-(trifluoromethyl) pyrimidin-4-ylamino)-5-methylphenyl)acrylamide (compound 1-90 in the '230 publication, 11 mg, 0.025 mmol), 1.5 mL of DMSO, 3 mL of water, and 100 mg of glutathione. The pH of the resulted suspension was then adjusted to pH 8.5 using about 500 of 1 M Tris-base. The mixture was placed on a shaker at 37 °C for 2 days. The resulting mixture was then concentrated and purified on prep-HPLC, affording ca. 5 mg of product as white powder after lyophilization. LC-MS: m/z 766.3 (ES+, M+H). Example 6. (R)-2-(2-amino-3-((3-((2-((2-((2-methoxy-5-methylpyridin-4-yl)amino)-5- (trifluoromethyl) pyrimidin-4-yl)amino)-5-methylphenyl)amino)-3- oxopropyl)thio)pr anamido)acetic acid (Compound 1-34)

Compound 1-34

[00416] Compound 1-34 was made in the same way as described for Compound 1-33 above, substituting Cys-Gly di-peptide for glutathione with. LC-MS: m/z 637.3 (ES+, M+H).

Example 7. Synthesis of (R)-2-amino-3-((3-((2-((2-((2-methoxy-5-methylpyridin-4- yl)amino)-5-(trifluoromethyl) pyrimidin-4-yl)amino)-5-methylphenyl)amino)-3- oxopropyl)thio)propanoic acid (Compound 1-35)

Compound 1-35

[00417] Compound 1-35 was made in the same way as described for Compound 1-33, substituting glutathione with cysteine. LC-MS: m/z 580.3 (ES+, M+H).

Example 8. In Vitro Assays

Protein Mass Modification Assay [00418] Intact protein: Erkl from Millipore (Cat. No. 14-439) was incubated for 60 min. at room temperature with a 10-fold excess of test compound to protein. 4 μΐ_^ aliquots of the resulting mixture were diluted with 15 μΐ_^ of 0.2% TFA prior to micro C4 ZipTipping directly onto the MALDI target using sinapinic acid as the desorption matrix (10 mg/ml in 0.1% TFA:Acetonitrile 50:50, v/v). The centroid mass of the target protein in the control sample was compared with the centroid mass of the target protein incubated with compound. A shift in the centroid mass of the treated protein compared to the untreated protein was divided by the molecular weight of the compound. This number corresponds to the percentage of modified protein (a measure of the proportion of protein covalently modified by a test compound) after one hour incubation. Results from this assay are reported in Table A in the row labelled "ERK1 Mass Mod (%)."

Omnia Assay Protocol for Potency Assessment against MEK1 activated ERK1:

[00419] The protocol below describes continuous-read kinase assays to measure potency of compounds against activated ERK1 enzyme. The mechanics of the assay platform are best described by the vendor (Invitrogen, Carlsbad, CA) on their website at the following URL: invitrogen.com/site/us/en/home.html.

[00420] Briefly, a 1.25X stock of ERK1 enzyme (14-439-K) from Millipore (Billerica, MA), 5X ATP (AS001A) and ST17-Sox conjugated peptide substrate (KNZ1 171C) were prepared in IX kinase reaction buffer consisting of 20 mM Tris, pH 7.5, 5 mM MgCl₂, 1 mM EGTA, 5 mM β-glycerophosphate, 5% glycerol (10X stock, KB001A) and 0.2 mM DTT. 10 μΕ of ATP/ST 17- sox peptide substrate mix was combined with 0.5 volume of 100% DMSO and serially diluted compounds were prepared in 100% DMSO in a Corning (#3574) 384-well, white, non- binding surface microtiter plate (Corning, NY). Kinase reactions were started with the addition of 40 of ERK1 solution and monitored every 71 seconds for 30-240 minutes at _ex360/ _em485 in a Synergy plate reader from BioTek (Winooski, VT). At the conclusion of each assay, progress curves from each well were examined for linear reaction kinetics and fit statistics (R², 95%) confidence interval, absolute sum of squares). Initial velocity (0 minutes to -30+ minutes) from each reaction was determined from the slope of a plot of relative fluorescence units vs time (minutes or seconds) and then plotted against inhibitor concentration to estimate ^AppICso from log[Inhibitor] vs Response, Variable Slope model in GraphPad Prism from GraphPad Software (San Diego, CA).

[Reagent] used in optimized protocol:

[ERK1] = 4 nM, [ATP] = 50 μΜ, [ST17-Sox] = 10 μΜ (ATP ^appK_M 48 μΜ)

[00421] The results of this assay show the degree of inhibition of ER activity, which is a direct measurement of inhibition of ERK activity. Results from this assay are reported in Table A in the row labelled "IC₅₀ (nM)." pRSK MSD Assay

Detection of total and phosphor-RSK by MSD ELISA (HT-29)

[00422] The protocol below describes an assay to measure the kinase activity of ERKl/2 to phosphorylate a substrate, p90RSK, in the presence or absence of a test compound. This experiment was conducted using a Mesoscale Discovery plate. The day before the assay, HT29 cells were split and plated at 50,000 cells/well in complete growth media. After allowing cells to adhere, the media was removed and replaced with media containing 0.1% FBS and incubated overnight. Blank MSD plates (Mesoscale Discovery, Cat # L15XA3) were coated with 25 μΐ/well RSK capture antibody (BD Biosciences, Cat. # 610226) and incubated at 4 °C overnight, then blocked with 150 μΐ of 3% BSA solution. The next day, the media from the cell culture plate was removed and replaced with 100 μΐ of media containing a test compound and incubated for 120 minutes at 37 °C. The media was removed and replaced with 55 μΐ per well of lysis buffer with protease inhibitors (Roche Biosciences, Cat. # 11836170001) and phosphatase inhibitors (Sigma-Aldrich, Cat. # P-0044 and P-5726), followed by incubation at 4 °C for 30 minutes. 50 μΐ of lysate was transferred to a blocked MSD plate, followed by incubation at room temperature for 2 hours under constant shaking. The plate was washed 3 times with MSD wash buffer (Mesoscale Discovery, Cat. # R617TX), and 25 μΐ/well phospho-RSK (pRSK) detection antibody (Cell Signaling Technology, Cat. # 9335) was added with Sulfo-tagged detection antibody (Mesoscale Discovery, Cat. # R32AB-1) diluted in 1% BSA in MSD wash buffer. This mixture was incubated for 1 hour at room temperature under constant shaking. The plate was washed 3 times, and 150 μΐ IX MSD read buffer was added, followed by signal detection in an MSD plate reader. Curve fitting analysis was done with variable slope in GraphPad software to generate EC50 based on DMSO control (untreated) being 100% pRSK signal and maximum inhibition with a reference compound provided by the manufacturer as a positive control. Results from this assay, showing EC₅₀ (i.e., the concentration at which a test compound inhibits phosphorylation of RSK by 50%) are reported in Table A in the row labelled "EC₅o MSD (nM) / HT29".

Detection of total and phosphor-RSK by MSD ELISA (A375)

[00423] The protocol below describes an assay to measure the kinase activity of ER 1/2 to phosphorylate a substrate, p90RSK, in the presence or absence of a test compound.

Cell Treatment

[00424] A375 cells were grown in DMEM/10% FBS. Twenty four hours prior to the assay, 50,000 cells per well were plated in a 96 well flat bottom plate. Once cells attached to the plate, the medium was replaced with 100 μΐ of DMEM/0.1% FBS. Cells were cultured overnight in an incubator at 37 °C.

Compound dilution

[00425] Compound stock solutions of 10 mM in DMSO were prepared. lOOOx dilutions were then prepared in DMSO. 1 μΕ of DMSO solution was then transfered to 1 ml DMEM/0.1%FBS in a deep well plate. Cell plate media was discarded, followed by addition of 100 of the compound-containing media. The preparation was incubated at 37 °C for 2 hrs.

[00426] Cell lysates were prepared as described below.

pRSK or total RSK MSD Assay

DAY 1

[00427] MSD plates: Blank MSD plates were coated with 30 μΕ capture antibody (BD 610226) at a final concentration of 1 μg/mL in PBS. Both pRSK and total RSK MSD assays used the same capture antibody at the same concentration. Antibody stock concentration was 250 μg/mL. Once antibody solution was added to the MSD plate, the sides were tapped to be certain it was coated completely (visual inspection). It was then covered and placed at 4 °C overnight on a level surface.

DAY 2

[00428] Block MSD Plate: The coating antibody was removed and the plate was washed on a plate washer in MSD wash buffer. The last bit of wash solution was tapped out and 150 μΕΛνεΙΙ of 3% BSA in MSD wash buffer was added in. The preparation was placed on a shaker at room temperature for at least an hour.

[00429] Add samples: Media was removed from compound-treated cells and replaced with 55 μίΛνεΙΙ MSD cell lysis buffer containing protease and phosphatase I & II inhibitors. The preparation was incubated on a shaker in a cold room for 30-45 min. The blocked MSD plate was washed a on plate washer, tapping out the last bit of wash solution, followed by addition of 50 (of the 55 μΐ) cell lysate in a well-well transfer. The preparation was covered and incubated on a shaker at room temp for 2 hours. The lysate was removed, washed on a plate washer 3 times, and the last bit of wash buffer was tapped out and replace with 25 μΙ,ΛνεΙΙ detection antibody (described below).

Detection antibody:

[00430] For pRSK detection, a pRSK antibody stock of 21 μg/mL was prepared as follows: 1 μg/mL pRSK Ser380 antibody (Cell Signaling Technology, Cat. # 9335) + 1 :750 anti-rabbit SulfoTag (Mesoscale Discovery, Cat. # R32AB-1) in 1% BSA in MSD wash buffer.

[00431] For total RSK detection, a total RSK antibody stock of 200 μg/mL was prepared as follows: 1 μg/mL total RSK (Santa Cruz sc-231G) antibody + 1 :750 anti-goat SulfoTag (from MSD, R32AG-1) in 1% BSA in MSD wash buffer.

[00432] The plate was incubated for 1 hr at room temperature on a shaker, followed by three washings. The last bit of wash buffer was tapped out. 150 μΙ,ΛνεΙΙ IX MSD Read buffer was added and the plate was then analyzed. Curve fitting analysis was done with variable slope in Graph Pad software to generate EC50 based on DMSO control (untreated) being 100% pRSK signal and maximum inhibition with a reference compound provided by the manufacturer as a positive control. Results from this assay, showing EC50 (i.e., the concentration at which a test compound inhibits phosphorylation of RSK by 50%) are reported in Table A in the row "EC50 MSD (nM) A375".

Detection of total and phospho-RSK by MSD ELISA (HCT116)

[00433] The protocol below describes an assay to measure the kinase activity of ERKl/2 to phosphorylate a substrate, p90RSK, in the presence or absence of a test compound.

Cell Treatment [00434] HCT116 cells were grown in RPMI/10%FBS. Prior to the assay, 50,000 cells per well were plated in a 96 well flat bottom plate. Cells were cultured overnight in an incubator at 37 °C.

Compound dilution

[00435] Compound stocks were 10 mM in DMSO. A lOOOx dilution was made in DMSO. 1 μΙ_, of the DMSO solution was transfered to 1 ml RPMI/10%FBS in a deep well plate. Media in the cell plate was discarded, and 100 μΐ, of the compound-containing media was added. The preparation was incubated at 37 °C for 2 hrs.

Preparation of cell lysates are described below.

pRSK or total RSK MSD Assay

DAY 1

[00436] MSD plates: Blank MSD plates were coated with 30 μΐ. capture antibody (BD 610226) at a final concentration of 1 μg/mL in PBS. Both pRSK and total RSK MSD assays used the same capture antibody at the same concentration. Antibody stock concentration was 250 μg/mL. Once antibody solution was added to MSD plate, the sides were tapped to be certain it was coated completely (visual inspection). It was then covered and placed at 4 °C overnight on a level surface.

DAY 2

[00437] Block MSD Plate: The coating antibody was removed and the plate was washed on a plate washer in MSD wash buffer. The last bit of wash solution was tapped out and 150 μΙ,ΛνεΙΙ 3% BSA (MSD Blocker A) in MSD wash buffer was added. The preparation was placed on a shaker at room temperature for at least an hour.

[00438] Add samples: Media was removed from compound-treated cells and replaced with 55 μίΛνεΙΙ MSD cell lysis buffer containing protease and phosphatase I & II inhibitors. The preparations was incubate on a shaker in a cold room for 30-45 min. The blocked MSD plate was washed on a plate washer, and the last bit of wash solution was tapped out. Next was added 50 μί (of the 55 μί) cell lysate in a well-well transfer. The preparation was covered and incubated on shaker at room temp for 2 hours. The lysate was then removed and washed on plate washer 3 times. The last bit of wash buffer was then tapped out and replaced with 25 μΙ,ΛνεΙΙ detection antibody (described below).

Detection antibody: [00439] For pRSK detection, a pRSK antibody stock of 21 μ§/ηιΙ, was prepared as follows: 1 μΒ/mL pRSK Ser380 antibody (Cell Signaling Technology, Cat. # 9335) + 1 :750 anti-rabbit SulfoTag (from MSD, R32AB-1) in 1% BSA in MSD wash buffer.

[00440] For total RSK detection, a total RSK antibody stock of 200 μg/mL was prepared as follows: 1 μg/mL total RSK (Santa Cruz sc-231G) antibody + 1 :750 anti-goat SulfoTag (from MSD, R32AG-1) in 1% BSA in MSD wash buffer.

[00441] The plate was incubated for 1 hr at room temperature on a shaker, followed by three washes. The last bit of wash buffer was tapped out. Next, 150 μΙ,ΛνεΙΙ IX MSD Read buffer was added and the plate was analyzed by the MSD reader. Curve fitting analysis was done with variable slope in Graph Pad software to generate EC₅₀ based on DMSO control (untreated) being 100% pRSK signal and maximum inhibition with a reference compound provided by the manufacturer as a positive control. Results from this assay, showing EC₅₀ (i.e., the concentration at which a test compound inhibits phosphorylation of RSK by 50%) are reported in Table A in the row labelled "EC₅₀ MSD (nM) / HCT116."

[00442] Table A shows data for selected compounds in various assays. Compound numbers in Table A correspond to Compound numbers indicated in the Tables and Examples above. Compounds having an activity designated as "A" provided an EC₅₀/IC₅₀ of 100-1000 nM; compounds having an activity designated as "B" provided an EC₅₀/IC₅₀ of 1001-5000 nM; compounds having an activity designated as "C" provided an EC₅₀/IC₅₀ of >5000 nM.

Table A.

NM: No significant protein modification

[00443] While we have described a number of embodiments of this invention, it is apparent that our basic examples may be altered to provide other embodiments that utilize the compounds and methods of this invention. Therefore, it will be appreciated that the scope of this invention is to be defined by the appended claims rather than by the specific embodiments that have been represented by way of example.

Claims

We claim:

1. A compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

X, X', and X" are each independently O;

p, q, and r are each independently 0 or 1;

R¹ is -OR';

R' is -H, -CH₃, -S0₃H or -Glu;

each Glu is a glucuronyl moiety;

R² and R³ are each independently -H, -OH, -OS0₃H, -OGlu, -SR⁵, or:

R² and R³ are taken together to form a double bond; or:

R² and R^2' are taken together to form =0;

R³ and R³ are each -H, or:

R^3' and R^3" are taken together to form =0;

R⁴ is -H, -OH, -OS0₃H or -OGlu; eac ⁵ is independently selected from:

u III IV

R^x and R^y are each independently -OH, -OSO₃H, or -OGlu;

R^z, R^z' and R^z" are each independently -H, -CH₃, -OH, -OSO3H, or -OGlu; or

R^z" and R³ are taken together to form -0-;

m is 0, 1, 2, or 3;

n is 0, 1, or 2;

wherein the compound is other than:

2. A compound of Formula II :

pharmaceutically acceptable salt thereof, wherein:

X, X', and X" are each independently O;

p, q, and r are each independently 0 or 1;

R¹ is -OR';

R' is -H, -CH₃, -S0₃H or -Glu;

each Glu is a glucuronyl moiety;

R² and R³ are each independently -H, -OH, -OS0₃H, -OGlu, -SR⁵, or:

R² and R³ are taken together to form a double bond; or:

R² and R^2' are taken together to form =0;

R³ and R³ are each -H, or:

R^3' and R^3" are taken together to form =0;

R⁴ is -H, -OH, -OS0₃H or -OGlu;

eac ⁵ is independently selected from:

i ii Hi iv

R⁶ is -CH₃, -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu;

R⁷ is -CH₃, -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu;

R^x and R^y are each independently -OH, -OS0₃H, or -OGlu;

R^z" and R³ are taken together to form -0-;

m is 0, 1, 2, or 3;

n is 0, 1, or 2;

wherein the compound is other than:

3. The compound according to claim 1 or 2, wherein R^w is H.

4. The compound according to claim 1 or 2, wherein R^w is

5. The compound according to claim 1 or 2, wherein R is -OH.

6. The compound according to claim 1 or 2, wherein R¹ is -OCH₃.

7. The compound according to claim 1 or 2, wherein R¹ is -OSO₃H.

8. The compound according to claim 1 or 2, wherein R¹ is -OGlu.

9. The compound according to claim 1 or 2, wherein R⁴ is -H.

10. The compound according to claim 1 or 2, wherein R⁴ is -OH.

11. The compound according to claim 1 or 2, wherein R⁴ is -OSO₃H.

12. The compound according to claim 1 or 2, wherein R⁴ is -OGlu.

13. The compound according to claim 1 or 2, wherein R^x is -OH.

14. The compound according to claim 1 or 2, wherein R^x is -OSO₃H.

15. The compound according to claim 1 or 2, wherein R^x is -OGlu.

16. The compound according to claim 1 or 2, wherein m is 0.

17. The compound according to claim 1 or 2, wherein m is 1.

18. The compound according to claim 1 or 2, wherein m is 2.

19. The compound according to claim 1 or 2, wherein m is 3.

20. The compound according to claim 1 or 2, wherein R^y is -OH.

21. The compound according to claim 1 or 2, wherein R^y is -OSO₃H.

22. The compound according to claim 1 or 2, wherein R^y is -OGlu.

23. The compound according to claim 1 or 2, wherein n is 0.

24. The compound according to claim 1 or 2, wherein n is 1.

25. The compound according to claim 1 or 2, wherein n is 2.

26. The compound according to claim 1 or 2, wherein p is 1.

27. The compound according to claim 1 or 2, wherein q is 1.

28. The compound according to claim 1 or 2, wherein r is 1.

29. The compound according to claim 1 or 2, wherein R^z, R^z and R^z are each -H.

30. The compound according to claim 1 or 2, wherein one of R^z, R^z or R^z is -CH₃, -OH, - OS0₃H, or -OGlu.

31. The compound according to claim 2, wherein R⁶ is selected from -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu.

32. The compound according to claim 2, wherein R⁷ is selected from -CH₂OH, -CH₂OS0₃H, or -CH₂OGlu. The compound according to claim 2, wherein the compound has the structure of Formula