WO2010075290A1 - Unsaturated heterocyclic inhibitors of necroptosis - Google Patents

Abstract

The invention features new small molecule inhibitors of necroptosis. The compounds of the invention are described by Formulas (I) and (II). The invention also features pharmaceutical compositions that include compounds of Formula (I) and (II). The compounds and compositions of the invention are also featured in kits and in methods of treatment of conditions that include neurodegenerative diseases, ischemic brain and heart injuries, and head trauma.

Description

UNSATURATED HETEROCYCLIC INHIBITORS OF NECROPTOSIS

Cross-Reference to Related Applications

This application claims benefit of U.S. Provisional Application No. 61/139,904, filed December 22, 2008, which is hereby incorporated by reference.

Statement as to Federally Sponsored Research

This invention was made with government support under NS050560 awarded by the National Institutes of Health. The U.S. government has certain rights to this invention.

Field of the Invention

The invention relates to heterocyclic compounds and to cell death, in particular through necrosis and necroptosis, and regulation thereof by heterocyclic compounds.

Background of the Invention

In many diseases, cell death is mediated through apoptotic and/or necrotic pathways. While much is known about the mechanisms of action that control apoptosis, control of necrosis is not as well understood. Understanding the mechanisms regulating both necrosis and apoplosis in cells is essential to being able to treat conditions, such as neurodegenerative diseases, stroke, coronary heart disease, kidney disease, and liver disease. A thorough understanding of necrotic and apoptotic cell death pathways is also crucial to treating AIDS and the conditions associated with AIDS, such as retinal necrosis. Cell death has traditionally been categorized as either apoptotic or necrotic based on morphological characteristics (Wyllie et al., Int. Rev. Cytol. 68: 251 (1980)). These two modes of cell death were also initially thought to occur via regulated (caspase-dependent) and non-regulated processes, respectively. More recent studies, however, demonstrate that the underlying cell death mechanisms resulting in these two phenotypes are much more complicated and, under some circumstances, interrelated. Furthermore, conditions that lead to necrosis can occur by either regulated caspase- independent or non-regulated processes.

One regulated caspase- independent cell death pathway with morphological features resembling necrosis, called necroptosis, has recently been described (Degterev et al., Nat. Chem. Biol. 1 :112 (2005)). This manner of cell death can be initiated with various stimuli (e.g., ΪNF-α and Fas ligand) and in an array of cell types (e.g., monocytes, fibroblasts, lymphocytes, macrophages, epithelial cells and neurons). Necroptosis may represent a significant contributor to and, in some cases, predominant mode of cellular demise under pathological conditions involving excessive cell stress, rapid energy loss, and massive oxidative species generation, where the highly energy-dependent apoptosis process is not operative.

The identification and optimization of low molecular weight molecules capable of inhibiting necroptosis will assist in elucidating its role in disease patho-physiology and could provide compounds (i.e., necrostatins) for anti- necroptosis therapeutics. The discovery of compounds that prevent caspase- independent cell death (e.g., necrosis or necroptosis) would also provide useful therapeutic agents for treating or preventing conditions in which necrosis occurs. These compounds and methods would be particularly useful for the treatment of neurodegenerative diseases, ischemic brain and heart injuries, and head trauma.

Summary of the Invention

The invention features new compounds, pharmaceutical compositions, kits, and methods for treating a condition, e.g., a neurodegenerative condition, in a subject. In one aspect, the invention features a compound of the formula

(I), where each X¹, X², X³, X⁴, X⁵, and X⁶ is selected, independently, from N or

CR^X1; each Y , Y , and Y is selected, independently, from O, S, NR , or

CR^Y2R^Y3; each Z¹ and Z² is selected, independently, from O₅ S, or NR^Z!; each R^Y1 and R^Z1 is selected, independently, from H, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, - C(=O)R^5A, -C(-O)OR^5A, or -C(-O)NR^5AR^6A; each R^X1, R^Y2, and R^Y3 is selected, independently, from H, halogen, CN, NC, NO₂, N₃, OR³, SR³, NR³R⁴, -C(=O)R^SA, -C(=O)OR^5A, -C(=O)NR^5AR^6A ₅ - S(=O)R^5A, -S(=O)₂R^5A, -S(=O)₂OR^5A, -S(=O)₂NR^5AR^6A, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C₂_₆ alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each R¹, R², R^5A, R^5B, R^6A, and R^6B is selected from H, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R^5A and R^6A, or R^5B and R^6B combine to form a heterocyclyl; and each R³ and R⁴ is selected from H, optionally substituted C_{1 -6} alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, -C(=O)R^5B, -C(-S)R^5B, -C(-NR^6B)R^5B, -C(-O)OR^5B, -C(-O)NR^5BR^6B, -S(=O)R^5B, -S(-O)₂R^5B, - S(=O)₂OR^5B, or -S(=O)₂NR^5BR^6B; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In some embodiments, Formula (I) does not include compounds where when R¹ is H, X¹, X², and X⁴ are each CH, X³ and X⁵ are each N, Y¹ is S, Y² is NH, Z¹ is NH, and Z² is O, R² is not 4-fluorophenyl, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In some embodiments, Formula (I) does not include compounds where when R¹ is H, X¹, X², and X⁴ are each CH, X³, X⁵, and X⁶ are each N, Y¹ and Y³ are each S, Y² is NH, Z¹ is NH, and Z² is O, R² is not 4-fluorophenyl, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In some embodiments, R¹ is unsubstituted phenyl or substituted phenyl. In some embodiments, R¹ is phenyl having 1, 2, 3, 4, or 5 substituents as defined herein.

In some embodiments, the compound of Formula (I) has the following structure:

(II), where

X¹, X², X⁴, X⁵, R¹, Y², and R^Z1 are as defined for Formula (I); each R^2A, R^2B, R^2C, R^2D, and R^2E is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyL optionally substituted C₂.₆ alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, CN, NC, NO₂, N₃, OR⁷, SR⁷, S(K))R¹², S(O)₂R¹², S(K))OR¹². S(=O)₂OR¹², NR⁷R⁸, C(^O)R¹². C(K))OR¹², C(K))NR¹²R¹³, C(^S)R¹², C(=S)OR¹², C(=S)NR¹²R¹³, C(=NR⁹)R¹², C(=NR⁹)OR¹², or C(=NR⁹)NR^!2R³³, or R^2A and R^2B, R^2B and R^2C, R^2C and R^2D, or R^2D and R^2E combine to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyi; each R , R , and R is selected, independently, from H, optionally substituted Cj-,, alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, SrO)R¹⁰, Sf- O)₂R³⁰, C(O)R¹⁰, C(O)OR¹⁰. C( -OjNR¹⁰R³¹, C(=S)R¹⁰, C(-S)()R¹⁰, Cf-S)NR¹⁰R¹¹. C(=NR¹⁴)R¹⁰, C(-NR¹⁴)OR¹⁰. or

CC-NR¹⁴JNR⁵⁰R¹¹. or R⁷ and R⁸ combine io form an optionally substituted heterocyclyl; and each R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ is selected, independently, from H, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R¹⁰ and R¹¹ or R¹² and R¹³ combine to form an optionally substituted heterocyclyl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In some embodiments, Formula (II) does not include compounds where when R¹ is H, X¹, X², and X⁴ are each CH, X³ and X⁵ are each N, Y¹ is S, Y² is NH, Z¹ is O, and Z² is NH, R² is not 4-fluorophenyl, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof. In other embodiments, the compound of ihe indention has the following formula:

(H-A), where each R¹, R^M, R^2B, R^2C, R^2D. R^2F, R^YI, R⁷¹, R³, R⁴, R⁵\ R^6A, R⁷, R⁸, R⁹, and R¹² is as defined in Formula (II); and where each R^X1A and R^{X1 B} is selected, independently, from H, halogen,

CN, NC, NO₂, N₃, OR³, NR³R⁴, -C(O)R^5Λ, -C(=O)OR^5A. -C( U)NR^5AR^oA, -S(O)₂R^5Λ, -S(=O)₂OR^5A, -S(-O)₂NR^5AR^6A, optionally substituted C_{1 -6} alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof. In any of the compounds described herein, R^Y1 is H.

In any of the compounds described herein, or any pharmaceutically

71 acceptable salt or solvate thereof, or stereoisomer thereof, R is H.

In any of the compounds described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, R^X1Λ is H and R^X1B is H or optionally substituted C_{.₆ alkyl.

In any of the compounds described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, R^X1A and R^X1B is H.

In any of the compounds described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, R¹ is H. In any of the compounds described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, R^2Λ, R^lO, and R^2E are H.

In any of the compounds described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, R^2C is selected from halogen, OR⁷, SR⁷, S(^O)₂R¹², NR⁷R⁸, C(=O)R¹², C(^O)OR¹², C(=O)NR^l2R¹³, optionally substituted C_1-6 alkyl, optionally substituted heterocyclyl, or optionally substituted aryl.

In any of the compounds described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, R^{" '} is halogen. In any of the compounds described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, R^2B is also halogen. In some embodiments, the compound is of the formula

where

R_XIB is H or optionally substituted C_1-6 alkyl; each R^2A, R^2B, R^2C, R^2D, and R^2E is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, OR⁷, S(=O)R¹², S(O)₂R¹², NR⁷R⁸, C(=O)R¹², C(=O)OR¹², or C(=O)NR¹²R¹³, or R^2A and R^2B, R^2B and R^2C ₅ R^2C and R^2D, or R^2D and R^2E combine to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl; each R and R is selected, independently, from H, optionally substituted Ci_-6 alkyl. optionally substituted cycloalkyl , optionally substituted heterocyclyl, optionally substituted aryl, optional]}' substituted heteroaryl, S(O)R.'⁰, SC=O)₂R¹⁰, C(O)R¹⁰, C(O)OR¹⁰, C(O)NR¹⁰R¹¹, or R⁷ and R⁸ combine to form an optionally substituted heterocyclyl; and each R¹⁰, R^{1 1}, R¹², and R¹³ is selected, independently, from H, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R¹⁰ and R¹¹ or R¹² and R¹³ combine to form an optionally substituted heterocyclyl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In some embodiments, R^XIB is H. In other embodiments, R^2Λ, R^;'D, and R^2E arc H. In still other embodiments, R^2C is halogen. In other embodiments, R^iB is halogen. In some embodiments, R^2C is optionally substituted CV₆ alkyl. In certain embodiments, R^2C is optionally substituted heterocyclyl, NR R⁸, or C(=O)NR¹²R¹³. In further embodiments R^2C is NR⁷R⁸. In some embodiments, R⁷ and R⁸ are each C_1-6 alkyl.

In still other embodiments, the compound is of the formula

independently, an integer between 0-4; G is a bond, O, S, or NR ; R is H or optionally substituted C_1-6 alkyl; and where (n+m) is at least 2, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In other embodiments, the compound is of the formula

where each n and m is, independently, an integer between 0-4; G is a bond, O, S, or NR^2J; R^2J is H or optionally substituted C_1-6 alkyl; and where (n+m) is at least 2, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In other embodiments, the compound is of the formula

where G¹ is a covalent bond or S(=O)₂; each R^2K, R^2L, R^2M, R^2N, and R²⁰ is selected, independently, from H, halogen, optionally substituted Cj._o alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl. optionally substituted heteroaryl, CN, NC, NO₂, N₃, OR^2P, SR^2P, S(-O)R^2P, S(-O)₂R^2P, S(=O)OR^2P, S(=O)₂OR^2P, NR^?QR ^"R, C{ =0 S R^2P. C(=O)OR^2P, C(=O)NR^2QR^2R, C(=S)R^2P, C(=S)OR^2P, C(-S)NR^2QR^2R, C(=NR^2S)R^2Q, C(=NR^2S)OR^2Q, or C(=NR^2S)NR^2QR^2R, or R^2K and R^2L, R^2L and R^2M, R^2M and R^2N, R^2N and R²⁰, or R^2Q and R^2R combine to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl; and each R^2P, R , R , and R is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C₂-₆ alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted hcteroaryl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof. In some embodiments, G¹ Is S(~O)a- In other embodiments, G¹ Ls a bond. Jn still other embodiments, each R^"κ, ϊl^2L. R^m, R^2N, and R²⁰ is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, and 0-(optionally substituted C_1-6 alkyl). In certain embodiments, R and R are H. In siil! other embodiments, at least one of R^, R^?N, and R ^>o is O~(oplionally substituted C_1-6 alkyl). In other embodiments, at least one of

R^2M. R^"1V, and R²° is optionally substituted C_{f -6} alkyl. In some embodiments, at least one of R^2M, R^2N, and R²⁰ is halogen.

In a second aspect, the invention features a pharmaceutical composition that includes a pharmaceutically acceptable excipient and any of the compounds of the invention described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In a third aspect the invention features a method of treating a condition in a subject, where the method includes the step of administering any of the compounds of the invention, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, to the subject in a dosage sufficient to decrease necroptosis. In some embodiments, the condition is a neurodegenerative disease of the central or peripheral nervous system, the result of retinal neuronal cell death, the result of cell death of cardiac muscle, the result of cell death of cells of the immune system; stroke, liver disease, pancreatic disease, the result of cell death associated with renal failure; heart, mesenteric, retinal, hepatic or brain ischemic injury, ischemic injury during organ storage, head trauma, septic shock, coronary heart disease, cardiomyopathy, myocardial infarction, bone avascular necrosis, sickle cell disease, muscle wasting, gastrointestinal disease, tuberculosis, diabetes, alteration of blood vessels, muscular dystrophy, graft-versus-host disease, viral infection, Crohn's disease, ulcerative colitis, asthma, or any condition in which alteration in cell proliferation, differentiation or intracellular signaling is a causative factor.

In some embodiments, the condition is one in which alteration in cell proliferation, differentiation or intracellular signaling is a causative factor. In some embodiments, these conditions are cancer or infection by viruses (acute, latent and persistent), bacteria, fungi, or by other microbes. In some embodiments, the infection is caused by a virus selected from human immunodeficiency virus (HIV), Epstein- Barr virus (EBV), cytomegalovirus (CMV)5 human herpesviruses (HHV), herpes simplex viruses (HSV), human T-CeIl leukemia viruses (HTL V) 5 Varicella-Zoster virus (VZV), measles virus, papovaviruses (JC and BK), hepatitis viruses, adenovirus, parvoviruses, and human papillomaviruses. In other embodiments, the infection is caused by bacteria selected from Campylobacter jejuni, Enterobacter species, Enterococcus faecium, ϋnterococcus faecal is, Escherichia coli (e.g., E. coli O157:H7), Group Λ streptococci, Haemophilus influenzae, Helicobacter pylori, listeria, Mycobacterium tuberculosis, Pseudomonas aeruginosa, S. pneumoniae, Salmonella, Shigella, Staphylococcus aureus, and Staphylococcus epidermidis.

In a third aspect, the invention features a method of decreasing necroptosis comprising contacting a cell with any of the compounds of the invention described herein, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In a fourth aspect, the invention features a kit including (a) a pharmaceutically acceptable composition comprising any of the compounds of the invention described herein , or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof; and (b) instructions for the use of the pharmaceutical composition of (a) to treat a condition in a subject. In any of the compositions, methods, or kits of the invention, the

compound is

(3), or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In any of the compounds, compositions, methods, or kits of the invention, the compound is any of Compounds (16)-(21), (22)-(36), (37)-(43), (44)-(52), or (53)-(65), or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof. In some embodiments, the compound is selected from the group consisting of:

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

By "C]_-4 alkaryl" is meant a C]_-4 alkyl group having an optionally substituted aryl or an optionally substituted heteroaryl located at any position of the carbon chain. The C_{1 -4} alkyl group may be linear or branched and may also be substituted with, for example, 1, 2, 3, 4, or 5 additional substituents as described herein.

By "alkoxy"' is meant a group having the structure -O(optionally substituted C_{] -6} alkyl), where the optionally substituted C_1-6 alkyl may be branched, linear, or cyclic. The C_1-6 alkyl may be substituted or unsubstituted. A substituted Cj.₆ alkyl can have, for example, 1 , 2, 3_? 4, 5, or 6 suhstituerrts located at any position. Exemplary alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, tert-butoxy, and the like. By "C_2-6 alkenyl" or "alkenyl" is meant an optionally substituted unsaturated C_2-6 hydrocarbon group having one or more carbon-carbon double bonds. Exemplary C_2-6 alkenyl groups include, but are not limited to -CH=CH (ethenyl), propenyl, 2-propenyl, 2-methyl-l-propenyl, 1-butenyl, 2-butenyl, and the like. A C₂.₆ alkenyl may be linear or branched and may be unsubstituted or substituted. A substituted C_2-6 alkenyl may have, for example, 1, 2, 3, 4, 5, or 6 substituents located at any position.

By "C_1-6 alkyl" or "alkyl" is meant an optionally substituted C_1-6 saturated hydrocarbon group. An alkyl group may be linear, branched, or cyclic ("cycloalkyl"). Examples of alkyl radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, sec pentyl, iso-pentyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl, n- heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, and the like, which may bear one or more substituents. Substituted alkyl groups may have, for example, 1, 2, 3, 4, 5, or 6 substituents located at any position. Exemplary substituted alkyl groups include, but are not limited to, optionally substituted C_1-4 alkaryl groups.

By ''C_2-6 alkynyl" or "alkynyl" is meant an optionally substituted unsaturated C₂.₆ hydrocarbon group having one or more carbon-carbon triple bonds. Exemplary C_2-6 alkynyl groups include, but are not limited to ethynyl. 1-propynyl, and the like

By "amino" is meant a group having a structure -NR' R", where each R' and R" is selected, independently, from H, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or R' and R" combine to form an optionally substituted heterocyclyl. When R' is not H or R" is not H, R' and R" may be unsubstituted or substituted with, for example, 1, 2, 3, 4, 5, or 6 substituents.

By "aryl" is meant is an optionally substituted C₆-Ci₄ cyclic group with [4ft + 2] π electrons in conjugation and where n is 1, 2, or 3. Non-limiting examples of aryls include heteroaryls and, for example, benzene, naphthalene, anthracene, and phenanthrene. Aryls also include bi- and tri-cyclic ring systems in which a non-aromatic saturated or partially unsaturated carbocyclic ring (e.g., a cycloalkyl or cycloalkenyl) is fused to an aromatic ring such as benzene or naphthalene. Exemplary aryls fused to a non-aromatic ring include indanyl, tetrahydronaphthyl,. Any aryls as defined herein may be unsubstituted or substituted. A substituted aryl may be optionally substituted with, for example, 1, 2, 3, 4, 5, or 6 substituents located at any position of the ring. By "aryloxy" is meant a group having the structure -O(optionally substituted aryl >, where aryl is as defined herein. By "azido" is meant a group having the structure -Nv

By "carbamate" or "carbamoyl" is meant a group having the structure -OCONR' R" or - NR⁵CO₂R", where each R' and R" is selected, independently, from II, optionally substituted Ci.₆ alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or R' and R" combine to form an optionally substituted heterocyclyl. When R' a ^' > nut Il or R" is not H, R' and R" may be unsubstituted or substituted with, for example, 1, 2, 3, 4, 5. or 6 substhuents.

By "carbonate" is meant a group having a the structure -OCO^R', where R' is selected from H, optionally substituted C_1^6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl. When R' is not H, R may be unsubstituted or substituted with, for example, 1, 2, 3, 4, 5, or 6 substituents.

By '"earboxarnido^5" or '^"amido'^" is meant a group having the structure -CONR'R" or -NR'C(-0)R", where each R' and R" is selected, independently, from H, optionally substituted Ci_-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted hetεroaryl, or R' and R" combine io form an optionally substituted heterocyclyl. When R' is not f-ϊ or R"' is not H, K" and R"' may be unsubstituted or substituted with, for example, 1, 2, 3, 4, 5, or 6 substituents.

By "carboxylic group" is meant a group having the structure -CO₂R', where R' is selected from H, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl. When R' is not H, R may be unsubstituted or substituted with, for example, I₅ 2. 3, -4, 5, or 6 substituents. By "cyano" is meant a group having the structure -CN. By "C₃..₁₍) cycloalkyl" or "cycloalkyl" is meant an optionally substituted, saturated or partially unsaturated 3- to 10-membered monocyclic or poly cyclic (e.g., bicyclic, or tricyclic) hydrocarbon ring system. Where a cycloalkyl is polycyclic, the constituent cycloalkyl rings may be fused together, form a spirocyclic structure, or the polycyclic cycloalkyl may be a bridged cycloalkyl (e.g., adamantyl or norbonanyl) . Exemplary cycloalkyls induce cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Cycloalkyls may be unsubstituted or substituted. A substituted cycloalkyl can have, for example, 1 , 2, 3, 4, 5, or 6 substituents.

By "cycloalkenyl" is meant a non-aromatic, optionally substituted 3- to 10-membered monocyclic or bicyclic hydrocarbon ring system having at least one carbon-carbon double bound. For example, a cycloalkenyl may have 1 or 2 carbon-carbon double bonds. Cycloalkenyls may be unsubstituted or substituted. A substituted cycloalkenyl can have, for example, 1, 2, 3, 4, 5, or 6 substituents. Exemplary cycloalkenyls include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, and the like.

By "effective amount" or "therapeutically effective amount" of an agent, as used herein, is that amount sufficient to effect beneficial or desired results, such as clinical results, and, as such, an effective amount depends upon the context in which it is being applied. For example, in the context of administering an agent that is an inhibitor of necroptosis, an effective amount of an agent is, for example, an amount sufficient to achieve a reduction in necroptosis as compared to the response obtained without administration of the agent.

By "ester" is meant a group having a structure selected from -OCOR', where R' is selected from H, optionally substituted C₁^ alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl. When R' is not H, R may be unsubstituted or substituted with, for example, 1, 2, 3, 4, 5, or 6 substituents. By "haJogcfT or "halo" is meant fluorine (-F), chlorine (-Cl), bromine (-Br), or iodine (-1).

By "heteroaryl" is mean an aryl group that contains 1, 2, or 3 heteroatoms in the cyclic framework. Exemplary heteroaryls include, but are not limited to, furan, thiophene, pyrrole, thiadiazole (e.g., 1,2,3-thiadiazole or 1,2,4-thiadiazole), oxadiazole (e.g., 1,2,3-oxadiazole or 1,2,5-oxadiazole), oxazole, benzoxazole, isoxazolc, isothiazole, pyrazole, thiazole, benzthiazole, triazole (e.g., 1 ,2,4-triazole or 1,2,3-triazole), benzotriazole, pyridines, pyrimidines, pyrazines, quinoline, isoquinoline, purine, pyrazine, pteridine, triazine (ε.g, LzJ-triazine, 1 ,2,4-triazine, or U.S-tπazine )indoles, 1 ,2,4,5- tetrazine, befS7ϋ[Z>jthiophene, benzo[cj thiophene, bcnzo furan, isobenzofuran, and benzimidazole. Heteroaryls may be unsubstituted or substituted.

Subsiiuied heteroaryls can have, for example, 1 , 2, 3, 4, 5, or 6 subsitutents. By "heterocyclic" or "heterocyclyi'" is meant an optionally substituted non-aromatic, partially unsaturated or fully saturated, 3- to 10 membered ring system, which includes single rings of 3 to 8 atoms in size, and polycyclic ring systems (e.g., bi- and tri-cyclic ring systems) which may include an aryl (e.g., phenyl or naphthyl) or heteroaryl group that is fused to a non-aromatic ring (e.g., cycloalkyl, cycloalkenyl, or heterocyclyl), where the ring system contains at least one heterotom. Heterocyclic rings include those having from one to three heteroatoms independently selected from oxygen, sulfur, and nitrogen, in which the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized or substituted. In certain embodiments, the term heterocylic refers to a non-aromatic 5-, o-, or 7- membered monocyclic ring wherein at least one ring atom is a heteroatom selected from O, S, and N (wherein the nitrogen and sulfur heteroatoms may be optionally oxidized), and the remaining ring atoms are carbon, the radical being joined to the rest of the molecule via any of the ring atoms. Where a heterocycle is polycyclic, the constituent rings may be fused together, form a spirocyclic structure, or the polycyclic heterocycle may be a bridged heterocycle (e.g., quinuclidyl or . Exemplary heterocyclics include, but arc not limited to, aziridinyl, azetindinyl, 1,3-diazatidinyl, pyrrolidinyl, piperidinyl, piperazinyl, thiranyl, thietanyl, tetrahydrothiophenyl, dithiolanyl, tetrahydrothiopyranyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyranonyl, 3,4-dihydro-2H-pyranyl, chromenyl, 2H-chromen-2-onyl, chromanyl, dioxanyl (e.g.. 1,3-dioxanyl or 1,4-dioxanyl), 1 ,4-benzodioxanyl, oxazinyl, oxathiolanyl, morpholinyl, thiomorpholinyl, thioxanyl, quinuclidinyl, and also derivatives of said exemplary heterocyclics where the heterocyclic is fused to an aryl (e.g., a benzene ring) or a heteroaryl (e.g., a pyridine or pyrimidine) group. Any of the heterocyclic groups described herein may be imsubsiituted or substituted. A substituted heterocycle may have, for example, 1, 2, 3, 4, 5, or 6 substituents. By "ketone" or "acyl" is meant a group having the structure -COR', where R' is selected from H, optionally substituted Ci_-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl. When R' is not H, R may be unsubstituted or substituted with, for example, 1, 2, 3, 4, 5, or 6 substituents. By '"nitrυ" is meant a group having the structure NO₂.

A "pharmaceutically acceptable excipient" as used herein refers any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being nontoxic and non-infiaπiπiaiorv in a patient, Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintεgrants, dyes (colors K emollients, cniulsificrs, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, or waters of hydration. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycoiate, sorbitol, starch (com), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. The term "pharmaceutically acceptable salt," as used herein, represents those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and 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, 1911 , 66:1-19. The salts can be prepared in situ during the final isolation and purification of the compounds of the invention or separately by reacting the free base group with a suitable organic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphersulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy- ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine. triethylamine, ethyl amine and the like.

The term "pharmaceutically acceptable solvates," as used herein, refers to compounds that retain non-covalent associations to residual solvent molecules in the solid state. For example, solvates may be prepared by crystallization, recrystallization, or precipitation from a solution that includes organic solvents, water, or a mixture thereof. Solvates include, but are not limited to, compounds that include solvent molecules in the crystal lattice following recrystallization. The molecular stoichiometry of solvation can vary from, for example, 1:1 solventxompound to 10:1 solvent xompound. These ratios can include a mixture of associated solvent molecules. Exemplary, non- limiting examples of solvents that can form solvates with the compounds of the invention include water (for example, mono-, di-, and tri-hydrates), N- methylpyrrolidinone (ΝMP), dimethyl sulfoxide (DMSO), NN'- dimethylformamide (DMF), N₅N '-dimethylacetamide (DMAC), 1 ,3-dimethyl- 2-imidazolidinone (DMEU), l,3-dimethyl-3,4,5,6-tetrahydro-2-(lH)- pyrimidinone (DMPU), acetonitrile (ACΝ), propylene glycol, ethyl acetate, benzyl alcohol, 2-pyrrolidone, benzyl benzoate, or any combination thereof. By "pharmaceutical composition" is meant a composition containing a compound of the invention, formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Excipients consisting of DMSO are specifically excluded. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or any other formulation described herein. By "stereoisomer" is meant a diastereomer, enantiomer, or epimer of a compound. A chiral center in a compound may have the ^-configuration or the ^-configuration. Enantiomers may also be described by the direction in which they rotate polarized light (i.e., (+) or (-)). Diastereomers of a compound include stereoisomers in which some, but not all, of the chiral centers have the opposite configuration as well as those compounds in which substituents are differently oriented in space (for example, frans versus cis).

Where a group is substituted, the group may be substituted with L 2, 3, 4. 5, or 6 substituents. Optional substituents include, but are not limited to: C_{1- 6} alkyl. C₂-₆ alkenvl, C₂.₆ alkynyl, cycloalkyL cycloalkenyl, heterocyclyl, aryl, heteroaryl, halogen; azido(-N₃), nitro (-NO₂), cyano (-CN), acyloxy(-OC(=O)R'), acyl (-C(=O)R'), alkoxy (-OR'), amido (-NR'C(=0)R" or -C(O)NRR'), amino (-NRR'), carboxylic acid (-CO₂H), carboxylic ester (- CO₂R'), carbamoyl (-OC(=O)NR'R" or -NRC(=0)0R'), hydroxy (-OH), isocyano (-NC), sulfonate (-S(O)₂OR), sulfonamide (-S(=O)₂NRR' or - NRS(O)₂R^" I or sulfonyl (-5(-O)Jl), where each R or R^" Is selected, independently, from H, C_1-6 alkyl, C₂.₆ alkenyl, C_2-6 alkynyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl. A substituted group may have, for example, L 2, 3, 4, 5, 6, 7, 8, or 9 substituents. In some embodiments, each hydrogen in a group may be replaced by a substituent group (e.g., perhaloalkyl groups such as -CF₃ or -CF₂CF₃ or perhaloaryls such as -C₆F₅). In other embodiments, a substituent group may itself be further substituted by replacing a hydrogen of said substituent group with another substituent group such a& those described herein. Substituents may be further substituted with, for example, 1 , 2, 3, 4. 5, or 6 substituents as defined herein, tor example, a lower CY* alkyl or an aryl substituent group (e.g., heteroaryl, phenyl, or naphthyl) may be further substituted with 1, 2, 3, 4, 5, or 6 substituents as described herein. Brief Description of the Drawings

Fig. 1 shows that Compound (3) does not inhibit recombinant RIPl kinase.

Detailed Description of the Invention

We have discovered a series of heterocyclic derivatives that inhibit tumor necrosis factor alpha (TNF-α)-induced necroptosis. The heterocyclic compounds of the invention are described by Formulas (I) and (II) and are shown to inhibit TNF-α induced necroptosis in FADD-deficient variant of human Jurkat T cells. Pharmaceutical compositions including the compounds of the invention arc also described. The invention also features kits and methods of treatment featuring the compounds and compositions of the invention.

Compounds of the invention can be described generally by Formula (I):

(I), where each X¹, X², X³, X⁴, X⁵, and X⁶ is selected, independently, from N or

CR XI , each Y „ Y . and Y" is selected, independently, from O, S, NR Yl , or

CR^Y2R^Y3; each Z¹ and Z² is selected, independently, from O, S, or NR^ZI ; each R^γl and R^Z1 is selected, independently, from H, optionally substituted Ci_-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, -

-C(=O)OR^5A, or -C(-O)NR^5AR^6A; each R^X1, R^Y2, and R^Y3 is selected, independently, from H, halogen, CN, NC, NO₂, N₃, OR³, SR³, NR³R⁴, -Cf ^" O)R⁵ \ -C0-O/JR⁵\ -C(=O)NR^5AR^6A, - S(O)R^5A, -S(=O)₂R^5A, -S(=O)₂OR^5A, -S(=O)₂NR^5AR^6A, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each R¹, R², R^5A, R^5B, R^6A, and R^6B is selected from H, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R^5A and R^6A, or R^5B and R^6B combine to form a heterocyclyl; and each R³ and R⁴ is selected from H, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, -C(=O)R^5B, ~C(=S)R^5B, -C(-NR^6B)R^5B, -C(O)OR⁵⁰, -C(=O)NR^5BR^6B, -S(O)R^5B, -S(O)₂R^5B, - S(0)₂OR^5B, or -S(=O)₂NR^5BR^6B; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In some embodiments, when R¹ is H₅ X^!, X², and X⁴ are each CH, X³ and X- are each N, Y¹ is S, Y² is NH, Z¹ is NH, and Z² is O, R² is not 4- fluorophenyl.

A further embodiment is described by Formula (II):

(II), where

X¹, X², X⁴, X⁵, R¹, Y², and R^Z1 are as defined for Formula (I); each R^2A, R^2B, R^2C, R^2D, and R^2E is selected, independently, from H, halogen, optionally substituted Ci_-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, CN, NC, NO₂, N₃, OR⁷, SR⁷, S(-O)R¹², SC-O)₂R¹², S(O)OR¹², S(O)₂OR¹I NR⁷R⁸, C(=O)R¹², C(O)OR¹², C(=OjNR^{f 2}R³³, Q=S)R¹², CY=S)OR¹², Cl-SjNR¹²R¹I C(=NR⁹)R¹², C(-NR⁹)OR¹², or C(=NR⁹)NR¹²R¹³, or R²⁴ and R^2B. R^2β and R^2C, R~^c and R^2D _; or R^2D and R^2E combine to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl; each R⁷, R⁸, and R⁹ is selected, independently, from H, optionally substituted Ci. «₅ alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, S(O)R ^{! 0}, S(O)^R¹". C(O)R^{f ft}, CC=O)OR¹⁰, CrO)NR¹⁰R¹¹. C(- SjR¹I C(=SjOR¹⁰, C(=S)NR¹⁰R^π. C(-NR¹⁴)R¹⁰, C(=NR^1J)OR¹⁰, or C(-^:-NR¹⁴)NR^iϋR^{! i} _; or R'' and R⁸ combine to form an optionally substituted hctcrocycM; and each R¹⁰, R¹¹, R¹², R¹', and R¹⁴ is selected, independently, from H, optionally substituted C^₀ alkyl. optionally substituted C₂^. alkenyL optionally substituted Q->_-0 alkynyl, optionally substituted cycloalkyL optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R¹⁰ and R¹¹ or R¹² and R^!3 combine to form an optionally substituted heterocyclyl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

Compounds of the invention may also be described by Formula (H-A):

(II- A), where each R¹, R^2Λ, R^2B, R^2C, R^2D, R^2E, R^Y1, R^Z1 _; R\ R⁴, R^5A R^6A. R⁷, R\ R⁹ _; and R¹² is as defined in Formula (II); and where each R^X1A and R^X1B is selected, independently, from H, halogen, CN, NC, NO₂, Ni. OR³, NR³R⁴, -C(O)R^5A, -C(O)0R^5A, -C( O)NR⁵V, -S(O)_?R^5Λ, -S(=O)_?OR^5A, -S(=O)>MR^5ΛK^6A, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

In a further embodiment, the compounds of the invention may be described according to Formula (H-B):

(II-B), where

RX^1B is selected from H, halogen, CN, NC, NO₂, N₃, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl. - C(=O)R^5A, -C(=O)OR^5A, or -C(-O)NR^5ΛR^6A; each R^2A, R^2B, R^2C, R^2D, and R^2E is selected, independently, from H, halogen, CN, NC, NO₂, N₃, OR⁷, NR⁷R⁸, SR⁷, Q=O)R¹ °, C(=O)OR¹⁰, C(O)NR¹⁰R¹¹, S(=O)R¹², S(=O)₂R¹², optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R^2A and R²³, R^2B and R^2C, R^2C and R^?D, or R^2D and R^2E combine to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl; and where each R^5A. R^6A, R⁷, K\ R¹⁰, R¹¹, and R¹² is as defined in Formula

(H) or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

Exemplary compounds of the invention include compounds of Formula (II-C) such as those shown in Table 1: Table 1

Additional exemplary compounds include those of Formula (II-B-1) such as those described in Table 2

Table 2

21

Additional exemplary compounds include those of Formula (II-B-2) such as those described in Table 3.

Table 3

30 Still further compounds are those of Formula (II-B-3) as shown in Table

4.

Table 4

Other compounds of the invention include those of Formula (II-D), including the compounds shown in Table 5.

Table 5

Compounds of the invention can be synthesized according to methods known in the art or by the methods provided in the examples below. The synthesis of (3) is outlined in Scheme 1 and the procedures described herein can be used to prepare the other compounds of the invention. 2-Aminothiazole was acylated with 2-chloroacetyl chloride followed by treatment with potassium thiocyanate to form the thiazolidone (1) (Kwon et al., J. Heterocyclic Chem. 20: 1725 (1983)). The semicarbazone of 4-fluoroacetophenone was treated with Vilsmeier-Haack reagent to afford pyrrazolc (2) (Kira et al., J Heterocycl. Chem. 2: 25 (1970) and Baraldi et al. Synthesis, 10: 1140 (1997)). Compound (3) was derived from a Knoevenagel condensation of (1) and (2) with the aid of sodium acetate anhydrous in glacial acetic acid (Kwon et al., J. Heterocyclic Chem. 20: 1725 (1983)).

Scheme 1

Reagents and conditions: (a) 2-chloroacetyi chloride, Ei ₅N, CH7CI₂, 0° to room temperature. 53%: (b) KSCN, EtOH, reflux, 85%; (c) H,NHNCONH₂-HC1, NaOAc, CH₁OHZH-A 98%; (d) (I)POCl₃-DMF^'', (H) 1O⁰ZoNaOH₅ (Hi) I O N HCl 86%; (ε) NaOAc, HOAc, reflux, 91%.

Pharmaceutical Compositions

Compounds of the invention (e.g., compounds of Formulas (I) and (H)) can be formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo. Accordingly, the present invention provides a pharmaceutical composition comprising a compound of the invention in admixture with a pharmaceutically acceptable excipient. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2003 - 20^th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF 19), published in 1999.

The compounds of the invention may be used in the form of the free base, in the form of salts, solvates, and as prodrugs. All forms are within the scope of the invention, In accordance with the methods of the invention, the described compounds or salts, solvates, or prodrugs thereof may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds of the invention may be administered, for example, by oral, parenteral, buccal, sublingual, nasal rectal, patch, pump, or transdermal administration and the pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

Pharmaceutically Acceptable Excipients

Pharmaceutically acceptable excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, dϊsintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, or waters of hydration. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

Oral Administration

A compound of the invention may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsules, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet. For oral therapeutic administration, a compound of the invention may be incorporated with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.

Parenteral Administration A compound of the invention may also be administered parenterally.

The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that may be easily administered via syringe.

Nasal Administration

Compositions for nasal administration may conveniently be formulated as aerosols, drops, gels, and powders. Aerosol formulations typically include a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomizing device. Alternatively, the sealed container may be a unitary dispensing device, such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it will contain a propellant, which can be a compressed gas, such as compressed air or an organic propellant, such as fluorochlorohydrocarbon. The aerosol dosage forms can also take the form of a pump-atomizer.

Buccal or Sublingual Administration

Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, where the active ingredient is formulated with a carrier, such as sugar, acacia, tragacanth, or gelatin and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base, such as cocoa butter. The compounds of the invention may be administered to an animal alone or in combination with pharmaceutically acceptable carriers, as noted above, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration, and standard pharmaceutical practice.

Dosage Amounts

The amount of active ingredient in the compositions of the invention can be varied. One skilled in the art will appreciate that the exact individual dosages may be adjusted somewhat depending upon a variety of factors, including the protein being administered, the time of administration, the route of administration, the nature of the formulation, the rate of excretion, the nature of the subject's conditions, and the age, weight, health, and gender of the patient. Generally, dosage levels of between 0.1 μg/kg to 100 mg/kg of body weight are administered daily as a single dose or divided into multiple doses. Desirably, the general dosage range is between 250 μg/kg to 5.0 mg/kg of body- weight per day. Wide variations in the needed dosage are to be expected in view of the differing efficiencies of the various routes of administration. For instance, oral administration generally would be expected to require higher dosage levels than administration by intravenous injection. Variations in these dosage levels can be adjusted using standard empirical routines for optimization, which are well known in the art. In general, the precise therapeutically effective dosage will be determined by the attending physician in consideration of the above identified factors.

Therapeutic Uses and Screening Methods

Compounds disclosed herein can be used to treat disorders where necroptosis is likely to play a substantial role, e.g., cerebral ischemia, traumatic brain injury, a neurodegenerative disease of the central or peripheral nervous system, the result of retinal neuronal cell death, ihc result of cell death of cardiac muscle, the result of cell death of cells of the immune system; stroke, liver disease, pancreatic disease, the result of cell death associated with renal failure; heart, mesenteric, retinal, hepatic or brain ischemic injury, ischemic injury during organ storage, head trauma, septic shock, coronary heart disease, cardiomyopathy, myocardial infarction, bone avascular necrosis, sickle cell disease, muscle wasting, gastrointestinal disease, tuberculosis, diabetes, alteration of blood vessels, muscular dystrophy, graft-versus-host disease, viral infection, Crohn's disease, ulcerative colitis, asthma, or any condition in which alteration in cell proliferation, differentiation or intracellular signaling is a causative factor. Compounds of the invention can also be used in screening methods to identify targets of necroptosis and to identify additional inhibitors of necroptosis, as well as in assay development.

Compounds disclosed herein can be evaluated for their pharmacological properties in animal models of disease. The compounds identified to decrease necrosis or necroptosis may be structurally modified and subsequently used to decrease necrosis or necroptosis, or to treat a subject with a condition in which necrosis or necroptosis occurs. The methods used to generate structural derivatives of the small molecules that decrease necrosis or necroptosis are readily known to those skilled m the fields of organic and medicinal chemistry. Therapy according to the invention may be performed alone or in conjunction with another therapy, for example in combination with apopiosis inhibitors, and may be provided at home, the doctor's office, a clinic, a hospital's outpatient department, or a hospital. Treatment generally begins at a hospital so that the doctor can observe the therapy's effects closely and make any adjustments that are needed. The duration of the therapy depends on the age and condition of the patient, as well as how the patient responds to the treatment. Additionally, a person having a greater risk of developing a condition may receive prophylactic treatment to inhibit or delay symptoms of the disease.

In some embodiments, the compounds and methods of the invention can be used to treat any of the following disorders where necroptosis is likely to play a substantial role: a neurodegenerative disease of the central or peripheral nervous system, the result of retinal neuronal cell death, the result of cell death of cardiac muscle, the result of cell death of cells of the immune system; stroke, liver disease, pancreatic disease, the result of cell death associated with renal lailυre; heart, mesenteric, retinal, hepatic or brain ischemic injury, ischemic injury during organ storage, head trauma, septic shock, coronary heart disease, cardiomyopathy, myocardial infarction, bone avascular necrosis, sickle cell disease, muscle wasting, gastrointestinal disease, tuberculosis, diabetes, alteration of blood vessels, muscular dystrophy, graft- versus-host disease, viral infection, bacterial infection. Crohn's disease, ulcerative colitis, asthma, and any condition in which alteration in cell proliferation, differentiation or intracellular signaling is a causative factor.

Conditions Caused by Alteration in Cell Proliferation. Differentiation, or Intracellular Signaling Conditions in which alteration in cell proliferation, differentiation or intracellular signaling is a causative factor include cancer and Infeαloru e.g., by viruses (e.g., acute, latent and persistent), bacteria, fungi, or other microbes.

Exemplary viruses are human immunodeficiency virus (HIV), Epstein- Barr virus (EBV), cytomegalovirus (CMV)5 human herpesviruses (HHV), herpes simplex viruses (HSV), human T-CeIl leukemia viruses (HTL V)5 Varicella-Zoster virus (VZV), measles virus, papovaviruses (JC and BK), hepatitis viruses, adenovirus, parvoviruses, and human papillomaviruses. Exemplary diseases caused by viral infection include, but are not limited to, chicken pox. Cytomegalovirus infections, genital herpes, Hepatitis B and C, influenza, and shingles.

Exemplary bacteria include, but are not limited to Campylobacter jejuni, Enterobacter species, Enterococcus faecium, Enterococcus faecalis, Escherichia coli (e.g., F. coli O157:H7), Group A streptococci, Haemophilus influenzae, Helicobacter pylori, listeria, Mycobacterium tuberculosis, Pseudomonas aeruginosa, S. pneumoniae, Salmonella, Shigella, Staphylococcus aureus, and Staphylococcus epidermidis. Exemplary diseases caused by bacterial infection include, but are not limited to, anthrax, cholera, diphtheria, foodborne illnesses, leprosy, meningitis, peptic ulcer disease, pneumonia, sepsis, tetanus, tuberculosis, typhoid fever, and urinary tract infection.

Neurodegenerative Diseases

Exemplary neurodegenerative diseases are Alzheimer's disease, Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis, HIV- associated dementia, cerebral ischemia, amyotropic lateral sclerosis, multiple sclerosis, Lewy body disease, Menke's disease, Wilson's disease, Creutzfeldt- Jakob disease, and Fahr disease. Exemplary muscular dystrophies or related diseases are Becker's muscular dystrophy, Duchenne muscular dystrophy, myotonic dystrophy, limb-girdle muscular dystrophy, Landouzy-Dejerine muscular dystrophy, facioscapulohumeral muscular dystrophy (Steinert's disease), myotonia congenita, Thomsen's disease, and Pompe's disease. Muscle wasting can be associated with cancer, AIDS, congestive heart failure, and chronic obstructive pulmonary disease, as well as include necrotizing myopathy of Intensive care.

Compounds and methods of the invention can additionally be used to boost the immune system, whether or not the patient being treated has an immunocompromising condition. For example, the compounds described herein can be used in a method to strengthen the immune system during immunization, e.g., by functioning as an adjuvant, or by being combined with an adjuvant.

Kits

Any of the compounds or pharmaceutical compositions of the invention (e.g., compounds of Formulas (I) and (II)) can be used together with a set of instructions, i.e., to form a kit. The kit may include instructions for use of the compounds of the invention in a screening method or as a therapy as described herein. The following non-limiting examples are illustrative of the present invention.

EXAMPLES Example 1: Synthesis of Compound (3)

Synthesis ofThiazolidone (1)

Scheme 2 K_SCN

(11) (12) (1)

The solution of 2~aminothiazole (Ii) (5 rnrnol) and triethylamine (0.7 mL) in distilled THF (20 niL) was cooled with an ice- water bath. A solution of 2-chloroacetyl chloride (0.4 mL) was added dropwise to above solution with stirring. The resulting mixture was stirred for another 3 hours ai room temperature. The solvent was removed in vacuo and dichloromethane (30 mL) and water (30 mL) were added to the residue. The organic phase was separated and the aqueous phase was extracted with dichloromethane (20 mL * 3). The combine organic phases were washed sequentially with 2N HCl, saturated aqueous NaHCO₃, brine (saturated aqueous NaCl), and then dried over Na₂SO₄. Removal of the solvent afforded the solid (12) (630 mg, 72%). Further purification can be conducted through crystallization from elhanol/water. The solution of amide (12) (2 mmol) and potassium thiocyanate (2 mmol) in ethanol (15 mL) was heated to reflux and reacted for 8 hours. The reaction was then cooled to room temperature. The precipitate was collected by suction filtration and washed with water (10 mL x 3) and ethanol. After drying, thiazolidone (1) was obtained (320 mg, 80%). Melting point: 205-207 ° C ; ¹H NMR(DMSO, 300MHz) 63.97(S,

2H₅ -CH₂-), 7.36(d, IH, Thiazole, J=3.6Hz), 7.56(d, IH, Thiazole, J=3.6Hz), 12.0(brs, IH, -NH); EIMS 200(M⁺ i l)(6.10), 199(M⁺)(47.74), 166(47.76),

126(54.37), 125(61.55), 58(100, Base): IR(Cm^'1) 31 15, 3087, 2826, 1716, 1596, 1141; Anal. Calcd for C₆H₅N₃OS₂ : C, 36.17; H, 2.53; N, 21.09; Found : C, 36.39; H, 2.47; N, 21.43.

Synthesis ofDiazole (2) Scheme 3

(13)

A solution of acetophenone (13) (10 mmol) in methanol (15 mL) was added dropwise to a solution of hydrochloride aminocarbonhydrazine (10 mmol) and sodium acetate (10 mmol) in water (10 mL) at room temperature and with stirring. The resulting mixture was stirred for another 3 hours. The precipitate was filtered, washed with water (10 mL x 2) and anhydrous ethyl ether (5 mL). The white solid (14) was dried under air (1.9 g, 98%) and the product was sufficiently pure for the next step. The ketone semicarbazone (14) (5 mmol) was added, portionwise with stirring, to a mixture of phosphorus oxychloride-dimethylformamide. The latter was prepared by the slow addition of POCl₃ (1 1 mmol) to DMF (22mmol) at 0 ° C. The reaction mixture was heated at 60-70 ^ϋ C for about 4 hours and then poured onto cracked ice (10 g). The resulting mixture was neutralized with NaOH (2 g in 8 mL water), heated at 50-60 ° C for 5 minutes, and cooled. Impurities were then removed by filtration. The filtrate was acidified to pH - 5-6 using IO N HCl. The resulting precipitated solid was collected by filtration and washed with water. The product (2) was dried in the air (712 mg, 75%). If needed, the solid was further purified by crystallization from ethanol/water. Melting point 163-165 ^c C; ¹H NMR(CDCl₃, 300MHz) 87.17-7.28(m, 3H, 0-ArH, -NH-), 7.69-7.74(m, 2H, m-ArH), 8.18(s, IH, Pyrazole), 9.97(s, IH, -CHO); EIMS 191(M⁺+1)(12.62), 190(M⁺)(IOO, Base), 189(M⁺-H)(93.84); IR(Cm^"1) 3165, 2897, 1646, 1517, 1481, 1250, 931; Anal. Calcd for C₁₀H₇FN₂O : C, 63.16; H, 3.71 ; N, 14.73; Found : C, 63.16; H, 3.71; N, 15.06.

Synthesis of Compound (3)

Scheme 4

(1) (2) (3) A mixture of compound (1) (0.7 mmol), compound (2) (0.71 mmol), sodium acetate (2 mmol), and acetic acid (4 mL) was heated to 1 10 ° C and stirred for 14 hours at this temperature. Solid precipitated during the course of the reaction. After cooling to room temperature, the system was poured into 50 mL of water. The precipitate was collected by suction filtration and washed with water and ethanol. The crude product was dried and 1OmL ethanol were added. The mixture was heated to 80 ° C for several minutes and the cooled. Compound (3) was collected by filtration and dried to afford a yellow solid (236mg, 91%).

Melting point >300 ° C; ^!H NMR(DMSO, 300MHz) δ7.42-7.50(m, 4H), 7.60-7.65(m, 2H), 7.73(d, IH, J =3.6Hz)₅ 7.91+7.19(brs, IH), 12.54(brs, IH, =NH), 13.68+13.86(brs, IH, -NH-); EIMS:371(M⁺)(4.87), 370(17.68), 234(24.34), 209(21.97), 207(21.96), 178(19.86), 165(40.10), 91(100, base), 44(37.30), 41(20.31); IR(Cm^'1) 3200, 1713, 1593, 1498, 1438, 1304, 1 192, 1 132, 846; Anal Calcd for C₁₆H₁₀FN₅OS₂ : C, 51.74; H, 2.71 ; N, 18.86; Found : C, 51.68; H, 2.87; N, 19.06. Synthesis of Related Compounds

This procedure described for the synthesis of compound (3) can be used to prepare the other compounds of the invention by variation of the reagents.

In order to study of the influence of pyrazole ring of (3), derivatives with other aromatic heterocycles in place of pyrazole were synthesized, such as triazole and isoxazole (see Schemes 5-7). For example, derivatives (10) were prepared through condensation of these various carboxaldehyde derivatives (7). (8) or (9) with thiazolidone (1).

The requisite phenylpropiolaldehyde starting material was prepared from the corresponding p-substituted benzaldehyde through a Corey-Fuchs reaction (Journet et al., Tetrahedron Lett. 39: 6427 (1998)) and then cyclized with sodium azide in DMSO at room temperature to form 5-aryl-4- carbaldehyde-l,2,3-trizazoles (7a)-(7d) (Scheme 5; Journet et al., Tetrahedron Lett. 42: 9117 (2001)), where, respectively, R is H, Cl, OCII₃, or F.

Scheme 5

Reagents and conditions: (a) CBr₄, PPh₃, CH₂Cl₂, 0 ⁰C -rt, 81 -88%; (b) i) n-BuLi, -78°C, ii) DMF, iii) 10%KH₃PO₄, 64-71%; (c) i) NaN-,. DMSO,rt, n) 15%KH₂PO₄ 41-61%

The iV-hydroxybenzimidoyl chloride derived from chlorination of benzaldehyde oxime by N-chlorosuccinimide (NCS; Shang et al., Synthesis 12: 1663 (2002)) was treated with 3-dimethylaminoacrolein in the presence of triethylamine to generate the isoxazole carboxaldehydes (8a)-(8d) via a 1,3- dipolar cycloaddition (Scheme 6; Balsamo et al., Eur. J. Med. Chem. Chim. Ther. 38: 157 (2003)), where, respectively, R is CL F, Me, or NO₂. Scheme 6

-CHO —5-»- ^R-\ /," .

Λ

3a-d

Reagents and conditions: (a) HO-NH₂-HCl, K₂CO₃, EtOH, rf, 80-94%; (b) NCS, CH₂Cl₂, rt, 50-54%; (c) 3-dimethylaminoacrolein, Et₃N, THF, rt, 42-93%

The N atom of pyrazole can easily react with electrophilic reagents in the present of potassium carbonate at room temperature, such as iodom ethane, bromoacetonitrile and benzyl bromide. The main products were N-substituted 3-aryl-4-carboxaldehydepyrazoles (9) (Scheme 7). Scheme 7

Reagents and conditions: (a) RX, K₂CO₃, CH₃CN. ri, 65-91%.

Example 2: Evaluation of Necroptosis Inhibitory Activity

For EC₅₀ value determinations, cells were treated with 10 ng/mL of human TNF-α in the presence of increasing concentration of test compounds for 24 hours followed by ATP -based viability assessment.

Modification of the tiήazole of Compound (3)

The thiazole of Compound (3) was replaced with a 1 ,3,4-thiadiazole and with the addition of methyl substituents to various positions on the thiazole. As shown in Table 6, the thiadiazole derivatives were inactive. Adding a methyl at the thiazole 4-position increased the activity slightly, but adding this substituent to the 5-ρosition had a negative effect. Table 6

Compounds (16)-(21) compound X^s X² R EC₅₀(μM)

16 CH N H Inactive

17 CH N 4-F Inactive

18 CH N 3 -F Inactive

19 CH N 2-F Inactive

20 CH CMe 4-F 5.25

21 CMe CH 4-F 23.07

Influence of substituents on the phenyl ring of Compound (3)

For the study of the influence of substituents on the phenyl ring, a series of derivatives of compound (3) were prepared according to Scheme 1. Results of the biological studies arc shown in Table 7. As shown in Table 7, the fluoro group at the para-position could be replaced with a variety of electron- donating or electron- withdrawing substituents with retention of necroptosis inhibitory activity. It is worth noting that replacing the fluoro group by morpholine (29) or phenyl (33) increased necroptosis inhibitory activity. When the fluoro group in the para-position was retained, adding an additional fluoro group to the 3-position of phenyl ring enhanced the activity slightly ((34) vs. (3) and (35) vs. (29)).

Table 7

compound R EC₅₀(μM)

3 4-F 10.6

22 3-F Inactive

23 2-F inactive

24 H 4(16

25 4-CH₃O 14.9

26 4-CH₃ 7.77

27 4-Cl 15 63

28 4-Br 34.51

29 4-N(C₂H₄)₂O 2.93

30 4-NO₂ 22.96

31 4-SCH₃ —

32 4-OH 32.67

33 4-Ph 1.29

34 3,4-F₂ 6.32

36 3,4-O₂(CH₂) 34.83

65 3_F_4_N< C₂H₄)₂O 2.90

Since (29) (EC₅₀ =2.93 μm) with a rnorpholine group at the 4-posiτion of the phenyl ring displayed increased inhibitory activity, several derivatives with substituting grouos containing a N atom at the phenyl para-position were examined (Table 8). These derivatives were also synthesized according to Scheme L The acetophenones for compounds (37)-(39) were prepared by a SN _Ar reaction of N-contained heterocycles with 4-fluoroacetophenone in the presence of potassium carbonate while the staring material for (40) and (41) was derived from alkylation of 4-aminoacetophenone by iodomethane or iodoethane. Table 8

Compounds (37)-(43) compound _RA _RH _Z S EC₅₀(UM)

37 -(CH₂),- 6.36

38 -(CHz)₄- 9.53

39 -(C₂H₄)₂S 3.30

40 Me Me -

41 Et Et -

42 -(C₂H₄)₂O C-O 7.19

As shown in Table 8, heterocyclic groups without an oxygen atom ((37) and (38)) also increased activity, but less so compared to the morpholine ring. A derivative with a 6-membered ring displayed better activity compared to a 5- membered ring ((37) vs. (38)). Thiomorpholine substitution improved activity similar to that of morpholine ((39) vs. (29)). Finally, addition of the N- containing heterocycles to the phenyl ring through an acyl moiety, e.g. piperidine ring, resulted in further improvement in activity ((42) vs. (43)).

Compound (33) (EC₅₀ =1.29μM) with a biphenyl in place of the phenyl ring was also active. Various other replacements were also examined (Table 9). As shown in Table 9, most replacements were well tolerated. However, the disruption of the biphenyl structure had little influence on the activity (52). Thus, the conjugated structure is not important for optimal activity of the biphenyl. Notably, another conjugated bi-cyclic derivative with naphthyl group displayed greatly decreased activity (EC₅₀ =56.8 μM). Table 9

Compounds (44)-{52) compound Z' R EC₅₀(UM)

44 - o-OMe 2.41

45 - p-OMe 2.59

46 - p-Me 4.38

47 - p-Cl >100

48 - p-F 3.87

49 - o-F 47.97

50 -0- -H -

51 -S- -H _

52 -SO₂- -H 1.78

Finally, derivatives (53)-(64) were evaluated and the experimental results are shown in Table 10.

Table 10

Compounds (53)-(64j compound R EC₅₀(UM)

53 N NH H Inactive

54 N NH Cl Inactive

55 N NH OCH₃ Inactive

56 N NH F Inactive

57 CH O Cl Inactive

58 CH O F Inactive

59 CH O CH₃ Inactive

60 CH O NO₂ Inactive

61 CH C-Me F Inactive

62 CH C-CH₂CONH₂ F Inactive compound r R EC₅₀(μM)

63 CH C-Bn F Inactive 64 CH C-Ph F Inactive

In conclusion, a number of derivatives of (3) were found to inhibit TNF- α-induced necroptosis in FADD-deficient variant of human Jurkat T cells.

Example 3: RIPl Kinase Assay

RIPl kinase autophosphorylation reactions were performed in the presence of the indicated concentrations of Nec-1 and compound (3) for 30 min at 30 ⁰C. Reactions were performed as described in Degterev et al., Not. Chem. Biol, 4: 313-321 (2008). Briefly, recombinant human GST-RIPl (1-375 a.a.) was expressed using Baculogold system in Sf9 cells and purified using glutathione-sepharose beads. Protein was eluted in 50 mM Tris-HCl, pH 8.0. Two micrograms of protein were used in each reaction. Reactions were performed in the presence of 20 mM ATP and 3 mCi γ-³²P-ATP. Following the reaction, products were separated on 8% SDS-PAGE and visualized by autoradiography. As shown in Figure 1 and in contrast to the results obtained using Nee- \ , Compound (3) does not inhibit recombinant RlPl kinase. These data suggest the possibility that (3) may target an additional regulatory molecule in the pathway.

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

While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth, and follows in the scope of the claims.

Other embodiments are within the claims.

What is claimed is:

Claims

1. A compound of the formula

(I), wherein each X¹, X², X , X⁴, X⁵, and X⁶ is selected, independently, from N or

CR xi. each Y¹, Y², and Y³ is selected, independently, from O, S, NR^Y1, or

CR^Y2R^Y?; each Z and Z is selected, independently, from O, S, or NR Zl . each R^Y1 and R^Z1 is selected, independently, from H, optionally substituted C]_-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted hctcrocyciyl, optionally substituted aryl, optionally substituted heteroaryl, - ( X=C ))R^. -C(-O)OR^5A, or -C(=O)NR^5AR⁶⁴; each R , R , and R is selected, independently, from H, halogen, CN, NC, NO₂, N₃. OR³, SR³, NR³R⁴, -C(-Q)R^5A, -C(-O)OR^5A, -C(=O)NR^5AR^6A -

optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heierocycly. !, optionally substituted aryl, or optionally substituted heteroaryl; each R¹, Rl R^5A, R^5B, R^6A, and R^6B is selected from H, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl. optionally substituted aryl, or optionally substituted heteroaryl. or R^5A and R^6A, or R^5B and R^6B combine to form a heterocyclyl; and each R³ and R⁴ is selected from H, optionally substituted C_]-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, -C(=O)R^5B, -C(=S)R^5B, -C(=NR^6B)R^5B, -C(=O)OR^5B, -C(=O)NR^5BR^6B, -S(=O)R^5B, -S(=O)₂R^5B, - S(=O)₂OR^5B, or -S(=O)₂NR^5BR^6B; wherein when R¹ is H, X¹, X², and X⁴ are each CH, X³, X⁵, and X⁶ are each N, Y¹ and Y³ are each S, Y² is NH, Z¹ is NH, and Z² is O, R² is not 4- fluorophenyl, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

2. The compound of claim 1, wherein said compound is of the formula

(II), wherein

X¹, X², X⁴, X⁵, R¹, Y², and R^Z1 are as defined for Formula (I); each R^2A, R^2B, R^2C, R^2D, and R^2E is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, CN, NC, NO₂, N₃, OR⁷, SR⁷, S(=O)R¹², S(=O)₂R¹², S(=O)OR¹², S(O)₂OR¹², NR⁷R⁸, C(=O)R¹², C(O)OR¹², C(=O)NR¹²R¹³, Q=S)R¹², C(=S)OR¹², C(^S)NR¹²R¹³, C(=NR⁹)R¹², C(-NR⁹)OR¹², or C(=NR⁹)NR¹²R¹³, or R^2A and R^2β _; R^2B and R^2C, R^2C and R^2D, or R^2D and R^2E combine to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl; each R , R , and R is selected, independently, from H, optionally substituted C₁ ^ alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heieroaryl, S(=O)R¹⁰, S(-O)₂R¹⁰, CC-O)R¹⁰, CC=O)OR¹⁰, C(O)NR¹⁰R¹¹, C(-S)R¹⁰, C(=S)OR¹⁰, CC=S)NR¹⁰R¹¹, C(=NR¹⁴)R¹⁰, C(=NR¹⁴)OR¹⁰, or C(=NR^H)NR¹⁰Rⁿ, or K⁷ and R^s combine to form an optionally substituted heterocyclyl; and each R¹⁰, R¹¹, R¹², R^ϋ, and R¹⁴ is selected, independently, from H₅ optionally substituted C|__ft alkyl. optionally substituted C₂.β alkenyl, optionally substituted C₂^ alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R¹⁰ and R¹¹ or R¹² and R¹³ combine to form an optionally substituted heterocyclyl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

3. The compound of claim 2, wherein said compound of the formula

(II- A), wherein each R¹, R^2A, R^2B, R^2C, R²¹I R^2r, R^Y1, R⁷¹, R¹, R⁴, R⁵\ R^6A, R\ R⁸, R⁹, and R¹² is as defined in Formula (II); and where each R^X1A and R^X1B is selected, independently, from lϊ, halogen, CN, NC, NO₂, N,, OR³. NR³R⁴, -C(-Q)R^5A, -C(=O)OR^5A, -C(=O)NR^? V^A, -SC-O J₂R' *, -S(=O);OR⁵\ -S(=ObNR^iAR"^Λ, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof,

4. The compound of claim 3, wherein R^{v ι} is II, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

5. The compound of claim 3, wherein R_Zi is H, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

6. The compound of claim 3, wherein R^X1A is H and R^X1B is H or optionally substituted C_1-6 alkyl, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

7. The compound of claim 6, wherein R^X1A and R^X1B are H, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

8. The compound of claim 3, wherein R¹ is H, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

9. The compound of claim 3, wherein R^2A, R^2D, and R^JE are H, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

or^"" ^*7

10. The compound of claim 3, wherein R is selected from halogen, OR ,

SR⁷, S(=O)₂R¹², NR⁷R⁸, CC-O)R¹², C(KJ)OR¹², C(=O)NR¹²R¹³, optionally substituted C_{1 -6} alkyl, optionally substituted heterocyclyl, or optionally substituted aryl, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

OC

1 1. The compound of claim 10, wherein R is halogen. or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

12. The compound of claim 11, wherein R^2B is also halogen, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

13. The compound of claim 3, wherein said compound is of the formula

, wherein

R_X1B is H or optionally substituted C₁^ alkyl; each R^2A, R^2B, R^2C, R^2D, and R^2E is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, OR⁷, S(=O)R¹², S(=O)₂R¹², NR⁷R⁸, C(=O)R¹². C(=O)OR¹², or C(-O)NR¹²R¹³, or R^2A and R^2B, R^2B and R^2C, R^2C and R^2D, or R^2D and R^2E combine to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl; each R⁷ and R^s is selected, independently, from H. optionally substituted C]_-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, S(=O)R¹⁰, S(=O)₂R¹⁰, C(O)R¹⁰, C(-O)OR¹⁰, C(O)NR¹⁰R¹¹, or R⁷ and R⁸ combine to form an optionally substituted heterocyclyl; and each R¹⁰, R¹¹, R¹², and R¹³ is selected, independently, from H, optionally substituted C_1-6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R¹⁰ and Rⁿ or R¹² and R¹³ combine to form an optionally substituted heterocyclyl; or any pharmaceutical!} acceptable salt or solvate thereof, or stereoisomer thereof. YI Pl

14. The compound of claim 13, wherein R is H, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

Yl Tl

15. The compound of claim 13, wherein R is C_1-6 alkyl, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

16. The compound of claim 15, wherein said compound is

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

17. The compound of claim 13, wherein R^2A, R^2D, and R^2E are H, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

18. The compound of claim 13, wherein R is halogen, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

19. The compound of claim 18, wherein said compound is

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

20. The compound of claim 17 or 18, wherein R^2B is halogen, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

21. The compound of claim 20, wherein said compound is

(65), or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

22. The compound of claim 13, wherein R^2C is optionally substituted C]_-6 alkyl or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

23. The compound of claim 22, wherein said compound is

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

24. The compound of claim 13, wherein R is optionally substituted heterocyclyl, NR⁷R⁸, or C(-O)NR^!-R¹³, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

25. The compound of claim 24, wherein R^2C is NR⁷R⁸, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

26. The compound of claim 25, wherein R and R are each C_1-6 alky 1, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

27. The compound of claim 24, wherein said compound is of the formula

wherein each n and m is, independently, an integer between 0-4; G is a bond, O, S, or NR^2J; R is H or optionally substituted C_1-6 alkyl; and wherein (n+m) is at least 2, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

28. The compound of claim 27, wherein said compound is selected from the group consisting of:

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

29. The compound of claim 24, wherein said compound is of the formula

n each n and m is, independently, an integer between 0-4; G is a bond, O, S, or NR^2J; R^2J is H or optionally substituted C_1-6 alkyl; and wherein (n+m) is at least 2, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

30. The compound of claim 29, wherein said compound is

H (42). or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof,

31. The compound of claim 13, wherein said compound is of the formula

wherein

G¹ is a covalent bond or S(^O)₂; each R^2K, R^2L, R^2M, R^2N, and R²⁰ is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, optionally substituted C_2-6 alkenyl, optionally substituted C_2^6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, CN, NC, NO₂, N₃, OR^2P, SR^2P, S(=O)R^2P, S(-O)₂R^2P, S(-O)OR^2P, S(-O)₂OR^2P, NR^2QR^2R, C(=O)R^2P, C(=O)OR^2P, C(-O)NR^2QR^2R, C(=S)R^2P, C(=S)OR^2P, C(=S)NR^2QR^2R, C(=NR^2S)R^2Q, C(-NR^2S)OR^2Q, or C(-NR^2S)NR^2QR^2R, or R^2K and R^2L, R^2L and R^2M, R^2M and R^2N, R^2N and R²⁰, or R^2Q and R^2R combine to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl; and each R^2P ₅ R^2Q, R^2R, and R^2S is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, optionally substituted C₂,g alkenyl, optionally substituted C_2-6 alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

32. The compound of claim 31, wherein G¹ is S(=O)₂, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

33. The compound of claim 32, wherein said compound is

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

34. The compound of claim 31, wherein G¹ is a bond, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

35. The compound of claim 34, wherein each R^2K R¹, R^2M, R^2N, and R²⁰ is selected, independently, from H, halogen, optionally substituted C_1-6 alkyl, and O-(optionally substituted C_1-6 alkyl), or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

36. The compound of claim 35, wherein R^2K and R^2L are H, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

37. The compound of claim 35, wherein at least one of R^2M, R^2N, and R²⁰ is 0-(optionally substituted Ci_-6 alkyl), or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

38. The compound of claim 37, wherein said compound is

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

39. The compound of claim 35, wherein at least one of R^2M, R^2N, and R²⁰ is optionally substituted C_1^6 alkyl or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

40. The compound of claim 39, wherein said compound is

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

41. The compound of claim 35, wherein at least one of R^2M, R^2N, and R²⁰ is halogen, or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

42. The compound of claim 41, wherein said compound is

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

43. The compound of claim 35, wherein said compound is

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

44. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and the compound of any of claims 1-43, or a compound having the following structure

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

45. A method of treating a condition in a subject, said method comprising the step of administering the compound of any of claims 1 -43, or a compound having the following structure

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof, to said subject in a dosage sufficient to decrease necroptosis.

46. The method of claim 45, wherein said condition is a neurodegenerative disease of the central or peripheral nervous system, the result of retinal neuronal cell death, the result of cell death of cardiac muscle, the result of cell death of cells of the immune system; stroke, liver disease, pancreatic disease, the result of cell death associated with renal failure; heart, mesenteric, retinal, hepatic or brain ischemic injury, ischemic injury during organ storage, head trauma, septic shock, coronary heart disease, cardiomyopathy, myocardial infarction, bone avascular necrosis, sickle cell disease, muscle wasting, gastrointestinal disease, tuberculosis, diabetes, alteration of blood vessels, muscular dystrophy, graft-versus-host disease, viral infection, Crohn's disease, ulcerative colitis, asthma, or any condition in which alteration in cell proliferation, differentiation or intracellular signaling is a causative factor.

47. The method of claim 46, wherein said condition is a neurodegenerative disease of the central or peripheral nervous system.

48. The method of claim 46, wherein said condition is hepatic or brain ischemic injury, or ischemic injury during organ storage, head trauma, septic shock, or coronary heart disease.

49. The method of claim 46, wherein said condition is stroke.

50. The method of claim 46, wherein said condition is myocardial infarction.

51. A method of decreasing necroptosis comprising contacting a cell with the compound of any of claims 1-43, or a compound having the following structure

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof.

52. A kit comprising

(a) a pharmaceutically acceptable composition comprising the compound of any of claims 1 -43 or a compound having the following structure

or any pharmaceutically acceptable salt or solvate thereof, or stereoisomer thereof; and

(b) instructions for the use of the pharmaceutical composition of (a) to treat a condition in a subject.