WO2014152182A1 - Inhibiteurs hétérocycliques deutérés de nécroptose - Google Patents

Inhibiteurs hétérocycliques deutérés de nécroptose Download PDF

Info

Publication number
WO2014152182A1
WO2014152182A1 PCT/US2014/027045 US2014027045W WO2014152182A1 WO 2014152182 A1 WO2014152182 A1 WO 2014152182A1 US 2014027045 W US2014027045 W US 2014027045W WO 2014152182 A1 WO2014152182 A1 WO 2014152182A1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
compound
optionally substituted
alkyl
deuterium
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2014/027045
Other languages
English (en)
Inventor
Junying Yuan
Alexei Degterev
Gregory D. Cuny
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Institute of Organic Chemistry of CAS
Brigham and Womens Hospital Inc
Tufts University
Harvard University
Original Assignee
Shanghai Institute of Organic Chemistry of CAS
Brigham and Womens Hospital Inc
Tufts University
Harvard University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Institute of Organic Chemistry of CAS, Brigham and Womens Hospital Inc, Tufts University, Harvard University filed Critical Shanghai Institute of Organic Chemistry of CAS
Publication of WO2014152182A1 publication Critical patent/WO2014152182A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/42—Oxazoles
    • A61K31/422—Oxazoles not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4178—1,3-Diazoles not condensed 1,3-diazoles and containing further heterocyclic rings, e.g. pilocarpine, nitrofurantoin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms

Definitions

  • necrotic and/or necrotic pathways In many diseases, cell death is mediated through apoptotic and/or necrotic pathways. While m uch 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 apoptosis 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 (1 980)) . These two modes of cell death were also initially thought to occur via regulated (caspase-dependent) and non-regulated processes, respectively. Subsequent 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.
  • necroptosis One regulated caspase-independent cell death pathway with morphological features resembling necrosis, called necroptosis, has been described (Degterev et al., Nat. Chem. Biol. 1 :1 12 (2005)). This manner of cell death can be initiated with various stimuli (e.g., TNF-a and Fas ligand) and in an array of cell types (e.g., monocytes, fibroblasts, lymphocytes, macrophages, epithelial cells and neurons).
  • stimuli e.g., TNF-a and Fas ligand
  • 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.
  • necrostatins for anti-necroptosis therapeutics.
  • the discovery of compounds that prevent caspase- independent cell death 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.
  • the invention features new compounds, pharmaceutical compositions, kits, and methods for treating a condition in which necrosis or necroptosis is likely to play a substantial role, or those in which RIP1 and/or RI P3 protein is a contributing factor.
  • the invention features a composition that includes a compound of the formula
  • X 1 is 0 or NR 10 ;
  • each of X 2 and X 3 is, independently, 0 or S;
  • R 1 is H , D, or optionally substituted C1 -C6 alkyl ;
  • R 2 is H or D
  • each of R 3A and R 3B is, independently, H , D, or optionally substituted C1 -C6 alkyl ;
  • each of R 4 , R 5 , and R 6 is, independently, H or D ;
  • R 7 is H , D, halogen, optionally substituted C1 -C6 alkyl, or optionally substituted C1 -C6 alkoxy; each of R 8 , R 9 , and R 10 is, independently, H , D, or optionally substituted C1 -C6 alkyl ; and where at least one of R' -R 10 is D or includes a deuterium group, and
  • composition has an isotopic enrichment factor for deuterium of at least 5.
  • the invention features an isolated compound having a structure according to the following formula,
  • X 1 is 0 or NR 10 ;
  • each of X 2 and X 3 is, independently, 0 or S;
  • R 1 is H , D, or optionally substituted C1 -C6 alkyl ;
  • R 2 is H or D
  • each of R 3A and R 3B is, independently, H , D, or optionally substituted C1 -C6 alkyl ;
  • each of R 4 , R 5 , and R 6 is, independently, H or D ;
  • R 7 is H , D, halogen, optionally substituted C1 -C6 alkyl, or optionally substituted C1 -C6 alkoxy; each of R 8 , R 9 , and R 10 is, independently, H , D, or optionally substituted C1 -C6 alkyl ; and where at least one of R' -R 10 is D or includes a deuterium group, and
  • compositions and compounds described herein where said composition has an isotopic enrichment factor for deuterium of at least 5.
  • R 1 is H .
  • X 1 is 0.
  • X 1 is NR 10 .
  • R 10 is H .
  • X 2 and X 3 are both
  • one, two, or three of R 4 , R 5 , and R 6 is D.
  • one of R 4 , R 5 , and R 6 can be D, and two of R 4 , R 5 , and R 6 can be H .
  • R 4 is D
  • R 5 is D
  • R 6 is D.
  • R 2 is D.
  • R 2 is H
  • one of R and R 3B is D, or both R 3A and R 3B are D, or both R 3A and R 3B are H .
  • R 8 is H or D.
  • the optionally substituted C1 -C6 alkyl or said optionally substituted C1 -C6 alkoxy includes 0, 1 , 2, or 3 deuterium atoms.
  • R 7 is halogen, optionally substituted C1 alkyl, or optionally substituted C1 alkoxy.
  • the C1 alkyl or said C1 alkoxy includes 1 , 2, or 3 deuterium atoms.
  • R 7 is CI, CH 3 , OCH 3 , CD 3 ,
  • R 9 is optionally substituted C1 alkyl (e.g. , R 9 is CH 3 , CD 3 , CH D 2 , or CH 2 D) .
  • the compound has a structure according to one of the following formulas,
  • X 1 is N H or O
  • R 7 is halogen, optionally substituted C1 -C2 alkyl, or optionally substituted C1 -C2 alkoxy; and R 9 is optionally substituted C1 -C2 alkyl.
  • X 1 is NH .
  • X 1 is 0.
  • the optionally substituted C1 -C2 alkyl or said optionally substituted C1 -C2 alkoxy includes 0, 1 , 2, or 3 deuterium atoms.
  • R 9 is CH 3 .
  • R 9 is CD 3 .
  • R 7 is CI, CH 3 , OCH 3 , CD 3 , OCD 3 , or CF 3 .
  • R 1 -R 10 include 1 , 2, 3, 4, or 5 deuterium atoms.
  • the isotopic enrichment factor for deuterium is at least 1 0, 50, 1 00, 500, 1000, or 3000.
  • the carbon bearing the R 8 and X 1 groups has the (S)-configuration.
  • the carbon bearing the R 8 and X 1 groups has the (/ ⁇ -configuration.
  • the invention features a pharmaceutical composition that includes
  • the invention features a method of treating a condition in a subject, said method including the step of contacting any of the compositions or compounds described herein, or any pharmaceutically acceptable salt thereof, or stereoisomer thereof, to the subject in a dosage sufficient to decrease necroptosis.
  • the invention features a method of treating a condition in a subject, said method comprising the step of contacting any of the compounds (e.g., a compound according to any of formulas (l)-(VI I I)) or compositions described herein, or any pharmaceutically acceptable salt thereof, or stereoisomer thereof, to said subject in a dosage sufficient to modulate RI P1 and/or RI P3 activity, and wherein said condition is one in which RI P1 and/or RIP3 protein is a contributing factor.
  • any of the compounds e.g., a compound according to any of formulas (l)-(VI I I)
  • compositions described herein or any pharmaceutically acceptable salt thereof, or stereoisomer thereof
  • the methods of the invention can include administering to a subject any of the compounds (e.g. , a compound according to any of formulas (l)-(VI I I)) or compositions described herein, or any pharmaceutically acceptable salt thereof, or stereoisomer thereof.
  • any of the compounds e.g. , a compound according to any of formulas (l)-(VI I I)
  • compositions described herein or any pharmaceutically acceptable salt thereof, or stereoisomer thereof.
  • 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, atherosclerosis, a chronic or acute inflammatory condition, pain, or any condition in which alteration in cell proliferation , differentiation or intracellular signaling is a causative factor, or any condition where RIP1 and/or RI P3 protein is a contributing factor.
  • the condition is a neurodegenerative disease of the central or peripheral nervous system .
  • the condition is hepatic or brain ischemic injury, or ischemic injury during organ storage, head trauma, septic shock, or coronary heart disease.
  • the condition is stroke.
  • the condition is myocardial infarction.
  • the condition is pain (e.g., inflammatory pain, diabetic pain, pain associated with a burn, or pain associated with trauma) .
  • pain e.g., inflammatory pain, diabetic pain, pain associated with a burn, or pain associated with trauma.
  • the condition is atherosclerosis.
  • the condition is a chronic or acute inflammatory condition (e.g., rheumatoid arthritis, psoriasis, or Stevens-Johnson syndrome).
  • a chronic or acute inflammatory condition e.g., rheumatoid arthritis, psoriasis, or Stevens-Johnson syndrome.
  • the invention features a method of decreasing necroptosis including contacting a cell with any of the compositions or compounds described herein, or any pharmaceutically acceptable salt thereof, or stereoisomer thereof.
  • the invention features a kit that includes:
  • compositions or compounds described herein that includes any of the compositions or compounds described herein, or any pharmaceutically acceptable salt thereof, or stereoisomer thereof;
  • C 1- alkaryl is meant a 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.
  • C1 -C6 alkoxy is meant a group having the structure -O(optionally substituted C1 -C6 alkyl) , where the optionally substituted C1 -C6 alkyl may be branched, linear, or cyclic.
  • the C1 -C6 alkyl may be substituted or unsubstituted.
  • a substituted C1 -C6 alkyl can have, for example, 1 , 2, 3, 4, 5, or 6 substituents-including deuterium-located at any position.
  • Exemplary alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, tert-butoxy, and the like.
  • C2-C6 alkenyl or “alkenyl” is meant an optionally substituted unsaturated C2-C6 hydrocarbon group having one or more carbon-carbon double bonds.
  • a C2-C6 alkenyl may be linear or branched and may be unsubstituted or substituted.
  • a substituted C2-C6 alkenyl may have, for example, 1 , 2, 3, 4, 5, or 6 substituents located at any position.
  • C1 -C6 alkyl or “alkyl” is meant an optionally substituted C1 -C6 saturated hydrocarbon group.
  • An alkyl group may be linear, branched, or cyclic ("cycloalkyl”).
  • alkyl radicals include, but are not lim ited 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 Ci. 4 alkaryl groups.
  • Substituted C1 -C6 alkyl groups also encompass deuterated C1 -C6 alkyl groups (e.g., C1 -C6 alkyl groups that include 1 , 2, 3, 4, 5, or 6 deuterium atoms) .
  • C2-C6 alkynyl or “alkynyl” is meant an optionally substituted unsaturated C2-C6 hydrocarbon group having one or more carbon-carbon triple bonds.
  • exemplary C2-C6 alkynyl groups include, but are not limited to ethynyl, 1 -propynyl, and the like.
  • amino is meant a group having a structure -N R' R", where each R' and R" is selected , independently, from H, optionally substituted C1 -C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or R' and R" combine to form an optionally substituted heterocyclyl.
  • R' and R" may be unsubstituted or substituted with , for example, 1 , 2, 3, 4, 5, or 6 substituents.
  • aryl is meant is an optionally substituted C 6 -Ci 4 cyclic group with [4n + 2] ⁇ electrons in conjugation and where n is 1 , 2, or 3.
  • 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.
  • aryloxy is meant a group having the structure -O(optionally substituted aryl), where aryl is as defined herein.
  • azido is meant a group having the structure -N 3 .
  • carboxylate or “carbamoyl” is meant a group having the structure -OCON R' R" or -N R'C0 2 R", where each R' and R" is selected, independently, from H, optionally substituted C1 -C6 alkyl, optionally substituted cycloalkyi, optionally substituted heterocyclyl , optionally substituted aryl, optionally substituted heteroaryl, or R' and R" combine to form an optionally substituted heterocyclyl.
  • R' and R" may be unsubstituted or substituted with, for example, 1 , 2, 3, 4, 5, or 6 substituents.
  • carbonate is meant a group having a the structure -OC0 2 R', where R' is selected from H, optionally substituted C1 -C6 alkyl, optionally substituted cycloalkyi, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • R' is not H , R may be unsubstituted or substituted with, for example, 1 , 2, 3, 4, 5, or 6 substituents.
  • R' and R" may be unsubstituted or substituted with, for example, 1 , 2, 3, 4, 5, or 6 substituents.
  • Carboxylic group is meant a group having the structure -C0 2 R', where R' is selected from H , optionally substituted C1 -C6 alkyl, optionally substituted cycloalkyi, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • R' is not H , R may be unsubstituted or substituted with, for example, 1 , 2, 3, 4, 5, or 6 substituents.
  • cyano is meant a group having the structure -CN.
  • C3-C10 cycloalkyi or “cycloalkyi” is meant an optionally substituted, saturated or partially unsaturated 3- to 1 0-membered monocyclic or polycyclic (e.g. , bicyclic, or tricyclic) hydrocarbon ring system .
  • a cycloalkyi is polycyclic, the constituent cycloalkyi rings may be fused together, form a spirocyclic structure, or the polycyclic cycloalkyi may be a bridged cycloalkyi (e.g. , adamantyl or norbonanyl) .
  • cycloalkyls induce cyclopropyl, cyclobutyl, cyclopentyl , cyclohexyl, and cycloheptyl.
  • Cycloalkyls may be unsubstituted or substituted.
  • a substituted cycloalkyi can have, for example, 1 , 2, 3, 4, 5, or 6 substituents.
  • cycloalkenyl is meant a non-aromatic, optionally substituted 3- to 1 0-membered monocyclic or bicyclic hydrocarbon ring system having at least one carbon-carbon double bound.
  • 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.
  • D refers to deuterium .
  • a composition of the invention has a m inimum isotopic enrichment factor of at least 5 (0.075% deuterium incorporation) , e.g., at least 1 0 (0.15% deuterium incorporation) .
  • a composition has an isotopic enrichment factor of at least 50 (0.75% deuterium incorporation) , at least 1 00 (1 .5% deuterium incorporation) , at least 500 (7.5% deuterium incorporation) , at least 2000 (30% deuterium incorporation) , at least 3000 (45% deuterium incorporation), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation) , at least 5500 (82.5% deuterium incorporation) , at least 6000 (90% deuterium incorporation) , at least 6333.3 (95% deuterium incorporation) , at least 6466.7 (97% deuterium incorporation) , or at least 6600 (99% deuterium incorporation) .
  • chem ical groups, functional groups, or substituents described herein may be deuterated if the chem ical group, functional group, or substituent has -H .
  • Methods for the preparation of deuterated compounds are known in the art, and exemplary methods are described herein.
  • an effective amount or “therapeutically effective amount” of an agent 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.
  • 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.
  • ester is meant a group having a structure selected from -OCOR', where R' is selected from H , optionally substituted C1 -C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • R' is not H , R may be unsubstituted or substituted with, for example, 1 , 2, 3, 4, 5, or 6 substituents.
  • halogen or halo is meant fluorine (-F) , chlorine (-CI) , brom ine (-Br) , or iodine (-I).
  • 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.
  • oxazole benzoxazole, isoxazole, 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 (e.g, 1 ,2,3-triazine, 1 ,2,4-triazine, or 1 ,3,5-triazine) , indoles, 1 ,2,4,5-tetrazine, benzo[£>]thiophene, benzo[c]thiophene, benzofuran, isobenzofuran, and benzimidazole.
  • Heteroaryls may be unsubstidazole, pyrazole, thiazole, benzthiazole, triazole (
  • heterocyclic or “heterocyclyl” 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 .
  • polycyclic ring systems e.g., bi- and tri-cyclic ring systems
  • aryl e.g., phenyl or naphthyl
  • heteroaryl group e.g., cycloalkyl, cycloalkenyl, or heterocyclyl
  • 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.
  • the term heterocylic refers to a non-aromatic 5-, 6-, 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.
  • 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 are 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.
  • isotopic enrichment factor refers to the ratio of the isotopic abundance of a composition to the natural abundance of the specified isotope.
  • deuterium has a natural abundance of 0.01 5%.
  • a compound with , for example, 45% deuterium incorporation at a specified position has an isotopic enrichment factor of 3000 at that site relative to the natural abundance of deuterium .
  • inflammatory condition refers to medical disorders in which inflammation is a causative factor, or in which inflammation is a result (e.g., inflammatory pain associated with rheumatoid arthritis, psoriatic arthritis, psoriatic arthritis, lupus, or other diseases associated with tissue damage) .
  • Inflammatory conditions can be chronic or acute, and non-limiting causes of inflammatory conditions include pathogens (e.g. , bacterial pathogens or viral infections) , tissue injury, persistent foreign bodies, and autoimmune responses. As described herein, inflammation can be related to necrosis or necroptosis, or inflammation can be independent of necrosis or necroptosis.
  • pathogens e.g. , bacterial pathogens or viral infections
  • ketone or "acyl” is meant a group having the structure -COR', where R' is selected from H, optionally substituted C1 -C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • R' is not H , R may be unsubstituted or substituted with, for example, 1 , 2, 3, 4, 5, or 6 substituents.
  • nitro is meant a group having the structure -N0 2 .
  • 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-inflammatory in a patient.
  • Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors) , emollients, em ulsifiers, 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.
  • 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 palm itate, shellac, silicon dioxide, sodium carboxym ethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn) , stearic acid, stearic acid,
  • salts represent 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, 1 977, 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
  • 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 lim ited to ammonium , tetramethylammonium, tetraethylammonium, methylamine, dimethylam ine, trimethylam ine, triethylam ine, ethylamine and the like.
  • solvates refers to compounds that retain non-covalent associations to residual solvent molecules in the solid state.
  • 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 solvent:compound to 10:1 solvent:compound. These ratios can include a m ixture of associated solvent molecules.
  • N P N- methylpyrrolidinone
  • DMSO dimethyl sulfoxide
  • DM F ⁇ /, ⁇ /'-d
  • composition 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 adm inistration (e.g., as a cream , gel, lotion, or ointment) ; for intravenous adm inistration (e.g.
  • stereoisomer is meant a diastereomer, enantiomer, or epimer of a compound.
  • a chiral center in a compound may have the S-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, trans versus cis).
  • a substituted group may have, for example, 1 , 2, 3, 4, 5, 6, 7, 8, or 9 substituents.
  • each hydrogen in a group may be replaced by a substituent group (e.g. , perhaloalkyl groups such as -CF 3 or -CF 2 CF 3 or perhaloaryls such as -C 6 F 5 ).
  • a substituent group may itself be further substituted by replacing a hydrogen of said substituent group with another substituent group such as those described herein.
  • Substituents may be further substituted with, for example, 1 , 2, 3, 4, 5, or 6 substituents as defined herein.
  • a lower C1 -C6 alkyl or an aryl substituent group e.g., heteroaryl, phenyl, or naphthyl
  • 1 , 2, 3, 4, 5, or 6 substituents e.g., deuterium
  • heterocyclic derivatives that can inhibit tumor necrosis factor alpha (TNF-a)-induced necroptosis.
  • the heterocyclic compounds of the invention are described by, e.g. , any of Formulas (l)-(VI II) , and can inhibit TN F-a induced necroptosis in FADD-deficient variant of human Jurkat T cells.
  • Pharmaceutical compositions including the compounds of the invention are also described.
  • the invention also features kits and methods of treatment featuring the compounds and compositions of the invention.
  • the present invention features compounds, pharmaceutical compositions, kits, and methods for treating a range of conditions, e.g., those in which cell or tissue necrosis is a causative factor or result, those in which loss of proliferative capacity is a causative factor or a result, those in which cytokines of the TNFD family are a causative factor or a result, and those in which RIP1 and/or RIP3 protein is a contributing factor.
  • the compounds of the present invention can be used, for example, as therapeutics to decrease necrosis in a desired cell, to increase cell proliferation, to stimulate immune response, or to modulate inflammatory conditions.
  • the compounds of the present invention can also be used, for example, to treat conditions where necroptosis is likely to play a substantial role, including, but not lim ited to those described herein.
  • Exemplary a conditions in which the compounds of the invention can be useful for treatment include, but are not limited to: neurodegenerative diseases 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 ischem ic injury; ischemic injury during organ storage; head trauma; septic shock; coronary heart disease; cardiomyopathy; bone avascular necrosis; sickle cell disease; m uscle wasting ; gastrointestinal disease; tuberculosis; diabetes; alteration of blood vessels; muscular dystrophy; graft-versus-host disease ; viral infection; Crohn's disease; ulcerative colitis; asthma; atherosclerosis; pain (e.g.
  • inflammatory pain inflammatory pain, diabetic pain, or pain associated from trauma or burn
  • chronic or acute inflammatory conditions such as rheumatoid arthritis, psoriasis, and Stevens-Johnson syndrome
  • any condition in which cell or tissue necrosis is a causative factor or result any condition in which alteration in cell proliferation, differentiation or intracellular signaling is a causative factor
  • RIP1 and/or RIP3 protein is a contributing factor.
  • Other conditions are described herein.
  • the invention features isolated compounds that can be described generally by Form ula (I) and isotopically enriched compositions that include compounds that can be described generally by Form ula (I) .
  • Form ula (I) isotopically enriched compositions that include compounds that can be described generally by Form ula (I) .
  • X 1 is O or NR
  • each of X 2 and X 3 is, independently, 0 or S;
  • R 1 is H , D, or optionally substituted C1 -C6 alkyl ;
  • R 2 is H or D
  • each of R and R is, independently, H , D, or optionally substituted C1 -C6 alkyl ;
  • each of R 4 , R 5 , and R 6 is, independently, H or D ;
  • R 7 is H , D, halogen, optionally substituted C1 -C6 alkyl, or optionally substituted C1 -C6 alkoxy; each of R 8 , R 9 , and R 10 is, independently, H , D, or optionally substituted C1 -C6 alkyl ; and where at least one of R' -R 10 is D or includes a deuterium group.
  • compositions that include compounds according to formula (I) can have an isotopic enrichment factor for deuterium of at least 5 (e.g., at least 5, 1 0, 50, 1 00, 500, 1000, or 6000) .
  • Certain embodiments of Formula (I) include compounds according to any of Formulas (l l)-(VI I I) :
  • X 1 is N H or O
  • Fl 7 is halogen, optionally substituted C1 -C2 alkyl, or optionally substituted C1 -C2 alkoxy; and R 9 is optionally substituted C1 -C2 alkyl.
  • the carbon that bears the X 1 group has the ( ⁇ -configuration. In other embodiments, the carbon that bears the X 1 group has the (/ ⁇ -configuration.
  • Compounds of the invention may include, for example, 1 -10 deuterium atoms.
  • R 1 -R 10 can include, e.g. , 1 , 2, 3, 4, 5, or 6 deuterium groups.
  • at least one hydrogen has been replaced with deuterium ; e.g., 1 , 2, 3, 4, 5, or 6 hydrogens are replaced with deuterium.
  • all hydrogens have been replaced with deuterium .
  • a substituent is not further substituted. Additional optional substituents for various groups are described herein.
  • deuterated compounds and compositions described herein e.g., an isolated compound having a structure according to any of Formulas (l)-(VI I I) , or an isotopically enriched composition that includes a compound according to any of Formulas (l)-(VII I)
  • can show improved physicochemical properties e.g. , improved metabolic stability
  • Compounds of the invention can be formulated into pharmaceutical compositions for adm inistration to human subjects in a biologically compatible form suitable for adm inistration in vivo.
  • 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 N F1 9) , published in 1 999.
  • 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.
  • the described compounds or salts, solvates, or prodrugs thereof may be administered to or taken by 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 adm inistered by, e.g. , oral, parenteral, buccal, sublingual , nasal, rectal, patch, pump, or transdermal adm inistration and the pharmaceutical compositions form ulated 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 may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, 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.
  • 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 palm itate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn) , stearic acid, stearic acid, sucrose, talc, titanium
  • a compound of the invention may be orally administered, for example, with an inert diluent or with an assim ilable 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.
  • 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.
  • 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.
  • the form m ust be sterile and must be fluid to the extent that may be easily administered via syringe.
  • 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.
  • 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.
  • the dosage form comprises an aerosol dispenser
  • 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.
  • 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.
  • a carrier such as sugar, acacia, tragacanth, or gelatin and glycerine.
  • Compositions for rectal adm inistration 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 adm inistration, and standard pharmaceutical practice.
  • 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 adm inistration, 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.
  • dosage levels of between 0.1 ⁇ g kg to 1 00 mg/kg of body weight are administered daily as a single dose or divided into multiple doses.
  • the general dosage range is between 250 ⁇ g 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 adm inistration. 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 optim ization, 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
  • necroptosis One regulated caspase-independent cell death pathway with morphological features resembling necrosis, called necroptosis, has been described (Degterev et al., Nat. Chem. Biol. 1 :1 2 (2005)). This manner of cell death can be initiated with various stimuli (e.g., TNF-a and Fas ligand) and in an array of cell types (e.g., monocytes, fibroblasts, lymphocytes, macrophages, epithelial cells and neurons) .
  • stimuli e.g., TNF-a and Fas ligand
  • Necroptosis may represent a significant contributor to and in some cases predominant mode of cellular dem ise 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.
  • necrostatins are compounds that prevent caspase- independent cell death (e.g. , necrosis or necroptosis) for anti-necroptosis therapeutics.
  • the discovery of compounds that prevent caspase- independent cell death would also provide useful therapeutic agents for treating or preventing conditions in which necrosis occurs.
  • necrostatins can suppress necroptosis by specifically inhibiting receptor interacting protein 1 (RIP1 ) activity (e.g., Xie et al., Structure, 21 (3) :493-499, 2013).
  • RIP1 receptor interacting protein 1
  • RI P3 which is a RI P1 family member, has also been implicated in necroptosis (see, e.g. , Christofferson et al., Curr. Opin. Cell Biol. 22(2) :263-268, 2010) . Accordingly, methods by which R IP1 and/or RI P3 activity can be modulated can also be useful for the treatment of conditions in which RIP1 and/or RI P3 protein is a contributing factor.
  • the compounds and compositions disclosed herein can be used to treat disorders where necroptosis is likely to play a substantial role or where RI P1 and/or RIP3 protein is a contributing factor.
  • Exemplary conditions that can be treated using the methods described herein include: cerebral ischemia, traumatic brain injury (Gennarelli et al. In Textbook of Traumatic Brain Injury; Silver et al. , Eds.; American Psychiatric Publishing Inc. : Washington DC, 2005; p 37) , a neurodegenerative disease of the central or peripheral nervous system (Martin et al. Brain Res. Bull.
  • Atherosclerosis e.g, Lin et al., Cell Reports, 3:200-21 0, 2013
  • inflammatory conditions e.g., Wallach et al., Trends in Immunology, 32(1 1 ) :505-509, 201 1 ; Kang et al., Immunity, 38:27-40, 2013; and Chan , Cold Spring Harb. Perspect. Biol., 1 -12, 2012.
  • 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.
  • the compounds and compositions 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 in the fields of organic and medicinal chemistry.
  • Treatment may be performed alone or in conjunction with another therapy, for example in combination with apoptosis 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.
  • the compounds and compositions described herein 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 imm une 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, m uscular 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
  • Conditions in which alteration in cell proliferation, differentiation or intracellular signaling is a causative factor include cancer and infection, e.g. , by viruses (e.g. , acute, latent and persistent) , bacteria, fungi, or other microbes.
  • viruses e.g. , acute, latent and persistent
  • bacteria e.g. , fungi, or other microbes.
  • Exemplary viruses are human imm unodeficiency virus (H IV) , Epstein- Barr virus (EBV), cytomegalovirus (C V)5 human herpesviruses (HHV) , herpes simplex viruses (HSV), human T-Cell leukemia viruses (HTLV)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 lim ited to, chicken pox, Cytomegalovirus infections, genital herpes, Hepatitis B and C, influenza, and shingles.
  • Exemplary bacteria include, but are not lim ited to Campylobacter jejuni, Enterobacter species, Enterococcus faecium , Enterococcus faecalis, Escherichia coli (e.g., E. coli 0157: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 lim ited to, anthrax, cholera, diphtheria, foodborne illnesses, leprosy, meningitis, peptic ulcer disease, pneumonia, sepsis, tetanus, tuberculosis, typhoid fever, and urinary tract infection.
  • neurodegenerative diseases are Alzheimer's disease, Huntington's disease,
  • Parkinson's disease amyotrophic lateral sclerosis, H IV-associated dementia, cerebral ischemia, amyotropic lateral sclerosis, m ultiple sclerosis, Lewy body disease, Menke's disease, Wilson's disease, Creutzfeldt-Jakob disease, Fahr disease, and progressive supranuclear palsy.
  • Exemplary muscular dystrophies or related diseases are Becker's muscular dystrophy, Duchenne m uscular dystrophy, myotonic dystrophy, limb-girdle m uscular 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, AI DS, congestive heart failure, and chronic obstructive pulmonary disease, as well as include necrotizing myopathy of intensive care.
  • Exemplary neurodegenerative conditions are Alzheimer's disease, Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis, H IV-associated dementia, cerebral ischemia, amyotropic lateral sclerosis, m ultiple sclerosis, Lewy body disease, Menke's disease, Wilson's disease, Creutzfeldt-Jakob disease, Fahr disease, and progressive supranuclear palsy.
  • Exemplary muscular dystrophies or related diseases are Becker's muscular dystrophy, Duchenne m uscular dystrophy, myotonic dystrophy, limb-girdle muscular dystrophy, Landouzy-Dejerine m uscular dystrophy, facioscapulohumeral m uscular 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
  • the compounds and compositions described herein can additionally be used to boost the imm une system , whether or not the patient being treated has an imm unocompromising condition.
  • the compounds described herein can be used in a method to strengthen the imm une system during immunization, e.g., by functioning as an adjuvant, or by being combined with an adjuvant.
  • the compounds and compositions described herein can also be used to treat inflammatory conditions, which may be, e.g., chronic or acute.
  • Exemplary inflammatory conditions include: alkylosing spondylitis, arthritis (e.g., osteoarthritis, rheumatoid arthritis (RA), and psoriatic arthritis) , asthma, atherosclerosis, Crohn's disease, colitis, dermatitis, diverticulitis, fibromyalgia, hepatitis, irritable bowel syndrome (I BS), psoriasis, Stevens-Johnson syndrome, system ic lupus erythematous (SLE) , nephritis, and ulcerative colitis.
  • arthritis e.g., osteoarthritis, rheumatoid arthritis (RA), and psoriatic arthritis
  • asthma atherosclerosis
  • Crohn's disease colitis
  • dermatitis e.g., diverticulitis
  • fibromyalgia e.g., hepatitis
  • I BS irritable bowel syndrome
  • SLE system ic l
  • Still other inflammatory conditions include: immunoinflammatory disorders such as acne vulgaris; acute respiratory distress syndrome; Addison's disease; allergic rhinitis; allergic intraocular inflammatory diseases, ANCA-associated small-vessel vasculitis; ankylosing spondylitis; arthritis, asthma; atherosclerosis; atopic dermatitis; autoimmune hemolytic anemia; autoimm une hepatitis; Behcet's disease; Bell's palsy; bullous pemphigoid ; cerebral ischaem ia; chronic obstructive pulmonary disease; cirrhosis; Cogan's syndrome; contact dermatitis; COPD ; Crohn's disease; Cushing's syndrome;
  • immunoinflammatory disorders such as acne vulgaris; acute respiratory distress syndrome; Addison's disease; allergic rhinitis; allergic intraocular inflammatory diseases, ANCA-associated small-vessel vasculitis; ankylosing spondylitis; arthritis, asthma; atherosclerosis; atopic derma
  • hirsutism idiopathic cerato-scleritis; idiopathic pulmonary fibrosis; idiopathic thrombocytopenic purpura; inflammatory bowel or gastrointestinal disorders, inflammatory dermatoses; lichen planus; lupus nephritis; lymphomatous tracheobronchitis; macular edema; multiple sclerosis; myasthenia gravis; myositis;
  • osteoarthritis pancreatitis; pemphigoid gestationis; pemphigus vulgaris; polyarteritis nodosa; polymyalgia rheumatica; pruritus scroti ; pruritis/inflammation, psoriasis; psoriatic arthritis; rheumatoid arthritis;
  • glomerulosclerosis glomerulosclerosis; septic shock syndrome; shoulder tendinitis or bursitis; Sjogren's syndrome; Still's disease; stroke-induced brain cell death ; Sweet's disease; systemic lupus erythematosus; system ic sclerosis; Takayasu's arteritis; temporal arteritis ; toxic epidermal necrolysis; tuberculosis; type-1 diabetes; ulcerative colitis; uveitis; vasculitis; and Wegener's granulomatosis.
  • the compounds and compositions described herein can also be used in the treatment or prevention of pain, including nociceptive pain, inflammatory pain, functional pain and neuropathic pain, all of which may be acute or chronic.
  • the subject e.g., a human
  • the subject may be diagnosed as having peripheral diabetic neuropathy, compression neuropathy, post herpetic neuralgia, trigem inal or glossopharyngeal neuralgia, post traumatic or post surgical nerve damage, lumbar or cervical radiculopathy, AIDS neuropathy, metabolic neuropathy, drug induced neuropathy, complex regional pain syndrome, arachnoiditis, spinal cord injury, bone or joint injury, tissue injury, psoriasis, scleroderma, pruritis, cancer (e.g.
  • cardiovascular disease e.g. , myocardial infarction, angina, ischemic or thrombotic cardiovascular disease, peripheral vascular occlusive disease, or peripheral arterial occlusive disease
  • sickle cell anemia e.g. , myocardial infarction, angina, ischemic or thrombotic cardiovascular disease, peripheral vascular occlusive disease, or peripheral arterial occlusive disease
  • sickle cell anemia e.g. , myocardial infarction, angina, ischemic or thrombotic cardiovascular disease, peripheral vascular occlusive disease, or peripheral arterial occlusive disease
  • sickle cell anemia e.g. , myocardial infarction, angina, ischemic or thrombotic cardiovascular disease, peripheral vascular occlusive disease, or peripheral arterial occlusive disease
  • sickle cell anemia e.g. , myocardial infarction, angina, ischemic or thrombotic cardiovascular disease, peripheral vascular occlusive disease, or peripheral
  • the pain may be the result of or associated with trauma (e.g., a traumatic injury) , diabetes, surgery, burn of the cutaneous tissue (caused by a thermal, chem ical, or radiation stim ulus) , or a sunburn.
  • trauma e.g., a traumatic injury
  • the cutaneous tissue caused by a thermal, chem ical, or radiation stim ulus
  • a sunburn e.g., a traumatic injury
  • Additional conditions that can be treated using the compounds provided herein include those described in, e.g. : U .S. Patent Nos. 6,756,394; 7,253,201 ; 7,491 ,743; 8,1 43,300; 8,278,344; and 8,324,262; U .S. Patent Application Publication Nos. 20100087453, 2012/0309795, and 201 20122889; and International Publication Nos. WO 201 1 /133964 and WO/2012/061045; each of which is hereby incorporated by reference in its entirety.
  • treatment with the compounds and compositions described herein can be combined with therapies for the treatment of any of the conditions described herein.
  • Such treatments include surgery, radiotherapy, chemotherapy, or the administration of one or more additional compounds.
  • the compounds and compositions described herein can be adm inistered in combination with compounds that are apoptosis inhibitors, i .e. , compounds that inhibit apoptosis, including but not lim ited to reversible and irreversible caspase inhibitors.
  • apoptosis inhibitor includes zVAD, IETD, YVAD, DEVD, and LEH D.
  • the compounds of the invention are administered in combination with PARP poly(ADP-ribose) polymerase inhibitors.
  • PARP inhibitors include 6(5H)- phenanthridinone, 4-Amino-1 ,8-naphthalimide, 1 ,5-lsoquinolinediol, and 3-Am inobenzamide.
  • Src proteins are mammalian cytoplasmic tyrosine kinases that play an extensive role in signal transduction.
  • Src inhibitors include but are not lim ited to: PP1 (1 -(1 , 1 -dimethylethyl)-1 -(4-methylphenyl)- 1 H-pyrazolo[3,4-d]pyrimidin-4-am ine), PP2 (3-(4-chlorophenyl)-1 -(1 , 1 -dimethylethyl)-1 H-pyr- azolo[3,4- d]pyrim idin-4-amine) , damnacanthal (3-hydroxy-1 -methoxy-2-anthra-quinonecarboxaldehyde), and SU- 5565.
  • the methods of the invention involve, in some aspects, combinations of compounds that are inhibitors of cellular necrosis (e.g. , heterocyclic thiohydantoin, hydantoin, oxazolidinone, thioxo- oxazolidinone, pyrim idinone, or oxazinanone compounds, or combinations thereof) with agents for the treatment of cardiovascular disorders.
  • agents include anti-inflammatory agents, anti-thrombotic agents, anti-platelet agents, fibrinolytic agents, lipid reducing agents, direct thrombin inhibitors, glycoprotein II b/l lla receptor inhibitors, agents that bind to cellular adhesion molecules and inhibit the ability of white blood cells to attach to such molecules (e.g. anti-cellular adhesion molecule antibodies), calcium channel blockers, beta-adrenergic receptor blockers, cyclooxygenase-2 inhibitors, angiotensin system inhibitors, and any combinations thereof.
  • One preferred agent is aspirin.
  • Anti-inflammatory agents include alclofenac; alclometasone dipropionate; algestone acetonide; alpha amylase; amcinafal ; amcinafide; amfenac sodium ; amiprilose hydrochloride; anakinra; anirolac; anitrazafen; apazone ; balsalazide disodium ; bendazac; benoxaprofen; benzydamine hydrochloride; bromelains; broperamole; budesonide; carprofen ; cicloprofen; cintazone; cliprofen; clobetasol propionate; clobetasone butyrate; clopirac; cloticasone propionate; cormethasone acetate; cortodoxone; deflazacort; desonide; desoximetasone; dexamethasone dipropionate; diclofenac potassium ; diclofenac sodium ; dif
  • flufenamic acid flumizole; flunisolide acetate; flunixin ; flunixin meglumine; fluocortin butyl ;
  • fluorometholone acetate fluquazone; flurbiprofen; fluretofen; fluticasone propionate; furaprofen; furobufen ; halcinonide; halobetasol propionate; halopredone acetate; ibufenac; ibuprofen ; ibuprofen aluminum ; ibuprofen piconol; ilonidap; indomethacin ; indomethacin sodium ; indoprofen ; indoxole;
  • intrazole intrazole; isoflupredone acetate; isoxepac; isoxicam ; ketoprofen ; lofemizole hydrochloride; lomoxicam ; loteprednol etabonate; meclofenamate sodium ; meclofenamic acid; meclorisone dibutyrate; mefenamic acid ; mesalamine; meseclazone; methylprednisolone suleptanate; morniflumate; nabumetone; naproxen; naproxen sodium ; naproxol; nimazone; olsalazine sodium ; orgotein ; orpanoxin; oxaprozin;
  • prifelone prodolic acid ; proquazone; proxazole; proxazole citrate; rimexolone; romazarit; salcolex;
  • salnacedin salsalate; salycilates; sanguinarium chloride; seclazone; sermetacin ; sudoxicam ; sulindac; suprofen; talmetacin ; talniflumate; talosalate; tebufelone; tenidap; tenidap sodium ; tenoxicam ; tesicam ; tesim ide; tetrydamine; tiopinac; tixocortol pivalate; tolmetin ; tolmetin sodium ; triclonide; triflum idate;
  • zidometacin zidometacin; glucocorticoids; and zomepirac sodium .
  • Anti-thrombotic and fibrinolytic agents include plasm inogen (to plasmin via interactions of prekallikrein, kininogens, factors XI I, XI I la, plasminogen proactivator, and tissue plasm inogen activator
  • TPA streptokinase
  • urokinase anisoylated plasminogen-streptokinase activator complex
  • Anti-platelet agents include clopridogrel ; sulfinpyrazone; aspirin ; dipyridamole; clofibrate; pyridinol carbamate; PG E; glucagon; antiserotonin drugs; caffeine; theophyllin ; pentoxifyllin ; ticlopidine; and anagrelide.
  • Lipid reducing agents include gemfibrozil, cholystyramine, colestipol, nicotinic acid, probucol, lovastatin, fluvastatin, simvastatin, atorvastatin, pravastatin, and cirivastatin.
  • Direct thrombin inhibitors include hirudin, hirugen, hirulog, agatroban, PPACK, and thrombin aptamers.
  • Glycoprotein ⁇ bl ⁇ ⁇ a receptor inhibitors include both antibodies and non-antibodies, and include but are not limited to ReoPro (abcixamab) , lamifiban, and tirofiban.
  • Calcium channel blockers are a chemically diverse class of compounds having important therapeutic value in the control of a variety of diseases including several cardiovascular disorders, such as hypertension , angina, and cardiac arrhythmias (Fleckenstein, Cir. Res. 52 :13-16 (1 983) ; Fleckenstein, Experimental Facts and Therapeutic Prospects, John Wiley, New York (1983) ; McCall, D., Curr. Pract. Cardiol. 1 0:1 -1 1 (1 985)). Calcium channel blockers are a heterogenous group of drugs that prevent or slow the entry of calcium into cells by regulating cellular calcium channels. (Rem ington, The Science and Practice of Pharmacy, Nineteenth Edition, Mack Publishing Company, Eaton , Pa., p. 963 (1 995)) .
  • calcium channel blockers useful according to the invention include, but are not limited to, amrinone, am lodipine, bencyclane, felodipine, fendiline, flunarizine, isradipine, nicardipine, nimodipine, perhexylene, gallopamil, tiapamil and tiapamil analogues (such as 1993RO-1 1 -2933) , phenyloin, barbiturates, and the peptides dynorphin, omega-conotoxin, and omega-agatoxin, and pharmaceutically acceptable salts thereof.
  • Beta-adrenergic receptor blocking agents are a class of drugs that antagonize the cardiovascular effects of catecholam ines in angina pectoris, hypertension, and cardiac arrhythmias.
  • Beta-adrenergic receptor blockers include, but are not limited to, atenolol, acebutolol, alprenolol, befunolol, betaxolol, bunitrolol, carteolol, celiprolol, hedroxalol, indenolol, labetalol, levobunolol, mepindolol, methypranol, metindol, metoprolol , metrizoranolol, oxprenolol, pindolol, propranolol, practolol, practolol, sotalolnadolol, tiprenolol, tomalolol, ti
  • Cyclooxygenase-2 (COX-2) is an enzyme complex present in most tissues that produces various prostaglandins and thromboxanes from arachidonic acid.
  • COX-2 inhibitors include, but are not lim ited to, those described in U .S. Patent Nos.
  • COX-2 inhibitors are prodrugs of selective COX-2 inhibitors and exert their action by conversion in vivo to the active and selective COX-2 inhibitors.
  • Angiotensin system inhibitors are capable of interfering with the function , synthesis or catabolism of angiotensin I I .
  • agents include, but are not limited to, angiotensin-converting enzyme (ACE) inhibitors, angiotensin I I antagonists, angiotensin I I receptor antagonists, agents that activate the catabolism of angiotensin I I, and agents that prevent the synthesis of angiotensin I from which angiotensin I I is ultimately derived.
  • ACE angiotensin-converting enzyme
  • the renin-angiotensin system is involved in the regulation of hemodynamics and water and electrolyte balance. Factors that lower blood volume, renal perfusion pressure, or the concentration of Na + in plasma tend to activate the system , while factors that increase these parameters tend to suppress its function.
  • Angiotensin I and angiotensin I I are synthesized by the enzymatic renin-angiotensin pathway. The synthetic process is initiated when the enzyme renin acts on angiotensinogen, pseudoglobulin in blood plasma, to produce the decapeptide angiotensin I. Angiotensin I is converted by angiotensin converting enzyme (ACE) to angiotensin I I (angiotensin-[1 -8] octapeptide) . The latter is an active pressor substance which has been implicated as a causative agent in several forms of hypertension in various mammalian species, e.g., humans.
  • ACE angiotensin converting enzyme
  • Angiotensin (renin-angiotensin) system inhibitors are compounds that act to interfere with the production of angiotensin II from angiotensinogen or angiotensin I or interfere with the activity of angiotensin I I.
  • Such inhibitors are well known to those of ordinary skill in the art and include compounds that act to inhibit the enzymes involved in the ultimate production of angiotensin I I , including renin and ACE. They also include compounds that interfere with the activity of angiotensin I I, once produced.
  • classes of such compounds include antibodies (e.g. , to renin) , am ino acids and analogs thereof (including those conjugated to larger molecules), peptides (including peptide analogs of angiotensin and angiotensin I) , pro-renin related analogs, etc.
  • renin inhibitors renin inhibitors
  • ACE inhibitors angiotensin I I antagonists
  • the renin-angiotensin system inhibitors are renin inhibitors, ACE inhibitors, and angiotensin I I antagonists.
  • Angiotensin I I antagonists are compounds which interfere with the activity of angiotensin I I by binding to angiotensin I I receptors and interfering with its activity.
  • Angiotensin I I antagonists are well known and include peptide compounds and non-peptide compounds.
  • Most angiotensin I I antagonists are slightly modified congeners in which agonist activity is attenuated by replacement of phenylalanine in position 8 with some other am ino acid ; stability can be enhanced by other replacements that slow degeneration in vivo.
  • angiotensin I I antagonists include: peptidic compounds (e.g., saralasin, [(San' )(Val 5 )(Ala 8 )] angiotensin-(1 -8) octapeptide and related analogs) ; N-substituted im idazole-2-one (U.S. Patent No. 5,087,634) ; imidazole acetate derivatives including 2-N-butyl-4-chloro-1 -(2- chlorobenzile) imidazole-5-acetic acid (see Long et al. , J . Pharmacol. Exp. Ther.
  • peptidic compounds e.g., saralasin, [(San' )(Val 5 )(Ala 8 )] angiotensin-(1 -8) octapeptide and related analogs
  • N-substituted im idazole-2-one U.S. Patent No. 5,087,63
  • peptides e.g., U.S. Patent No. 4,772,684
  • antibodies to angiotensin I I e.g. , U.S. Patent No. 4,302,386
  • aralkyl im idazole compounds such as biphenyl-methyl substituted imidazoles (e.g. , EP Number 253,31 0, Jan.
  • Angiotensin converting enzyme is an enzyme which catalyzes the conversion of angiotensin I to angiotensin I I.
  • ACE inhibitors include amino acids and derivatives thereof, peptides, including di and tri peptides and antibodies to ACE which intervene in the renin-angiotensin system by inhibiting the activity of ACE thereby reducing or elim inating the formation of pressor substance angiotensin I I.
  • ACE inhibitors have been used medically to treat hypertension, congestive heart failure, myocardial infarction and renal disease.
  • Classes of compounds known to be useful as ACE inhibitors include acylmercapto and mercaptoalkanoyl prolines such as captopril (U.S. Patent No. 4, 1 05,776) and zofenopril (U .S. Patent No. 4,31 6,906), carboxyalkyl dipeptides such as enalapril (U.S. Patent No.
  • Renin inhibitors are compounds which interfere with the activity of renin. Renin inhibitors include amino acids and derivatives thereof, peptides and derivatives thereof, and antibodies to renin. Examples of renin inhibitors that are the subject of United States patents are as follows: urea derivatives of peptides (U.S. Patent No. 5,1 1 6,835) ; am ino acids connected by nonpeptide bonds (U .S. Patent No. 5,1 14,937) ; di and tri peptide derivatives (U .S. Patent No. 5, 1 06,835) ; amino acids and derivatives thereof (U .S. Patent Nos.
  • polypeptide agents that bind to cellular adhesion molecules and inhibit the ability of white blood cells to attach to such molecules include polypeptide agents.
  • polypeptides include polyclonal and monoclonal antibodies, prepared according to conventional methodology. Such antibodies already are known in the art and include anti-ICA 1 antibodies as well as other such antibodies.
  • anti-ICA 1 antibodies as well as other such antibodies.
  • the paratrope only a small portion of an antibody molecule, the paratrope, is involved in the binding of the antibody to its epitope (see, in general, Clark, W. R. (1 986) The Experimental Foundations of Modern Immunology, Wiley & Sons, Inc., New York; Roitt, I . (1 991 ) Essential Imm unology, 7th Ed. , Blackwell Scientific Publications, Oxford).
  • the pFc' and Fc regions are effectors of the complement cascade but are not involved in antigen binding.
  • an antibody from which the Fc region has been enzymatically cleaved, or which has been produced without the Fc region designated an antibody from which the Fc region has been enzymatically cleaved, or which has been produced without the Fc region, designated an Fab fragment, retains one of the antigen binding sites of an intact antibody molecule.
  • Fab fragments consist of a covalently bound antibody light chain and a portion of the antibody heavy chain denoted Fd.
  • the Fd fragments are the major determinant of antibody specificity (a single Fd Fragment may be associated with up to ten different light chains without altering antibody specificity
  • CDRs complementarity determining regions
  • Frs framework regions
  • CDR1 through CD R3 complementarity determining regions
  • non-CDR regions of a mammalian antibody may be replaced with sim ilar regions of conspecific or heterospecific antibodies while retaining the epitopic specificity of the original antibody.
  • This is most clearly manifested in the development and use of "humanized” antibodies in which non-human CDRs are covalently joined to human FR and/or Fc/pFc' regions to produce a functional antibody.
  • PCT International Publication N umber WO 92/04381 teaches the production and use of humanized murine RSV antibodies in which at least a portion of the m urine FR regions have been replaced by FR regions of human origin.
  • Such antibodies, including fragments of intact antibodies with antigen-binding ability, are often referred to as "chimeric" antibodies.
  • the present invention also provides for F(ab') 2 , Fab, Fv and Fd fragments; chimeric antibodies in which the Fc and/or Fr and/or CDR1 and/or CDR2 and/or light chain CDR3 regions have been replaced by homologous human or non-human sequences; chimeric F(ab') 2 fragment antibodies in which the FR and/or CDR1 and/or CDR2 and/or light chain CDR3 regions have been replaced by homologous human or non-human sequences; chimeric Fab fragment antibodies in which the FR and/or CDR1 and/or CDR2 and/or light chain CDR3 regions have been replaced by homologous human or non-human sequences; and chimeric Fd fragment antibodies in which the FR and/or CDR1 and/or CDR2 regions have been replaced by homologous human or nonhuman sequences.
  • the present invention also includes so-called single chain antibodies.
  • polypeptides of numerous size and type that bind specifically to cellular adhesion molecules may be derived also from sources other than antibody technology.
  • polypeptide binding agents can be provided by degenerate peptide libraries which can be readily prepared in solution , in immobilized form or as phage display libraries.
  • Combinatorial libraries also can be synthesized of peptides containing one or more am ino acids.
  • Phage display can be particularly effective in identifying binding peptides useful according to the invention. Briefly, one prepares a phage library (using, e.g., m 13, fd, or lambda phage), displaying inserts from 4 to about 80 amino acid residues using conventional procedures.
  • the inserts may represent, for example, a completely degenerate or biased array.
  • the mtrel linear portion of the sequence that binds to the cellular adhesion molecule can be determined. One can repeat the procedure using a biased library containing inserts containing part of all of the mtrel linear portion plus one or more additional degenerate residues upstream or downstream thereof. Yeast two-hybrid screening methods also may be used to identify polypeptides that bind to the cellular adhesion molecules. Thus, cellular adhesion molecules, or a fragment thereof, can be used to screen peptide libraries, including phage display libraries, to identify and select peptide binding partners of the cellular adhesion molecules.
  • any of the compounds or pharmaceutical compositions of the invention 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.
  • deuterated indole compounds can be prepared according to methods described in, e.g.: Vining et al., "Deuterium exchange labelling of biologically important phenols, indoles and steroids," Journal of Labelled Compounds and Radiopharmaceuticals, 18(1 1 ):1683-1692, 1981 ; Magnus et al., "Synthesis of 4,5,6,7 and 2,4,5,6,7 deuterium-labeled indole-3-acetic acid for use in mass spectrometric assays," Plant Physiology, 66(4):775-781 , 1980; and Kiado et al., "Synthesis of L-tryptophan labeled with hydrogen isotopes in the indole ring," Journal of Radioanalytical and Nuclear Chemistry, 279(2) :675-678, 2009.
  • Scheme 1 provides an exemplary method for the synthesis of the compounds of the invention.
  • a starting material such as toluene- ⁇ 3 ⁇ 4 (compound A) can be nitrated in order to afford compound B as a mixture of isomers.
  • Indole formation can be accomplished by treating compound B with, e.g., vinyl magnesium bromide to afford compound C.
  • Formylation under Vilsmeier-Haack conditions followed by cyanide addition can afford the C3-aminonitrile compound D.
  • Partial reduction of the nitrile followed by cyclization then affords the desired deuterated compound E.
  • Compounds such as Compound G can be prepared according to methods in the art (see, e.g. : U.S. Patent Nos. 7,491 ,743, 8,143,300, and 8,324,262; U.S. Patent Application Publication No.
  • Scheme 3 provides an exemplary synthetic scheme for compounds of the invention where, e.g.,
  • R and R are both deuterium, and R is a deuterated alkyl group.
  • necroptosis inhibitory activity can be performed using a FADD-deficient variant of human Jurkat T cells treated with TNF-a as previously described (Degterev et al., Nat. Chem. Biol.1:112 (2005) and Jagtap et al., J. Med. Chem.50: 1886 (2007)).
  • cells can be treated with 10 ng/mL of human TNF-a in the presence of increasing concentration of test compounds for 24 hours followed by ATP-based viability assessment.
  • necroptosis activity can be performed using a FADD- deficient variant of human Jurkat T cells treated with TNF-a.
  • cells 500,000 cells/mL, 100 ⁇ per well in a 96-well plate
  • 10 ng/mL of human TNF-a in the presence of increasing concentration of test compounds for 24 hours at 37 S C in a humidified incubator with 5% C0 2 followed by ATP-based viability assessment.
  • Stock solutions (30 m ) in DMSO can be prepared and then diluted with DMSO to give testing solutions, which were added to each test well. The final DMSO concentration can be 0.5%.
  • Eleven compound test concentrations (0.030 - 100 ⁇ ) can be used, and each concentration can be done in duplicate.
  • Cell viability assessments can be performed using a commercial luminescent ATP-based assay kit (CellTiter-Glo, Promega, Madison, Wl) according to the manufacturer's instructions. Briefly, 40 ⁇ of the cell lysis/ATP detection reagent can be added to each well. Plates can be incubated on a rocking platform for 10 minutes at room temperature and lum inescence was measured using a Wallac Victor 3 plate-reader (Perkin Elmer, Wellesley, MA) . Cell viability can be expressed as a ratio of the signal in the well treated with TNF-a and compound to the signal in the well treated with compound alone in order to account for nonspecific toxicity. EC 50 values can be calculated using nonlinear regression analysis of sigmoid dose-response (variable slope) curves from plots of log[l] verses viability values.
  • necroptosis inhibition can be assayed.
  • Cells are seeded at the density of 50,000 cell/well in 100 ⁇ _ of RPM I media supplemented with 1 0% Fetal Bovine Serum into 96-well plates and treated with a range of necrostatin concentrations (30 nM to 1 00 ⁇ ; 1 1 dose points) in the presence and absence of 10 ng m l "1 human TN Fa for 24 hours.
  • compound stocks in DMSO
  • compound stocks are diluted to appropriate concentrations in DMSO before addition to the cells to maintain final concentration of DMSO for all samples at 0.5%.
  • Cell viability is determined using CellTiter-Glo luminescent cell viability assay (Promega). The ratio of luminescence in compound and TN F-treated wells to compound-treated, TNF-untreated wells was calculated (viability, %) and used to calculate EC 5 o by nonlinear regression in GraphPad Prizm .
  • Sf9 cells are grown in a T1 75 flask at 27°C in Sf-900 I I SFM media. Cells are passaged (1 :3 or 1 :5) when they approach confluence or every 3 to 4 days.
  • Transfection Buffer A & B Set according to the manufacturer's protocol (BD Biosciences) is used. After 4 days, media from the 60 mm dish is transferred to a 1 5 ml conical tube and centrifuged at 1200 rpm for 5 m inutes to pellet any cells. The supernatant is transferred to a new conical tube. This is passage 1 (P1 ) baculovirus.
  • the cells are pelleted in a refrigerated centrifuge at 5,000 rpm for 15 minutes.
  • Cell lysis buffer 40 mM H EPES pH 7.3, 1 50 m M NaCI, 5 mM EDTA, 0.5 mM NaF, 0.2 mM NaV0 3 , 10 mM sodium pyrophosphate, 17.5 mM ⁇ -glycerolphosphate, 1 pg/ml aprotonin, 1 g/m l leupeptin, 1 ⁇ / ⁇ pepstatin, and 50 g/ml PMSF.
  • the cell lysate is sonicated in Sonifier Cell Disruptor 200 (Branson) using an output of 7 and 40% duty cycle for 30 seconds followed by 2 minute incubation on ice, repeated 3 times.
  • the cell lysate is centrifuged at 4 °C for 60 minutes at 12,000 rpms.
  • the filtered supernatant is loaded onto a 5 ml glutathione 4B sepharose colum n equilibrated with cell lysis buffer.
  • column is washed with at 6 colum n volumes (CVs) of GST wash buffer.
  • GST wash buffer 50 mM Tris pH 8.0, 150 mM NaCI.
  • GST-RIP1 8-327 protein is eluted with 6 CVs of GST elution buffer collecting 2 ml fractions using the ATKA purifier.
  • GST elution buffer 50 mM Tris pH 8.0, 1 50 mM NaCI, 20 mM reduced glutathione
  • SEC buffer 50 mM Tris pH 8.0, 150 mM NaCI, 2 mM ⁇ -mercaptoethanol.
  • GST-RIP1 8-327 100 nM protein, 2% DMSO, 50 ⁇ ATP, 1 ⁇ substrate 3 from HTRF KinEASE kit (Cisbio), three hour room temperature reaction, duplicate reactions.
  • Fluorescence is measured at 620 nm (cryptate) and 665 nm (XL-665) .
  • the thermal shift of recombinant RI P1 is measured in clear Light Cycler 480 m ultiwell plate 96 (Roche) with a final volume of 20 ⁇ . All components are diluted in thermal shift buffer (50 mM Tris pH 8.0, 1 50 m M NaCI, 1 mM MgCI 2 ) with final concentrations of RI P1 9 ⁇ , necrostatins 1 80 ⁇ , Sypro Orange (Life Technologies) 5X, and DMSO 3%. In duplicate wells, RIP1 and the compounds are added to the microplate and incubated at room temperature for 5 minutes.
  • Sypro Orange dye is added and the plate is measured using a LightCycler 480 instrument (Roche) with excitation at 465 nm and emission at 580 nm .
  • the protein denaturation fluorescence data is collected using melting curves analysis mode from 25 to 85 °C in continuous acquisition mode with 10 acquisitions per °C and a ramp rate of 0.06 °C/s.
  • the data is analyzed to calculate T m using nonlinear regression with Boltzmann sigmoidal equation in GraphPad 5 using approximately two degrees of pre- and post-transition baseline data.
  • Microsome stability can be determined in pooled mouse liver m icrosomes.
  • a test compound (3 DM final concentration) along with 0.5 mg/m L microsome protein and 1 mM NADPH can be incubated for 0, 5, 1 5, 30 and 60 m inutes. Incubation of test compound and microsomes in the absence of NAD PH can serve as a negative control.
  • the metabolic stability of deuterated compound H was measured and compared to the metabolic stability of compound P (Table 2) using a mouse liver microsomal assay. Data relating to inhibition of necroptosis are shown in Table 3. As shown in the table, the deuterated compound H showed both improved metabolic stability and improved potency as a necroptosis inhibitor.
  • the deuterated compounds and compositions described herein e.g., an isolated compound having a structure according to any of Formulas (l)-(VIII), or an isotopically enriched composition that includes a compound according to any of Formulas (l)-(VIII)
  • can show improved physicochemical properties e.g. , improved metabolic stability

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne des composés deutérés isolés (notamment des composés représentés par la formule (I)) et des sels pharmaceutiquement acceptables correspondants. La présente invention concerne également des compositions isotopiquement enrichies qui comprennent des composés répondant à la formule (I) et des sels pharmaceutiquement acceptables correspondants. L'invention porte également sur des compositions pharmaceutiques qui comprennent ces composés et leur usage thérapeutique pour le traitement de pathologies dans lesquelles la nécroptose joue probablement un rôle important, ou des pathologies dans lesquelles la protéine RIP1 et/ou RIP3 est un facteur contributif. (stucture indiquée)
PCT/US2014/027045 2013-03-15 2014-03-14 Inhibiteurs hétérocycliques deutérés de nécroptose Ceased WO2014152182A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201361789689P 2013-03-15 2013-03-15
US61/789,689 2013-03-15

Publications (1)

Publication Number Publication Date
WO2014152182A1 true WO2014152182A1 (fr) 2014-09-25

Family

ID=51581143

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2014/027045 Ceased WO2014152182A1 (fr) 2013-03-15 2014-03-14 Inhibiteurs hétérocycliques deutérés de nécroptose

Country Status (1)

Country Link
WO (1) WO2014152182A1 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016100166A1 (fr) * 2014-12-15 2016-06-23 Bristol-Myers Squibb Company Dihydro-1h-pyrrolo[3,2-c]pyridin-4(5h)-ones substituées en tant qu'inhibiteurs de ripk3
US9499521B2 (en) 2014-12-11 2016-11-22 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
US9586880B2 (en) 2008-12-23 2017-03-07 President And Fellows Of Harvard College Small molecule inhibitors of necroptosis
US9643977B2 (en) 2011-03-11 2017-05-09 President And Fellows Of Harvard College Necroptosis inhibitors and methods of use therefor
CN106619619A (zh) * 2017-01-06 2017-05-10 浙江大学 化合物hubin‑1在制备肝脏炎症性疾病预防和/或治疗药物中的用途
WO2017096301A1 (fr) * 2015-12-04 2017-06-08 Denali Therapeutics Inc. Inhibiteurs dérivés d'isoxazolidine de protéine kinase 1 interagissant avec un récepteur (ripk 1)
US9725452B2 (en) 2013-03-15 2017-08-08 Presidents And Fellows Of Harvard College Substituted indoles and pyrroles as RIP kinase inhibitors
WO2021075870A1 (fr) * 2019-10-17 2021-04-22 주식회사 스템모어 Composition comprenant un inhibiteur de la kinase rip, pour prévenir la chute des cheveux ou pour favoriser la repousse des cheveux
US11242318B2 (en) 2020-05-19 2022-02-08 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
EP4486743A4 (fr) * 2022-02-28 2026-01-28 Sironax Ltd Modulateurs de rip1, préparations et utilisations associées

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050119260A1 (en) * 2003-08-29 2005-06-02 Brigham And Women's Hospital, Inc. Inhibitors of cellular necrosis
US20090012112A1 (en) * 2007-02-26 2009-01-08 Concert Pharmaceuticals, Inc. Alpha 1A-adrenoceptor antagonists

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050119260A1 (en) * 2003-08-29 2005-06-02 Brigham And Women's Hospital, Inc. Inhibitors of cellular necrosis
US20090012112A1 (en) * 2007-02-26 2009-01-08 Concert Pharmaceuticals, Inc. Alpha 1A-adrenoceptor antagonists

Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9586880B2 (en) 2008-12-23 2017-03-07 President And Fellows Of Harvard College Small molecule inhibitors of necroptosis
US9643977B2 (en) 2011-03-11 2017-05-09 President And Fellows Of Harvard College Necroptosis inhibitors and methods of use therefor
US9725452B2 (en) 2013-03-15 2017-08-08 Presidents And Fellows Of Harvard College Substituted indoles and pyrroles as RIP kinase inhibitors
US10508102B2 (en) 2014-12-11 2019-12-17 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
US9499521B2 (en) 2014-12-11 2016-11-22 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
CN107530350A (zh) * 2014-12-11 2018-01-02 哈佛理事会 细胞坏死抑制剂及相关方法
US9944628B2 (en) 2014-12-11 2018-04-17 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
US20190023690A1 (en) * 2014-12-11 2019-01-24 President And Fellows Of Harvard College Inhibitors of cellular necrosis and related methods
US10301306B2 (en) 2014-12-15 2019-05-28 Bristol-Myers Squibb Company Substituted dihydro-1H-pyrrolo[3,2-c]pyridin-4(5H)-ones as RIPK3 inhibitors
WO2016100166A1 (fr) * 2014-12-15 2016-06-23 Bristol-Myers Squibb Company Dihydro-1h-pyrrolo[3,2-c]pyridin-4(5h)-ones substituées en tant qu'inhibiteurs de ripk3
CN108602809B (zh) * 2015-12-04 2022-09-30 戴纳立制药公司 异噁唑烷衍生的受体相互作用蛋白激酶1(ripk 1)的抑制剂
WO2017096301A1 (fr) * 2015-12-04 2017-06-08 Denali Therapeutics Inc. Inhibiteurs dérivés d'isoxazolidine de protéine kinase 1 interagissant avec un récepteur (ripk 1)
CN108602809A (zh) * 2015-12-04 2018-09-28 戴纳立制药公司 异噁唑烷衍生的受体相互作用蛋白激酶1(ripk 1)的抑制剂
US10709692B2 (en) 2015-12-04 2020-07-14 Denali Therapeutics Inc. Isoxazolidine derived inhibitors of receptor interacting protein kinase 1 (RIPK1)
CN106619619A (zh) * 2017-01-06 2017-05-10 浙江大学 化合物hubin‑1在制备肝脏炎症性疾病预防和/或治疗药物中的用途
WO2021075870A1 (fr) * 2019-10-17 2021-04-22 주식회사 스템모어 Composition comprenant un inhibiteur de la kinase rip, pour prévenir la chute des cheveux ou pour favoriser la repousse des cheveux
US11242318B2 (en) 2020-05-19 2022-02-08 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
US11724985B2 (en) 2020-05-19 2023-08-15 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
US11746088B2 (en) 2020-05-19 2023-09-05 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
US11834410B2 (en) 2020-05-19 2023-12-05 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
US11958807B2 (en) 2020-05-19 2024-04-16 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
US12110272B2 (en) 2020-05-19 2024-10-08 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
US12240813B2 (en) 2020-05-19 2025-03-04 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
US12291499B2 (en) 2020-05-19 2025-05-06 Cybin Irl Limited Deuterated tryptamine derivatives and methods of use
EP4486743A4 (fr) * 2022-02-28 2026-01-28 Sironax Ltd Modulateurs de rip1, préparations et utilisations associées

Similar Documents

Publication Publication Date Title
US9725452B2 (en) Substituted indoles and pyrroles as RIP kinase inhibitors
WO2014152182A1 (fr) Inhibiteurs hétérocycliques deutérés de nécroptose
CA2666060C (fr) Composes de thieno-[2,3-d]-pyrimidin-4-one et procedes d'utilisation de ceux-ci
US8324262B2 (en) Tricyclic necrostatin compounds
JP6178777B2 (ja) ネクロトーシスの小分子阻害剤
RU2593756C2 (ru) Конденсированные производные аминодигидротиазина, пригодные в качестве ингибиторов васе
US10383880B2 (en) Carbazole-containing sulfonamides as cryptochrome modulators
JP5433418B2 (ja) 多環式化合物
JP2014506564A (ja) 縮合アミノジヒドロチアジン誘導体
EA024933B1 (ru) Фенил-3-азабицикло[3.1.0]гекс-3-илметаноны и их применение в качестве лекарственного средства
AU2006206654A1 (en) Tertiary carbinamines having substituted heterocycles, which are active as inhibitors of beta-secretase, for the treatment of Alzheimer's disease
JP7071400B6 (ja) 免疫プロテアソーム阻害剤
CA2961424A1 (fr) Inhibiteurs d'une ssao derives d'une imidazo[4,5-c]pyridine
JP2018538334A (ja) Ｔｒｐｍ８拮抗薬としての４−ヒドロキシ−２−フェニル−１，３−チアゾール−５−イルメタノン誘導体

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 14769306

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 14769306

Country of ref document: EP

Kind code of ref document: A1