WO2009094205A2 - 3-aryl-6-aryl-7h-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazines and analogs as activators of caspases and inducers of apoptosis and the use thereof - Google Patents

3-aryl-6-aryl-7h-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazines and analogs as activators of caspases and inducers of apoptosis and the use thereof Download PDF

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WO2009094205A2
WO2009094205A2 PCT/US2009/000471 US2009000471W WO2009094205A2 WO 2009094205 A2 WO2009094205 A2 WO 2009094205A2 US 2009000471 W US2009000471 W US 2009000471W WO 2009094205 A2 WO2009094205 A2 WO 2009094205A2
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triazolo
thiadiazine
methoxyphenyl
methylphenyl
amino
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PCT/US2009/000471
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French (fr)
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WO2009094205A3 (en
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Sui Xiong Cai
Han-Zhong Zhang
William E. Kemnitzer
John A. Drewe
Nilantha Sudath Sirisoma
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Cytovia, Inc.
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems

Definitions

  • This invention is in the field of medicinal chemistry.
  • the invention relates to 3-aryl-6-aryl-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazines and analogs, and the discovery that these compounds are activators of caspases and inducers of apoptosis.
  • the invention also relates to the use of these compounds as therapeutically effective anti-cancer agents.
  • Organisms eliminate unwanted cells by a process variously known as regulated cell death, programmed cell death or apoptosis. Such cell death occurs as a normal aspect of animal development, as well as in tissue homeostasis and aging (Glucksmann, A., Biol. Rev. Cambridge Philos. Soc. 25:59-86 (1951); Glucksmann, A., Archives de Biologie 75:419-437 (1965); Ellis, et al, Dev. 772:591-603 (1991); Vaux, et al, Cell 76:111-119 (1994)). Apoptosis regulates cell number, facilitates morphogenesis, removes harmful or otherwise abnormal cells and eliminates cells that have already performed their function. Additionally, apoptosis occurs in response to various physiological stresses, such as hypoxia or ischemia (PCT published application WO96/20721).
  • Apoptosis is achieved through an endogenous mechanism of cellular suicide (Wyllie, A.H., in Cell Death in Biology and Pathology, Bowen and Lockshin, eds., Chapman and Hall (1981), pp. 9-34).
  • a cell activates its internally encoded suicide program as a result of either internal or external signals.
  • the suicide program is executed through the activation of a carefully regulated genetic program (Wyllie, et al, Int. Rev. Cyt. 68:25 ⁇ (1980); Ellis, et al, Ann. Rev. Cell Bio. 7:663 (1991)).
  • Apoptotic cells and bodies are usually recognized and cleared by neighboring cells or macrophages before lysis. Because of this clearance mechanism, inflammation is not induced despite the clearance of great numbers of cells (Orrenius, S., J. Internal Medicine 237:529-536 (1995)).
  • caspase family of cysteine proteases comprises 14 different members, and more may be discovered in the future. All known caspases are synthesized as zymogens that require cleavage at an aspartyl residue prior to forming the active enzyme. Thus, caspases are capable of activating other caspases, in the manner of an amplifying cascade.
  • Apoptosis and caspases are thought to be crucial in the development of cancer ⁇ Apoptosis and Cancer Chemotherapy, Hickman and Dive, eds., Humana Press (1999)).
  • cancer cells while containing caspases, lack parts of the molecular machinery that activates the caspase cascade. This makes the cancer cells lose their capacity to undergo cellular suicide and the cells become cancerous.
  • control points are known to exist that represent points for intervention leading to activation.
  • CED-9-BCL-like and CED-3-ICE-like gene family products are intrinsic proteins regulating the decision of a cell to survive or die and executing part of the cell death process itself, respectively (see, Schmitt, et al, Biochem. Cell. Biol. 75:301- 314 (1997)).
  • BCL-like proteins include BCL-xL and BAX-alpha, which appear to function upstream of caspase activation.
  • BCL-xL appears to prevent activation of the apoptotic protease cascade, whereas BAX-alpha accelerates activation of the apoptotic protease cascade.
  • chemotherapeutic drugs can trigger cancer cells to undergo suicide by activating the dormant caspase cascade. This may be a crucial aspect of the mode of action of most, if not all, known anticancer drugs (Los, et al, Blood 90:3118-3129 (1997); Friesen, et al, Nat. Med. 2:574 (1996)).
  • the mechanism of action of current antineoplastic drugs frequently involves an attack at specific phases of the cell cycle.
  • the cell cycle refers to the stages through which cells normally progress during their lifetime. Normally, cells exist in a resting phase termed G 0 . During multiplication, cells progress to a stage in which DNA synthesis occurs, termed S.
  • Antineoplastic drugs such as cytosine arabinoside, hydroxyurea, 6-mercaptopurine, and methotrexate are S phase specific, whereas antineoplastic drugs, such as vincristine, vinblastine, and paclitaxel are M phase specific.
  • Many slow growing tumors e.g. colon cancers, exist primarily in the G 0 phase, whereas rapidly proliferating normal tissues, for example bone marrow, exist primarily in the S or M phase.
  • a drug like 6-mercaptopurine can cause bone marrow toxicity while remaining ineffective for a slow growing tumor.
  • C-Myc is a proto-oncogene and encodes the c-Myc transcription factor.
  • cMyc expression correlates with cell proliferation in various cells and tissues of the body.
  • CMyc is implicated in various biological processes including cell growth, proliferation, loss of differentiation and apoptosis.
  • Deregulated expression of c-Myc occurs in a wide range of cancers and is often associated with poor prognosis suggesting an important role for this oncogene in tumor progression. Initially it was discovered in Burkitt's lymphoma as causative for the progression of the disease due to a translocation between chromosome 8 and the antibody-containing genes.
  • cMyc has been detected in a wide range of cancers that include breast, colon, cervical, small-cell lung carcinomas, osteocarcomas, glioblastomas, melanoma and myeloid leukemias (Nesbit, CE et al. Oncogene, 18, 3004-3016 (1999); Blackwood, E.M et al. Science 251, 1211-1217 (1991); Mo H. & Henriksson M. PNAS, 103: 6344-6349 (2006))
  • Inactivation of c-Myc was found to cause tumor regression with rapid proliferation arrest and apoptosis in hematopoietic malignancies and osteosarcoma. Therefore inactivating cMyc or downstream targets of cMyc may provide important therapeutic advantages.
  • the present invention is related to the discovery that 3-aryl-6-aryl-7//-
  • [l,2,4]triazolo[3,4-6][l,3,4]thiadiazines and analogs, as represented in Formulae I-V, are activators of the caspase cascade and inducers of apoptosis.
  • an aspect of the present invention is directed to the use of compounds of Formulae I-V as inducers of apoptosis.
  • a second aspect of the present invention is to provide a method for treating, preventing or ameliorating neoplasia and cancer by administering a compound of one of the Formulae I-V to a mammal in need of such treatment.
  • a third aspect of the present invention is to provide novel compounds of Formulae I-V, and to also provide for the use of these novel compounds for treating, preventing or ameliorating neoplasia and cancer.
  • a fourth aspect of the present invention is to provide a pharmaceutical composition useful for treating disorders responsive to the induction of apoptosis, containing an effective amount of a compound of one of the Formulae I-V in admixture with one or more pharmaceutically acceptable carriers or diluents.
  • a fifth aspect of the present invention is directed to methods for the preparation of novel compounds of Formulae I-V. DETAILED DESCRIPTION OF THE INVENTION
  • [l,2,4]triazolo[3,4-6][l,3,4]thiadiazines and analogs, as represented in Formulae I-V, are potent and highly efficacious activators of the caspase cascade and inducers of apoptosis. Therefore, compounds of Formulae I-V are useful for treating disorders responsive to induction of apoptosis.
  • Ar 1 and Ar 2 independently are optionally substituted carbocyclic, heterocyclic, aryl or heteroaryl;
  • X is S, O or NR 3 , wherein R 3 is hydrogen or an optionally substituted alkyl or aryl.
  • Preferred compounds of Formula I include compounds wherein Ari is phenyl, naphthyl, indolinyl, benzo[ ⁇ /][l,3]dioxol-5-yl, benzo[ ⁇ ][l,4]dioxin-6- yl, benzofuranyl, benzothiophenyl, benzimidazolyl, indolyl, imidazopyridinyl, pyridyl, quinolyl, isoquinolyl, isoxazolyl, pyrazolyl, imidazolyl, thienyl, furyl or pyrrolyl, each of which is optionally substituted.
  • Ar 2 is an optionally substituted aryl or heteroaryl. More preferably, Ar 2 is an optionally phenyl, 1,4,5,6- tetrahydrocyclopenta[c]pyrazol-3-yl, imidazopyridinyl, pyridyl, pyrazolyl, imidazolyl, thienyl, furyl or pyrrolyl.
  • Another group of preferred compounds of Formula I include compounds wherein Ri and R 2 are hydrogen.
  • Another group of preferred compounds of Formula I include compounds wherein X is S or O.
  • Another group of preferred compounds of Formula I include compounds wherein X is S.
  • Ari and Ar 2 independently are optionally substituted aryl or optionally substituted heteroaryl.
  • Preferred compounds of Formula II include compounds wherein An and Ar 2 are phenyl, naphthyl, indolinyl, benzo[d][l,3]dioxol-5-yl, benzo[6][l,4]dioxin-6-yl, benzofuranyl, benzothiophenyl, benzimidazolyl, indolyl, l,4,5,6-tetrahydrocyclopenta[c]pyrazol-3-yl, pyridyl, imidazopyridinyl, quinolyl, isoquinolyl, isoxazolyl, pyrazolyl, imidazolyl, thienyl, furyl or pyrrolyl, each of which is optionally substituted.
  • Ari is phenyl, indolinyl, benzo[ ⁇ /][l,3]dioxol-5-yl, benzo[ ⁇ ][l,4]dioxin-6-yl, benzofuranyl, benzothiophenyl, benzimidazolyl, indolyl, or pyridyl
  • Ar 2 is phenyl, pyridyl, 1,4,5,6- tetrahydrocyclopenta[c]pyrazol-3-yl, imidazopyridinyl, thienyl or furyl.
  • Ar 2 is an optionally substituted aryl or optionally substituted heteroaryl
  • R 9 -R 13 independently are hydrogen, halo, amino, di(C 1-1 o alkyl)amino, alkoxy, C MO alkyl, haloalkyl, aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl, hydroxyalkoxy, aminoalkyl, aminoalkoxy, carboxyalkyl, nitro, cyano, acylamido, hydroxy, thiol, acyloxy, azido, carboxy, carbonylamido, alkylsulfonyl, aminosulfonyl, dialkylaminosulfonyl, alkylsulfiniyl, or alkyl
  • R 9 and Rio taken together with the atoms to which they are attached to form an aryl, heteroaryl, partially saturated carbocyclic or partially saturated heterocyclic group, wherein said group is optionally substituted.
  • Another group of preferred compounds of Formula III include compounds wherein Rio or Ri 2 is an amino, and Rn is an alkyl or alkoxy.
  • Another group of preferred compounds of Formula III include compounds wherein Ar 2 is phenyl, pyridyl, l,4,5,6-tetrahydrocyclopenta[c]pyrazol-3-yl, imidazopyridinyl, thienyl, furyl or pyrrolyl. More preferably Ar 2 is phenyl, pyridyl, or furyl.
  • Especially preferred compounds of Formula III include compounds wherein Rio or R 12 is an amino, and Rj 1 is an alkoxy or methyl.
  • R 4 -Ri 3 independently are hydrogen, halo, amino, di(Ci-io alkyl)amino, alkoxy, C 1-1O alkyl, haloalkyl, aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl, hydroxyalkoxy, aminoalkyl, aminoalkoxy, carboxyalkyl, nitro, cyano, acylamido, hydroxy, thiol, acyloxy, azido, carboxy, carbonylamido, alkylsulfonyl, aminosulfonyl, dialkylaminosulfonyl, alkylsulfiniyl, or
  • Another group of preferred compounds of Formula IV include compounds wherein R 1O or Ri 2 is an amino, and Rn is an alkyl or alkoxy.
  • Another group of preferred compounds of Formula IV include compounds wherein R 4 or R 8 is subsututited. More preferably R 4 or R 8 is an alkyl, alkoxy, thioalkoxy, halo or amino.
  • Another group of preferred compounds of Formula IV include compounds wherein R 4 and R 6 are both substituted, or R 4 and R 7 are both substituted, or R 5 and R 7 are both substituted.
  • Especially preferred compounds of Formula IV include compounds wherein Rio or R 12 is an amino, and R 1 ] is an alkoxy.
  • Ar 2 is an optionally substituted aryl or optionally substituted heteroaryl
  • R 9 -Rn and R 13 independently are hydrogen, halo, amino, di(Ci -10 alkyl)amino, alkoxy, C 1- Io alkyl, haloalkyl, aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl, hydroxyalkoxy, aminoalkyl, aminoalkoxy, carboxyalkyl, nitro, cyano, acylamido, hydroxy, thiol, acyloxy, azido, carboxy, carbonylamido, alkylsulfonyl, aminosulfonyl, dialkylaminosulfonyl, alkylsulfiniyl,
  • Rj 4 is the side chain of a natural or non-natural amino acid.
  • One group of preferred compounds of Formula V include compounds wherein Ar 2 is phenyl, pyridyl, l,4,5,6-tetrahydrocyclopenta[c]pyrazol-3-yl, imidazopyridinyl, thienyl, furyl or pyrrolyl. More preferably Ar 2 is phenyl, pyridyl, or furyl.
  • Another group of preferred compounds of Formula V include compounds wherein Ri 4 is hydrogen or methyl.
  • Exemplary preferred compounds of Formulae I-V that may be employed in the method of the invention include, without limitation:
  • alkyl refers to both straight and branched chain radicals of up to ten carbons.
  • Useful alkyl groups include straight-chained and branched C 1-I o alkyl groups, more preferably Cj -6 alkyl groups.
  • Typical Ci -10 alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, /er/-butyl, 3-pentyl, hexyl and octyl groups, which may be optionally substituted.
  • amino as employed herein by itself or as part of another group is -NH 2 , -NHR 3 , or -NR 3 R b , wherein R 3 and R b are independently alkyl groups or together, with the nitrogen, form a 5 or 6 membered heterocyclo group optionally containing an additional N or O atom.
  • alkenyl as employed herein by itself or as part of another group means a straight or branched chain radical of 2-10 carbon atoms, unless the chain length is limited thereto, including at least one double bond between two of the carbon atoms in the chain.
  • Typical alkenyl groups include ethenyl, 1-propenyl, 2-propenyl, 2-methyl-l-propenyl, 1-butenyl and 2-butenyl.
  • alkynyl is used herein to mean a straight or branched chain radical of 2-10 carbon atoms, unless the chain length is limited thereto, wherein there is at least one triple bond between two of the carbon atoms in the chain.
  • Typical alkynyl groups include ethynyl, 1-propynyl, l-methyl-2- propynyl, 2-propynyl, 1-butynyl and 2-butynyl.
  • Useful alkoxy groups include oxygen substituted by one of the Ci -I0 alkyl groups mentioned above, which may be optionally substituted.
  • Alkoxy substituents include, without limitation, halo, morpholino, amino including alkylamino and dialkylamino, and carboxy including esters thereof.
  • Useful alkylthio groups include sulfur substituted by one of the Ci -I0 alkyl groups mentioned above, which may be optionally substituted. Also included are the sulfoxides and sulfones of such alkylthio groups.
  • Useful amino and optionally substituted amino groups include -NH 2 , -
  • NHRi 5 and -NRi 5 Ri 6 wherein Ri 5 and Ri 6 are Ci -10 alkyl or cycloalkyl groups, or R 15 and Ri 6 are combined with the N to form a ring structure, such as a piperidine, or Ri 5 and Ri 6 are combined with the N and other group to form a ring, such as a piperazine.
  • the alkyl group may be optionally substituted.
  • Optional substituents on the alkyl, alkoxy, alkylthio, alkenyl, alkynyl, cycloalkyl, carbocyclic and heterocyclic groups include one or more halo, hydroxy, carboxyl, amino, nitro, cyano, Ci-C 6 acylamino, Ci-C 6 acyloxy, Ci-C 6 alkoxy, aryloxy, alkylthio, C 6 -Ci 0 aryl, C 4 -C 7 cycloalkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 6 -Ci 0 aryl(C 2 -C 6 )alkenyl, C 6 -Ci 0 aryl(C 2 -C 6 )alkynyl, saturated and unsaturated heterocyclic or heteroaryl.
  • Optional substituents on the aryl, arylalkyl, arylalkenyl, arylalkynyl and heteroaryl and heteroarylalkyl groups include one or more halo, methylenedioxy, Ci-C 6 haloalkyl, C 6 -Ci 0 aryl, C 4 -C 7 cycloalkyl, Cj-C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 6 -C 10 aiyl(d-C 6 )alkyl, C 6 -Ci 0 aryl(C 2 -C 6 )alkenyl, C 6 -Ci 0 aryl(C 2 -C 6 )alkynyl, C 1 -C 6 hydroxyalkyl, nitro, amino, ureido, cyano, C 1 -C 6 acylamino, hydroxy, thiol, Ci-C
  • aryl as employed herein by itself or as part of another group refers to monocyclic, bicyclic or tricyclic aromatic groups containing from 6 to 14 carbons in the ring portion.
  • Useful aryl groups include C 6- I 4 aryl, preferably C 6-I0 aryl. Typical
  • C 6-I4 aryl groups include phenyl, naphthyl, phenanthrenyl, anthracenyl, indenyl, azulenyl, biphenyl, biphenylenyl and fluorenyl groups.
  • the term "carbocycle” as employed herein include cycloalkyl and partially saturated carbocyclic groups. Useful cycloalkyl groups are C 3-8 cycloalkyl. Typical cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
  • Useful saturated or partially saturated carbocyclic groups are cycloalkyl groups as described above, as well as cycloalkenyl groups, such as cyclopentenyl, cycloheptenyl and cyclooctenyl.
  • Useful halo or halogen groups include fluorine, chlorine, bromine and iodine.
  • arylalkyl is used herein to mean any of the above- mentioned C 1-10 alkyl groups substituted by any of the above-mentioned C 6-I4 aryl groups.
  • the arylalkyl group is benzyl, phenethyl or naphthylmethyl.
  • arylalkenyl is used herein to mean any of the above- mentioned C 2-10 alkenyl groups substituted by any of the above-mentioned C 6-I4 aryl groups.
  • arylalkynyl is used herein to mean any of the above- mentioned C 2-10 alkynyl groups substituted by any of the above-mentioned C 6-I4 aryl groups.
  • aryloxy is used herein to mean oxygen substituted by one of the above-mentioned C 6-I4 aryl groups, which may be optionally substituted.
  • Useful aryloxy groups include phenoxy and 4-methylphenoxy.
  • arylalkoxy is used herein to mean any of the above mentioned Cj -1O alkoxy groups substituted by any of the above-mentioned aryl groups, which may be optionally substituted.
  • Useful arylalkoxy groups include benzyloxy and phenethyloxy.
  • Useful haloalkyl groups include Ci -I0 alkyl groups substituted by one or more fluorine, chlorine, bromine or iodine atoms, e.g., fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, 1,1-difluoroethyl, chloromethyl, chlorofluoromethyl and trichloromethyl groups.
  • Useful acylamino (acylamido) groups are any Ci -6 acyl (alkanoyl) attached to an amino nitrogen, e.g., acetamido, chloroacetamido, propionamido, butanoylamido, pentanoylamido and hexanoylamido, as well as aryl-substituted Ci -6 acylamino groups, e.g., benzoylamido, and pentafluorobenzoylamido .
  • Useful acyloxy groups are any Cj -6 acyl (alkanoyl) attached to an oxy
  • (-O-) group e.g., formyloxy, acetoxy, propionoyloxy, butanoyloxy, pentanoyloxy and hexanoyloxy.
  • heterocycle is used herein to mean a saturated or partially saturated 3-7 membered monocyclic, or 7-10 membered bicyclic ring system, which consists of carbon atoms and from one to four heteroatoms independently selected from the group consisting of O, N, and S, wherein the nitrogen and sulfur heteroatoms can be optionally oxidized, the nitrogen can be optionally quaternized, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring, and wherein the heterocyclic ring can be substituted on carbon or on a nitrogen atom if the resulting compound is stable.
  • Useful saturated or partially saturated heterocyclic groups include tetrahydrofuranyl, pyranyl, piperidinyl, piperazinyl, pyrrolidinyl, imidazolidinyl, imidazolinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl, isochromanyl, chromanyl, pyrazolidinyl pyrazolinyl, tetronoyl and tetramoyl groups.
  • heteroaryl refers to groups having 5 to
  • Useful heteroaryl groups include thienyl (thiophenyl), benzo[6]thienyl, naphtho[2,3-6]thienyl, thianthrenyl, furyl (furanyl), pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxanthiinyl, pyrrolyl, including without limitation pyrrolyl, including 7H-pyrrolyl, 2H-pyrrolyl, and 3H-pyrrolyl, imidazolyl, pyrazolyl, pyridyl (pyridinyl), including without limitation 2-pyridyl, 3- pyridyl, and 4-pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quino
  • heteroaryl group contains a nitrogen atom in a ring
  • nitrogen atom may be in the form of an N-oxide, e.g., a pyridyl N-oxide, pyrazinyl N-oxide and pyrimidinyl N-oxide.
  • heteroaryloxy is used herein to mean oxygen substituted by one of the above-mentioned heteroaryl groups, which may be optionally substituted.
  • Useful heteroaryloxy groups include pyridyloxy, pyrazinyloxy, pyrrolyloxy, pyrazolyloxy, imidazolyloxy and thiophenyloxy.
  • heteroarylalkoxy is used herein to mean any of the above- mentioned C J-IO alkoxy groups substituted by any of the above-mentioned heteroaryl groups, which may be optionally substituted.
  • Some of the compounds of the present invention may exist as stereoisomers including optical isomers.
  • the invention includes all stereoisomers and both the racemic mixtures of such stereoisomers as well as the individual enantiomers that may be separated according to methods that are well known to those of ordinary skill in the art.
  • Examples of pharmaceutically acceptable addition salts include inorganic and organic acid addition salts, such as hydrochloride, hydrobromide, phosphate, sulphate, citrate, lactate, tartrate, maleate, fumarate, mandelate and oxalate; and inorganic and organic base addition salts with bases, such as sodium hydroxy, Tris(hydroxymethyl)aminomethane (TRIS, tromethane) and iV-methyl-glucamine.
  • inorganic and organic acid addition salts such as hydrochloride, hydrobromide, phosphate, sulphate, citrate, lactate, tartrate, maleate, fumarate, mandelate and oxalate
  • bases such as sodium hydroxy, Tris(hydroxymethyl)aminomethane (TRIS, tromethane) and iV-methyl-glucamine.
  • Examples of prodrugs of the compounds of the invention include the simple esters of carboxylic acid containing compounds (e.g., those obtained by condensation with a Ci -4 alcohol according to methods known in the art); esters of hydroxy containing compounds (e.g., those obtained by condensation with a Ci -4 carboxylic acid, C 3-6 dioic acid or anhydride thereof, such as succinic and fumaric anhydrides according to methods known in the art); imines of amino containing compounds (e.g., those obtained by condensation with a C i- 4 aldehyde or ketone according to methods known in the art); carbamate of amino containing compounds, such as those described by Leu, et. al, (J. Med. Chem.
  • Amino acid protecting groups are well known and include, for example, t-BOC and FMOC groups.
  • the compounds of this invention may be prepared using methods known to those skilled in the art, or the novel methods of this invention.
  • the compounds of this invention with Formulae I-V can be prepared as illustrated by the exemplary reaction in Scheme 1.
  • Reaction of 4- amino-5-(2-methoxyphenyl)-3-mercapto-(4H)-l ,2,4-triazole with 2-bromo- 1 - (4-methoxy-3-nitrophenyl)ethanone in isopropanol produced 3-(2- methoxyphenyl)-6-(4-methoxy-3 -nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,A- 6][l,3,4]thiadiazine.
  • the nitro group was reduced by treatment with tin (II) chloride dihydrate to produce the amino compound 6-(3-amino-4- methoxyphenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- ⁇ ][l,3,4]thiadiazine.
  • the HCl salt was prepared via treatment with HCl in ether/methanol.
  • An amino acid prodrug of compounds of this invention can be prepared as illustrated by the exemplary reaction in Scheme 3.
  • An important aspect of the present invention is the discovery that compounds having Formulae I-V are activators of caspases and inducers of apoptosis. Therefore, these compounds are useful in a variety of clinical conditions in which there is uncontrolled cell growth and spread of abnormal cells, such as in the case of cancer.
  • Another important aspect of the present invention is the discovery that compounds having Formulae I-V are potent and highly efficacious activators of caspases and inducers of apoptosis in drug resistant cancer cells, such as breast and prostate cancer cells, which enables these compounds to kill these drug resistant cancer cells.
  • drug resistant cancer cells such as breast and prostate cancer cells
  • most standard anti-cancer drugs are not effective in killing drug resistant cancer cells under the same conditions. Therefore, compounds of this invention are useful for the treatment of drug resistant cancer, such as breast cancer in animals.
  • the present invention includes a therapeutic method useful to modulate in vivo apoptosis or in vivo neoplastic disease, comprising administering to a subject in need of such treatment an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis.
  • the present invention also includes a therapeutic method comprising administering to an animal an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of said compound of Formulae I- V, wherein said therapeutic method is useful to treat cancer, which is a group of diseases characterized by the uncontrolled growth and spread of abnormal cells.
  • Such diseases include, but are not limited to, Hodgkin's disease, non- Hodgkin's lymphoma, acute lymphocytic leukemia, chronic lymphocytic leukemia, multiple myeloma, neuroblastoma, breast carcinoma, ovarian carcinoma, lung carcinoma, Wilms 1 tumor, cervical carcinoma, testicular carcinoma, soft-tissue sarcoma, primary macroglobulinemia, bladder carcinoma, chronic granulocytic leukemia, primary brain carcinoma, malignant melanoma, small-cell lung carcinoma, stomach carcinoma, colon carcinoma, malignant pancreatic insulinoma, malignant carcinoid carcinoma, choriocarcinoma, mycosis fungoides, head or neck carcinoma, osteogenic sarcoma, pancreatic carcinoma, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary carcinoma, thyroid carcinoma,
  • compositions containing therapeutically effective concentrations of the compounds formulated for oral, intravenous, local and topical application, for the treatment of neoplastic diseases and other diseases in which caspase cascade mediated physiological responses are implicated are administered to an individual exhibiting the symptoms of one or more of these disorders.
  • the amounts are effective to ameliorate or eliminate one or more symptoms of the disorders.
  • An effective amount of a compound for treating a particular disease is an amount that is sufficient to ameliorate, or in some manner reduce, the symptoms associated with the disease.
  • Such amount may be administered as a single dosage or may be administered according to a regimen, whereby it is effective.
  • the amount may cure the disease but, typically, is administered in order to ameliorate the symptoms of the disease. Typically, repeated administration is required to achieve the desired amelioration of symptoms.
  • a pharmaceutical composition comprising a compound, or a pharmaceutically acceptable salt of said compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis in combination with a pharmaceutically acceptable vehicle is provided.
  • Another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of said compound of Formulae I- V, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known cancer chemotherapeutic agent, or a pharmaceutically acceptable salt of said agent.
  • cancer chemotherapeutic agents which may be used for combination therapy include, but not are limited to alkylating agents, such as busulfan, cis-platin, mitomycin C, and carboplatin; antimitotic agents, such as colchicine, vinblastine, paclitaxel, and docetaxel; topo I inhibitors, such as camptothecin and topotecan; topo II inhibitors, such as doxorubicin and etoposide; RNA/DNA antimetabolites, such as 5-azacytidine, 5-fluorouracil and methotrexate; DNA antimetabolites, such as 5-fluoro-2'-deoxy-uridine, ara-C, hydroxyurea and thioguanine; antibodies, such as campath, Herceptin® or Rituxan®.
  • alkylating agents such as busulfan, cis-platin, mitomycin C, and carboplatin
  • antimitotic agents such as colchicine, vinblastine, paclitaxel
  • cancer chemotherapeutic agents which may be used for combination therapy include melphalan, chlorambucil, cyclophosamide, ifosfamide, vincristine, mitoguazone, epirubicin, aclarubicin, bleomycin, mitoxantrone, elliptinium, fludarabine, octreotide, retinoic acid, tamoxifen, Gleevec® and alanosine.
  • the compound of the invention may be administered together with at least one known chemotherapeutic agent as part of a unitary pharmaceutical composition.
  • the compound of the invention may be administered apart from at least one known cancer chemotherapeutic agent.
  • the compound of the invention and at least one known cancer chemotherapeutic agent are administered substantially simultaneously, i.e. the compounds are administered at the same time or one after the other, so long as the compounds reach therapeutic levels in the blood at the same time.
  • the compound of the invention and at least one known cancer chemotherapeutic agent are administered according to their individual dose schedule, so long as the compounds reach therapeutic levels in the blood.
  • alpha- 1 -adrenoceptor antagonists such as doxazosin, terazosin, and tamsulosin can inhibit the growth of prostate cancer cell via induction of apoptosis (Kyprianou, N., et ah, Cancer Res 60:4550- 4555, (2000)).
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known alpha- 1 -adrenoceptor antagonists, or a pharmaceutically acceptable salt of said agent.
  • known alpha- 1 -adrenoceptor antagonists which can be used for combination therapy include, but are not limited to, doxazosin, terazosin, and tamsulosin.
  • sigma-2 receptors are expressed in high densities in a variety of tumor cell types (Vilner, B. J., et al., Cancer Res. 55: 408-413 (1995)) and that sigma-2 receptor agonists, such as CB-64D, CB- 184 and haloperidol activate a novel apoptotic pathway and potentiate antineoplastic drugs in breast tumor cell lines. (Kyprianou, N., et al., Cancer Res. (52:313-322 (2002)).
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known sigma-2 receptor agonist, or a pharmaceutically acceptable salt of said agonist.
  • known sigma-2 receptor agonists which can be used for combination therapy include, but are not limited to, CB-64D, CB-184 and haloperidol.
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known HMG-CoA reductase inhibitor, or a pharmaceutically acceptable salt of said agent.
  • HMG-CoA reductase inhibitors which can be used for combination therapy include, but are not limited to, lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin and cerivastatin.
  • Another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known HIV protease inhibitor, or a pharmaceutically acceptable salt of said agent.
  • HIV protease inhibitors which can be used for combination therapy include, but are not limited to, amprenavir, abacavir, CGP-73547, CGP-61755, DMP-450, indinavir, nelfinavir, tipranavir, ritonavir, saquinavir, ABT-378, AG 1776, and BMS-232,632.
  • retinoids such as fenretinide (N-(4- hydroxyphenyl)retinamide, 4HPR)
  • fenretinide N-(4- hydroxyphenyl)retinamide, 4HPR
  • 4HPR also was reported to have good activity in combination with gamma-radiation on bladder cancer cell lines (Zou, C, et al, Int. J. Oncol. 75:1037-1041 (1998)).
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known retinoid and synthetic retinoid, or a pharmaceutically acceptable salt of said agent.
  • retinoids and synthetic retinoids which can be used for combination therapy include, but are not limited to, bexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid, ⁇ -difluoromethylornithine, ILX23-7553, fenretinide, and N-4-carboxyphenyl retinamide.
  • proteasome inhibitors such as lactacystin
  • lactacystin exert anti-tumor activity in vivo and in tumor cells in vitro, including those resistant to conventional chemotherapeutic agents.
  • proteasome inhibitors may also prevent angiogenesis and metastasis in vivo and further increase the sensitivity of cancer cells to apoptosis (Almond, J. B., et al., Leukemia 75:433-443 (2002)).
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known proteasome inhibitor, or a pharmaceutically acceptable salt of said agent.
  • known proteasome inhibitors which can be used for combination therapy include, but are not limited to, lactacystin, MG- 132, and PS-341.
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known tyrosine kinase inhibitor, or a pharmaceutically acceptable salt of said agent.
  • tyrosine kinase inhibitors which can be used for combination therapy include, but are not limited to, Gleevec®, ZDl 839 (Iressa), SH268, genistein, CEP2563, SU6668, SUl 1248, and EMD121974.
  • prenyl-protein transferase inhibitors such as farnesyl protein transferase inhibitor Rl 15777
  • Rl 15777 preclinical antitumor activity against human breast cancer
  • Synergy of the protein farnesyltransferase inhibitor SCH66336 and cisplatin in human cancer cell lines also has been reported (Adjei, A. A., et al, Clin. Cancer. Res. 7:1438-1445 (2001)).
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known prenyl-protein transferase inhibitor, including farnesyl protein transferase inhibitor, inhibitors of geranylgeranyl-protein transferase type I (GGPTase-I) and geranylgeranyl-protein transferase type-II, or a pharmaceutically acceptable salt of said agent.
  • known prenyl- protein transferase inhibitors which can be used for combination therapy include, but are not limited to, Rl 15777, SCH66336, L-778,123, BAL9611 and TAN-1813.
  • CDK cyclin-dependent kinase
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known cyclin-dependent kinase inhibitor, or a pharmaceutically acceptable salt of said agent.
  • known cyclin-dependent kinase inhibitor which can be used for combination therapy include, but are not limited to, flavopiridol, UCN-01, roscovitine and olomoucine.
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known COX-2 inhibitor, or a pharmaceutically acceptable salt of said inhibitor.
  • known COX-2 inhibitors which can be used for combination therapy include, but are not limited to, celecoxib, valecoxib, and rofecoxib.
  • Another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a bioconjugate of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in bioconjugation with at least one known therapeutically useful antibody, such as Herceptin ® or Rituxan ® , growth factors, such as DGF, NGF; cytokines, such as IL-2, IL-4, or any molecule that binds to the cell surface.
  • the antibodies and other molecules will deliver a compound described herein to its targets and make it an effective anticancer agent.
  • the bioconjugates could also enhance the anticancer effect of therapeutically useful antibodies, such as Herceptin ® or Rituxan ® .
  • another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with radiation therapy.
  • the compound of the invention may be administered at the same time as the radiation therapy is administered or at a different time.
  • Yet another embodiment of the present invention is directed to a composition effective for post-surgical treatment of cancer, comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis.
  • the invention also relates to a method of treating cancer by surgically removing the cancer and then treating the animal with one of the pharmaceutical compositions described herein.
  • a wide range of immune mechanisms operates rapidly following exposure to an infectious agent. Depending on the type of infection, rapid clonal expansion of the T and B lymphocytes occurs to combat the infection.
  • the elimination of the effector cells following an infection is one of the major mechanisms for maintaining immune homeostasis.
  • the elimination of the effector cells has been shown to be regulated by apoptosis.
  • Autoimmune diseases have lately been determined to occur as a consequence of deregulated cell death.
  • the immune system directs its powerful cytotoxic effector mechanisms against specialized cells, such as oligodendrocytes in multiple sclerosis, the beta cells of the pancreas in diabetes mellitus, and thyrocytes in Hashimoto's thyroiditis (Ohsako, S. & Elkon, K.B., Cell Death Differ. 6:13-21 (1999)).
  • specialized cells such as oligodendrocytes in multiple sclerosis, the beta cells of the pancreas in diabetes mellitus, and thyrocytes in Hashimoto's thyroiditis (Ohsako, S. & Elkon, K.B., Cell Death Differ. 6:13-21 (1999)).
  • lymphocyte apoptosis receptor Fas/APO-l/CD95 are reported to be associated with defective lymphocyte apoptosis and autoimmune lymphoproliferative syndrome (ALPS), which is characterized by chronic, histologically benign splenomegaly, generalized lymphadenopathy, hypergammaglobulinemia, and autoantibody formation.
  • APS autoimmune lymphoproliferative syndrome
  • Fas-Fas ligand (FasL) interaction is known to be required for the maintenance of immune homeostasis.
  • Experimental autoimmune thyroiditis (EAT) characterized by autoreactive T and B cell responses and a marked lymphocytic infiltration of the thyroid, is a good model to study the therapeutic effects of FasL. Batteux, F., et al, (J. Immunol. 752:603-608 (1999)) reported that by direct injection of DNA expression vectors encoding FasL into the inflamed thyroid, the development of lymphocytic infiltration of the thyroid was inhibited and induction of infiltrating T cells death was observed. These results show that FasL expression on thyrocytes may have a curative effect on ongoing EAT by inducing death of pathogenic autoreactive infiltrating T lymphocytes.
  • Bisindolylmaleimide VIII is known to potentiate Fas-mediated apoptosis in human astrocytoma 132 INl cells and in Molt-4T cells; both of which were resistant to apoptosis induced by anti-Fas antibody in the absence of bisindolylmaleimide VIII. Potentiation of Fas-mediated apoptosis by bisindolylmaleimide VIII was reported to be selective for activated, rather than non-activated, T cells, and was Fas-dependent. Zhou T., et al, (Nat. Med.
  • Psoriasis is a chronic skin disease that is characterized by scaly red patches.
  • Psoralen plus ultraviolet A (PUVA) is a widely used and effective treatment for psoriasis vulgaris.
  • an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis is an effective treatment for hyperproliferative skin diseases, such as psoriasis.
  • Synovial cell hyperplasia is a characteristic of patients with rheumatoid arthritis (RA). It is believed that excessive proliferation of RA synovial cells, as well as defects in synovial cell death, may be responsible for synovial cell hyperplasia. Wakisaka, et al., Clin. Exp. Immunol. 774:119-128 (1998), found that although RA synovial cells could die via apoptosis through a Fas/FasL pathway, apoptosis of synovial cells was inhibited by proinflammatory cytokines present within the synovium. Wakisaka, et al.
  • an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis is an effective treatment for rheumatoid arthritis.
  • an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis, is an effective treatment for inflammation.
  • Caspase cascade activators and inducers of apoptosis may also be a desirable therapy in the elimination of pathogens, such as HIV, Hepatitis C and other viral pathogens.
  • pathogens such as HIV, Hepatitis C and other viral pathogens.
  • the long lasting quiescence, followed by disease progression, may be explained by an anti-apoptotic mechanism of these pathogens leading to persistent cellular reservoirs of the virions. It has been reported that HIV-I infected T leukemia cells or peripheral blood mononuclear cells (PBMCs) underwent enhanced viral replication in the presence of the caspase inhibitor Z-VAD-fmk.
  • PBMCs peripheral blood mononuclear cells
  • Z-VAD-fmk also stimulated endogenous virus production in activated PBMCs derived from HIV-I- infected asymptomatic individuals (Chinnaiyan, A., et al., Nat. Med. J:333 (1997)). Therefore, apoptosis serves as a beneficial host mechanism to limit the spread of HIV and new therapeutics using caspase/apoptosis activators are useful to clear viral reservoirs from the infected individuals.
  • HCV infection also triggers anti-apoptotic mechanisms to evade the host's immune surveillance leading to viral persistence and hepatocarcinogenesis (Tai, D.I., et al. Hepatology 3:656-64 (2000)).
  • apoptosis inducers are useful as therapeutics for HIV and other infectious disease.
  • Stent implantation has become the new standard angioplasty procedure.
  • in-stent restenosis remains the major limitation of coronary stenting.
  • New approaches have been developed to target pharmacological modulation of local vascular biology by local administration of drugs. This allows for drug applications at the precise site and time of vessel injury.
  • Numerous pharmacological agents with antiproliferative properties are currently under clinical investigation, including actinomycin D, rapamycin or paclitaxel coated stents (Regar E., et al., Br. Med. Bull. 59:227- 248 (2001)). Therefore, apoptosis inducers, which are antiproliferative, are useful as therapeutics for the prevention or reduction of in-stent restenosis.
  • compositions within the scope of this invention include all compositions wherein the compounds of the present invention are contained in an amount that is effective to achieve its intended purpose. While individual needs vary, determination of optimal ranges of effective amounts of each component is within the skill of the art.
  • the compounds may be administered to animals, e.g., mammals, orally at a dose of 0.0025 to 50 mg/kg of body weight, per day, or an equivalent amount of the pharmaceutically acceptable salt thereof, to a mammal being treated. Preferably, approximately 0.01 to approximately 10 mg/kg of body weight is orally administered.
  • the dose is generally approximately one-half of the oral dose.
  • a suitable intramuscular dose would be approximately 0.0025 to approximately 25 mg/kg of body weight, and most preferably, from approximately 0.01 to approximately 5 mg/kg of body weight.
  • a known cancer chemotherapeutic agent is also administered, it is administered in an amount that is effective to achieve its intended purpose.
  • the amounts of such known cancer chemotherapeutic agents effective for cancer are well known to those skilled in the art.
  • the unit oral dose may comprise from approximately 0.01 to approximately 50 mg, preferably approximately 0.1 to approximately 10 mg of the compound of the invention.
  • the unit dose may be administered one or more times daily, as one or more tablets, each containing from approximately 0.1 to approximately 10 mg, conveniently approximately 0.25 to 50 mg of the compound or its solvates.
  • the compound In a topical formulation, the compound may be present at a concentration of approximately 0.01 to 100 mg per gram of carrier.
  • the compounds of the invention may be administered as part of a pharmaceutical preparation containing suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the compounds into preparations that may be used pharmaceutically.
  • suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the compounds into preparations that may be used pharmaceutically.
  • the preparations particularly those preparations which may be administered orally and that may be used for the preferred type of administration, such as tablets, dragees, and capsules, and also preparations that may be administered rectally, such as suppositories, as well as suitable solutions for administration by injection or orally, contain from approximately 0.01 to 99 percent, preferably from approximately 0.25 to 75 percent of active compound(s), together with the excipient.
  • nontoxic pharmaceutically acceptable salts of the compounds of the present invention are included within the scope of the present invention.
  • Acid addition salts are formed by mixing a solution of the compounds of the present invention with a solution of a pharmaceutically acceptable non-toxic acid, such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid, and the like.
  • Basic salts are formed by mixing a solution of the compounds of the present invention with a solution of a pharmaceutically acceptable non-toxic base, such as sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, Tris, N-methyl- glucamine and the like.
  • compositions of the invention may be administered to any animal, which may experience the beneficial effects of the compounds of the invention.
  • animals are mammals, e.g., humans and veterinary animals, although the invention is not intended to be so limited.
  • compositions of the present invention may be administered by any means that achieve their intended purpose.
  • administration may be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal or topical routes.
  • administration may be by the oral route.
  • the dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment, and the nature of the effect desired.
  • compositions of the present invention are manufactured in a manner, which is itself known, e.g., by means of conventional mixing, granulating, dragee-making, dissolving, or lyophilizing processes.
  • pharmaceutical preparations for oral use may be obtained by combining the active compounds with solid excipients, optionally grinding the resulting mixture and processing the mixture of granules, after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores.
  • Suitable excipients are, in particular: fillers, such as saccharides, e.g. lactose or sucrose, mannitol or sorbitol; cellulose preparations and/or calcium phosphates, e.g. tricalcium phosphate or calcium hydrogen phosphate; as well as binders, such as starch paste, using, e.g., maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone.
  • fillers such as saccharides, e.g. lactose or sucrose, mannitol or sorbitol
  • cellulose preparations and/or calcium phosphates e.g. tricalcium phosphate or calcium hydrogen phosphate
  • binders such as starch paste, using, e.g., maize starch, wheat starch, rice starch, potato
  • disintegrating agents may be added, such as the above-mentioned starches and also carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate.
  • Auxiliaries are, above all, flow-regulating agents and lubricants, e.g., silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium stearate, and/or polyethylene glycol.
  • Dragee cores are provided with suitable coatings which, if desired, are resistant to gastric juices.
  • concentrated saccharide solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures.
  • suitable cellulose preparations such as acetylcellulose phthalate or hydroxypropylmethyl-cellulose phthalate, are used.
  • Dye stuffs or pigments may be added to the tablets or dragee coatings, e.g., for identification or in order to characterize combinations of active compound doses.
  • Other pharmaceutical preparations which may be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
  • the push-fit capsules may contain the active compounds in the form of: granules, which may be mixed with fillers, such as lactose; binders, such as starches; and/or lubricants, such as talc or magnesium stearate and, optionally, stabilizers.
  • the active compounds are preferably dissolved or suspended in suitable liquids, such as fatty oils, or liquid paraffin.
  • suitable liquids such as fatty oils, or liquid paraffin.
  • stabilizers may be added.
  • Possible pharmaceutical preparations which may be used rectally include, e.g., suppositories, which consist of a combination of one or more of the active compounds with a suppository base.
  • Suitable suppository bases are, e.g., natural or synthetic triglycerides, or paraffin hydrocarbons.
  • gelatin rectal capsules which consist of a combination of the active compounds with a base.
  • Possible base materials include, e.g., liquid triglycerides, polyethylene glycols, or paraffin hydrocarbons.
  • Suitable formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form, e.g., water-soluble salts and alkaline solutions.
  • suspensions of the active compounds as appropriate oily injection suspensions may be administered.
  • Suitable lipophilic solvents or vehicles include fatty oils, e.g., sesame oil, or synthetic fatty acid esters, e.g., ethyl oleate or triglycerides or polyethylene glycol-400 (the compounds are soluble in PEG-400), or cremophor, or cyclodextrins.
  • Aqueous injection suspensions may contain substances which increase the viscosity of the suspension include, e.g., sodium carboxymethyl cellulose, sorbitol, and/or dextran.
  • the suspension may also contain stabilizers.
  • compounds of the invention are employed in topical and parenteral formulations and are used for the treatment of skin cancer.
  • the topical compositions of this invention are formulated preferably as oils, creams, lotions, ointments and the like by choice of appropriate carriers.
  • Suitable carriers include vegetable or mineral oils, white petrolatum (white soft paraffin), branched chain fats or oils, animal fats and high molecular weight alcohol (greater than Ci 2 ).
  • the preferred carriers are those in which the active ingredient is soluble.
  • Emulsifiers, stabilizers, humectants and antioxidants may also be included, as well as agents imparting color or fragrance, if desired.
  • transdermal penetration enhancers may be employed in these topical formulations. Examples of such enhancers are found in U.S. Patent Nos. 3,989,816 and 4,444,762.
  • Creams are preferably formulated from a mixture of mineral oil, self- emulsifying beeswax and water in which mixture of the active ingredient, dissolved in a small amount of an oil, such as almond oil, is admixed.
  • An oil such as almond oil
  • a typical example of such a cream is one which includes approximately 40 parts water, approximately 20 parts beeswax, approximately 40 parts mineral oil and approximately 1 part almond oil.
  • Ointments may be formulated by mixing a solution of the active ingredient in a vegetable oil, such as almond oil, with warm soft paraffin and allowing the mixture to cool.
  • a vegetable oil such as almond oil
  • a typical example of such an ointment is one which includes approximately 30 % almond oil and approximately 70 % white soft paraffin by weight.
  • the black solution was heated at 60 0 C for 4 h and then cooled to room temperature.
  • the solution was diluted with cold water (10 mL) then extracted with CHCl 3 (100 mL).
  • the organic layer was washed with saturated aqueous NaHCO 3 (20 mL), water (20 mL), brine (20 mL), dried over MgSO 4 , filtered and concentrated to give a brown oil. It was purified by flash chromatography (gradient elution: hexanes, EtOAc; 1 :1 to 100% EtOAc) to give 0.017 g (5%) of the title compound as a yellow solid.
  • reaction solution was diluted with CHCI 3 (100 mL) and the organic layer was then washed with saturated NaHCO 3 (25 mL), brine (25 mL), dried over MgSO 4 , filtered and was concentrated to give a yellow solid. It was purified by flash chromatography (elution: hexanes, EtOAc; 1 :1), gave 0.136 g (62%) of the title compound as a yellow solid.
  • the mixture was cooled and diluted with water (30 mL), neutralized with 2N hydrochloride to produce precipitates.
  • the mixture was filtered and the solids were washed with aqueous sodium carbonate, dried to give a crude product which was used without further purification.
  • the aqueous solution was then extracted with CHCl 3 (2 x 250 mL).
  • the extracts were washed with brine (50 mL), dried over MgSO 4 , filtered and concentrated to an orange solid. It was purified by flash chromatography (silica gel, gradient elution with EtOAc :hexanes, 1 :4 to 1 :1) to give 0.130 g (13%) of yellow solid.
  • the yellow suspension was equilibrated to rt and then heated to 80 0 C, followed by addition of isoamyl nitrite (0.064 mL, 0.48 mmol). It was heated at 80 °C for 17 h, and then cooled to rt. The suspension was quenched with NaHCO 3 ( aq) (10 mL) and then extracted with CHCl 3 (2 x 50 mL), dried over MgSO 4 , filtered and concentrated. The residue was purified by flash chromatography (silica gel, gradient elution with EtOAc :hexanes, 1 :1 to MeOHiCH 2 Cl 2 , 4:96) to give 0.003 g (1%) of white solid.
  • the yellow suspension was equilibrated to rt and then heated to 80 0 C, followed by addition of isoamyl nitrite (0.051 mL, 0.38 mmol). It was heated for 17 h at 80 0 C, and then cooled to rt.
  • the suspension was neutralized with NaHCO 3(aq) (10 mL) and then extracted with CHCl 3 (2 x 50 mL), washed with brine (20 mL), dried over MgSO 4 , filtered and concentrated.
  • the residue was purified by flash chromatography (silica gel, gradient elution with EtOAc:hexanes, 1 :1 to MeOH:CH 2 Cl 2 , 4:96) to give 0.001 g ( ⁇ 1%) of white solid.
  • the title compound was prepared in a manner similar to Example 10c. From 5-fluoro-2-methoxybenzoyl-2-dithiocarboxyhydrazide, potassium salt (4.50 g, 15.1 mmol) and hydrazine (6.60 mL, 181 mmol) was obtained 2.75 g (76%) of the crude product as a white solid. Recrystallization (EtO ⁇ rwater, 50:5 mL) gave of the title compound as a white solid.
  • the title compound was prepared in a manner similar to Example Ib. From 2-bromo-l-(3-fluoro-4-methyl-5-nitrophenyl)ethanone (380 mg, 1.38 mmol) and 4-amino-5-(2-methoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (315 mg, 1.42 mmol) was obtained the title compound as a light yellow solid (182 mg, 0.456 mmol, 33%).
  • the title compound was prepared in a manner similar to Example Ib. From 2-bromo-l-(4-methyl-3-nitrophenyl)ethanone (516 mg, 2 mmol) and 4- amino-5-(2-ethoxyphenyl)-3-mercapto-(4//)-l,2,4-triazole (437 mg, 1.69 mmol) in isopropanol was obtained 580 mg (87%) of the title compound.
  • the title compound was prepared in a manner similar to Example 2. From a mixture of 6-(4-methyl-3-nitrophenyl)-3-(2-ethoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (530 mg, 1.34 mmol) and tin chloride (1.51 g, 6.69 mmol) in ethanol (25 mL) was obtained 520 mg (89%) of the title compound.
  • the title compound was prepared in a manner similar to Example 75 in two steps. From 6-(3-amino-4-methylphenyl)-3 -(2-methoxypheny 1)-7H- [l,2,4]triazolo[3,4- ⁇ ][l,3,4]thiadiazine (420 mg, 1.20 mmol) and Boc-Gly (231 mg, 1.32 mmol) was obtained 200 mg (35%) of the title compound.
  • the title compound was prepared in a manner similar to Example 75 in two steps. From 6-(3-amino-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-b][l,3,4]thiadiazine (1.0 g, 2.84 mmol) and Boc- Lys(Boc)OH (1.18 g, 3.42 mmol) was obtained 328 mg (20%) of the title compound.
  • the title compound was prepared in a manner similar to Example Ib. From 2-bromo-l-(4-methoxy-3-nitrophenyl)ethanone (438 mg, 1.6 mmol) and 4-amino-5-(2,5-dimethylfuryl)-3-mercapto-(4H)-l,2,4-triazole (336 mg, 1.6 mmol) in isopropanol was obtained 680 mg (98%) of the title compound.
  • the title compound was prepared in a manner similar to Example Ib. From 4-amino-5-(l,4,5,6-tetrahydro-3-methylcyclopenta[c]pyrazolyl)-3- mercapto-(4H)-l,2,4-triazole (151 mg, 0.679 mmol) and 2-bromo- 1 -(4- methyl-3-nitrophenyl)ethanone (189 mg, 0.733 mmol) in isopropanol was obtained the title compound as a yellow solid (242 mg, 0.634 mmol, 93%).
  • the title compound was prepared in a manner similar to Example 21. From 3-(l,4,5,6-tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3- nitrophenyl)-7H-[l,2,4]triazolo[3,4- ⁇ ][l,3,4]thiadiazine (217 mg, 0.569 mmol) was obtained the title compound as a light yellow solid (28 mg, 0.08 mmol, 14%).
  • Human breast cancer cell lines T-47D, human colon carcinoma cell line HCTl 16, human hepatocellular carcinoma cell line SNU398, human Burkitt's lymphoma cell line Namalwa were grown according to media component mixtures designated by American Type Culture Collection + 10% FCS (Invitrogen Corporation), in a 5 % CO 2 -95 % humidity incubator at 37 0 C.
  • T-47D cells were maintained at a cell density between 50 and 80 % confluency at a cell density of 0.1 to 0.6 x 10 6 cells/mL. Cells were harvested at 600xg and resuspended at 0.65 x 10 6 cells/mL into appropriate media + 10 % FCS.
  • REU Relative Fluorescence Unit values
  • the activity of caspase cascade activation was determined by the ratio of the net RFU value for 6-(3-amino-4-methoxyphenyl)-3-(2-methoxyphenyl)- 7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (Example 2) or other test compound to that of control samples.
  • the EC 50 (nM) was determined by a sigmoidal dose-response calculation (Prism 3.0, GraphPad Software Inc.).
  • 6-(3 -amino-4-methoxyphenyl)-3 -(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4- ⁇ ][l,3,4]thiadiazine (Example 2) and analogs are identified as potent caspase cascade activators and inducers of apoptosis in several solid tumor cells.
  • Human breast cancer cell lines T-47D, human colon carcinoma cell line HCTl 16, human hepatocellular carcinoma cell line SNU398, human Burkitt's lymphoma cell line Namalwa were grown and harvested as in Example 86.
  • An aliquot of 90 ⁇ L of cells (4.4 x 10 4 cells/mL) was added to a well of a 96-well microtiter plate containing 5 ⁇ L of a 10 % DMSO in RPMI- 1640 media solution containing 10 nM to 100 ⁇ M of 6-(3-amino-4- methoxyphenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine (1 nM to 10 ⁇ M final).
  • Baseline for GI 50 dose for 50% inhibition of cell proliferation
  • GI 50 dose for 50% inhibition of cell proliferation
  • the samples were mixed by agitation and then incubated at 37 0 C for 0.5 h in a 5% CO 2 -95% humidity incubator. After incubation, the samples were removed from the incubator and 25 ⁇ L of CellTiter-Glo TM reagent (Promega) was added.

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Abstract

Disclosed are 3-aryl-6-aryl-7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazines and analogs thereof, represented by the Formula I wherein Ar1, Ar2, R1, R2, and X are defined herein. The present invention relates to the discovery that compounds having Formula I are activators of caspases and inducers of apoptosis. Therefore, the activators of caspases and inducers of apoptosis of this invention may be used to induce cell death in a variety of clinical conditions in which uncontrolled growth and spread of abnormal cells occurs.

Description

3-ARYL-6-ARYL-7H- [ 1 ,2,4]TRIAZOLO[3 ,4- b][l,3,4]TΗIADIAZINES AND ANALOGS AS ACTIVATORS OF
CASPASES AND INDUCERS OF APOPTOSIS AND THE USE
THEREOF
BACKGROUND OF THE INVENTION
Field of the Invention
[0001] This invention is in the field of medicinal chemistry. In particular, the invention relates to 3-aryl-6-aryl-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazines and analogs, and the discovery that these compounds are activators of caspases and inducers of apoptosis. The invention also relates to the use of these compounds as therapeutically effective anti-cancer agents.
Related Art
[0002] Organisms eliminate unwanted cells by a process variously known as regulated cell death, programmed cell death or apoptosis. Such cell death occurs as a normal aspect of animal development, as well as in tissue homeostasis and aging (Glucksmann, A., Biol. Rev. Cambridge Philos. Soc. 25:59-86 (1951); Glucksmann, A., Archives de Biologie 75:419-437 (1965); Ellis, et al, Dev. 772:591-603 (1991); Vaux, et al, Cell 76:111-119 (1994)). Apoptosis regulates cell number, facilitates morphogenesis, removes harmful or otherwise abnormal cells and eliminates cells that have already performed their function. Additionally, apoptosis occurs in response to various physiological stresses, such as hypoxia or ischemia (PCT published application WO96/20721).
[0003] There are a number of morphological changes shared by cells experiencing regulated cell death, including plasma and nuclear membrane blebbing, cell shrinkage (condensation of nucleoplasm and cytoplasm), organelle relocalization and compaction, chromatin condensation and production of apoptotic bodies (membrane enclosed particles containing intracellular material) (Orrenius, S., J. Internal Medicine 237:529-536 (1995)).
[0004] Apoptosis is achieved through an endogenous mechanism of cellular suicide (Wyllie, A.H., in Cell Death in Biology and Pathology, Bowen and Lockshin, eds., Chapman and Hall (1981), pp. 9-34). A cell activates its internally encoded suicide program as a result of either internal or external signals. The suicide program is executed through the activation of a carefully regulated genetic program (Wyllie, et al, Int. Rev. Cyt. 68:25\ (1980); Ellis, et al, Ann. Rev. Cell Bio. 7:663 (1991)). Apoptotic cells and bodies are usually recognized and cleared by neighboring cells or macrophages before lysis. Because of this clearance mechanism, inflammation is not induced despite the clearance of great numbers of cells (Orrenius, S., J. Internal Medicine 237:529-536 (1995)).
[0005] It has been found that a group of proteases are a key element in apoptosis (see, e.g., Thornberry, Chemistry and Biology 5:R97-R103 (1998); Thornberry, British Med. Bull. 53:478-490 (1996)). Genetic studies in the nematode Caenorhabditis elegans revealed that apoptotic cell death involves at least 14 genes, 2 of which are the pro-apoptotic (death -promoting) ced (for cell death abnormal) genes, ced-3 and ced-4. CED-3 is homologous to interleukin 1 beta-converting enzyme, a cysteine protease, which is now called caspase-1. When these data were ultimately applied to mammals, and upon further extensive investigation, it was found that the mammalian apoptosis system appears to involve a cascade of caspases, or a system that behaves like a cascade of caspases. At present, the caspase family of cysteine proteases comprises 14 different members, and more may be discovered in the future. All known caspases are synthesized as zymogens that require cleavage at an aspartyl residue prior to forming the active enzyme. Thus, caspases are capable of activating other caspases, in the manner of an amplifying cascade.
[0006] Apoptosis and caspases are thought to be crucial in the development of cancer {Apoptosis and Cancer Chemotherapy, Hickman and Dive, eds., Humana Press (1999)). There is mounting evidence that cancer cells, while containing caspases, lack parts of the molecular machinery that activates the caspase cascade. This makes the cancer cells lose their capacity to undergo cellular suicide and the cells become cancerous. In the case of the apoptosis process, control points are known to exist that represent points for intervention leading to activation. These control points include the CED-9-BCL-like and CED-3-ICE-like gene family products, which are intrinsic proteins regulating the decision of a cell to survive or die and executing part of the cell death process itself, respectively (see, Schmitt, et al, Biochem. Cell. Biol. 75:301- 314 (1997)). BCL-like proteins include BCL-xL and BAX-alpha, which appear to function upstream of caspase activation. BCL-xL appears to prevent activation of the apoptotic protease cascade, whereas BAX-alpha accelerates activation of the apoptotic protease cascade. It has been shown that chemotherapeutic (anti-cancer) drugs can trigger cancer cells to undergo suicide by activating the dormant caspase cascade. This may be a crucial aspect of the mode of action of most, if not all, known anticancer drugs (Los, et al, Blood 90:3118-3129 (1997); Friesen, et al, Nat. Med. 2:574 (1996)). The mechanism of action of current antineoplastic drugs frequently involves an attack at specific phases of the cell cycle. In brief, the cell cycle refers to the stages through which cells normally progress during their lifetime. Normally, cells exist in a resting phase termed G0. During multiplication, cells progress to a stage in which DNA synthesis occurs, termed S. Later, cell division, or mitosis occurs, in a phase called M. Antineoplastic drugs, such as cytosine arabinoside, hydroxyurea, 6-mercaptopurine, and methotrexate are S phase specific, whereas antineoplastic drugs, such as vincristine, vinblastine, and paclitaxel are M phase specific. Many slow growing tumors, e.g. colon cancers, exist primarily in the G0 phase, whereas rapidly proliferating normal tissues, for example bone marrow, exist primarily in the S or M phase. Thus, a drug like 6-mercaptopurine can cause bone marrow toxicity while remaining ineffective for a slow growing tumor. Further aspects of the chemotherapy of neoplastic diseases are known to those skilled in the art (see, e.g., Hardman, et al, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, Ninth Edition, McGraw-Hill, New York (1996), pp. 1225-1287). Thus, it is clear that the possibility exists for the activation of the caspase cascade, although the exact mechanisms for doing so are not clear at this point. It is equally clear that insufficient activity of the caspase cascade and consequent apoptotic events are implicated in various types of cancer. The development of caspase cascade activators and inducers of apoptosis is a highly desirable goal in the development of therapeutically effective antineoplastic agents. Moreover, since autoimmune disease and certain degenerative diseases also involve the proliferation of abnormal cells, therapeutic treatment for these diseases could also involve the enhancement of the apoptotic process through the administration of appropriate caspase cascade activators and inducers of apoptosis.
[0008] C-Myc is a proto-oncogene and encodes the c-Myc transcription factor.
Physiologically, cMyc expression correlates with cell proliferation in various cells and tissues of the body. CMyc is implicated in various biological processes including cell growth, proliferation, loss of differentiation and apoptosis. Deregulated expression of c-Myc occurs in a wide range of cancers and is often associated with poor prognosis suggesting an important role for this oncogene in tumor progression. Initially it was discovered in Burkitt's lymphoma as causative for the progression of the disease due to a translocation between chromosome 8 and the antibody-containing genes. More recently, cMyc has been detected in a wide range of cancers that include breast, colon, cervical, small-cell lung carcinomas, osteocarcomas, glioblastomas, melanoma and myeloid leukemias (Nesbit, CE et al. Oncogene, 18, 3004-3016 (1999); Blackwood, E.M et al. Science 251, 1211-1217 (1991); Mo H. & Henriksson M. PNAS, 103: 6344-6349 (2006)) Inactivation of c-Myc was found to cause tumor regression with rapid proliferation arrest and apoptosis in hematopoietic malignancies and osteosarcoma. Therefore inactivating cMyc or downstream targets of cMyc may provide important therapeutic advantages.
[0009] Nadkarni et al. {Arzneimittel-Forschung 51 :569-573 (2001)) reported the synthesis of a series of 3,6-disubstituted-7H-s-triazolo(3,4- b)(l,3,4)thiadiazines I (X = F,C1, R = H, 2-F, 3-F, 4-F, 3-CF3, etc.) by the condensation of the appropriate 3-substituted-4-amino-5- mercapto(l,2,4)triazoles II (X = F, Cl) with substituted phenacyl bromides in alcohol medium. These compounds have been studied for their in vivo anthelmintic activity in albino mice. A number of compounds showed promising activity when given by the oral route.
Figure imgf000006_0001
SUMMARY OF THE INVENTION
[0010] The present invention is related to the discovery that 3-aryl-6-aryl-7//-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazines and analogs, as represented in Formulae I-V, are activators of the caspase cascade and inducers of apoptosis. Thus, an aspect of the present invention is directed to the use of compounds of Formulae I-V as inducers of apoptosis.
[0011] A second aspect of the present invention is to provide a method for treating, preventing or ameliorating neoplasia and cancer by administering a compound of one of the Formulae I-V to a mammal in need of such treatment.
[0012] Many of the compounds within the scope of the present invention are novel compounds. Therefore, a third aspect of the present invention is to provide novel compounds of Formulae I-V, and to also provide for the use of these novel compounds for treating, preventing or ameliorating neoplasia and cancer.
[0013] A fourth aspect of the present invention is to provide a pharmaceutical composition useful for treating disorders responsive to the induction of apoptosis, containing an effective amount of a compound of one of the Formulae I-V in admixture with one or more pharmaceutically acceptable carriers or diluents.
[0014] A fifth aspect of the present invention is directed to methods for the preparation of novel compounds of Formulae I-V. DETAILED DESCRIPTION OF THE INVENTION
[0015] The present invention arises out of the discovery that 3-aryl-6-aryl-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazines and analogs, as represented in Formulae I-V, are potent and highly efficacious activators of the caspase cascade and inducers of apoptosis. Therefore, compounds of Formulae I-V are useful for treating disorders responsive to induction of apoptosis.
[0016] Specifically, compounds of the present invention are represented by
Formula I:
Figure imgf000007_0001
or pharmaceutically acceptable salts or prodrugs or tautomers thereof, wherein:
Ar1 and Ar2 independently are optionally substituted carbocyclic, heterocyclic, aryl or heteroaryl;
R1 and R2 independently are hydrogen, halo, optionally substituted amino, optionally substituted alkoxy, optionally substituted CMO alkyl, haloalkyl, aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl, aminoalkyl, carboxyalkyl, nitro, cyano, acylamido, hydroxy, thiol, acyloxy, azido, carboxy, carbonylamido or optionally substituted alkylthiol; or Ri and R2 are combined as =0; and
X is S, O or NR3, wherein R3 is hydrogen or an optionally substituted alkyl or aryl.
[0017] Preferred compounds of Formula I include compounds wherein Ari is phenyl, naphthyl, indolinyl, benzo[^/][l,3]dioxol-5-yl, benzo[ό][l,4]dioxin-6- yl, benzofuranyl, benzothiophenyl, benzimidazolyl, indolyl, imidazopyridinyl, pyridyl, quinolyl, isoquinolyl, isoxazolyl, pyrazolyl, imidazolyl, thienyl, furyl or pyrrolyl, each of which is optionally substituted. Another group of preferred compounds are wherein Ar2 is an optionally substituted aryl or heteroaryl. More preferably, Ar2 is an optionally phenyl, 1,4,5,6- tetrahydrocyclopenta[c]pyrazol-3-yl, imidazopyridinyl, pyridyl, pyrazolyl, imidazolyl, thienyl, furyl or pyrrolyl. Another group of preferred compounds of Formula I include compounds wherein Ri and R2 are hydrogen. Another group of preferred compounds of Formula I include compounds wherein X is S or O. Another group of preferred compounds of Formula I include compounds wherein X is S.
[0018] One group of preferred compounds of the present invention are represented by Formula II:
Ari (H)
Figure imgf000008_0001
or pharmaceutically acceptable salts, prodrugs or tautomers thereof, wherein:
Ari and Ar2 independently are optionally substituted aryl or optionally substituted heteroaryl.
[0019] Preferred compounds of Formula II include compounds wherein An and Ar2 are phenyl, naphthyl, indolinyl, benzo[d][l,3]dioxol-5-yl, benzo[6][l,4]dioxin-6-yl, benzofuranyl, benzothiophenyl, benzimidazolyl, indolyl, l,4,5,6-tetrahydrocyclopenta[c]pyrazol-3-yl, pyridyl, imidazopyridinyl, quinolyl, isoquinolyl, isoxazolyl, pyrazolyl, imidazolyl, thienyl, furyl or pyrrolyl, each of which is optionally substituted. More preferably, Ari is phenyl, indolinyl, benzo[</][l,3]dioxol-5-yl, benzo[ό][l,4]dioxin-6-yl, benzofuranyl, benzothiophenyl, benzimidazolyl, indolyl, or pyridyl, and Ar2 is phenyl, pyridyl, 1,4,5,6- tetrahydrocyclopenta[c]pyrazol-3-yl, imidazopyridinyl, thienyl or furyl.
[0020] Another group of preferred compounds of the present invention are represented by Formula III:
Figure imgf000009_0001
or pharmaceutically acceptable salts, prodrugs or tautomers thereof, wherein:
Ar2 is an optionally substituted aryl or optionally substituted heteroaryl;
R9-R13 independently are hydrogen, halo, amino, di(C1-1o alkyl)amino, alkoxy, CMO alkyl, haloalkyl, aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl, hydroxyalkoxy, aminoalkyl, aminoalkoxy, carboxyalkyl, nitro, cyano, acylamido, hydroxy, thiol, acyloxy, azido, carboxy, carbonylamido, alkylsulfonyl, aminosulfonyl, dialkylaminosulfonyl, alkylsulfiniyl, or alkylthiol; or
R9 and Rio, or R10 and Rn, or Rn and Rj2, or R12 and Ri3, taken together with the atoms to which they are attached to form an aryl, heteroaryl, partially saturated carbocyclic or partially saturated heterocyclic group, wherein said group is optionally substituted. Preferred are compounds of Formula III, wherein R9 and Rio, or Rio and Rn, or Rn and Ri2, or Rj2 and Rj3, taken together to form a structure selected from the group consisting of -OCH2O-, -(CH2)3- -(CH2).,-, -OCH2CH2O-, -CH2N(Ri4)CH2-, -CH2CH2N(R14)CH2-, -CH2N(Ri4)CH2CH2-, -N(R14)- CH=CH-, -CH=CH-N(R,4)-, -N(R^)-CH2-CH2- -CH2-CH2-N(R14)-, -N(RM)-CH=N- -N=CH-N(RI4)-, -0-CH=CH-, -CH=CH-O-, -S- CH=CH-, -CH=CH-S-, -N-C(=O)-O-, -N-CH2-CH2-N- and -N=CH- CH=N-, wherein Ri4 is hydrogen, Ci-10 alkyl, haloalkyl, aryl, fused aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl or aminoalkyl. [0022] Another group of preferred compounds of Formula III include compounds wherein Rio or Ri2 is an amino, and Rn is an alkyl or alkoxy. Another group of preferred compounds of Formula III include compounds wherein Ar2 is phenyl, pyridyl, l,4,5,6-tetrahydrocyclopenta[c]pyrazol-3-yl, imidazopyridinyl, thienyl, furyl or pyrrolyl. More preferably Ar2 is phenyl, pyridyl, or furyl. Especially preferred compounds of Formula III include compounds wherein Rio or R12 is an amino, and Rj1 is an alkoxy or methyl.
[0023] Another group of preferred compounds of the present invention are represented by Formula IV:
or pharmaceutically acceptable salts, prodrugs or tautomers thereof, wherein:
R4-Ri3 independently are hydrogen, halo, amino, di(Ci-io alkyl)amino, alkoxy, C1-1O alkyl, haloalkyl, aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl, hydroxyalkoxy, aminoalkyl, aminoalkoxy, carboxyalkyl, nitro, cyano, acylamido, hydroxy, thiol, acyloxy, azido, carboxy, carbonylamido, alkylsulfonyl, aminosulfonyl, dialkylaminosulfonyl, alkylsulfiniyl, or alkylthiol; or
R4 and R5, or R5 and R6, or R6 and R7, or R7 and R8, or R9 and Rio, or Rio and Rn, or Rn and Ri2, or Ri2 and Ri3, taken together with the atoms to which they are attached to form an aryl, heteroaryl, partially saturated carbocyclic or partially saturated heterocyclic group, wherein said group is optionally substituted.
[0024] Preferred are compounds of Formula IV, wherein R4 and R5, or R5 and
R6, or R6 and R7, or R7 and R8, or R9 and Rio, or Rio and Rn, or Rn and R]2, or Ri2 and Rj3, taken together to form a structure selected from the group consisting of -OCH2O-, -(CH2)3- -(CH2)4- -OCH2CH2O-, -CH2N(R14)CH2-, -CH2CH2N(R14)CH2-, -CH2N(Ri4)CH2CH2-, -N(R14)- CH=CH-, -CH=CH-N(Ri4)-, -N(RH)-CH2-CH2-, -CH2-CH2-N(R14)-, -N(RH)-CH=N-, -N=CH-N(R14)-, -0-CH=CH-, -CH=CH-O-, -S- CH=CH-, -CH=CH-S-, -N-C(=0)-0-, -N-CH2-CH2-N- and -N=CH- CH=N-, wherein R14 is hydrogen, Ci-1O alkyl, haloalkyl, aryl, fused aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl or aminoalkyl.
[0025] Another group of preferred compounds of Formula IV include compounds wherein R1O or Ri2 is an amino, and Rn is an alkyl or alkoxy. Another group of preferred compounds of Formula IV include compounds wherein R4 or R8 is subsututited. More preferably R4 or R8 is an alkyl, alkoxy, thioalkoxy, halo or amino. Another group of preferred compounds of Formula IV include compounds wherein R4 and R6 are both substituted, or R4 and R7 are both substituted, or R5 and R7 are both substituted. Especially preferred compounds of Formula IV include compounds wherein Rio or R12 is an amino, and R1 ] is an alkoxy.
[0026] One group of especially preferred compounds of Formula IV are wherein: Rg is halo or H; each of R4, R5, R7, Rg, and R\ \ is independently H, methoxy, ethoxy, trifluoromethoxy, dimethylamino, or methyl; each of R9, Rj Q> R-12= and R-13 *s independently H, amino, dimethylamino, or nitro; with the proviso that: at least one of R9, Rj Q, K\ 2, and K\ 3 is not H; at least one of R4, R5, R7, and Rg is not H; and when R4 or Rg is methoxy or methyl and Rj \ is methyl, then none of R]O and Rj 2 ιs nitro or amino.
[0027] Another group of preferred compounds of the present invention are represented by Formula V:
Figure imgf000012_0001
or pharmaceutically acceptable salts or tautomers thereof, wherein:
Ar2 is an optionally substituted aryl or optionally substituted heteroaryl;
R9-Rn and R13 independently are hydrogen, halo, amino, di(Ci-10 alkyl)amino, alkoxy, C1-Io alkyl, haloalkyl, aryl, carbocyclic, a heterocyclic group, a heteroaryl group, alkenyl, alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, carbocycloalkyl, heterocycloalkyl, hydroxyalkyl, hydroxyalkoxy, aminoalkyl, aminoalkoxy, carboxyalkyl, nitro, cyano, acylamido, hydroxy, thiol, acyloxy, azido, carboxy, carbonylamido, alkylsulfonyl, aminosulfonyl, dialkylaminosulfonyl, alkylsulfiniyl, or alkylthiol;
Rj4 is the side chain of a natural or non-natural amino acid.
[0028] One group of preferred compounds of Formula V include compounds wherein Ar2 is phenyl, pyridyl, l,4,5,6-tetrahydrocyclopenta[c]pyrazol-3-yl, imidazopyridinyl, thienyl, furyl or pyrrolyl. More preferably Ar2 is phenyl, pyridyl, or furyl. Another group of preferred compounds of Formula V include compounds wherein Ri4 is hydrogen or methyl.
[0029] Exemplary preferred compounds of Formulae I-V that may be employed in the method of the invention include, without limitation:
3-(2-Methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3 ,4-Z)][1 ,3,4]thiadiazine;
3-(4-Chloro-2-methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo[3,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(4-chloro-2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-£»][l,3,4]thiadiazine; 6-(4- Aminophenyl)-3 -(3 , 5-dimethoxyphenyl)-7H- [ 1 ,2,4] triazolo[3 ,4- 6][l,3,4]thiadiazine;
6-(3 - Amino-4-methoxyphenyl)-3 -(2-methylfiiran-3 -yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3 -Dimethylamino-4-methoxyphenyl)-3 -(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
3-(4-Bromo-2-methoxyphenyl)-6-(4-methylphenyl)-7H- [l,2,4]triazolo[3,4-&][l,3,4]thiadiazine;
3-(2-Methoxy-4-methylphenyl)-6-(4-methylphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Methylphenyl)-3 -(3 -methylthiophen-2-yl)-7H-[ 1 ,2,4]triazolo [3 ,4- b] [ 1 ,3 ,4]thiadiazine;
3-(4-Bromo-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
3-(4-Chloro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(4-chloro-2-methoxyphenyl)-7//- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(4-bromo-2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(4-Methoxyphenyl)-3-(2-methylfuran-3-yl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine;
6-(4-Ηydroxyphenyl)-3-(2-methylfuran-3-yl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(2-methylfuran-3 -yl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methyl-3-nitrophenyl)-3-(2-methyl-H-imidazo[l,2-α]pyridin-3- yl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [1,3 ,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(2-methyl-//-imidazo[l,2-α]pyridin-3- yl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methylphenyl)-3-(l-methyl-lH-pyrazol-5-yl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [1,3 ,4]thiadiazine;
3-(4-Chloro-2-methylphenyl)-6-(4-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine; 3-(4-Chloro-2-methylphenyl)-6-(4-hydroxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Dimethylamino-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(2,5-Dimethylfuran-3-yl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- ό][l,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(l-methyl-lH-indol-5-yl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Acetamido-3-aminophenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(2-methyl-lH-benzo[i/]iniidazol-6-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methoxy-3-nitrophenyl)-3-(3,5-dimethoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(3,5-dimethoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3 - Amino-4-methoxyphenyl)-3 -(3 ,5-dimethoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3 -Dimethylamino-4-methoxyphenyl)-3-(3 ,5 -dimethoxyphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3,4]thiadiazine;
6-(Benzofuran-5 -yl)-3 -(2-methoxypheny 1)-7H- [ 1 ,2,4]triazolo [3 ,4- ό][l,3,4]thiadiazine;
6-(Benzo[6]thiophen-5-yl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(4-Fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(4-fluoro-2-methoxyphenyl)-7//- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methoxy-3-nitrophenyl)-3-(5-methyl- 1 H-imidazol-4-yl)-7/7- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
3-(2-Dimethylaminophenyl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(2-dimethylaminophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine; 6-(4-Methyl-3-nitrophenyl)-3-(2-trifluoromethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(2-trifluoromethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-( 1 -methylsulfonyl-2,3-dihydro- lH-indol-5- yl)-7H-[ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(2-Methyl-lH-imidazo[l,2-α]pyridin-3-yl)-6-(4-methoxy-3- nitrophenyl)-7H-[ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(2-methyl-lH-imidazo[l,2- α]pyridin-3-yl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2-Methoxy-5-methylphenyl)-6-(4-methylphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2-Methoxy-5-methylphenyl)-6-(2-oxoindolin-5-yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Acetamide-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-δ][l,3,4]thiadiazine;
6-(lH-l-Acetylindazol-6-yl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-N-Ηydroxyamidinophenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(5-Fluoro-2-methoxyphenyl)-6-(4-methylphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(4-(N-methylacetamido)methylphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(4-(iV-methylaniino)methylphenyl)-7H- [ 1 ,2,4] triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine; and
6-(4-(N,N-Dimethylamino)methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3 ,4-b][\, 3,4]thiadiazine;
3-(5-Fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(5-fluoro-2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(2,5-Dimethylfuran-3-yl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4] triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine; 6-(3-Amino-4-methylphenyl)-3-(2,5-dimethylfυran-3-yl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine; and
3-(2-Methoxyphenyl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazin-7-one ;
6-(N-Acetyl-indolin-5-yl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(Indolin-5-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- Z>][l,3,4]thiadiazine;
6-(3-(N-Boc-<9-'Bu-Serinyl-amino)-4-methylphenyl)-3-(2- methoxyphenyl)-7H-[l,2,4]triazolo-[3,4-ό][l,3,4]thiadiazine;
6-(3-Fluoro-4-methyl-5-nitrophenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Amino-5-fluoro-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Methyl-3-nitrophenyl)-3-(2-ethoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(2-ethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(3-(Glycinyl-amino)-4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3-(Alaninyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(Glycinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3-(Lysinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(4-Methoxy-3-nitrophenyl)-3-(2,5-dimethylfuran-3-yl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3 - Amino-4-methoxyphenyl)-3-(2,5 -dimethylfuran-3 -yl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-(Fmoc-Alaninyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)- 7H-[l,2,4]triazolo-[3,4-6][l,3,4]thiadiazine;
3-(l,4,5,6-Tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3- nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,A-b] [ 1 ,3 ,4]thiadiazine; 3-(l,4,5,6-Tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3- aminophenyl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(Alaninyl-amino)-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3 -(Alaninyl-amino)-4-methoxyphenyl)-3 -(2-methylftiran-3 -yl)-7H- [1 ,2,4]triazolo [3,4-6] [1 ,3,4]thiadiazine;
6-(3-(Serinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7//- [l,2,4]triazolo-[3,4-6][l,3,4]thiadiazine; and pharmaceutically acceptable salts or prodrugs thereof. The present invention is also directed to novel compounds within the scope of Formulae I- V. Exemplary preferred compounds that may be employed in this invention include, without limitation:
3-(2-Methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H- [1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(4-Chloro-2-methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(4-chloro-2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(3,5-Dimethoxyphenyl)-6-(4-nitrophenyl)-7H-[l,2,4]triazolo[3,4- b][l ,3,4]thiadiazine;
6-(4-Aminophenyl)-3-(3,5-dimethoxyphenyl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine;
6-(4-Methoxy-3 -nitrophenyl)-3 -(2-methylfuran-3 -yl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3-Dimethylamino-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
3-(4-Bromo-2-methoxyphenyl)-6-(4-methylphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
3-(2-Methoxy-4-methylphenyl)-6-(4-methylphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine ; 6-(4-Methylphenyl)-3-(3-methylthiophen-2-yl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine;
3-(4-Bromo-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine;
3-(4-Chloro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [l,2,4]triazolo[3,4-Z>][l,3,4]thiadiazine;
6-(3 - Amino-4-methy lphenyl)-3 -(4-chloro-2-methoxypheny 1)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(4-bromo-2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methylphenyl)-3-( 1 H-pyrazol-5-yl)-7H-[ 1 ,2,4]triazolo[3 ,4- b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methoxyphenyl)-3-(2-methylfiiran-3-yl)-7H-[l ,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine;
6-(4-Ηydroxyphenyl)-3 -(2-methylftiran-3 -yl)-7H-[ 1 ,2,4]triazolo [3 ,4- b] [ 1 ,3 ,4]thiadiazine;
3-(2-Methylfuran-3-yl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(2-methylfuran-3 -yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(4-Methyl-3-nitrophenyl)-3 -(2-methyl-H-imidazo [ 1 ,2-α]pyridin-3 - yl)-7H-[l ,2,4]triazolo[3 ,4-Z)][1 ,3,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(2-methyl-H-imidazo [ 1 ,2-α]pyridin-3 - yl)-7H-[ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3,4]thiadiazine;
6-(4-Methylphenyl)-3-(l-methyl-lH-pyrazol-5-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methyl-3-nitrophenyl)-3-(l-methyl-4-nitro-lH-pyrrol-2-yl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(4-Chloro-2-methylphenyl)-6-(4-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(4-Chloro-2-methylphenyl)-6-(4-hydroxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
3-(4-Methyl-2-moφholinothiazol-5-yl)-6-(4-methylphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine; 6-(3-Dimethylamino-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(4-Methyl- 1 H-imidazol-5-yl)-6-(4-methylphenyl)- 7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2,5-Dimethylfiiran-3-yl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(l-methyl-lH-indol-5-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Acetamido-3-aminophenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(2-methyl- 1 H-benzo[d]imidazol-6-yl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(3,5-Dimethoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(4-Methoxy-3-nitrophenyl)-3-(3,5-dimethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(3,5-dimethoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(3,5-dimethoxyphenyl)-7H- [l,2,4]triazolo[3,4-δ][l,3,4]thiadiazine;
6-(3-Dimethylamino-4-methoxyphenyl)-3-(3,5-dimethoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(Benzofuran-5-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3,4]thiadiazine;
6-(Benzo[ό]thiophen-5-yl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3 ,4-6][l, 3,4]thiadiazine;
3 -(4-Fluoro-2-methoxyphenyl)-6-(4-methy 1-3 -nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3 -Amino-4-methylphenyl)-3 -(4-fluoro-2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methoxy-3-nitrophenyl)-3-(5-methyl- 1 H-imidazol-4-yl)-7H- [ 1 ,2,4] triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(5-methyl-lH-imidazol-4-yl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine; 3 -(2-Dimethylaminophenyl)-6-(4-methyl-3 -nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3 - Amino-4-methy lphenyl)-3 -(2-dimethylaminopheny 1)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Methyl-3-nitrophenyl)-3-(2-trifluoromethoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [1,3 ,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(2-trifluoromethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(l-methylsulfonyl-2,3-dihydro-lH-indol-5- yl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]tWadiazine;
3-(2-Methyl-lH-imidazo[l,2-α]pyridin-3-yl)-6-(4-methoxy-3- nitrophenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3 - Amino-4-methoxyphenyl)-3 -(2 -methyl- 1 H-imidazo [1,2- α]pyridin-3-yl)-7H-[l ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
3-(2-Methoxy-5-methylphenyl)-6-(4-methylphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
3-(2-Methoxy-5-methylphenyl)-6-(2-oxoindolin-5-yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Acetamide-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-( 1 H- 1 - Acetylindazol-6-yl)-3 -(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-N-Ηydroxyamidinophenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
3-(5-Fluoro-2-methoxyphenyl)-6-(4-methylphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(4-(N-methylacetamido)methylphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(4-(N-methylamino)methylphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine; and
6-(4-(N,N-Dimethylamino)methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(5-Fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [l,2,4]triazolo[3,4-Z)][l,3,4]thiadiazine; 6-(3-Amino-4-methylphenyl)-3-(5-fluoro-2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-2>][l ,3,4]thiadiazine;
3-(2,5-Dimethylfuran-3-yl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(2,5-dimethylfuran-3-yl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine; and
3-(2-Methoxyphenyl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazin-7-one;
6-(N-Acetyl-indolin-5-yl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo[3,4-6] [ 1 ,3 ,4]thiadiazine;
6-(Indolin-5-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- Z>][l,3,4]thiadiazine;
6-(3-(N-Boc-O-'Bu-Serinyl-amino)-4-methylphenyl)-3-(2- methoxyphenyl)-7H-[l ,2,4]triazolo-[3,4-δ][l ,3,4]thiadiazine;
6-(3-Fluoro-4-methyl-5-nitrophenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(3-Amino-5-fluoro-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Methyl-3-nitrophenyl)-3-(2-ethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(2-ethoxyphenyl)-7//- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3 -(Glycinyl-amino)-4-methoxyphenyl)-3 -(2-methylfuran-3 -yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(Alaninyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3-(Glycinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(3-(Lysinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Methoxy-3 -nitrophenyl)-3 -(2,5-dimethylfiiran-3 -y 1)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(2,5-dimethylfiiran-3-yl)-7//- [ 1 ,2,4]triazolo [3 ,4-6] [1,3 ,4]thiadiazine; 6-(3-(Fmoc-Alaninyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)- 7H-[1 ,2,4]triazolo-[3,4-6][l ,3,4]thiadiazine;
3-(l,4,5,6-Tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3- nitrophenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3 -( 1 ,4,5 ,6-Tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3 - aminophenyl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(Alaninyl-amino)-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(Alaninyl-amino)-4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H- [ 1 ,2,4]triazolo[3,4-ό] [ 1 ,3 ,4]thiadiazine;
6-(3-(Serinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo-[3,4-6][l ,3,4]thiadiazine; and pharmaceutically acceptable salts or prodrugs thereof.
[0031] The term "alkyl" as employed herein by itself or as part of another group refers to both straight and branched chain radicals of up to ten carbons. Useful alkyl groups include straight-chained and branched C1-Io alkyl groups, more preferably Cj-6 alkyl groups. Typical Ci-10 alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, /er/-butyl, 3-pentyl, hexyl and octyl groups, which may be optionally substituted.
[0032] The term "amino" as employed herein by itself or as part of another group is -NH2, -NHR3, or -NR3Rb, wherein R3 and Rb are independently alkyl groups or together, with the nitrogen, form a 5 or 6 membered heterocyclo group optionally containing an additional N or O atom.
[0033] The term "alkenyl" as employed herein by itself or as part of another group means a straight or branched chain radical of 2-10 carbon atoms, unless the chain length is limited thereto, including at least one double bond between two of the carbon atoms in the chain. Typical alkenyl groups include ethenyl, 1-propenyl, 2-propenyl, 2-methyl-l-propenyl, 1-butenyl and 2-butenyl. [0034] The term "alkynyl" is used herein to mean a straight or branched chain radical of 2-10 carbon atoms, unless the chain length is limited thereto, wherein there is at least one triple bond between two of the carbon atoms in the chain. Typical alkynyl groups include ethynyl, 1-propynyl, l-methyl-2- propynyl, 2-propynyl, 1-butynyl and 2-butynyl. [0035] Useful alkoxy groups include oxygen substituted by one of the Ci-I0 alkyl groups mentioned above, which may be optionally substituted. Alkoxy substituents include, without limitation, halo, morpholino, amino including alkylamino and dialkylamino, and carboxy including esters thereof.
[0036] Useful alkylthio groups include sulfur substituted by one of the Ci-I0 alkyl groups mentioned above, which may be optionally substituted. Also included are the sulfoxides and sulfones of such alkylthio groups.
[0037] Useful amino and optionally substituted amino groups include -NH2, -
NHRi5 and -NRi5Ri6, wherein Ri5 and Ri6 are Ci-10 alkyl or cycloalkyl groups, or R15 and Ri6 are combined with the N to form a ring structure, such as a piperidine, or Ri5 and Ri6 are combined with the N and other group to form a ring, such as a piperazine. The alkyl group may be optionally substituted.
[0038] Optional substituents on the alkyl, alkoxy, alkylthio, alkenyl, alkynyl, cycloalkyl, carbocyclic and heterocyclic groups include one or more halo, hydroxy, carboxyl, amino, nitro, cyano, Ci-C6 acylamino, Ci-C6 acyloxy, Ci-C6 alkoxy, aryloxy, alkylthio, C6-Ci0 aryl, C4-C7 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C6-Ci0 aryl(C2-C6)alkenyl, C6-Ci0 aryl(C2-C6)alkynyl, saturated and unsaturated heterocyclic or heteroaryl.
[0039] Optional substituents on the aryl, arylalkyl, arylalkenyl, arylalkynyl and heteroaryl and heteroarylalkyl groups include one or more halo, methylenedioxy, Ci-C6 haloalkyl, C6-Ci0 aryl, C4-C7 cycloalkyl, Cj-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C6-C10 aiyl(d-C6)alkyl, C6-Ci0 aryl(C2-C6)alkenyl, C6-Ci0 aryl(C2-C6)alkynyl, C1-C6 hydroxyalkyl, nitro, amino, ureido, cyano, C1-C6 acylamino, hydroxy, thiol, Ci-C6 acyloxy, azido, Ci-C6 alkoxy, carboxy, di(Ci-io alkyl)amino, alkylsulfonyl, aminosulfonyl, dialkylaminosulfonyl, or alkylsulfiniyl.
[0040] The term "aryl" as employed herein by itself or as part of another group refers to monocyclic, bicyclic or tricyclic aromatic groups containing from 6 to 14 carbons in the ring portion.
[0041] Useful aryl groups include C6-I4 aryl, preferably C6-I0 aryl. Typical
C6-I4 aryl groups include phenyl, naphthyl, phenanthrenyl, anthracenyl, indenyl, azulenyl, biphenyl, biphenylenyl and fluorenyl groups. [0042] The term "carbocycle" as employed herein include cycloalkyl and partially saturated carbocyclic groups. Useful cycloalkyl groups are C3-8 cycloalkyl. Typical cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
[0043] Useful saturated or partially saturated carbocyclic groups are cycloalkyl groups as described above, as well as cycloalkenyl groups, such as cyclopentenyl, cycloheptenyl and cyclooctenyl.
[0044] Useful halo or halogen groups include fluorine, chlorine, bromine and iodine.
[0045] The term "arylalkyl" is used herein to mean any of the above- mentioned C1-10 alkyl groups substituted by any of the above-mentioned C6-I4 aryl groups. Preferably the arylalkyl group is benzyl, phenethyl or naphthylmethyl.
[0046] The term "arylalkenyl" is used herein to mean any of the above- mentioned C2-10 alkenyl groups substituted by any of the above-mentioned C6-I4 aryl groups.
[0047] The term "arylalkynyl" is used herein to mean any of the above- mentioned C2-10 alkynyl groups substituted by any of the above-mentioned C6-I4 aryl groups.
[0048] The term "aryloxy" is used herein to mean oxygen substituted by one of the above-mentioned C6-I4 aryl groups, which may be optionally substituted. Useful aryloxy groups include phenoxy and 4-methylphenoxy.
[0049] The term "arylalkoxy" is used herein to mean any of the above mentioned Cj-1O alkoxy groups substituted by any of the above-mentioned aryl groups, which may be optionally substituted. Useful arylalkoxy groups include benzyloxy and phenethyloxy.
[0050] Useful haloalkyl groups include Ci-I0 alkyl groups substituted by one or more fluorine, chlorine, bromine or iodine atoms, e.g., fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, 1,1-difluoroethyl, chloromethyl, chlorofluoromethyl and trichloromethyl groups.
[0051] Useful acylamino (acylamido) groups are any Ci-6 acyl (alkanoyl) attached to an amino nitrogen, e.g., acetamido, chloroacetamido, propionamido, butanoylamido, pentanoylamido and hexanoylamido, as well as aryl-substituted Ci-6 acylamino groups, e.g., benzoylamido, and pentafluorobenzoylamido .
[0052] Useful acyloxy groups are any Cj-6 acyl (alkanoyl) attached to an oxy
(-O-) group, e.g., formyloxy, acetoxy, propionoyloxy, butanoyloxy, pentanoyloxy and hexanoyloxy.
[0053] The term heterocycle is used herein to mean a saturated or partially saturated 3-7 membered monocyclic, or 7-10 membered bicyclic ring system, which consists of carbon atoms and from one to four heteroatoms independently selected from the group consisting of O, N, and S, wherein the nitrogen and sulfur heteroatoms can be optionally oxidized, the nitrogen can be optionally quaternized, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring, and wherein the heterocyclic ring can be substituted on carbon or on a nitrogen atom if the resulting compound is stable.
[0054] Useful saturated or partially saturated heterocyclic groups include tetrahydrofuranyl, pyranyl, piperidinyl, piperazinyl, pyrrolidinyl, imidazolidinyl, imidazolinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl, isochromanyl, chromanyl, pyrazolidinyl pyrazolinyl, tetronoyl and tetramoyl groups.
[0055] The term "heteroaryl" as employed herein refers to groups having 5 to
14 ring atoms; 6, 10 or 14 π electrons shared in a cyclic array; and containing carbon atoms and 1, 2 or 3 oxygen, nitrogen or sulfur heteroatoms.
[0056] Useful heteroaryl groups include thienyl (thiophenyl), benzo[6]thienyl, naphtho[2,3-6]thienyl, thianthrenyl, furyl (furanyl), pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxanthiinyl, pyrrolyl, including without limitation pyrrolyl, including 7H-pyrrolyl, 2H-pyrrolyl, and 3H-pyrrolyl, imidazolyl, pyrazolyl, pyridyl (pyridinyl), including without limitation 2-pyridyl, 3- pyridyl, and 4-pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, phthalzinyl, naphthyridinyl, quinozalinyl, cinnolinyl, pteridinyl, carbazolyl, β-carbolinyl, phenanthridinyl, acrindinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isoxazolyl, furazanyl, phenoxazinyl, l,4-dihydroquinoxaline-2,3-dione, 7-amino- isocoumarin, pyrido[l ,2-α]pyrimidin-4-one, tetrahydrocyclopenta[c]pyrazol-3- yl, pyrazolo[l,5-α]pyrimidinyl, including without limitation pyrazolo[l,5- α]pyrimidin-3-yl, l,2-benzoisoxazol-3-yl, benzimidazolyl, 2-oxindolyl, thiadiazolyl, including 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl,
1,2,4-thiadiazolyl, and 1,2,5-thiadiazolyl, and 2-oxobenzimidazolyl. Where the heteroaryl group contains a nitrogen atom in a ring, such nitrogen atom may be in the form of an N-oxide, e.g., a pyridyl N-oxide, pyrazinyl N-oxide and pyrimidinyl N-oxide.
[0057] The term "heteroaryloxy" is used herein to mean oxygen substituted by one of the above-mentioned heteroaryl groups, which may be optionally substituted. Useful heteroaryloxy groups include pyridyloxy, pyrazinyloxy, pyrrolyloxy, pyrazolyloxy, imidazolyloxy and thiophenyloxy.
[0058] The term "heteroarylalkoxy" is used herein to mean any of the above- mentioned CJ-IO alkoxy groups substituted by any of the above-mentioned heteroaryl groups, which may be optionally substituted.
[0059] Some of the compounds of the present invention may exist as stereoisomers including optical isomers. The invention includes all stereoisomers and both the racemic mixtures of such stereoisomers as well as the individual enantiomers that may be separated according to methods that are well known to those of ordinary skill in the art.
[0060] Examples of pharmaceutically acceptable addition salts include inorganic and organic acid addition salts, such as hydrochloride, hydrobromide, phosphate, sulphate, citrate, lactate, tartrate, maleate, fumarate, mandelate and oxalate; and inorganic and organic base addition salts with bases, such as sodium hydroxy, Tris(hydroxymethyl)aminomethane (TRIS, tromethane) and iV-methyl-glucamine.
[0061] Examples of prodrugs of the compounds of the invention include the simple esters of carboxylic acid containing compounds (e.g., those obtained by condensation with a Ci-4 alcohol according to methods known in the art); esters of hydroxy containing compounds (e.g., those obtained by condensation with a Ci-4 carboxylic acid, C3-6 dioic acid or anhydride thereof, such as succinic and fumaric anhydrides according to methods known in the art); imines of amino containing compounds (e.g., those obtained by condensation with a C i-4 aldehyde or ketone according to methods known in the art); carbamate of amino containing compounds, such as those described by Leu, et. al, (J. Med. Chem. 42:3623-3628 (1999)) and Greenwald, et. al, (J. Med. Chem. 42:3657-3667 (1999)); amide of amino containing compounds via reaction with a protected amino acid and optional cleavage of the protecting group; and acetals and ketals of alcohol containing compounds (e.g., those obtained by condensation with chloromethyl methyl ether or chloromethyl ethyl ether according to methods known in the art).
Amino acid protecting groups are well known and include, for example, t-BOC and FMOC groups.
[0062] The compounds of this invention may be prepared using methods known to those skilled in the art, or the novel methods of this invention. Specifically, the compounds of this invention with Formulae I-V can be prepared as illustrated by the exemplary reaction in Scheme 1. Reaction of 4- amino-5-(2-methoxyphenyl)-3-mercapto-(4H)-l ,2,4-triazole with 2-bromo- 1 - (4-methoxy-3-nitrophenyl)ethanone in isopropanol produced 3-(2- methoxyphenyl)-6-(4-methoxy-3 -nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,A- 6][l,3,4]thiadiazine. The nitro group was reduced by treatment with tin (II) chloride dihydrate to produce the amino compound 6-(3-amino-4- methoxyphenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- ό][l,3,4]thiadiazine. The HCl salt was prepared via treatment with HCl in ether/methanol.
Scheme 1
Figure imgf000027_0001
[0063] Other compounds of this invention may be prepared similarly as illustrated by the exemplary reaction in Scheme 2. Reaction of methyl 4- bromo-2-methoxybenzoate with hydrazine hydrate in ethanol produced 4- bromo-2-methylbenzohydrazide, which was reacted with carbon disulfide in ethanol in the presence of a base such as KOH to produce 4-bromo-2- methylbenzoyl-2-dithiocarboxyhydrazide as a potassium salt. Reaction of the potassium salt of 4-bromo-2-methylbenzoyl-2-dithiocarboxyhydrazide with hydrazine hydrate in ethanol produced 4-amino-5-(4-bromo-2-methylphenyl)- 3-mercapto-(4H)-l,2,4-triazole, which was reacted with 2-bromo-l-(4- methylphenyl)ethanone in isopropanol to produce 3-(4-bromo-2- methylphenyl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine.
Scheme 2
Figure imgf000028_0001
An amino acid prodrug of compounds of this invention can be prepared as illustrated by the exemplary reaction in Scheme 3. Reaction of 6- (3 -amino-4-methylphenyl)-3 -(2-methoxypheny 1)-7H- [ 1 ,2,4]triazolo [3 ,A- δ][l,3,4]thiadiazine with t-butoxycarbonyl-alanine (Boc-Ala) in the presence of benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), Ηydroxybenzotriazole (ΗOBt) and triethyl amine (Et3N) in acetonitrile produced 6-(3-(Boc-alaninyl-amino)-4-methylphenyl)-3-(2- methoxyphenyl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine. The Boc protecting group was removed by treatment with HCl in dioxane/CΗ2Cl2 to produce 6-(3-(alaninyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine, the alanine prodrug. It was found that the HCl salt of the alanine prodrug is highly water soluble with solubility of >14 mg/ml at a pΗ = 6 aqueous solution. Scheme 3
Figure imgf000029_0001
[0065] An important aspect of the present invention is the discovery that compounds having Formulae I-V are activators of caspases and inducers of apoptosis. Therefore, these compounds are useful in a variety of clinical conditions in which there is uncontrolled cell growth and spread of abnormal cells, such as in the case of cancer.
[0066] Another important aspect of the present invention is the discovery that compounds having Formulae I-V are potent and highly efficacious activators of caspases and inducers of apoptosis in drug resistant cancer cells, such as breast and prostate cancer cells, which enables these compounds to kill these drug resistant cancer cells. In comparison, most standard anti-cancer drugs are not effective in killing drug resistant cancer cells under the same conditions. Therefore, compounds of this invention are useful for the treatment of drug resistant cancer, such as breast cancer in animals.
[0067] The present invention includes a therapeutic method useful to modulate in vivo apoptosis or in vivo neoplastic disease, comprising administering to a subject in need of such treatment an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis.
[0068] The present invention also includes a therapeutic method comprising administering to an animal an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of said compound of Formulae I- V, wherein said therapeutic method is useful to treat cancer, which is a group of diseases characterized by the uncontrolled growth and spread of abnormal cells. Such diseases include, but are not limited to, Hodgkin's disease, non- Hodgkin's lymphoma, acute lymphocytic leukemia, chronic lymphocytic leukemia, multiple myeloma, neuroblastoma, breast carcinoma, ovarian carcinoma, lung carcinoma, Wilms1 tumor, cervical carcinoma, testicular carcinoma, soft-tissue sarcoma, primary macroglobulinemia, bladder carcinoma, chronic granulocytic leukemia, primary brain carcinoma, malignant melanoma, small-cell lung carcinoma, stomach carcinoma, colon carcinoma, malignant pancreatic insulinoma, malignant carcinoid carcinoma, choriocarcinoma, mycosis fungoides, head or neck carcinoma, osteogenic sarcoma, pancreatic carcinoma, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary carcinoma, thyroid carcinoma, esophageal carcinoma, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial carcinoma, polycythemia vera, essential thrombocytosis, adrenal cortex carcinoma, skin cancer, and prostatic carcinoma.
[0069] In practicing the therapeutic methods, effective amounts of compositions containing therapeutically effective concentrations of the compounds formulated for oral, intravenous, local and topical application, for the treatment of neoplastic diseases and other diseases in which caspase cascade mediated physiological responses are implicated, are administered to an individual exhibiting the symptoms of one or more of these disorders. The amounts are effective to ameliorate or eliminate one or more symptoms of the disorders. An effective amount of a compound for treating a particular disease is an amount that is sufficient to ameliorate, or in some manner reduce, the symptoms associated with the disease. Such amount may be administered as a single dosage or may be administered according to a regimen, whereby it is effective. The amount may cure the disease but, typically, is administered in order to ameliorate the symptoms of the disease. Typically, repeated administration is required to achieve the desired amelioration of symptoms.
[0070] In another embodiment, a pharmaceutical composition comprising a compound, or a pharmaceutically acceptable salt of said compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis in combination with a pharmaceutically acceptable vehicle is provided.
[0071] Another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of said compound of Formulae I- V, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known cancer chemotherapeutic agent, or a pharmaceutically acceptable salt of said agent. Examples of known cancer chemotherapeutic agents which may be used for combination therapy include, but not are limited to alkylating agents, such as busulfan, cis-platin, mitomycin C, and carboplatin; antimitotic agents, such as colchicine, vinblastine, paclitaxel, and docetaxel; topo I inhibitors, such as camptothecin and topotecan; topo II inhibitors, such as doxorubicin and etoposide; RNA/DNA antimetabolites, such as 5-azacytidine, 5-fluorouracil and methotrexate; DNA antimetabolites, such as 5-fluoro-2'-deoxy-uridine, ara-C, hydroxyurea and thioguanine; antibodies, such as campath, Herceptin® or Rituxan®. Other known cancer chemotherapeutic agents which may be used for combination therapy include melphalan, chlorambucil, cyclophosamide, ifosfamide, vincristine, mitoguazone, epirubicin, aclarubicin, bleomycin, mitoxantrone, elliptinium, fludarabine, octreotide, retinoic acid, tamoxifen, Gleevec® and alanosine.
[0072] In practicing the methods of the present invention, the compound of the invention may be administered together with at least one known chemotherapeutic agent as part of a unitary pharmaceutical composition. Alternatively, the compound of the invention may be administered apart from at least one known cancer chemotherapeutic agent. In one embodiment, the compound of the invention and at least one known cancer chemotherapeutic agent are administered substantially simultaneously, i.e. the compounds are administered at the same time or one after the other, so long as the compounds reach therapeutic levels in the blood at the same time. On another embodiment, the compound of the invention and at least one known cancer chemotherapeutic agent are administered according to their individual dose schedule, so long as the compounds reach therapeutic levels in the blood.
[0073] It has been reported that alpha- 1 -adrenoceptor antagonists, such as doxazosin, terazosin, and tamsulosin can inhibit the growth of prostate cancer cell via induction of apoptosis (Kyprianou, N., et ah, Cancer Res 60:4550- 4555, (2000)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known alpha- 1 -adrenoceptor antagonists, or a pharmaceutically acceptable salt of said agent. Examples of known alpha- 1 -adrenoceptor antagonists, which can be used for combination therapy include, but are not limited to, doxazosin, terazosin, and tamsulosin.
[0074] It has been reported that sigma-2 receptors are expressed in high densities in a variety of tumor cell types (Vilner, B. J., et al., Cancer Res. 55: 408-413 (1995)) and that sigma-2 receptor agonists, such as CB-64D, CB- 184 and haloperidol activate a novel apoptotic pathway and potentiate antineoplastic drugs in breast tumor cell lines. (Kyprianou, N., et al., Cancer Res. (52:313-322 (2002)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known sigma-2 receptor agonist, or a pharmaceutically acceptable salt of said agonist. Examples of known sigma-2 receptor agonists which can be used for combination therapy include, but are not limited to, CB-64D, CB-184 and haloperidol.
[0075] It has been reported that combination therapy with lovastatin, a HMG-
CoA reductase inhibitor, and butyrate, an inducer of apoptosis in the Lewis lung carcinoma model in mice, showed potentiating antitumor effects (Giermasz, A., et al., Int. J. Cancer 97: 746-750 (2002)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known HMG-CoA reductase inhibitor, or a pharmaceutically acceptable salt of said agent. Examples of known HMG-CoA reductase inhibitors, which can be used for combination therapy include, but are not limited to, lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin and cerivastatin.
[0076] It has been reported that HIV protease inhibitors, such as indinavir or saquinavir, have potent anti-angiogenic activities and promote regression of Kaposi sarcoma (Sgadari, C, et al., Nat. Med. 5:225-232 (2002)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known HIV protease inhibitor, or a pharmaceutically acceptable salt of said agent. Examples of known HIV protease inhibitors, which can be used for combination therapy include, but are not limited to, amprenavir, abacavir, CGP-73547, CGP-61755, DMP-450, indinavir, nelfinavir, tipranavir, ritonavir, saquinavir, ABT-378, AG 1776, and BMS-232,632.
[0077] It has been reported that synthetic retinoids, such as fenretinide (N-(4- hydroxyphenyl)retinamide, 4HPR), have good activity in combination with other chemotherapeutic agents, such as cisplatin, etoposide or paclitaxel in small-cell lung cancer cell lines (Kalemkerian, G. P., et al, Cancer Chemother. Pharmacol. 45:145-150 (1999)). 4HPR also was reported to have good activity in combination with gamma-radiation on bladder cancer cell lines (Zou, C, et al, Int. J. Oncol. 75:1037-1041 (1998)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known retinoid and synthetic retinoid, or a pharmaceutically acceptable salt of said agent. Examples of known retinoids and synthetic retinoids, which can be used for combination therapy include, but are not limited to, bexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid, α-difluoromethylornithine, ILX23-7553, fenretinide, and N-4-carboxyphenyl retinamide.
[0078] It has been reported that proteasome inhibitors, such as lactacystin, exert anti-tumor activity in vivo and in tumor cells in vitro, including those resistant to conventional chemotherapeutic agents. By inhibiting NF-kappaB transcriptional activity, proteasome inhibitors may also prevent angiogenesis and metastasis in vivo and further increase the sensitivity of cancer cells to apoptosis (Almond, J. B., et al., Leukemia 75:433-443 (2002)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known proteasome inhibitor, or a pharmaceutically acceptable salt of said agent. Examples of known proteasome inhibitors, which can be used for combination therapy include, but are not limited to, lactacystin, MG- 132, and PS-341.
[0079] It has been reported that tyrosine kinase inhibitors, such as STI571
(Imatinib mesilate, Gleevec®), have potent synergetic effect in combination with other anti-leukemic agents, such as etoposide (Liu, W.M., et al. Br. J. Cancer 56:1472-1478 (2002)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known tyrosine kinase inhibitor, or a pharmaceutically acceptable salt of said agent. Examples of known tyrosine kinase inhibitors, which can be used for combination therapy include, but are not limited to, Gleevec®, ZDl 839 (Iressa), SH268, genistein, CEP2563, SU6668, SUl 1248, and EMD121974.
[0080] It has been reported that prenyl-protein transferase inhibitors, such as farnesyl protein transferase inhibitor Rl 15777, possess preclinical antitumor activity against human breast cancer (Kelland, L. R., et. al., Clin. Cancer Res. 7:3544-3550 (2001)). Synergy of the protein farnesyltransferase inhibitor SCH66336 and cisplatin in human cancer cell lines also has been reported (Adjei, A. A., et al, Clin. Cancer. Res. 7:1438-1445 (2001)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known prenyl-protein transferase inhibitor, including farnesyl protein transferase inhibitor, inhibitors of geranylgeranyl-protein transferase type I (GGPTase-I) and geranylgeranyl-protein transferase type-II, or a pharmaceutically acceptable salt of said agent. Examples of known prenyl- protein transferase inhibitors, which can be used for combination therapy include, but are not limited to, Rl 15777, SCH66336, L-778,123, BAL9611 and TAN-1813.
[0081] It has been reported that cyclin-dependent kinase (CDK) inhibitors, such as flavopiridol, have potent synergetic effect in combination with other anticancer agents, such as CPT-11, a DNA topoisomerase I inhibitor in human colon cancer cells (Motwani, M., et al., Clin. Cancer Res. 7:4209-4219, (2001)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known cyclin-dependent kinase inhibitor, or a pharmaceutically acceptable salt of said agent. Examples of known cyclin- dependent kinase inhibitor, which can be used for combination therapy include, but are not limited to, flavopiridol, UCN-01, roscovitine and olomoucine.
[0082] It has been reported that in preclinical studies COX-2 inhibitors were found to block angiogenesis, suppress solid tumor metastases, and slow the growth of implanted gastrointestinal cancer cells (Blanke, C. D., Oncology (Huntingt) 16 (No. 4 Suppl. 3): 17-21 (2002)). Therefore, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with at least one known COX-2 inhibitor, or a pharmaceutically acceptable salt of said inhibitor. Examples of known COX-2 inhibitors which can be used for combination therapy include, but are not limited to, celecoxib, valecoxib, and rofecoxib.
[0083] Another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a bioconjugate of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in bioconjugation with at least one known therapeutically useful antibody, such as Herceptin® or Rituxan®, growth factors, such as DGF, NGF; cytokines, such as IL-2, IL-4, or any molecule that binds to the cell surface. The antibodies and other molecules will deliver a compound described herein to its targets and make it an effective anticancer agent. The bioconjugates could also enhance the anticancer effect of therapeutically useful antibodies, such as Herceptin® or Rituxan®.
[0084] Similarly, another embodiment of the present invention is directed to a composition effective to inhibit neoplasia comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis, in combination with radiation therapy. In this embodiment, the compound of the invention may be administered at the same time as the radiation therapy is administered or at a different time.
[0085] Yet another embodiment of the present invention is directed to a composition effective for post-surgical treatment of cancer, comprising a compound, or a pharmaceutically acceptable salt or prodrug of a compound described herein, which functions as a caspase cascade activator and inducer of apoptosis. The invention also relates to a method of treating cancer by surgically removing the cancer and then treating the animal with one of the pharmaceutical compositions described herein.
[0086] A wide range of immune mechanisms operates rapidly following exposure to an infectious agent. Depending on the type of infection, rapid clonal expansion of the T and B lymphocytes occurs to combat the infection. The elimination of the effector cells following an infection is one of the major mechanisms for maintaining immune homeostasis. The elimination of the effector cells has been shown to be regulated by apoptosis. Autoimmune diseases have lately been determined to occur as a consequence of deregulated cell death. In certain autoimmune diseases, the immune system directs its powerful cytotoxic effector mechanisms against specialized cells, such as oligodendrocytes in multiple sclerosis, the beta cells of the pancreas in diabetes mellitus, and thyrocytes in Hashimoto's thyroiditis (Ohsako, S. & Elkon, K.B., Cell Death Differ. 6:13-21 (1999)). Mutations of the gene encoding the lymphocyte apoptosis receptor Fas/APO-l/CD95 are reported to be associated with defective lymphocyte apoptosis and autoimmune lymphoproliferative syndrome (ALPS), which is characterized by chronic, histologically benign splenomegaly, generalized lymphadenopathy, hypergammaglobulinemia, and autoantibody formation. (Infante, A.J., et al, J. Pediatr. 133:629-633 (1998) and Vaishnaw, A.K., et al, J. Clin. Invest. 103:355-363 (1999)). It was reported that overexpression of Bcl-2, which is a member of the bcl-2 gene family of programmed cell death regulators with anti-apoptotic activity, in developing B cells of transgenic mice, in the presence of T cell dependent costimulatory signals, results in the generation of a modified B cell repertoire and in the production of pathogenic autoantibodies (Lopez-Hoyos, M., et al, Int. J. MoI. Med. 7:475-483 (1998)). It is therefore evident that many types of autoimmune disease are caused by defects of the apoptotic process. One treatment strategy for such diseases is to turn on apoptosis in the lymphocytes that are causing the autoimmune disease (O'Reilly, L. A. & Strasser, A., Inflamm. Res. 48:5-21 (1999)).
[0087] Fas-Fas ligand (FasL) interaction is known to be required for the maintenance of immune homeostasis. Experimental autoimmune thyroiditis (EAT), characterized by autoreactive T and B cell responses and a marked lymphocytic infiltration of the thyroid, is a good model to study the therapeutic effects of FasL. Batteux, F., et al, (J. Immunol. 752:603-608 (1999)) reported that by direct injection of DNA expression vectors encoding FasL into the inflamed thyroid, the development of lymphocytic infiltration of the thyroid was inhibited and induction of infiltrating T cells death was observed. These results show that FasL expression on thyrocytes may have a curative effect on ongoing EAT by inducing death of pathogenic autoreactive infiltrating T lymphocytes.
[0088] Bisindolylmaleimide VIII is known to potentiate Fas-mediated apoptosis in human astrocytoma 132 INl cells and in Molt-4T cells; both of which were resistant to apoptosis induced by anti-Fas antibody in the absence of bisindolylmaleimide VIII. Potentiation of Fas-mediated apoptosis by bisindolylmaleimide VIII was reported to be selective for activated, rather than non-activated, T cells, and was Fas-dependent. Zhou T., et al, (Nat. Med. 5:42-48 (1999)) reported that administration of bisindolylmaleimide VIII to rats during autoantigen stimulation prevented the development of symptoms of T cell-mediated autoimmune diseases in two models, the Lewis rat model of experimental allergic encephalitis and the Lewis adjuvant arthritis model. Therefore, the application of a Fas-dependent apoptosis enhancer, such as bisindolylmaleimide VIII, may be therapeutically useful for the more effective elimination of detrimental cells and inhibition of T cell-mediated autoimmune diseases. Therefore, an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I- V, which functions as a caspase cascade activator and inducer of apoptosis, is an effective treatment for autoimmune diseases.
[0089] Psoriasis is a chronic skin disease that is characterized by scaly red patches. Psoralen plus ultraviolet A (PUVA) is a widely used and effective treatment for psoriasis vulgaris. Coven, et al., Photodermatol. Photoimmunol. Photomed. 15:22-27 (1999), reported that lymphocytes treated with psoralen 8-MOP or TMP and UVA, displayed DNA degradation patterns typical of apoptotic cell death. Ozawa, et al, J. Exp. Med. 7S9:711-718 (1999) reported that induction of T cell apoptosis could be the main mechanism by which 312- nm UVB resolves psoriasis skin lesions. Low doses of methotrexate may be used to treat psoriasis to restore a clinically normal skin. Heenen, et al, Arch. Dermatol. Res. 290:240-245 (1998), reported that low doses of methotrexate may induce apoptosis and that this mode of action could explain the reduction in epidermal hyperplasia during treatment of psoriasis with methotrexate. Therefore, an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis, is an effective treatment for hyperproliferative skin diseases, such as psoriasis.
[0090] Synovial cell hyperplasia is a characteristic of patients with rheumatoid arthritis (RA). It is believed that excessive proliferation of RA synovial cells, as well as defects in synovial cell death, may be responsible for synovial cell hyperplasia. Wakisaka, et al., Clin. Exp. Immunol. 774:119-128 (1998), found that although RA synovial cells could die via apoptosis through a Fas/FasL pathway, apoptosis of synovial cells was inhibited by proinflammatory cytokines present within the synovium. Wakisaka, et al. also suggested that inhibition of apoptosis by the proinflammatory cytokines may contribute to the outgrowth of synovial cells, and lead to pannus formation and the destruction of joints in patients with RA. Therefore, an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis, is an effective treatment for rheumatoid arthritis.
[0091] There has been an accumulation of convincing evidence that apoptosis plays a major role in promoting resolution of the acute inflammatory response. Neutrophils are constitutively programmed to undergo apoptosis, thus limiting their pro-inflammatory potential and leading to rapid, specific, and non- phlogistic recognition by macrophages and semi-professional phagocytes (Savill, J., J. Leukoc. Biol. 67:375-380 (1997)). Boirivant, et al, Gastroenterology 116:557-565 (1999), reported that lamina propria T cells, isolated from areas of inflammation in Crohn's disease, ulcerative colitis, and other inflammatory states, manifest decreased CD2 pathway-induced apoptosis. In addition, studies of cells from inflamed Crohn's disease tissue indicate that this defect is accompanied by elevated Bcl-2 levels. Therefore, an effective amount of a compound, or a pharmaceutically acceptable salt or prodrug of the compound of Formulae I-V, which functions as a caspase cascade activator and inducer of apoptosis, is an effective treatment for inflammation.
[0092] Caspase cascade activators and inducers of apoptosis may also be a desirable therapy in the elimination of pathogens, such as HIV, Hepatitis C and other viral pathogens. The long lasting quiescence, followed by disease progression, may be explained by an anti-apoptotic mechanism of these pathogens leading to persistent cellular reservoirs of the virions. It has been reported that HIV-I infected T leukemia cells or peripheral blood mononuclear cells (PBMCs) underwent enhanced viral replication in the presence of the caspase inhibitor Z-VAD-fmk. Furthermore, Z-VAD-fmk also stimulated endogenous virus production in activated PBMCs derived from HIV-I- infected asymptomatic individuals (Chinnaiyan, A., et al., Nat. Med. J:333 (1997)). Therefore, apoptosis serves as a beneficial host mechanism to limit the spread of HIV and new therapeutics using caspase/apoptosis activators are useful to clear viral reservoirs from the infected individuals. Similarly, HCV infection also triggers anti-apoptotic mechanisms to evade the host's immune surveillance leading to viral persistence and hepatocarcinogenesis (Tai, D.I., et al. Hepatology 3:656-64 (2000)). Therefore, apoptosis inducers are useful as therapeutics for HIV and other infectious disease. [0093] Stent implantation has become the new standard angioplasty procedure. However, in-stent restenosis remains the major limitation of coronary stenting. New approaches have been developed to target pharmacological modulation of local vascular biology by local administration of drugs. This allows for drug applications at the precise site and time of vessel injury. Numerous pharmacological agents with antiproliferative properties are currently under clinical investigation, including actinomycin D, rapamycin or paclitaxel coated stents (Regar E., et al., Br. Med. Bull. 59:227- 248 (2001)). Therefore, apoptosis inducers, which are antiproliferative, are useful as therapeutics for the prevention or reduction of in-stent restenosis.
[0094] Pharmaceutical compositions within the scope of this invention include all compositions wherein the compounds of the present invention are contained in an amount that is effective to achieve its intended purpose. While individual needs vary, determination of optimal ranges of effective amounts of each component is within the skill of the art. Typically, the compounds may be administered to animals, e.g., mammals, orally at a dose of 0.0025 to 50 mg/kg of body weight, per day, or an equivalent amount of the pharmaceutically acceptable salt thereof, to a mammal being treated. Preferably, approximately 0.01 to approximately 10 mg/kg of body weight is orally administered. For intramuscular injection, the dose is generally approximately one-half of the oral dose. For example, a suitable intramuscular dose would be approximately 0.0025 to approximately 25 mg/kg of body weight, and most preferably, from approximately 0.01 to approximately 5 mg/kg of body weight. If a known cancer chemotherapeutic agent is also administered, it is administered in an amount that is effective to achieve its intended purpose. The amounts of such known cancer chemotherapeutic agents effective for cancer are well known to those skilled in the art.
[0095] The unit oral dose may comprise from approximately 0.01 to approximately 50 mg, preferably approximately 0.1 to approximately 10 mg of the compound of the invention. The unit dose may be administered one or more times daily, as one or more tablets, each containing from approximately 0.1 to approximately 10 mg, conveniently approximately 0.25 to 50 mg of the compound or its solvates. [0096] In a topical formulation, the compound may be present at a concentration of approximately 0.01 to 100 mg per gram of carrier.
[0097] In addition to administering the compound as a raw chemical, the compounds of the invention may be administered as part of a pharmaceutical preparation containing suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the compounds into preparations that may be used pharmaceutically. Preferably, the preparations, particularly those preparations which may be administered orally and that may be used for the preferred type of administration, such as tablets, dragees, and capsules, and also preparations that may be administered rectally, such as suppositories, as well as suitable solutions for administration by injection or orally, contain from approximately 0.01 to 99 percent, preferably from approximately 0.25 to 75 percent of active compound(s), together with the excipient.
[0098] Also included within the scope of the present invention are the nontoxic pharmaceutically acceptable salts of the compounds of the present invention. Acid addition salts are formed by mixing a solution of the compounds of the present invention with a solution of a pharmaceutically acceptable non-toxic acid, such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid, and the like. Basic salts are formed by mixing a solution of the compounds of the present invention with a solution of a pharmaceutically acceptable non-toxic base, such as sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, Tris, N-methyl- glucamine and the like.
[0099] The pharmaceutical compositions of the invention may be administered to any animal, which may experience the beneficial effects of the compounds of the invention. Foremost among such animals are mammals, e.g., humans and veterinary animals, although the invention is not intended to be so limited.
[00100] The pharmaceutical compositions of the present invention may be administered by any means that achieve their intended purpose. For example, administration may be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal or topical routes. Alternatively, or concurrently, administration may be by the oral route. The dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment, and the nature of the effect desired.
[00101] The pharmaceutical preparations of the present invention are manufactured in a manner, which is itself known, e.g., by means of conventional mixing, granulating, dragee-making, dissolving, or lyophilizing processes. Thus, pharmaceutical preparations for oral use may be obtained by combining the active compounds with solid excipients, optionally grinding the resulting mixture and processing the mixture of granules, after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores.
[00102] Suitable excipients are, in particular: fillers, such as saccharides, e.g. lactose or sucrose, mannitol or sorbitol; cellulose preparations and/or calcium phosphates, e.g. tricalcium phosphate or calcium hydrogen phosphate; as well as binders, such as starch paste, using, e.g., maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone. If desired, disintegrating agents may be added, such as the above-mentioned starches and also carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate. Auxiliaries are, above all, flow-regulating agents and lubricants, e.g., silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium stearate, and/or polyethylene glycol. Dragee cores are provided with suitable coatings which, if desired, are resistant to gastric juices. For this purpose, concentrated saccharide solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. In order to produce coatings resistant to gastric juices, solutions of suitable cellulose preparations, such as acetylcellulose phthalate or hydroxypropylmethyl-cellulose phthalate, are used. Dye stuffs or pigments may be added to the tablets or dragee coatings, e.g., for identification or in order to characterize combinations of active compound doses.
[00103] Other pharmaceutical preparations, which may be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules may contain the active compounds in the form of: granules, which may be mixed with fillers, such as lactose; binders, such as starches; and/or lubricants, such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds are preferably dissolved or suspended in suitable liquids, such as fatty oils, or liquid paraffin. In addition, stabilizers may be added.
[00104] Possible pharmaceutical preparations, which may be used rectally include, e.g., suppositories, which consist of a combination of one or more of the active compounds with a suppository base. Suitable suppository bases are, e.g., natural or synthetic triglycerides, or paraffin hydrocarbons. In addition, it is also possible to use gelatin rectal capsules, which consist of a combination of the active compounds with a base. Possible base materials include, e.g., liquid triglycerides, polyethylene glycols, or paraffin hydrocarbons.
[00105] Suitable formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form, e.g., water-soluble salts and alkaline solutions. In addition, suspensions of the active compounds as appropriate oily injection suspensions may be administered. Suitable lipophilic solvents or vehicles include fatty oils, e.g., sesame oil, or synthetic fatty acid esters, e.g., ethyl oleate or triglycerides or polyethylene glycol-400 (the compounds are soluble in PEG-400), or cremophor, or cyclodextrins. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension include, e.g., sodium carboxymethyl cellulose, sorbitol, and/or dextran. Optionally, the suspension may also contain stabilizers.
[00106] In accordance with one aspect of the present invention, compounds of the invention are employed in topical and parenteral formulations and are used for the treatment of skin cancer.
[00107] The topical compositions of this invention are formulated preferably as oils, creams, lotions, ointments and the like by choice of appropriate carriers. Suitable carriers include vegetable or mineral oils, white petrolatum (white soft paraffin), branched chain fats or oils, animal fats and high molecular weight alcohol (greater than Ci2). The preferred carriers are those in which the active ingredient is soluble. Emulsifiers, stabilizers, humectants and antioxidants may also be included, as well as agents imparting color or fragrance, if desired. Additionally, transdermal penetration enhancers may be employed in these topical formulations. Examples of such enhancers are found in U.S. Patent Nos. 3,989,816 and 4,444,762.
[00108] Creams are preferably formulated from a mixture of mineral oil, self- emulsifying beeswax and water in which mixture of the active ingredient, dissolved in a small amount of an oil, such as almond oil, is admixed. A typical example of such a cream is one which includes approximately 40 parts water, approximately 20 parts beeswax, approximately 40 parts mineral oil and approximately 1 part almond oil.
[00109] Ointments may be formulated by mixing a solution of the active ingredient in a vegetable oil, such as almond oil, with warm soft paraffin and allowing the mixture to cool. A typical example of such an ointment is one which includes approximately 30 % almond oil and approximately 70 % white soft paraffin by weight.
[00110] The following examples are illustrative, but not limiting, of the method and compositions of the present invention. Other suitable modifications and adaptations of the variety of conditions and parameters normally encountered in clinical therapy and which are obvious to those skilled in the art are within the spirit and scope of the invention.
EXAMPLE 1
3-(2-Methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00111] (a) 2-Bromo-l-(4-methoxy-3-nitrophenyl)ethanone. To a solution of
4-methoxy-3-nitroacetophenone (3.90 g, 20 mmol) in dichloromethane (30 mL) was added bromine (3.2 g, 20 mmol) dropwise. It was stirred at room temperature for 1.5 h until the red color disappeared. It was evaporated under vacuo and the residue was crystallized from ethyl acetate/hexane (10:1) to give 2.9 g (53%) of the title compound. 1H NMR (CDCl3) 8.48 (d, J= 2.4 Hz, IH), 8.23-8.19 (dd, J= 8.7 and 2.1 Hz, IH), 7.20 (d, J = 9.0 Hz, IH), 4.39 (s, 2H), 4.07 (s, 3H).
[00112] (b) 3-(2-Methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine. A mixture of 2-bromo-l-(4-methoxy- 3-nitrophenyl)ethanone (741 mg, 2.7 mmol) and 4-amino-5-(2- methoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (600 mg, 2.7 mmol) in isopropanol was refluxed for 4 h. It was cooled and neutralized with aqueous sodium carbonate. The precipitate was collected and dried to give 1.05 g (98%) of the title compound. 1H NMR (CDCl3) 8.36 (d, J= 2.4 Hz, IH), 8.04- 8.00 (dd, J = 9.0 and , 2.7 Hz, IH), 7.61 (d, J = 9.0 Hz, IH), 7.52 (t, J = 8.7 Hz, IH), 7.17 (d, J= 8.7 Hz, IH), 7.11 (t, J= 7.5 Hz, IH), 7.02 (d, J= 8.7 Hz, IH), 4.03 (s, 3H), 3.98 (s, 2H), 3.77 (s, 3H).
EXAMPLE 2
6-(3 - Amino-4-methoxyphenyl)-3 -(2-methoxyphenyl)-7H-[ 1 ,2,4]triazolo [3 ,4- 6][l,3,4]thiadiazine hydrochloride
[00113] A mixture of 3-(2-methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-b][l,3,4]thiadiazine (1.05 g, 2.64 mmol) and tin(II) chloride (2.69 g, 11.9 mmol) in methanol (25 mL) was refluxed under argon for 5 h. It was then diluted with water (30 mL) and neutralized with 2N sodium hydroxide to pH = 12. The mixture was extracted with ethyl acetate (3 x 20 mL). The extracts were dried, concentrated and the residue was purified by column chromatography (EtOAc/ MeOH 100:1). The product was dissolved in methanol (10 mL) and acidified with IN hydrochloride in ether (8 mL). The solution was evaporated to around 1 mL and ethyl ether (20 mL) was added into the solution to produce a precipitate. The solid was collected via filtration to give 860 mg (74%) of the title compound. 1H NMR (CD3OD) 8.15 (d, J = 8.7 Hz, IH), 8.07 (d, J = 2.1 Hz, IH), 7.84 (d, J = 7.8 Hz, IH), 7.75 (t, J = 7.2 Hz, IH), 7.43 (d, J = 8.4 Hz, IH), 7.35 (d, J = 8.7 Hz, IH), 7.25 (t, J= 7.5 Hz, IH), 4.50 (s, 2H), 4.09 (s, 3H), 3.96 (s, 3H).
EXAMPLE 3
3-(4-Chloro-2-methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00114] The title compound was prepared in a manner similar to Example Ib.
From 2-bromo-l-(4-methoxy-3-nitrophenyl)ethanone (170 mg, 0.62 mmol) and 4-amino-5-(4-chloro-2-methoxyphenyl)-3-mercapto-(4//)- 1 ,2,4-triazole (159 mg, 0.62 mmol) in isopropanol was obtained 230 mg (86%) of the title compound. 1H NMR (CD3OD) 8.30 (d, J = 2.7 Hz, IH), 8.04-8.00 (dd, J= 9.3 and , 2.7 Hz, 2H), 7.54 (d, J= 8.4 Hz, IH), 7.19 (d, J= 9.3 Hz, IH), 7.12-7.08 (m, IH), 7.0.2 (d, J= 2.1 Hz, IH), 4.04 (s, 3H), 3.99 (s, 2H), 3.77 (s, 3H).
EXAMPLE 4
6-(3 - Amino-4-methoxyphenyl)-3 -(4-chloro-2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine hydrochloride
[00115] The title compound was prepared in a manner similar to Example 2.
From 3-(4-chloro-2-methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-b][l,3,4]thiadiazine (230 mg, 0.53 mmol) and tin chloride (598 mg, 2.66 mmol) was obtained 167 mg (66%) of the title compound. 1H NMR (CD3OD) 8.10 (d, J = 8.7 Hz, IH), 8.01 (s, IH), 7.67 (d, J = 8.4 Hz, IH), 7.40 (d, J= 8.7 Hz, IH), 7.34 (s, IH), 7.25 (d, J= 8.4 Hz, IH), 4.32 (s, 2H), 4.10 (s, 2H), 3.89 (s, 3H).
EXAMPLE 5
3-(3,5-Dimethoxyphenyl)-6-(4-nitrophenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00116] The title compound was prepared in a manner similar to Example Ib.
From 2-bromo-l-(4-nitrophenyl)ethanone (513 mg, 2.1 mmol) and 4-amino-5- (3,5-dimethoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (530 mg, 2.1 mmol) in isopropanol was obtained 780 mg (94%) of the title compound. 1H NMR (DMSO-d6) 8.40 (d, J = 9.0 Hz, 2H), 8.22 (d, J = 9.0 Hz, 2H), 7.16 (s, 2H), 6.68 (s, IH), 4.47 (s, 2H), 3.80 (s, 6H).
EXAMPLE 6
6-(4-Aminophenyl)-3-(3,5-dimethoxyphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine hydrochloride
[00117] The title compound was prepared in a manner similar to Example 2.
From 3-(3,5-dimethoxyphenyl)-6-(4-nitrophenyl)-7H-[l,2,4]triazolo[3,4- ό][l,3,4]thiadiazine (564 mg, 1.41 mmol) and tin chloride (1.59 g, 7.1 mmol) was obtained 235 mg (38%) of the title compound. 1H NMR (CD3OD) 8.18 (d, J= 8.7 Hz, 2H), 7.40 (d, J= 2.1 Hz, IH), 7.33 (d, J= 8.7 Hz, 2H), 6.89 (t, J = 2.4 Hz, IH), 4.50 (s, 2H), 4.10 (s, 2H), 4.00 (s, 3H). EXAMPLE 7
6-(4-Methoxy-3-nitrophenyl)-3-(2-methylftιran-3-yl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00118] The title compound was prepared in a manner similar to Example Ib.
From 2-bromo-l-(4-methoxy-3-nitrophenyl)ethanone (140 mg, 0.51 mmol) and 4-amino-5-(2-methylfuan)-3-mercapto-(4H)-l,2,4-triazole (100 mg, 0.51 mmol) in isopropanol was obtained 178 mg (94%) of the title compound. 1H NMR (acetone-d6) 8.53 (d, J = 2.4 Hz, IH), 8.41-8.37 (dd, J = 9.0 and , 2.4 Hz, IH), 7.58 (d, J = 2.1 Hz, IH), 7.57 (d, J = 8.7 Hz, 2H), 7.02 (d, J = 2.1 Hz, IH), 4.46 (s, 2H), 4.11 (s, 3H), 2.62 (s, 3H).
EXAMPLE 8
6-(3-Amino-4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H-[l,2,4]triazolo[3,4-
Z>][l,3,4]thiadiazine hydrochloride
[00119] The title compound was prepared in a manner similar to Example 2.
From 6-(4-methoxy-3-nitrophenyl)-3-(2-methylfuran-3-yl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (100 mg, 0.27 mmol) and tin chloride (240 mg, 1.08 mmol) was obtained 107 mg (97%) of the title compound. 1H NMR (CD3OD) 8.16 (m, 2H), 7.62 (d, J = 1.8 Hz, 2H), 7.42 (d, J = 9.6 Hz, IH), 6.98 (d, J= 1.8 Hz, IH), 4.44 (s, 2H), 4.10 (s, 2H), 4.10 (s, 3H), 2.59 (s, 3H).
EXAMPLE 9
6-(3-Dimethylamino-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine hydrochloride
[00120] To a mixture of 6-(3-amino-4-methoxyphenyl)-3-(2-methoxyphenyl)-
7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (367 mg, 1 mmol) in methanol (10 mL) was added 5 drops of acetic acid and 37% aqueous formaldehyde (487 mg, 6 mmol). It was stirred at room temperature for 1 h and was then cooled to 0 0C. To it was added sodium borohydride cyanate (377 mg, 6 mmol), and it was stirred at room temperature overnight. It was evaporated and the residue was dissolved in ethyl acetate (30 mL). The solution was washed with water (30 mL), dried and evaporated. The residue was purified by column chromatography to give the product. It was dissolved in methanol (8 mL), and 2 mL of 2N hydrochloride in ether was added. After evaporation of most solvent, ether (10 mL) was added to produce the title compound as white solid (235 mg, 50%). 1H NMR (CD3OD) 8.46 (bs, IH), 8.26 (d, J = 6.9 Hz, IH), 7.99 (s, IH), 7.79 (t, J= 6.9 Hz, IH), 7.51 (d, J= 6.6 Hz, IH), 7.39 (d, J = 8.1 Hz, IH), 7.29 (bs, IH), 4.63 (bs, 2H), 4.16 (s, 3H), 4.02 (s, 3H), 3.35 (s, 6H).
EXAMPLE 10
3-(4-Bromo-2-methoxyphenyl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00121] (a) 4-Bromo-2-methoxybenzohydrazide. To an oven-dried round bottom flask charged with a magnetic stir bar at room temperature under argon was added methyl 4-bromo-2-methoxybenzoate (1.00 g, 4.08 mmol), ethanol (20 mL) and hydrazine hydrate (0.60 mL, 12 mmol). The clear solution was refluxed at 100 0C for 4 h and then cooled to room temperature. The resulting white precipitate was diluted with ethanol and then filtered through a buchner funnel to give 0.275 g (28%) of the title compound as a white solid. 1H NMR (DMSO-J6) 9.24 (br s, IH), 7.58 (d, J= 8.2 Hz, IH), 7.32 (d, J= 1.7 Hz, IH), 7.22 (dd, J= 8.2 and 1.9 Hz, IH), 4.53 (d, J= 4.4 Hz, 2H), 3.88 (s, 3H).
[00122] (b) 4-Bromo-2-methoxybenzoyl-2-dithiocarboxyhydrazide, potassium salt. To an oven-dried carousel round flask charged with a magnetic stir bar at room temperature under argon with 4-bromo-2-methoxybenzohydrazide (0.250 g, 1.02 mmol) and ethanol (4.0 mL) was added potassium hydroxide (0.086 g, 1.5 mmol). The resulting white suspension was stirred at room temperature for 20 min and then carbon disulfide (0.092 mL, 1.5 mmol) was added dropwise via syringe. The yellow suspension was stirred overnight, diluted with ethanol and then filtered through a buchner funnel to give 0.345 g (94%) of the title compound as a yellow solid. 1H NMR (DMSO-^6) 11.91 (d, J = 8.1 Hz, IH), 9.79 (d, J = 8.4 Hz, IH), 7.91 (dd, J = 8.1 and 1.8 Hz, IH), 7.39 (dd, J = 14.3 and 1.8 Hz, IH), 7.29 (td, J = 8.8 and 1.7 Hz, IH), 4.00 (s, 3H).
[00123] (c) 4-Amino-5-(4-bromo-2-methoxyphenyl)-3-mercapto-4H-[l ,2,4]- triazole. To an oven-dried round bottom flask charged with a magnetic stir bar at room temperature under argon with 4-bromo-2-methoxybenzoyl-2- dithiocarboxyhydrazide, potassium salt (0.300 g, 0.835 mmol) in ethanol (4.2 mL) and water (1.2 mL) was added hydrazine (0.36 mL, 10 mmol). The yellow suspension was heated at 100 0C for 9 h and then cooled to room temperature and stirred overnight. The resulting solution was diluted with water (7 mL), then acidified with concentrated HCl until the pH = 1. The precipitate that formed was filtered and collected on a buchner funnel, giving 0.375 g (>100%) of the crude product as a white solid. The crude product was recrytallized (EtOHiH2O; 10:1 mL) by heating to 110 0C, filtering hot, and then stirring the filtrate in an ice bath until a precipitate formed. The precipitate was then filtered through a buchner funnel to give 0.082 (33%) of the title compound as a white solid. 1H NMR (DMSO-J6) 13.88 (br s, IH), 7.41-7.29 (m, 3H), 5.45 (br s , 2H), 3.84 (s, 3H).
[00124] (d) 3-(4-Bromo-2-methoxyphenyl)-6-(4-methylphenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]-thiadiazine. To an oven-dried round bottom flask charged with a magnetic stir bar at room temperature under argon with 2-bromo-l-(4-methylphenyl)-ethanone (0.057 g, 0.27 mmol) and 4-amino-5- (4-bromo-2-methoxyphenyl)-3-mercapto-4H-[l,2,4]-triazole (0.080 g, 0.27 mmol) was added anhydrous isopropanol (1.4 mL). The resulting white suspension was heated to reflux at 130 0C for 7 h and then cooled to room temperature. The white suspension was diluted with isopropanol and then filtered through a buchner funnel to give 0.051 g (39%) of the title compound as a white solid. 1H NMR (DMSO-J6) 7.77 (d, J = 8.1 Hz, 2H), 7.47-7.44 (m, 2H), 7.35-7.32 (m, 3H), 4.41 (s, 2H), 3.77 (s, 3H), 2.36 (s, 3H).
EXAMPLE 11
3-(2-Methoxy-4-methylphenyl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00125] (a) 2-Methoxy-4-methylbenzohydrazide. The title compound was prepared in a manner similar to Example 10a. From methyl 2-methoxy-4- methylbenzoate (1.00 g, 5.55 mmol) and hydrazine hydrate (0.81 mL, 17 mmol) was obtained 0.557 g (56%) of the title compound as a white solid. 1H NMR (DMSO-J6) 9.10 (br s, IH), 7.63 (d, J = 7.7 Hz, IH), 6.94 (br s, IH), 6.85-6.82 (m IH), 4.49 (d, J= 3.3 Hz, 2H), 3.86 (s, 3H), 2.33 (s, 3H).
[00126] (b) 2-Methoxy-4-methylbenzoyl-2-dithiocarboxyhydrazide, potassium salt. The title compound was prepared in a manner similar to Example 10b. From 2-methoxy-4-methylbenzohydrazide (0.500 g, 2.77 mmol) and carbon disulfide (0.25 mL, 4.2 mmol) was obtained 0.834 g (>100%) of the title compound as a yellow solid. 1H NMR (DMSO-cfe) 11.94 (d, J = 8.4 Hz, IH), 9.72 (d, J= 8.4 Hz, IH), 7.89 (d, J = 7.7 Hz, IH), 7.02 (s, IH), 6.91 (d, J= 7.7 Hz, IH), 3.95 (s, 3H), 2.37 (s, 3H).
[00127] (c) 4-Amino-5-(2-methoxy-5-methylphenyl)-3-mercapto-4H-[l ,2,4]- triazole. The title compound was prepared in a manner similar to Example 10c. From 2-methoxy-4-methylbenzoyl-2-dithiocarboxyhydrazide, potassium salt (0.500 g, 0.170 mmol) and hydrazine (0.74 mL, 20 mmol) was obtained 0.102 (25%) of the title compound as a white solid. 1H NMR (DMSO-^6) 14.60 (br s, IH), 7.61 (d, J = 7.7 Hz, IH), 7.09 (s, IH), 6.94 (d, J = 8.1 Hz, IH), 5.42 (br s, 2H), 3.88 (s, 3H), 2.39 (s, 3H).
[00128] (d) 3-(2-Methoxy-4-methylphenyl)-6-(4-methylphenyl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]-thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methylphenyl)- ethanone (0.083 g, 0.39 mmol) and 4-amino-5-(2-methoxy-4-methylphenyl)- 3-mercapto-4H-[l,2,4]-triazole (0.092 g, 0.39 mmol) was obtained 0.010 g (7%) of the title compound as a white solid. 1H NMR (Acetone-d6) 7.85 (d, J= 8.5 Hz, 2H), 7.43 (d, J = 7.7 Hz, IH), 7.33 (d, J = 8.5 Hz, 2H), 7.02 (s, IH), 6.93 (d, J= 7.7 Hz, IH), 4.35 (s, 2H), 3.75 (s, 3H), 2.43 (s, 3H), 2.39 (s, 3H).
EXAMPLE 12
6-(4-Methylphenyl)-3-(3-methylthiophen-2-yl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00129] (a) 3-Methylthiophene-2-dithiocarboxyhydrazide, potassium salt. The title compound was prepared in a manner similar to Example 10b. From 3-methyl-thiophene-2-carboxyhydrazide (0.500 g, 3.20 mmol) and carbon disulfide (0.29 mL, 4.8 mmol) was obtained 0.660 g (76%) of the title compound as a white solid. 1H NMR (CD3OD) 7.48 (d, J= 5.1 Hz, IH), 6.95 (d, J= 4.8 Hz, IH), 2.56 (s, 3H).
[00130] (b) 4-Amino-5-(3-methylthiophen-2-yl)-3-mercapto-4H-[l ,2,4]- triazole. The title compound was prepared in a manner similar to Example 10c. From 3-methylthiophenyl-2-dithiocarboxyhydrazide, potassium salt (0.500 g, 0.185 mmol) and hydrazine (0.81 mL, 22 mmol) was obtained 0.117 (30%) of the title compound as a yellow solid. 1H NMR (DMSO-cfe) 13.89 (br s, IH), 7.71 (d, J = 5.1 Hz, IH), 7.07 (d, J= 5.1 Hz, IH), 5.75 (br s, 2H), 2.44 (S5 3H).
[00131] (c) 6-(4-Methylphenyl)-3-(3-methylthiophen-2-yl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]-thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methylphenyl)- ethanone (0.100 g, 0.471 mmol) and 4-amino-5-(3-methylthiophen-2-yl)-3- mercapto-4H-[l,2,4]-triazole (0.100 g, 0.471 mmol) was obtained 0.129 g (67%) of the title compound as a yellow solid. 1H NMR (DMSO-J6) 7.99 (d, J = 8.1 Hz, 2H), 7.80 (d, J = 5.1 Hz, IH), 7.42 (d, J= 8.4 Hz, 2H), 7.13 (d, J = 5.1 Hz, IH), 4.46 (s, 2H), 2.55 (s, 3H), 2.41 (s, 3H).
EXAMPLE 13
3 -(4-Bromo-2-methoxyphenyl)-6-(4-methy 1-3 -nitrophenyl)-7H- [ 1 ,2 ,4] triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00132] The title compound was prepared in a manner similar to Example 1Od.
From 2-bromo-l-(4-methyl-3-nitrophenyl)-ethanone (0.090 g, 0.35 mmol) and 4-amino-5-(4-bromo-2-methoxyphenyl)-3-mercapto-4H-[l ,2,4]-triazole (0.105 g, 0.349 mmol) was obtained 0.157 g (83%) of the title compound as a white solid. 1H NMR (DMSO-J6) 8.44-8.42 (m, IH), 7.70-7.67 (m, IH), 7.49-7.43 (m, 2H), 7.35-7.32 (m, IH), 4.49-4.47 (m, 2H), 3.79-3.76 (m, 3H), 2.58-2.55 (m, 3H).
EXAMPLE 14
3-(4-Chloro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine
[00133] The title compound was prepared in a manner similar to Example 1Od.
From 2-bromo-l-(4-methyl-3-nitrophenyl)-ethanone (0.252 g, 0.976 mmol) and 4-amino-5-(4-chloro-2-methoxyphenyl)-3-mercapto-4H-[l,2,4]-triazole (0.250 g, 0.976 mmol) was obtained 0.389 g (80%) of the title compound as a white solid. 1H NMR (DMSO-J6) 8.45 (d, J = 1.8 Hz, IH), 8.09 (dd, J = 8.1 and 1.8 Hz, IH), 7.68 (d, J= 8.4 Hz, IH), 7.5 (d, J= 8.1 Hz, IH), 7.35 (d, J = 1.8 Hz, IH), 7.21 (dd, J= 8.1 and 1.8 Hz, IH), 4.50 (s, 2H), 3.80 (s, 3H), 2.58 (s, 3H). EXAMPLE 15
6-(3-Amino-4-methylphenyl)-3-(4-chloro-2-methoxyphenyl)-7//- [1 ,2,4]triazolo[3,4-6] [1 ,3,4]thiadiazine
[00134] To an oven-dried round bottom reaction flask charged with a magnetic stir bar under argon at room temperature with 3-(4-chloro-2-methoxyphenyl)- 6-(4-methyl-3-nitrophenyl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (0.350 g, 0.704 mmol) and tin (II) chloride dihydrate (0.715 g, 3.17 mmol) was added anhydrous ethanol (3.5 mL). The resulting yellow suspension was heated at 95 0C for 9 h and then cooled to room temperature. The solvent was removed by rotary evaporation, diluted with H2O (25 mL) and then basified using 2N NaOH until the pH = 10. The aqueous layer was then extracted with CHCI3 (2 x 125 mL), washed with brine (20 mL), dried over MgSO4, filtered and concentrated to give 0.175 g (64%) of the title compound as a yellow solid. 1H NMR (DMSO-dβ) 7.52 (d, J = 8.4 Hz, IH), 7.31 (d, J = 2.2 Hz, IH), 7.19 (dd, J= 8.1 and 1.8 Hz, IH), 7.07-7.00 (m, 3H), 5.14 (br s, 2H), 4.30 (s, 2H), 3.78 (s, 3H), 2.09 (s, 3H).
EXAMPLE 16
6-(3-Amino-4-methylphenyl)-3-(4-bromo-2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00135] The title compound was prepared in a manner similar to Example 15.
From 3 -(4-bromo-2-methoxyphenyl)-6-(4-methy 1-3 -nitrophenyl)-7//-
[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (0.140 g, 0.259 mmol) and tin (II) chloride dihydrate (0.263 g, 1.16 mmol) was obtained 0.020 g (18%) of the title compound as a yellow solid. 1H NMR (DMSO-^6) 7.46-7.42 (m, 2H), 7.32 (dd, J = 8.2 and 1.6 Hz, IH), 7.07-6.98 (m, 3H), 5.15 (br s, 2H), 4.30 (s, 2H), 3.78 (s, 3H), 2.09 (s, 3H).
EXAMPLE 17
6-(4-Methylphenyl)-3-(lH-pyrazol-5-yl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00136] (a) 2H-Pyrazole-2-dithiocarboxyhydrazide, potassium salt. The title compound was prepared in a manner similar to Example 10b. From 2H-pyrazole-3-carboxyhydrazide (0.500 g, 4.00 mmol) and carbon disulfide (0.36 mL, 6.0 mmol) was obtained 0.509 g (53%) of the title compound as a yellow solid. 1H NMR (DMSCMO 13.28 (br s, IHO, 10.50 (br s, IH), 9.81 (br s, IH), 7.84 (s, IH), 6.68 (s, IH).
[00137] (b) 4-Amino-5-(lH-pyrazol-5-yl)-3-mercapto-4H-[l,2,4]-triazole. The title compound was prepared in a manner similar to Example 10c. From 2H- pyrazole-2-dithiocarboxyhydrazide, potassium salt (0.500 g, 2.09 mmol) and hydrazine (0.79 mL, 25 mmol) was obtained 0.051 (13%) of the title compound as a white solid. 1H NMR (DMSO-J6) 13.81 (br s, IH), 13.43 (br s, IH), 7.93 (s, IH), 6.90 (s, IH), 5.93 (br s, 2H).
[00138] (c) 3-(lH-Pyrazol-5-yl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4-
6][l,3,4]-thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methylphenyl)-ethanone (0.058 g, 0.27 mmol) and 4-amino-5-(lH-pyrazol-5-yl)-3-mercapto-4H-[l,2,4]-triazole (0.050 g, 0.27 mmol) was obtained 0.082 g (79%) of the title compound as a white solid. 1H NMR (DMSO-J6) 7.96 (d, J- 8.4 Hz, 2H), 7.94 (d, J= 1.8 Hz, IH), 7.41 (d, J = 8.4 Hz, 2H), 6.93 (d, J = 2.2 Hz, IH), 4.45 (s, 2H), 2.41 (s, 3H).
EXAMPLE 18
6-(4-Methoxyphenyl)-3 -(2-methylfuran-3 -y l)-7H-[ 1 ,2,4]triazolo [3 ,A- b] [ 1 ,3 ,4]thiadiazine
[00139] The title compound was prepared in a manner similar to Example 1Od.
From 2-bromo-4'-methoxyacetophenone (0.350 g, 1.53 mmol) and 4-amino-5- (2-methyl-3-furyl)-3-mercapto-4H-[l,2,4]-triazole (0.300 g, 1.53 mmol) was obtained 0.560 g (90%) of the title compound as a yellow solid. 1H NMR (DMSO-J6) 7.99 (d, J= 9.2 Hz, 2H), 7.73 (d, J= 1.8 Hz, IH), 7.14 (d, J= 9.2 Hz, 2H), 6.97 (d, J= 1.8 Hz, IH), 4.41 (s, 2H), 3.86 (s, 3H), 2.58 (s, 3H).
EXAMPLE 19
6-(4-Hydroxyphenyl)-3-(2-methylfuran-3-yl)-7H-[l,2,4]triazolo[3,4- b][\ ,3,4]thiadiazine
[00140] To an erlenmeyer flask was added 6-(4-methoxyphenyl)-3-(2- methylfuran-3-yl)-7H-[l ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine hydrobromide salt (0.560 g) and saturated aqueous NaHCO3 (20 mL). The mixture was stirred for 20 min, and was extracted with EtOAc (2 x 100 mL). The organic extracts were combined, washed with brine (15 mL), dried over MgSO4, filtered and concentrated to give 0.390 g of 6-(4-methoxyphenyl)-3-(2- methylfuran-3-yl)-7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine. To an oven- dried round bottom flask charged with a magnetic stir bar at room temperature under argon was added 6-(4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thia-diazine (0.380 g, 1.16 mmol) and anhydrous methylene chloride (7.8 mL). To the brown solution was added boron tribromide (1.0M, 3.49 mL, 3.49 mmol), tetrabutyl ammonium bromide (1.23 g, 3.49 mmol) and potassium iodide (0.580 g, 3.49 mmol). The black solution was heated at 60 0C for 4 h and then cooled to room temperature. The solution was diluted with cold water (10 mL) then extracted with CHCl3 (100 mL). The organic layer was washed with saturated aqueous NaHCO3 (20 mL), water (20 mL), brine (20 mL), dried over MgSO4, filtered and concentrated to give a brown oil. It was purified by flash chromatography (gradient elution: hexanes, EtOAc; 1 :1 to 100% EtOAc) to give 0.017 g (5%) of the title compound as a yellow solid. 1H NMR (DMSO-^6) 10.32 (br s, IH), 7.89 (d, J = 8.8 Hz, 2H), 7.71 (d, J= 1.8 Hz, IH), 6.97 (d, J= 1.8 Hz, IH), 6.93 (d, J = 8.8 Hz, 2H), 4.36 (s, 2H), 2.58 (s, 3H).
EXAMPLE 20
3 -(2-Methylfuran-3 -yl)-6-(4-methyl-3 -nitropheny l)-7H-[ 1 ,2,4]triazolo [3 ,4- b] [ 1 ,3 ,4]thiadiazine
[00141] The title compound was prepared in a manner similar to Example 1Od.
From 2-bromo-l-(4-methyl-3-nitrophenyl)-ethanone (0.395 g, 1.53 mmol) and 4-amino-5-(2-methyl-3-furyl)-3-mercapto-4H-[l,2,4]-triazole (0.300 g, 1.53 mmol) was obtained 0.604 g (90%) of the title compound as a yellow solid. 1H NMR (DMSO-^6) 8.56 (d, J= 1.8 Hz, 2H), 8.23 (dd, J = 8.2 and 2.0 Hz, IH), 7.75-7.72 (m, 2H), 6.96 (d, J= 1.8 Hz, IH), 4.49 (s, 2H), 2.60 (s, 3H), 2.59 (s, 3H).
EXAMPLE 21
6-(3 - Amino-4-methylphenyl)-3 -(2-methylfuran-3 -yl)-7H- [ 1 ,2,4] triazolo [3 ,4- b][l ,3,4]thiadiazine
[00142] A mixture of 3-(2-methylfuran-3-yl)-6-(4-methyl-3-nitrophenyl)-7H-
[l ,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (0.200 g, 0.458 mmol) in methanol (9.2 mL) was hydrogenated at 65 psi for 22 h using Palladium on carbon (10% Pd content, 0.200 g). The resulting mixture was filtered through a pad of Celite and the filtrate was concentrated under vacuum to obtain the crude product. It was purified by flash chromatography (gradient elution: hexanes, EtOAc; 9:1 to 1 :1 to 100% EtOAc) to give 0.045 g (30%) of the title compound as a yellow solid. 1H NMR (DMSO-^6): 7.71 (d, J = 2.2 Hz, IH), 7.25 (s, IH), 7.10-7.09 (m, 2H), 6.98 (d, J= 1.9 Hz, IH), 5.20 (br s, 2H), 4.32 (s, 2H), 2.58 (s, 3H), 2.13 (s, 3H).
EXAMPLE 22
6-(4-Methyl-3-nitrophenyl)-3-(2-methyl-H-imidazo[l,2-α]pyridin-3-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-δ] [ 1 ,3 ,4]thiadiazine
[00143] The title compound was prepared in a manner similar to Example 1Od.
From 2-bromo-l-(4-methyl-3-nitrophenyl)-ethanone (0.063 g, 0.24 mmol) and 4-amino-5-(2-methyl-H-imidazo[l,2-α]pyridin-3-yl)-3-mercapto-4H-[l,2,4]- triazole (0.060 g, 0.24 mmol) was obtained 0.037 g (37%) of the title compound as a white solid. 1H NMR (DMSO-cfe) 8.95-8.93 (m, IH), 8.44 (s, IH), 8.12 (d, J = 7.7 Hz, IH), 7.97-7.89 (m, 2H), 7.68 (d, J = 8.4 Hz, IH), 7.41 (t, J= 6.6 Hz, IH), 4.62 (s, 2H), 2.57 (s, 3H), 2.55 (s, 3H).
EXAMPLE 23
6-(3-Amino-4-methylphenyl)-3-(2-methyl-H-imidazo[l,2-fl]pyridin-3-yl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine
[00144] The title compound was prepared in a manner similar to Example 15.
From 6-(4-methyl-3-nitrophenyl)-3-(2-methyl-H-imidazo[l,2-α]pyridin-3-yl)- 7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (0.028 g, 0.058 mmol) and tin (II) chloride dihydrate (0.058 g, 0.26 mmol) was obtained 0.003 g (14%) of the title compound as a yellow solid. 1H NMR (DMSO-^6) 8.68 (m, IH), 7.64 (d, J= 8.4 Hz, IH), 7.40-7.37 (m, IH), 7.12-7.01 (m, 4H), 5.13 (br s, 2H), 4.43 (s, 2H), 2.41 (s, 3H), 2.09 (s, 3H).
EXAMPLE 24
6-(4-Methylphenyl)-3-(l-methyl-lH-pyrazol-5-yl)-7//-[l,2,4]triazolo[3,4- b][\ ,3,4]thiadiazine
[00145] (a) 1 -Methyl- lH-pyrazol-5-yl-2-dithiocarboxyhydrazide, potassium salt. The title compound was prepared in a manner similar to Example 10b. From 1 -methyl- lH-pyrazol-S-yl-S-carboxyhydrazide (0.500 g, 3.57 mmol) and carbon disulfide (0.32 mL, 5.4 mmol) was obtained 0.38 g (42%) of the title compound as a brown solid. 1H NMR (DMSO-^6) 10.16 (br s, IH), 9.71 (br s, IH), 7.45 (d, J= 2.2 Hz, IH), 6.89 (d, J= 1.8 Hz, IH), 4.04 (s, 3H).
[00146] (b) 4-Amino-5-(l -methyl-lH-pyrazol-5-yl)-3-mercapto-4H-[l ,2,4]- triazole. The title compound was prepared in a manner similar to Example 10c. From l-methyl-lH-pyrazol-5-yl-3-yl-2-dithiocarboxyhydrazide, potassium salt (0.350 g, 1.37 mmol) and hydrazine (0.52 mL, 16 mmol) was obtained 0.076 (28%) of the title compound as a white solid. 1H NMR (DMSO-^6) 14.14 (br s, IH), 7.59 (d, J = 2.2 Hz, IH), 7.07 (d, J = 2.2 Hz, IH), 4.01 (s, 3H).
[00147] (c) 6-(4-Methylphenyl)-3-(l-methyl-lH-pyrazol-5-yl)-7//-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methylphenyl)-ethanone (0.076 g, 0.36 mmol) and 4-amino-5-(l -methyl- lH-pyrazol-5-yl)-3-mercapto- 4H-[l,2,4]-triazole (0.070 g, 0.36 mmol) was obtained 0.014 g (13%) of the title compound as a white solid. 1H NMR (DMSO-^6) 7.93 (d, J = 8.4 Hz, 2H), 7.65 (d, J = 1.8 Hz, IH), 7.41 (d, J = 8.4 Hz, 2H), 6.95 (d, J = 1.8 Hz, IH), 4.45 (s, 2H), 4.17 (s, 3H), 2.40 (s, 3H).
EXAMPLE 25
6-(4-Methyl-3-mtrophenyl)-3-(l-methyl-4-nitro-l//-pyrrol-2-yl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine
[00148] (a) l-Methyl-4-nitro-lH-pyrrol-2-yl-2-dithiocarboxyhydrazide, potassium salt. The title compound was prepared in a manner similar to Example 10b. From l-methyl-4-nitro-lH-pyrrol-2-yl-3-carboxyhydrazide (0.515 g, 2.71 mmol) and carbon disulfide (0.25 mL, 4.1 mmol) was obtained 0.68 g (85%) of the title compound as a brown solid. 1H NMR (DMSO-cfe) 10.16 (br s, IH), 9.63 (br s, IH), 8.14 (s, IH), 7.46 (s, IH), 3.90 (s, 3H).
[00149] (b) 4-Amino-5-(l -methyl-4-nitro-lH-pyrrol-2-yl)-3-mercapto-4//-
[l,2,4]-triazole. The title compound was prepared in a manner similar to Example 10c. From l-methyl-4-nitro-lH-pyrrol-2-yl-2- dithiocarboxyhydrazide, potassium salt (0.683 g, 2.29 mmol) and hydrazine (1.00 mL, 27.5 mmol) was obtained 0.225 (41%) of the title compound as a white solid. 1H NMR (DMSCW6) 14.07 (br s, IH), 8.28 (d, J = 2.2 Hz, IH)5 7.61 (d, J= 2.2 Hz, IH) 5.81 (br s, 2H), 3.88 (s, 3H).
[00150] (c) 6-(4-Methyl-3-nitrophenyl)-3-(l-methyl-4-nitro-lH-pyrrol-2-yl)-
7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methyl-3- nitrophenyl)-ethanone (0.103 g, 0.400 mmol) and 4-amino-5-(l-methyl-4- nitro-lH-pyrrol-2-yl)-3-mercapto-4H-[l,2,4]-triazole (0.096 g, 0.40 mmol) was obtained 0.153 g (80%) of the title compound as a yellow solid. 1H NMR (DMSCW6) 8.58 (d, J = 1.8 Hz, IH), 8.34 (d, J = 2.2 Hz, IH), 8.22 (dd, J = 8.2 and 2.0 Hz, IH), 7.77 (d, J= 8.1 Hz, IH), 7.32 (d, J= 2.2 Hz, IH), 4.48 (s, 2H), 4.03 (s, 3H), 2,61 (s, 3H).
EXAMPLE 26
3-(4-Chloro-2-methylphenyl)-6-(4-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00151] The title compound was prepared in a manner similar to Example 1Od.
From 2-bromo-4'-methoxyacetophenone (0.152 g, 0.665 mmol) and 4-amino- 5-(4-chloro-2-methylphenyl)-3-mercapto-4H-[l,2,4]-triazole (0.160 g, 0.665 mmol) was obtained 0.131 g (43%) of the title compound as a white solid. 1H NMR (DMSCW6) 7.86 (d, J = 9.2 Hz, 2H), 7.55-7.52 (m, 2H), 7.46-7.43 (m, IH), 7.08 (d, J= 9.2 Hz, 2H), 4.42 (s, 2H), 3.82 (s, 3H), 2.37 (s, 3H).
EXAMPLE 27
3-(4-Chloro-2-methylphenyl)-6-(4-hydroxyphenyl)-7H-[l,2,4]triazolo[3,4-
6][l,3,4]thiadiazine
[00152] The title compound was prepared in a manner similar to Example 19.
From 3-(4-chloro-2-methylphenyl)-6-(4-methoxyphenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (0.010 g, 0.027 mmol) and boron tribromide (1.0M, 0.081 mL, 0.081 mmol) was obtained 0.005 g (52%) of the title compound as a white solid. 1H NMR (DMSO-J6) 7.75 (d, J= 8.8 Hz, 2H), 7.54-7.51 (m, 2H), 7.46-7.43 (m, IH), 6.87 (d, J = 8.4 Hz, 2H), 4.38 (s, 2H), 2.37 (s, 3H). EXAMPLE 28
3 -(4-Methyl-2-morpholinothiazol-5 -yl)-6-(4-methylphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine
[00153] (a) 4-Methyl-2-morpholinothiazol-5-yl-2-dithiocarboxyhydrazide, potassium salt. The title compound was prepared in a manner similar to Example 10b. From 4-methyl-2-morpholin-4-yl-l,3,-thiazole-5- benzohydrazide (0.500 g, 2.06 mmol) and carbon disulfide (0.19 mL, 3.1 mmol) was obtained 0.718 g (98%) of the title compound as a yellow solid. 1H NMR (DMSO-J6) 3.70-3.67 (m, 4H), 3.46-3.44 (m, 4H), 2.48 (s, 3H).
[00154] (b) 4-Amino-5-(4-methyl-2-morpholinothiazol-5-yl)-3-mercapto-4H-
[l,2,4]-triazole. The title compound was prepared in a manner similar to Example 10c. From 4-methyl-2-morpholinothiazol-5-yl-2- dithiocarboxyhydrazide, potassium salt (0.500 g, 1.40 mmol) and hydrazine (0.53 mL, 17 mmol) was obtained 0.144 (34%) of the title compound as a white solid. 1H NMR (DMSO-J6) 13.75 (s, IH), 5.71 (s, 2H), 3.72-3.69 (m, 4H), 3.45-3.41 (m, 4H), 2.43 (s, 3H).
[00155] (c) 3-(4-Methyl-2-morpholinothiazol-5-yl)-6-(4-methylphenyl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazines. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methylphenyl)-ethanone (0.093 g, 0.44 mmol) and 4-amino-5-(4-methyl-2-morpholinothiazol-5-yl)-3- mercapto-4H-[l,2,4]-triazole (0.130 g, 0.44 mmol) was obtained 0.065 g (36%) of the title compound as a yellow solid. 1H NMR (DMSO-J6) 7.96 (d, J = 7.7 Hz, 2H), 7.42 (d, J = 8.1 Hz, 2H), 4.43 (s, 2H), 3.73-3.72 (m, 4H), 3.47 (t, J= 4.8 Hz, 4H), 2.54 (s, 3H), 2.40 (s, 3H).
EXAMPLE 29
6-(3-Dimethylamino-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine
[00156] To a mixture of 6-(3-amino-4-methylphenyl)-3-(2-methoxyphenyl)-
7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (0.200 g, 0.569 mmol) in methanol (2.8 mL) was added 37% aqueous formaldehyde (0.250 mL, 2.85 mmol) and acetic acid (5 drops). The yellow solution was cooled to 0 0C and then sodium cyanoborohydride (0.179 g, 2.85 mmol) was added in portions. The solution was stirred at 0 0C for 1 h and then equilibrated to room temperature and stirred overnight. The reaction solution was diluted with CHCI3 (100 mL) and the organic layer was then washed with saturated NaHCO3 (25 mL), brine (25 mL), dried over MgSO4, filtered and was concentrated to give a yellow solid. It was purified by flash chromatography (elution: hexanes, EtOAc; 1 :1), gave 0.136 g (62%) of the title compound as a yellow solid. 1H NMR (DMSO-^6) 7.61-7.55 (m, IH), 7.52-7.48 (m, 2H), 7.44-7.41 (m, IH), 7.29-7.21 (m, 2H), 7.11 (t, J= 7.3 Hz, IH), 4.40 (s, 2H), 3.75 (s, 3H), 2.62 (s, 6H), 2.30 (s, 3H).
EXAMPLE 30
3-(4-Methyl-lH-imidazol-5-yl)-6-(4-methylphenyl)- 7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00157] (a) 4-Methyl-lH-imidazol-5-carbohydrazide. The title compound was prepared in a manner similar to Example 10a. From ethyl 4-methyl-5- imidazole carboxylate (1.00 g, 6.49 mmol) and hydrazine hydrate (1.90 mL, 38.9 mmol) was obtained 0.072 g (8%) of the title compound as a white solid. 1H NMR (OMSO-dβ) 12.23 (br s, IH), 8.73 (s, IH), 7.52 (s, IH), 4.24 (br s, 2H), 2.42 (s, 3H).
[00158] (b) 4-Methyl-lH-imidazol-5-yl-2-dithiocarboxyhydrazide, potassium salt. The title compound was prepared in a manner similar to Example 10b. From 4-methyl-lH-imidazol-5-carbohydrazide (0.290 g, 2.07 mmol) and carbon disulfide (0.23 mL, 3.1 mmol) was obtained 0.432 g (82%) of the title compound as a yellow solid. 1H NMR (DMSO-^6) 12.30 (br s, IH), 10.58 (d, J = 6.2 Hz, IH), 9.63 (d, J= 7.0 Hz, IH), 7.55 (s, IH), 2.44 (s, 3H).
[00159] (c) 4-Amino-5-(4-methyl-lH-imidazol-5-yl)-3-mercapto-4//-[l,2,4]- triazole. The title compound was prepared in a manner similar to Example 10c. From 4-methyl-lH-imidazol-5-yl-2-dithiocarboxyhydrazide, potassium salt (0.532 g, 2.09 mmol) in ethanol (10.5 mL) and hydrazine (0.91 mL, 25 mmol) was obtained 0.179 (44%) of the title compound as a white solid. 1H NMR (DMSO-^6) 14.21 (s, IH), 9.10 (s, IH), 2.45 (s, 3H).
[00160] (d) 3-(4-Methyl-l//-imidazol-5-yl)-6-(4-methylphenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]-thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methylphenyl)- ethanone (0.054 g, 0.26 mmol) and 4-amino-5-(4-methyl-lH-imidazol-5-yl)-3- mercapto-4H-[l,2,4]-triazole (0.050 g, 0.26 mmol) was obtained 0.005 g (6%) of the title compound as a white solid. 1H NMR (DMSO-J6) 7.89 (d, J - 8.4 Hz, 2H), 7.77 (s, IH), 7.33 (d, J= 8.1 Hz, 2H), 4.29 (s, 2H), 2.43 (s, 3H), 2.41 (s, 3H).
EXAMPLE 31
3-(2,5-Dimethylfuran-3-yl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b][l ,3 ,4]thiadiazine
[00161] (a) 2,5-Dimethylfuran-3-yl-2-dithiocarboxyhydrazide, potassium salt.
The title compound was prepared in a manner similar to Example 1 Ob. From 2,5-dimethylfuran-3-yl-carbohydrazide (0.418 g, 2.71 mmol) and carbon disulfide (0.25 mL, 4.1 mmol) was obtained 0.438 g (60%) of the title compound as a yellow solid. 1H NMR (DMSO-J6) 9.87 (br s, IH), 9.58 (br s, IH), 6.38 (s, IH), 2.47 (s, 3H), 2.22 (s, 3H).
[00162] (b) 4-Amino-5-(2,5-dimethylfuran-3-yl)-3-mercapto-4H-[l ,2,4]- triazole. The title compound was prepared in a manner similar to Example 10c. From 2,5-dimethylfuran-3-yl-2-dithiocarboxyhydrazide, potassium salt (0.300 g, 1.12 mmol) and hydrazine (0.49 mL, 13 mmol) was obtained 0.036 (15%) of the title compound as a white solid. 1H NMR (DMSO-J6) 13.77 (br s, IH), 6.66 (s, IH), 5.66 (s, 2H), 2.42 (s, 3H), 2.27 (s, 3H).
[00163] (c) 3-(2,5-Dimethylfuran-3-yl)-6-(4-methylphenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazines. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methylphenyl)-ethanone (0.036 g, 0.17 mmol) and 4-amino-5-(2,5-dimethyl-3-furyl)-3-mercapto-4H- [l,2,4]-triazole (0.036 g, 0.17 mmol) was obtained 0.020 g (29%) of the title compound as a brown solid. 1H NMR (DMSO-J6) 7.91 (d, J = 8.4 Hz, 2H), 7.40 (d, J = 8.1 Hz, 2H), 6.55 (s, IH), 4.41 (s, 2H), 2.52 (s, 3H), 2.40 (s, 3H), 2.31 (s, 3H).
EXAMPLE 32
3-(2-Methoxyphenyl)-6-(l-methyl-l//-indol-5-yl)-7H-[l,2,4]triazolo[3,4- b][\ ,3,4]thiadiazine
[00164] (a) 2-Bromo-l -(I -methyl- lH-indol-5-yl)-ethanone. To an oven-dried round bottom flask charged with a magnetic stir bar at room temperature under argon was added aluminum chloride (0.693 g, 5.20 mmol), 1 ,2-dichloroethane (20 mL) and bromoacetyl bromide (0.45 mL, 5.20 mmol). The orange solution was cooled to 0 0C and then 1 -methyl indole (0.262 g, 2.00 mmol) was added via addition funnel. The solution was stirred at 0 0C for 1 h and then allowed to equilibrate to room temperature. The solution was stirred at room temperature for 3 h, and was poured into a slurry of ice and water. The resulting solution was basified using saturated NaHCO3 until the pH = 7 and then extracted using EtOAc (2 x 100 mL). The combined organic extracts were washed with brine (25 mL), dried over MgSO4, filtered and concentrated to give a brown residue. It was purified by flash chromatography (gradient elution: hexanes, EtOAc; 4:1 to 1 :1) to give 0.135 g (27%) of the title compound as a purple solid. 1H NMR (DMSO-J6) 8.36-8.33 (m, IH), 7.84 (s, IH), 7.38-7.32 (m, 3H), 4.31 (s, 2H), 3.88 (s, 3H).
[00165] (b) 3-(2-Methoxyphenyl)-6-(l-methyl-lH-indol-5-yl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]-thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(l-methyl-lH-indol-5-yl)- ethanone (0.135 g, 0.535 mmol) and 4-amino-5-(2-methoxyphenyl)-3- mercapto-4H-[l,2,4]-triazole (0.119 g, 0.535 mmol) was obtained 0.055 g (23%) of the title compound as a brown solid. 1H NMR (DMSO-J6) 8.28 (s, IH), 7.81 (d, J = 8.4 Hz, IH), 7.66-7.61 (m, IH), 7.55 (d, J = 7.7 Hz, 2H), 7.29-7.23 (m, 2H), 7.15 (t, J = 7.1 Hz, IH), 7.03 (t, J = 7.9 Hz, IH), 4.34 (s, 2H), 3.87 (s, 3H), 3.71 (s, 3H).
EXAMPLE 33
6-(4-Acetamido-3-aminophenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4-
6][l,3,4]thiadiazine
[00166] A mixture of 6-(4-acetamido-3-nitrophenyl)-3-(2-methoxyphenyl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (80 mg, 0.19 mmol) and methanol (20 mL) was hydrogenated over Palladium on carbon (10% Pd/C, 80 mg, 65 psi). The resulting mixture was filtered through a pad of Celite and the filtrate was concentrated. The crude was purified by chromatography (5% methanol/dichloromethane) to give the title compound as a yellow solid (35 mg, 0.089 mmol, 47%). 1H NMR (CDCl3/Me0H-d4) 7.57 (dd, J = 7.8, 1.5 Hz, IH), 7.52 (m, IH), 7.41 (d, J = 8.4 Hz, IH), 7.29 (s, IH), 7.26 (d, J = 1.8 Hz, IH), 7.19 (dd, J= 8.4, 2.1 Hz, IH), 7.10 (dt, J= 7.8, 1.2 Hz, IH), 7.03 (d, J= 8.7 Hz, IH), 3.96 (s, 2H), 3.74 (s, 3H), 2.21 (s, 3H). EXAMPLE 34
3-(2-Methoxyphenyl)-6-(2-methyl-lH-benzo[-f]imidazol-6-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00167] A solution of 6-(4-acetamido-3-aminophenyl)-3-(2-methoxyphenyl)-
7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (33 mg, 0.084 mmol) in glacial acetic acid (8 mL) was refiuxed for 30 h. The reaction mixture was cooled to room temperature and the glacial acetic acid was removed under vacuum. The residue was dissolved in 50 mL of chloroform and washed with saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The crude was purified by chromatography (5% methanol/dichloromethane) to give the title compound as a white solid (29 mg, 0.077 mmol, 92%). 1H NMR (CDCl3) 7.98 (d, J= 0.9 Hz, IH), 7.68 (dd, J = 8.4, 1.5 Hz, IH), 7.60 (dd, J= 7.2, 1.5 Hz, IH), 7.55 (d, J= 8.4 Hz, IH), 7.47 (m, IH), 7.05 (dt, J = 7.8, 1.2 Hz, IH), 6.98 (dd, J= 8.1, 0.6 Hz, IH), 4.03 (s, 2H), 3.68 (s, 3H), 2.63 (s, 3H).
EXAMPLE 35
3-(3 ,5-Dimethoxyphenyl)-6-(4-methyl-3 -nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4- Z>][l,3,4]thiadiazine hydrobromide
[00168] A mixture of 2-bromo-l-(4-methyl-3-nitrophenyl)ethanone (175 mg,
0.678 mmol) and 4-amino-5-(3,5-dimethoxyphenyl)-3-mercapto-(4H)-l,2,4- triazole (179 mg, 0.709 mmol) in isopropyl alcohol (8 ml) was refiuxed for 3 h. The reaction mixture was cooled to room temperature, filtered and the solids were dried to give the title compound as light yellow solids (300 mg, 0.67 mmol, 99%). 1H NMR (DMSO-d6) 8.58 (d, J = 2.1 Hz, IH), 8.23 (dd, J = 7.8, 1.8 Hz, IH), 7.74 (d, J= 8.4 Hz, IH), 7.21 (d, J= 2.4 Hz, 2H), 6.70 (t, J = 2.7 Hz, IH), 4.48 (s, 2H), 3.82 (s, 6H), 2.61 (s, 3H).
EXAMPLE 36
6-(4-Methoxy-3-nitrophenyl)-3-(3,5-dimethoxyphenyl)-7H- [1 ,2,4]triazolo[3 ,4-6][l ,3,4]thiadiazine hydrobromide
[00169] The title compound was prepared in a manner similar to Example 35.
From 2-bromo-l-(4-methoxy-3-nitrophenyl)ethanone (175 mg, 0.64 mmol) and 4-amino-5-(3,5-dimethoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (170 mg, 0.67 mmol) was obtained the title compound as a yellow solid (270 mg, 0.63 mmol, 98%). 1H NMR (DMSO-d6) 8.53 (d, J = 2.7 Hz, IH), 8.29 (dd, J = 9.0, 2.4 Hz, IH), 7.61 (d, J= 9.0 Hz, IH), 7.21 (d, J= 2.7 Hz, 2H), 6.94 (t, J = 2.1 Hz, IH), 4.54 (s, 2H), 4.04 (s, 3H), 3.82 (s, 6H).
EXAMPLE 37
6-(3-Amino-4-methylphenyl)-3-(3,5-dimethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine
[00170] The title compounds was prepared in a manner similar to Example 33.
From 3-(3,5-dimethoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine hydrobromide (295 mg, 0.72 mmol) was obtained the title compound as a white solid (75 mg, 0.20 mmol, 27%). 1U NMR (CDCls/MeOΗ-cU) 7.31 (m, 4Η), 7.20 (m, 2H), 6.61 (t, J = 2.1 Hz, IH), 4.01 (s, 2H), 3.85 (s, 6H), 2.25 (s, 3H).
EXAMPLE 38
6-(3-Amino-4-methoxyphenyl)-3-(3,5-dimethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine
[00171] The title compound was prepared in a manner similar to example 33.
From 6-(4-methoxy-3-nitrophenyl)-3-(3,5-dimethoxyphenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine hydrobromide was obtained the title compound as a light yellow solid (58%). 1H NMR (CDCl3) 7.37 (m, 3H), 7.24 (dd, J = 8.1, 2.1 Hz, IH), 6.86 (d, J = 8.4, 2H), 6.59 (t, J = 2.1 Hz, IH), 3.94 (m, 5H), 3.85 (s, 6H).
EXAMPLE 39
6-(3 -Dimethylamino-4-methoxypheny l)-3 -(3,5 -dimethoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00172] To a solution of 6-(3-amino-4-methoxyphenyl)-3-(3,5- dimethoxyphenyl)-7Η-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (29 mg, 0.073 mmol) in methanol (5 mL) was added formaldehyde (35% aqoues, 1 mL) followed by two drops of glacial acetic acid. The reaction mixture was cooled to 0 0C, then sodium cyanoborohydride was added (25 mg x 3). The reaction mixture was stirred at 0 0C for 1 h and warmed to room temperature. The mixture was diluted with 50 mL of ethyl acetate and washed with saturated NaHCO3 (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by chromatography (90% ethylacetate/hexane, 0.1 mL methanol/ 10 mL solvent) to give the title compound (30 mg, 0.071 mmol, 97%). 1H NMR (CDCl3) 7.66 (d, J= 2.4 Hz, IH), 7.46 (dd, J = 8.7, 2.7 Hz, IH), 7.41 (d, J = 2.1 Hz, 2H), 6.94 (d, J = 8.7 Hz, 6.58 (t, J = 2.1 Hz, IH), 3.98 (s, 3H), 3.97 (s, 2H), 3.84 (s, 6H), 2.86 (s, 6H).
EXAMPLE 40
6-(Benzofuran-5-yl)-3 -(2-methoxyphenyl)-7H-[ 1 ,2,4]triazolo [3 ,4- 6][l,3,4]thiadiazine hydrobromide
[00173] The title compound was prepared in a manner similar to Example 35.
From l-(benzofuran-5-yl)-2-bromoethanone (100 mg, 0.418 mmol) and 4-amino-5-(2-methoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (98 mg, 0.44 mmol) was obtained the title compound as an off-white solid (126 mg, 0.35 mmol, 83%). 1H NMR (DMSOd6) 8.23 (m, IH), 8.12 (m, IH), 7.83 (m, IH), 7.74 (d, J= 9.0 Hz, IH), 7.53 - 7.63 (m, 2H), 7.24 (d, J= 8.7 Hz, IH), 7.14 (t, J= 7.5 Hz, IH), 7.09 (m, IH), 4.51 (s, 2H), 3.76 (s, 3H).
EXAMPLE 41
6-(Benzo[6]thiophen-5-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine hydrobromide
[00174] The title compound was prepared in a manner similar to Example 35.
From l-(benzo[ό]thiophen-5-yl)-2-bromoethanone (106 mg, 0.415 mmol) and 4-amino-5-(2-methoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (98 mg, 0.44 mmol) was obtained the title compound as an off-white solid (32 mg, 0.085 mmol, 20%). 1H NMR (DMSO-d6) 8.45 (d, J = 1.8 Hz, IH), 8.14 (d, J= 8.7 Hz, IH), 7.90 (d, J = 5.7 Hz, IH), 7.81 (dd, J = 8.4, 1.8 Hz, IH), 7.53 - 7.63 (m, 3H), 7.23 (d, J = 8.7 Hz, IH), 7.14 (dt, J= 6.6, 0.6 Hz, IH), 4.53 (s, 2H), 3.76 (s, 3H).
EXAMPLE 42
3-(4-Fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine
[00175] (a) Methyl 4-fluoro-2-methoxybenzoate. A mixture of 4-fluoro-2- methoxybenzoic acid (5.01 g, 29.4 mmol) in methanol (40 mL) and concentrated sulfuric acid (1.5 mL) was refluxed overnight. The reaction mixture was cooled to room temperature and most of methanol was evaporated under vacuum. The residue was dissolved in 50 mL of ethyl acetate and washed with saturated NaHCO3, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified by chromatography (10% ethyl acetate/hexane) to give the title compound (5.21 g, 28.3 mmol, 96%) as a clear liquid. 1H NMR (CDCl3) 7.89 (m, IH), 6.68 (m, 2H), 3.89 (s, 3H), 3.87 (s, 3H).
[00176] (b) 4-Fluoro-2-methoxybenzohydrazide. A solution of methyl 4- fluoro-2-methoxybenzoate (5.20 g, 28.2 mmol) and hydrazine hydrate (3.0 mL, 61.7 mmol) in absolute ethanol (60 mL) was heated at 85 0C for 18 h and cooled to room temperature. The resulting white precipitate was collected, washed with cold ethanol and dried under vacuum to give the title compound (3.55 g, 19.3 mmol, 69%). 1H NMR (DMSO-d6) 9.18 (s, broad, IH), 7.73 (dd, J= 8.4, 7.2 Hz, IH), 7.01 (dd, J= 11.7, 2.4 Hz, IH), 6.85 (dt, J= 10.8, 2.4 Hz, IH), 4.51 (s, 2H), 3.87 (s, 3H), 3.30 (s, 3H).
[00177] (c) 4-Fluoro-2-methoxy benzoyl-2-dithiocarboxyhydrazide, potassium salt. A mixture of 4-fiuoro-2-methoxybenzohydrazide (1.12 g, 6.10 mmol) in absolute ethanol and potassium hydroxide (0.38 g, 6.8 mmol) was stirred at room temperature for 15 min, and to the solution was added carbon disulfide (0.45 mL, 7.5 mmol) dropwise via syringe. A yellow precipitate was observed and the mixture was stirred for 4 h at room temperature. The mixture was filtered and the solids were washed with cold ethanol, and dried under vacuum to give the title compound as an off white solid (1.70 g, 5.70 mmol, 76%). 1H NMR (DMSO-d6) 7.79 (dd, J = 8.7, 6.6 Hz, IH), 7.22, (dd, J = 11.4, 2.4 Hz, IH), 6.98 (dt, J= 9.0, 2.7 Hz, IH), 5.44 (s, broad, 2H), 3.92 (s, 3H).
[00178] (d) 4-Amino-5-(4-fluoro-2-methoxyphenyl)-3-mercapto-(4H)- 1,2,4- triazole. A solution of 4-fluoro-2-methoxy benzoyl-2-dithiocarboxyhydrazide potassium salt (0.89 g, 2.97 mmol) and hydrazine (0.4 mL, 8.2 mmol) in ethanol (8 mL) and water (5 mL) was refluxed for 4 h and cooled to room temperature. The resulting solution was diluted with water (8 mL) and acidified to pΗ = 1 with concentrated HCl with cooling on an ice bath. The precipitated product was collected by filtration, washed with cold water and dried under vacuum to give the title compound (289 mg). The crude product was used for next reaction without further purification.
[00179] (e) 3-(4-Fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine. A mixture of 2-bromo-l-(4-methyl-3- nitrophenyl)ethanone (140 mg, 0.543 mmol) and 4-amino-5-(4-fluoro-2- methoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (123 mg, 0.512 mmol) in isopropyl alcohol (6 ml) was refluxed for 3 h and cooled to room temperature. The reaction mixture was diluted with chloroform and washed with saturated NaHCO3, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified by chromatography (80% ethyl acetate/hexane, 0.2 mL methanol/10 mL solvent) to give the title compound as a light yellow solid (68 mg, 0.17 mmol, 33%). 1H NMR (CDCl3) 8.41 (d, J= 2.1 Hz, IH), 7.94 (dd, J = 8.1, 2.1 Hz, IH), 7.58 (dd, J = 8.4, 6.6 Hz, IH), 7.46 (d, J = 8.4 Hz, IH), 6.78 (m, 2H), 4.05 (s, 2H), 3.76 (s, 3H), 2.67 (s, 3H).
EXAMPLE 43
6-(3 - Amino-4-methylphenyl)-3 -(4-fluoro-2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00180] The title compound was prepared in a manner similar to Example 33.
From 3-(4-fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine was obtained the title compound as a yellow solid (17%). 1H NMR (CDCl3) 7.58 (dd, J= 8.4, 6.6 Hz, IH), 7.09 (m, 3H), 6.77 (m, 2H), 3.94 (s, 2H), 3.73 (s, 3H), 2.21 (s, 3H).
EXAMPLE 44
6-(4-Methoxy-3-nitrophenyl)-3-(5-methyl-lH-imidazol-4-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-Z>] [ 1 ,3 ,4]thiadiazine
[00181] The title compound was prepared in a manner similar to Example Ib.
From 2-bromo-l-(4-methoxy-3-nitrophenyl)ethanone (70 mg, 0.25 mmol) and 4-amino-5-(5-methyl-lH-imidazol)-3-mercapto-(4H)-l,2,4-triazole (50 mg, 0.25 mmol) was obtained 75 mg (79%) of the title compound. 1H NMR (CD3OD) 8.45 (bs, IH), 8.15 (m, IH), 7.42 (d, J = 9.0 Hz, IH), 7.31 (d, J = 9.0 Hz, IH), 4.31 (s, 2H), 4.03 (s, 3H), 2.45 (s, 3H). EXAMPLE 45
6-(3-Amino-4-methoxyphenyl)-3-(5-methyl-lH-imidazol-4-yl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine
[00182] The title compound was prepared in a manner similar to Example 2.
From 6-(3-amino-4-methoxyphenyl)-3-(5-methyl-lH-imidazol-4-yl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (74 mg, 0.2 mmol) and tin(II) chloride (226 mg, 1 mmol) was obtained 16 mg (23%) of the title compound. 1H NMR (CD3OD): 7.76 (s, IH), 7.41 (s, IH), 7.32 (m, IH), 6.80 (m, IH), 4.21 (s, 2H), 3.91 (s, 3H), 2.42 (s, 3H).
EXAMPLE 46
3-(2-Methoxyphenyl)-6-( 1 -methylsulfonyl-2,3-dihydro- 1 H-indol-5-yl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine
[00183] The title compound was prepared in a manner similar to Example Ib.
From 2-chloro-l -[I -(methylsulfonyl)-2,3-dihydro-lH-indol-5-yl]ethanone (789 mg, 3.0 mmol) and 4-amino-5-(2-methoxyphenyl)-3-mercapto-(4H)- 1 ,2,4-triazole (666 mg, 3.0 mmol) in isopropanol was obtained 1.12 g (87%) of the title compound. 1H NMR (CDCl3): 7.68 (s, IH), 7.66-7.60 (m, 2H), 7.55-7.48 (m, IH), 7.44 (d, J= 8.4 Hz, IH), 7.13-7.08 (m, IH), 7.03 (t, J= 8.7 Hz, IH), 4.05 (t, J = 8.4 Hz, IH), 3.96 (s, 2H), 3.76 (s, 3H), 3.19 (t, J = 8.7 Hz, IH), 2.93 (s, 3H).
EXAMPLE 47
3-(2-Methyl-l//-imidazo[l,2-α]pyridin-3-yl)-6-(4-methoxy-3-nitrophenyl)-7//- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00184] The title compound was prepared in a manner similar to Example Ib.
From 2-bromo-l-(4-methoxy-3-nitrophenyl)ethanone (37 mg, 0.13 mmol) and 4-amino-5-(2 -methyl- 1 H-imidazo [ 1 ,2-a]pyridin-3 -yl)-4H-pyrazole-3 -thiol (33 mg, 0.13 mmol) was obtained 46 mg (81%) of the title compound. 1H NMR (CDCl3): 8.81 (d, J= 6.9 Hz, IH), 8.32 (d, J= 2.1 Hz, IH), 8.16-8.12 (dd, J = 9.0, 2.4 Hz, IH), 7.66 (d, J= 8.7 Hz, IH), 7.34 (t, J= 7.8 Hz, IH), 7.20 (d, J = 9.0 Hz, IH), 6.92 (t, J= 7.8 Hz, IH), 4.06 (s, 3H), 2.49 (s, 3H). EXAMPLE-48
6-(3-Amino-4-methoxyphenyl)-3-(2-methyl-l//-imidazo[l,2-α]pyridin-3-yl)- 7H-[1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine
[00185] A mixture of 3-(2-methyl-lH-imidazo[l,2-α]pyridin-3-yl)-6-(4- methoxy-3-nitrophenyl)-7H-[l ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine (42 mg, 0.10 mmol) and tin(II) chloride (102 mg, 0.45 mmol) in methanol (5 mL) was refluxed under argon for 5 h. It was then diluted with water (30 mL) and neutralized with 2N sodium hydroxide to pΗ = 12. The mixture was extracted with ethyl acetate (3 x 20 mL). The extracts were dried, concentrated and the residue was purified by column chromatography (EtOAc/ MeOH 10:1) to give 15 mg (38%) of the title compound. 1H NMR (CDCl3): 8.82 (d, J = 6.9Hz, IH), 7.65 (d, J = 9.0 Hz, IH), 7.34 (d, J = 76.6 Hz, IH), 7.19 (m, IH), 6.90 (m, IH), 6.82 (d, J= 8.7 Hz, IH), 4.00 (s, 2H), 3.92 (s, 3H), 2.51 (s, 3H).
EXAMPLE 49
3 -(2-Dimethylaminophenyl)-6-(4-methyl-3 -nitrophenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine
[00186] The title compound was prepared in a manner similar to Example Ib.
From 2-bromo-l-(4-methyl-3-nitrophenyl)ethanone (122 mg, 0.47 mmol) and 4-amino-5-(2-dimethylaminophenyl)-3-mercapto-(4H)-l,2,4-triazole (111 mg, 0.47 mmol) in isopropanol was obtained 171 mg (92%) of the title compound. 1K NMR (DMSOd6): 8.42 (s, 1Η), 8.06 (d, J = 8.1 Hz, IH), 7.67 (d, J = 8.1 Hz, IH), 7.46 (t, J= 7.2 Hz, IH), 7.39 (d, J = 7.5 Hz, IH), 7.12 (d, J= 8.14 Hz, IH), 7.00 (d, J= 7.5 Hz, IH), 4.47 (s, 2H), 2.56 (s, 3H), 2.50 (s, 6H).
EXAMPLE 50
6-(3 - Amino-4-methylphenyl)-3 -(2-dimethylaminophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine hydrochloride
[00187] The title compound was prepared in a manner similar to Example 2.
From 3-(2-dimethylaminophenyl)-6-(4-methyl-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (140 mg, 0.35 mmol) and tin chloride (360 mg, 1.6 mmol) was obtained 142 mg (85%) of the title compound as solid. 1H NMR (DMSO-d6): 7.95 (bs, IH), 7.91 (s, IH), 7.85-7.72 (m, 3H), 7.62 (t, J = 7.2 Hz, IH), 7.49 (d, J = 7.5 Hz, IH), 4.46 (s, 2H), 3.16 (s, 6H), 2.42 (s, 3H). EXAMPLE 51
6-(4-Methyl-3 -nitrophenyl)-3 -(2-trifluoromethoxyphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00188] (a) 4-Amino-5-(2-trifluoromethoxyphenyl)-3-mercapto-(4H)-l ,2,4- triazole. To a mixture of potassium hydroxide (764 mg, 13.6 mmol) in ethanol (30 mL) was added carbon disulfide (1.04 g, 13.6 mmol). It was stirred at room temperature for 3 h. The solution was diluted with water (50 mL) and extracted with ethyl acetate (3 x 30 mL). The organic extracts were dried and concentrated. To the residue was added water (2 mL) and hydrazine (727 mg, 14.5 mmol) and it was refluxed for 2 h. The mixture was cooled and diluted with water (30 mL), neutralized with 2N hydrochloride to produce precipitates. The mixture was filtered and the solids were washed with aqueous sodium carbonate, dried to give a crude product which was used without further purification.
[00189] (b) 6-(4-Methyl-3-nitrophenyl)-3-(2-trifluoromethoxyphenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine. The title compound was prepared in a manner similar to Example Ib. From 2-bromo-l-(4-methyl-3- nitrophenyl)ethanone (516 mg, 2.0 mmol) and 4-amino-5-(2- trifluoromethoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (550 mg, 2.0 mmol) in isopropanol was obtained 160 mg (18%) of the title compound. 1H NMR (CDCl3): 8.39 (d, J = 2.1 Hz, IH), 7.99-7.95 (dd, J = 8.4, 2.1 Hz, IH), 7.87- 7.84 (dd, J= 8.4, 2.1 Hz, IH), 7.62-7.59 (m, IH), 7.50-7.46 (m, 2H), 7.40 (m, IH), 4.02 (s, 2H), 2.68 (s, 3H).
EXAMPLE 52
6-(3 - Amino-4-methylphenyl)-3 -(2-trifluoromethoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00190] The title compound was prepared in a manner similar to Example 2.
From 3-(2-trifluoromethoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H-
[l,2,4]triazolo[3 ,4-&][l, 3,4]thiadiazine (140 mg, 0.32 mmol) and tin chloride (363 mg, 1.61 mmol) was obtained 122 mg (79%) of the title compound. 1H NMR (CD3OD): 7.97 (d, J= 7.8 Hz, IH), 7.83 (d, J= 8.1 Hz, IH), 7.79 (t, J = 7.8 Hz, IH), 7.64-7.56 (m, 3H), 4.44 (s, 2H), 2.48 (s, 3H). EXAMPLE 53
3-(2-Methoxy-5-methylphenyl)-6-(4-methylphenyl)-7//-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00191] The title compound was prepared in a manner similar to Example 1Od.
From 2-bromo-l-(4-methylphenyl)ethanone (0.180 g, 0.846 mmol) and 4-amino-5-(2-methoxy-5-methylphenyl)-4H-[l ,2,4]triazole-3-thiol (0.200 g, 0.846 mmol) was obtained 0.292 g (80%) of the title compound as a white solid. 1H NMR (DMSO-J6): 7.77 (d, J = 8.4 Hz, 2H), 7.63 (dd, J = 8.8 and 2.6 Hz, IH), 7.55 (d, J= 3.0 Hz, IH), 7.34 (d, J= 8.1 Hz, 2H), 7.26 (d, J= 9.1 Hz, IH), 4.41 (s, 2H), 3.74 (s, 3H), 2.36 (s, 3H).
EXAMPLE 54
3-(2-Methoxy-5-methylphenyl)-6-(2-oxoindolin-5-yl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00192] (a) 5-(2-Bromoacetyl)indolin-2-one. To an oven-dried round bottom flask charged with a magnetic stir bar at rt under argon was added aluminum chloride (1.30 g, 9.76 mmol) and CH2Cl2 (37 mL). The resulting suspension was cooled to 0 °C, bromoacetyl bromide (0.85 mL, 9.8 mmol) was added dropwise and then oxindole (0.500 g, 3.75 mmol) was added in portions over 2 min. The resulting brown solution was stirred at 0 0C and then equilibrated to rt. The reaction solution was poured into a slurry of ice:water (500 mL) and then neutralized with 1.0 M NaOH(aq) until the pH = 7. The aqueous solution was then extracted with CHCl3 (2 x 250 mL). The extracts were washed with brine (50 mL), dried over MgSO4, filtered and concentrated to an orange solid. It was purified by flash chromatography (silica gel, gradient elution with EtOAc :hexanes, 1 :4 to 1 :1) to give 0.130 g (13%) of yellow solid. 1H NMR (CDCl3): 7.95-7.90 (m, 2H), 7.84 (br s, IH), 6.95 (d, J= 7.8 Hz, IH), 4.39 (s, 2H), 3.61 (s, 2H).
[00193] (b) 3-(2-Methoxy-5-methylphenyl)-6-(2-oxoindolin-5-yl)-7//-
[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 5-(2-bromoacetyl)indolin-2-one (0.050 g, 0.20 mmol) and 4-amino-5-(2-methoxyphenyl)-4H-[l,2,4]triazole-3-thiol (0.200 g, 0.846 mmol) was obtained 0.102 g (100%) of the title compound as a yellow solid. 1H NMR (DMSO-^6): 10.74 (s, IH), 7.75 (d, J = 8.1 Hz, IH), 7.70 (s, IH), 7.61-7.55 (m, IH), 7.51 (dd, J= 7.5 and 1.6 Hz, IH), 7.22 (d, J = 8.4 Hz, IH), 7.12 (t, J = 7.5 Hz, IH), 6.93 (d, J = 8.1 Hz, IH), 4.39 (s, 2H), 3.74 (s, 3H), 3.55 (s, 2H).
EXAMPLE 55
6-(3-Acetamide-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-Z)] [ 1 ,3,4]thiadiazine
[00194] To an oven-dried round bottom flask charged with a magnetic stir bar at rt under argon with an addition funnel was added 6-(3-amino-4- methylphenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- Z?][l,3,4]thiadiazine (0.250 g, 0.711 mmol), potassium acetate (0.202 g, 2.06 mmol) and anhydrous DMF (2.8 mL). The resulting yellow suspension was cooled to 0 0C and then acetic anhydride (0.19 mL, 2.1 mmol) was added dropwise via the addition funnel. The yellow suspension was equilibrated to rt and then heated to 80 0C, followed by addition of isoamyl nitrite (0.064 mL, 0.48 mmol). It was heated at 80 °C for 17 h, and then cooled to rt. The suspension was quenched with NaHCO3(aq) (10 mL) and then extracted with CHCl3 (2 x 50 mL), dried over MgSO4, filtered and concentrated. The residue was purified by flash chromatography (silica gel, gradient elution with EtOAc :hexanes, 1 :1 to MeOHiCH2Cl2, 4:96) to give 0.003 g (1%) of white solid. 1H NMR (DMSO-^6): 9.44 (s, IH), 7.92 (s, IH), 7.59-7.57 (m, 2H), 7.49 (d, J= 7.3 Hz, IH), 7.35 (d, J= 8.4 Hz, IH), 7.20 (d, J= 8.4 Hz, IH), 7.11 (t, J= 7.5 Hz, IH), 4.38 (s, 2H), 3.75 (s, 3H), 2.25 (s, 3H), 2.06 (s, 3H).
EXAMPLE 56
6-(lH-l-Acetylindazol-6-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00195] To an oven-dried round bottom flask charged with a magnetic stir bar at rt under argon was added 6-(3-amino-4-methylphenyl)-3-(2- methoxyphenyl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (0.200 g, 0.569 mmol), potassium acetate (0.162 g, 1.65 mmol) and anhydrous CHCl3 (2.3 mL). The resulting yellow suspension was cooled to 0 0C and then acetic anhydride (0.155 mL, 1.65 mmol) was added dropwise. The yellow suspension was equilibrated to rt and then heated to 80 0C, followed by addition of isoamyl nitrite (0.051 mL, 0.38 mmol). It was heated for 17 h at 80 0C, and then cooled to rt. The suspension was neutralized with NaHCO3(aq) (10 mL) and then extracted with CHCl3 (2 x 50 mL), washed with brine (20 mL), dried over MgSO4, filtered and concentrated. The residue was purified by flash chromatography (silica gel, gradient elution with EtOAc:hexanes, 1 :1 to MeOH:CH2Cl2, 4:96) to give 0.001 g (<1%) of white solid. 1H NMR (DMSO-βfe): 8.80 (s, IH), 8.57 (d, J = 0.7 Hz, IH), 8.05 (d, J = 8.4 Hz, IH), 7.88 (d, J= 8.4 Hz, IH), 7.64-7.57 (m, IH), 7.52 (dd, J= 7.9 and 1.3 Hz, IH), 7.24 (d, J = 8.1 Hz, IH), 7.12 (t, J = 7.5 Hz, IH), 4.54 (s, 2H), 3.81 (s, 3H), 2.74 (s, 3H).
EXAMPLE 57
6-(4-N-Hydroxyamidinophenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00196] To an oven-dried round bottom flask at rt under argon charged with a magnetic stir bar and 6-(4-cyanophenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-Z>][l,3,4]thiadiazine (0.140 g, 0.403 mmol) was added anhydrous ethanol (2.0 mL) and hydroxylamine (0.027 mL, 0.44 mmol). The resulting yellow suspension was heated to 80 0C for 3 h and then cooled to rt. The solvent was removed by rotary evaporation to give the crude product as a yellow solid. It was purified by flash chromatography (silica gel, gradient elution with EtOAc :hexanes, 1 :1 to MeOΗ:CΗ2Cl2, 4:96 to 1:1) to give 0.037 g (24%) of white solid. 1H NMR (DMSO-<fc): 10.00 (br s, IH), 8.09 (br s, IH), 8.01-7.79 (m, 4H), 7.60-7.55 (m, IH), 7.51 (d, J = 7.3 Hz, IH), 7.22 (d, J = 8.4 Hz, IH), 7.12 (t, J= 7.5 Hz, IH), 6.22 (br s, 2H), 4.45 (d, J= 5.7 Hz, 2H), 3.74 (s, 3H).
EXAMPLE 58
3-(5-Fluoro-2-methoxyphenyl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
[00197] (a) 5-Fluoro-2-methoxybenzoyl-2-dithiocarboxyhydrazide, potassium salt. The title compound was prepared in a manner similar to Example 10b. From 5-fluoro-2-methoxybenzohydrazide (2.95 g, 16.0 mmol) and ethanol (80 mL) and carbon disulfide (1.44 mL, 24.0 mmol) was obtained 4.70 g (>100%) of the title compound as a yellow solid. 1H NMR (DMSO-cfe): 12.04 (s, IH), 9.77 (s, IH), 7.73-7.69 (m, IH), 7.42-7.35 (m, IH), 7.26-7.21 (m, IH), 3.97 (s, 3H).
[00198] (b) 4-Amino-5-(5-fluoro-2-methoxyphenylHH-[l ,2,4]triazole-3-thiol.
The title compound was prepared in a manner similar to Example 10c. From 5-fluoro-2-methoxybenzoyl-2-dithiocarboxyhydrazide, potassium salt (4.50 g, 15.1 mmol) and hydrazine (6.60 mL, 181 mmol) was obtained 2.75 g (76%) of the crude product as a white solid. Recrystallization (EtOΗrwater, 50:5 mL) gave of the title compound as a white solid. 1H NMR (DMSO-J6): 13.91 (br s, IH), 7.45-7.38 (m, IH), 7.35-7.31 (m, IH), 7.22-7.18 (m, IH), 5.46 (s, 2H), 3.80 (s, 3H).
[00199] (c) 3-(5-Fluoro-2-methoxyphenyl)-6-(4-methylphenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine. The title compound was prepared in a manner similar to Example 1Od. From 2-bromo-l-(4-methylphenyl)ethanone (0.089 g, 0.42 mmol) and 4-amino-5-(5-fiuoro-2-methoxyphenyl)-4H- [l,2,4]triazole-3 -thiol (0.100 g, 0.416 mmol) was obtained 0.130 g (72%) of the title compound as a white solid. 1H NMR (DMSO-J6): 7.80-7.75 (m, 2H), 7.46-7.33 (m, 4H), 7.26-7.22 (m, IH), 4.41 (t, J = 4.2 Hz, 2H), 3.72 (t, J= 4.4 Hz, 3H), 2.36 (t, J= 4.4 Hz, 3H).
EXAMPLE 59
3-(2-Methoxyphenyl)-6-(4-(N-methylacetamido)methylphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-b] [ 1 ,3 ,4]thiadiazine
[00200] The title compound was prepared in a manner similar to Example 42e.
From N-(4-(2-chloroacetyl)benzyl)-N-methylacetamide (238 mg, 0.993 mmol) and 4-amino-5-(2-methoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (220 mg, 0.99 mmol) was obtained the title compound as an off-white solid (358 mg, 0.878 mmol, 89%). 1H NMR (CDCl3): 7.38 - 7.82 (m, 2H), 7.61 (dd, J= 7.5, 1.5 Hz, IH), 7.51 (m, IH), 7.25 - 7.34 (m, 2H), 7.10 (m, IH), 7.02 (m, IH), 4.60 (m, 2H), 3.98 (m, 2H), 3.76 (m, 3H), 2.95 (s, 3H), 2.14 (m, 3H).
EXAMPLE 60
3-(2-Methoxyphenyl)-6-(4-(N-methylamino)methylphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine
[00201] A mixture of 3-(2-methoxyphenyl)-6-(4-(N- methylacetamido)methylphenyl)-7H-[l,2,4]triazolo[3,4-Z>][l,3,4]thiadiazine (49 mg, 0.12 mmol) in aquoues hydrochloric acid (3N, 15 mL) was heated at 100 0C overnight and cooled to room temperature. The reaction mixture was basified by addition of solid NaHCO3 in small portions and the resulting solution was extracted with chloroform (20 mL x 2). The combined organic extracts were washed with saturated NaHCO3, dried over anhydrous Na2SO4, filtered and concentrated to give the product (42 mg, 0.114 mmol, 96%). 1H NMR (CDCl3): 7.76 (m, 2H), 7.62 (dd, J = 7.5, 1.8 Hz, IH), 7.50 (ddd, J = 8.1, 7.2, 1.5 Hz, IH), 7.40 (m, 2H), 7.09 (dt, J= 7.5, 0.9 Hz, IH), 7.02 (d, J =
8.1 Hz, IH), 3.99 (s, 2H), 3.81 (s, 2H), 3.74 (s, 3H), 2.46 (s, 3H).
EXAMPLE 61
6-(4-(N,iV-Dimethylamino)methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00202] The title compound was prepared in a manner similar to Example 39.
From 3 -(2-methoxyphenyl)-6-(4-(N-methylamino)methylphenyl)-7H-
[l,2,4]triazolo[3,4-&][l,3,4]thiadiazine (36 mg, 0.099 mmol), formaldehyde (37% aqueous, 0.20 mL) and sodium caynoborohydride (50 mg, 0.8 mmol) was obtained the title compound as an off-white solid (36 mg, 0.095 mmol, 96%). 1U NMR (CDCl3): 7.75 (m, 2Η), 7.62 (dd, J = 7.8, 2.1 Hz, IH), 7.50 (ddd, J = 8.4, 7.2, 1.8 Hz, IH), 7.41 (m, 2H), 7.09 (dt, J = 7.2, 0.6 Hz, IH),
7.02 (d, J= 8.1 Hz, IH), 3.99 (s, 2H), 3.75 (s, 3H), 3.47 (s, 2H), 2.25 (s, 6H).
EXAMPLE 62
3-(5-Fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine
[00203] The title compound was prepared in a manner similar to Example 1 Od.
From 2-bromo-l-(4-methyl-3-nitrophenyl)-ethanone (0.537 g, 2.08 mmol) and 4-amino-5-(5-fluoro-2-methoxyphenyl)-4H-[l ,2,4]triazole-3-thiol (0.500 g, 2.08 mmol) was obtained 0.815 g (81%) of the title compound as a white solid. 1H NMR (DMSO-J6) 8.45 (d, J= 1.8 Hz, IH), 8.08 (dd, J= 8.1 and 1.8 Hz, IH), 7.69 (d, J = 8.1 Hz, IH), 7.48-7.38 (m, 2H), 7.28-7.23 (m, IH), 4.49 (s, 2H), 3.74 (s, 3H), 2.58 (s, 3H). EXAMPLE 63
6-(3-Amino-4-methylphenyl)-3-(5-fluoro-2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine
[00204] The title compound was prepared in a manner similar to Example 15.
From 3-(5-fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]-thiadiazine (0.800 g, 1.66 mmol) and tin (II) chloride dihydrate (1.69 g, 7.50 mmol) was obtained 0.410 g (67%) of the title compound as a yellow solid. 1H NMR (DMSO-J6) 7.46-7.35 (m, 2H), 7.24- 7.20 (m, IH), 7.08-7.05 (m, 2H), 7.00-6.98 (m, IH), 5.14 (br s, 2H), 4.30 (s, 2H), 3.72 (s, 3H), 2.09 (s, 3H).
EXAMPLE 64
3-(2,5-Dimethylfuran-3-yl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00205] The title compound was prepared in a manner similar to Example 1Od.
From 2-bromo-l-(4-methyl-3-nitrophenyl)-ethanone (0.083 g, 0.323 mmol) and 4-amino-5-(2,5-dimethylfuran-3-yl)-3-mercapto-4H-[l ,2,4]-triazole
(0.068 g, 0.323 mmol) was obtained 0.079 g (55%) of the title compound as a brown solid. 1H NMR (DMSO-J6) 8.56 (d, J = 1.8 Hz, 2H), 8.23 (dd, J= 8.1 and 1.8 Hz, IH), 7.73 (d, J = 8.1 Hz, IH), 6.56 (s, IH), 4.48 (s, 2H), 2.61 (s, 3H), 2.30 (s, 3H).
EXAMPLE 65
6-(3-Amino-4-methylphenyl)-3-(2,5-dimethylfuran-3-yi)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
[00206] The title compound was prepared in a manner similar to Example 15.
From 3-(2,5-dimethylfuran-3-yl)-6-(4-methyl-3-nitrophenyl)-7H-
[l,2,4]triazolo[3,4-δ][l,3,4]-thiadiazine (0.067 g, 0.149 mmol) and tin (II) chloride dihydrate (0.151 g, 0.669 mmol) was obtained 0.012 g (24%) of the title compound as a yellow solid. 1H NMR (DMSO-J6) 7.24 (d, J = 1.5 Hz, IH), 7.10-7.08 (m, 2H), 6.56 (d, J= 1.1 Hz, IH), 5.22 (br s, 2H), 4.31 (s, 2H), 2.53 (s, 3H), 2.32 (s, 3H), 2.13 (s, 3H). EXAMPLE 66
3-(2-Methoxyphenyl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b][l ,3,4]thiadiazin-7-one
[00207] To an oven-dried round bottom flask charged with a magnetic stir bar at room temperature under argon was added (4-methylphenyl)(oxo) acetic acid (0.500 g, 3.04 mmol), 4-amino-5-(2-methoxyphenyl)-3-mercapto-4H-[l,2,4]- triazole (0.723 g, 3.25 mmol) and POCl3 (15.2 mL). The yellow suspension was refluxed at 130 0C for 20 h and then cooled to room temperature. The resulting green solution was concentrated by rotary evaporation and then diluted with water. The reaction mixture was then neutralized using NaHCO3 until pH = 7, followed by extraction with CHCl3 (2 x 100 mL). The combined organic extracts were washed with brine (20 mL), dried over MgSO4, filtered, and concentrated to a brown solid. It was purified by flash chromatography (silica gel, elution with EtOAc :hexanes, 1 :1) to give 0.018 g (2%) of a brown solid. 1H NMR (DMSO-J6) 8.21 (d, J= 8.1 Hz, 2H), 7.83 (d, J= 8.1 Hz, IH), 7.65 (t, J = 7.7 Hz, IH), 7.46 (d, J = 8.1 Hz, 2H), 7.30 (d, J = 8.4 Hz, IH), 7.16 (t, J= 7.5 Hz, IH), 3.91 (s, 3H), 2.43 (s, 3H).
EXAMPLE 67
6-(N-Acetyl-indolin-5-yl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
(a) TV-Acetyl-indoline. To an oven-dried round bottom flask charged with a magnetic stir bar at room temperature under argon was added indoline (1.00 g, 8.39 mmol) and acetic acid (14.0 mL, 0.6 M). The yellow solution was cooled to 0 0C and acetyl chloride (3.5 mL, 2.4 M) was added dropwise via syringe. The orange solution was heated to reflux at 90 0C for 3 h and then was cooled to room temperature. The solvent was removed by rotary evaporation, followed by vacuum pump to give 1.58 g (> 100%) of the title compound as a brown solid. 1H NMR (DMSO-^6) 8.03 (d, J = 8.1 Hz, IH), 7.22 (d, J= 7.3 Hz, IH), 7.13 (t, J= 7.9 Hz, IH), 6.97 (X, J= 7.5 Hz, IH), 4.07 (X, J= 8.6 Hz, 2H), 3.13 (t, J= 8.6 Hz, 2H), 2.15 (s, 3H).
(b) l-(//-Acetyl-indolin-5-yl)-2-bromo-ethanone. The title compound was prepared in a manner similar to Example 32. From aluminum chloride (0.728 g, 5.46 mmol), 1,2-dichloroethane (27 mL), bromoacetyl bromide (0.47 mL, 5.46 mmol) and JV-acetyl-indoline (0.338 g, 2.10 mmol) was obtained 0.229 g (39%) of the title compound as yellow solid. 1H NMR (CDCl3) 8.27 (d, J = 9.2 Hz, IH), 7.86-7.84 (m, 2H), 4.42 (s, 2H), 4.15 (t, J= 8.6 Hz, 2H), 3.27 (t, J= 8.6 Hz, 2H), 2.27 (s, 3H).
(c) 6-(iV-Acetyl-indolin-5-yl)-3-(2-methoxyphenyl)-7H-
[l,2,4]triazolo[3,4-Z)][l,3,4]thiadiazine. The title compound was prepared in a manner similar to Example Ib. From l-(iV-acetyl-indolin-5-yl)-2-bromo- ethanone (0.050 g, 0.18 mmol) and 4-amino-5-(2-methoxyphenyl)-4H- [l,2,4]triazole-3-thiol (0.038 g, 0.18 mmol) in isopropanol (0.90 mL) was obtained 0.079 g (93%) of the title compound as a brown solid. 1H-NMR (OMSO-(I6) 8.09 (d, J = 9.5 Hz, IH), 7.72-7.71 (m, 2H), 7.60-7.54 (m, IH), 7.50 (dd, J= 7.5 and 1.7 Hz, IH), 7.22 (d, J= 8.1 Hz, IH), 7.14-7.09 (m, IH), 4.38 (s, 2H), 4.14 (t, J = 8.2 Hz, 2H), 3.74 (s, 3H), 3.17 (t, J = 8.2 Hz, 2H), 2.18 (s, 3H).
EXAMPLE 68
6-(Indolin-5-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
The title compound was prepared in a manner similar to Example 60. From 6-(N-acetyl-indolin-5-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- £][l,3,4]thiadiazine (0.050 g, 0.10 mmol) and 6N HCl (1.2 mL, 0.085 M) was obtained 0.020 g (55%) of the title compound as orange solid 1H NMR (CDCl3) 7.62 (d, J= 7.7 Hz, IH), 7.58 (s, IH), 7.49-7.46 (m, 2H), 7.09 (t, J = 7.5 Hz, IH), 7.02 (d, J = 8.4 Hz, IH), 6.57 (d, J = 8.4 Hz, IH), 3.91 (s, 2H), 3.76 (s, 3H), 3.66 (t, J= 8.4 Hz, 2H), 3.05 (t, J= 8.4 Hz, 2H).
EXAMPLE 69
6-(3-(N-Boc-O-'Bu-Serinyl-amino)-4-methylphenyl)-3-(2- methoxyphenyl)-7H-[ 1 ,2 ,4]triazolo-[3 ,A-b] [ 1 ,3 ,4]thiadiazine
To an oven-dried round bottom reaction flask charged with a magnetic stir bar at room temperature under argon with 6-(3-amino-4-methylphenyl)-3- (2-methoxyphenyl)-7//-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (0.200 g, 0.569 mmol), Boc-Ser('Bu)OH DCHA (0.482 g, 1.09 mmol), PyBOP (0.554 g, 1.25 nimol), HOBt (0.231 g, 1.71 mmol) and CH3CN (5.7 mL) was added triethylamine (0.100 mL, 0.716 mmol). The yellow solution was heated at 50 0C for 19 h and then was cooled to room temperature The solvent was removed by rotary evaporation to give a residue that was diluted with EtOAc (150 mL). The organic extracts were washed with saturated NaHCO3 (20 mL), IM citric acid (20 mL), brine (20 mL), dried over MgSO4, filtered and concentrated to give the crude product. Purification by flash chromatography (12 g pre-packed silica gel, elution with EtOAc:hexanes, 1 :1) gave 0.282 g (83%) of the title compound as a white solid. 1H NMR (DMSO-^6) 9.45 (s, IH), 7.83 (d, J- 1.8 Hz, IH), 7.63-7.60 (m, IH), 7.57-7.51 (m, IH), 7.48 (dd, J = 7.7 and 1.8 Hz, IH), 7.37 (d, J = 8.4 Hz, IH), 7.17 (d, J = 8.1 Hz, IH), 7.12-7.07 (m, IH), 6.75(d, J= 8.1 Hz, IH), 4.38 (s, 2H), 4.20 (m, IH), 3.74 (s, 3H), 3.53 (d, J= 5.9 Hz, 2H), 2.24 (s, 3H), 1.40 (s, 9H), 1.11 (s, 6H).
EXAMPLE 70
6-(3-Fluoro-4-methyl-5-nitrophenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo[3,4-b] [ 1 ,3,4]thiadiazine
The title compound was prepared in a manner similar to Example Ib. From 2-bromo-l-(3-fluoro-4-methyl-5-nitrophenyl)ethanone (380 mg, 1.38 mmol) and 4-amino-5-(2-methoxyphenyl)-3-mercapto-(4H)-l,2,4-triazole (315 mg, 1.42 mmol) was obtained the title compound as a light yellow solid (182 mg, 0.456 mmol, 33%). 1H NMR (CDCl3) 7.93 (d, J = 8.7 Hz, IH), 7.58 (dd, J= 7.8, 1.8 Hz, IH), 7.47 (ddd, J = 8.4, 7.5, 1.8 Hz, IH, 7.38 (d, J= 6.9 Hz, IH), 7.07 (dt, J = 7.8, 1.2 Hz, IH), 6.96 (d, J= 8.1 Hz, IH), 3.88 (s, 2H), 3.75 (s, 3H), 2.38 (d, J= 2.1 Hz, 3H).
EXAMPLE 71
6-(3-Amino-5-fluoro-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine The title compound was prepared in a manner similar to Example 21. From 6-(3-fiuoro-4-methyl-5-nitrophenyl)-3-(2-methoxyphenyl)-7H-
[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (170 mg, 0.426 mmol) was obtained the title compound as yellow solid (114 mg, 0.309 mmol, 73%). 1H NMR (CDCl3) 7.57 (dd, J = 7.8, 1.8 Hz, IH), 7.45 (m, IH), 7.25 - 7.28 (m, 2H), 7.08 (dt, J = 7.5, 0.9 Hz, IH), 6.99 (d, J = 8.7 Hz, IH), 3.98 (s, 2H), 3.75 (s, 3H), 2.18 (d, J= 1.2 Hz, 3H).
EXAMPLE 72
6-(4-Methyl-3-nitrophenyl)-3-(2-ethoxyphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine
The title compound was prepared in a manner similar to Example Ib. From 2-bromo-l-(4-methyl-3-nitrophenyl)ethanone (516 mg, 2 mmol) and 4- amino-5-(2-ethoxyphenyl)-3-mercapto-(4//)-l,2,4-triazole (437 mg, 1.69 mmol) in isopropanol was obtained 580 mg (87%) of the title compound.
EXAMPLE 73
6-(3-Amino-4-methylphenyl)-3-(2-ethoxyphenyl)-7H-[l,2,4]triazolo[3,4- ό][l,3,4]thiadiazine hydrochloride
The title compound was prepared in a manner similar to Example 2. From a mixture of 6-(4-methyl-3-nitrophenyl)-3-(2-ethoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (530 mg, 1.34 mmol) and tin chloride (1.51 g, 6.69 mmol) in ethanol (25 mL) was obtained 520 mg (89%) of the title compound. 1H NMR (DMSO-d6): 7.81 (bs, IH), 7.79 (bs, IH), 7.73- 7.66(m, 2H), 7.56 (d, J= 8.4 Hz, IH), 7.32 (d, J= 8.4 Hz, IH), 7.24 (t, J= 8.1 Hz, IH), 4.50 (s, 2H), 4.15 (q, J = 6.9 Hz, 2H), 2.48 (s, 3H), 1.16 (t, J = 6.9 Hz, 2H).
EXAMPLE 74
6-(3-(Glycinyl-amino)-4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H- [l,2,4]triazolo[3,4-b][l,3,4]thiadiazine (a) 6-(3 -(Boc-glycinyl-amino)-4-methoxyphenyl)-3 -(2-methylfuran-3 - yl)-7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine. A mixture of 6-(3-amino-4- methoxyphenyl)-3-(2-methylfuran-3-yl)-7H-[l,2,4]triazolo[3,4- ό][l,3,4]thiadiazine (342 mg, 1 mmol), Boc-Gly (192.5 mg, 1.1 mmol), PyBOP (530 mg, 1.2 mmol), ΗOBt (230 mg, 1.7 mmol) and triethyl amine (0.5 mL) in acetonitrile (20 mL) was stirred at 6O0C for 8 h. It was then cooled and diluted with water (100 mL), the mixture was extracted with ethyl acetate (3x 30 mL) and the organic layer was combined, washed with brine, dried and concentrated. The residue was purified by column chromatography to give 288 mg (58%) of the title compound. 1H NMR (CDCl3): 8.99 (d, J = 2.1 Hz, IH), 8.53 (s, IH), 7.72-7.68 (dd, J= 8.7, 2.4 Hz, IH), 7.39 (t, J = 2.1 Hz, IH), 7.01 (s, IH), 7.00 (t, J = 8.4 Hz, IH), 5.20 (bs, IH), 3.98 (s, 3H), 3.97 (s, 3H), 2.70 (s, 3H), 1.50 (s, 9H):
(b) 6-(3 -(Glycinyl-amino)-4-methoxyphenyl)-3 -(2-methylfuran-3 -yl)- 7H-[l,2,4]triazolo[3,4-b][l,3,4]thiadiazine. To a solution of 6-(3-(Boc- glycinyl-amino)-4-methoxyphenyl)-3 -(2-methylfuran-3 -yl)-7H- [l,2,4]triazolo[3,4-b][l,3,4]thiadiazine (280 mg, 0.56 mmol) in dichloromethane (8 mL) was added 4N hydrochloride in dioxane (5 mL) at room temperature and it was stirred for 1 h and become cloudy. The mixture was diluted with ether (30 mL) and produced a precipitate, which was filtered and dried to give 230 mg (93%) of the title compound. 1H NMR (DMSO-d6): 10.10 (bs, IH), 8.95 (bs, IH), 8.30 (bs, 3H), 7.93 (d, J = 8.7 Hz, IH), 7.87 (d, J= 1.8 Hz, IH), 7.41 (d, J= 9.0 Hz, IH), 7.41 (d, J= 9.0Hz, IH), 7.16 (d, J = 1.5 Hz, IH), 4.54 (s, 2H), 4.10 (s, 3H), 4.02 (m, 2H), 2.73 (s, 3H).
EXAMPLE 75
6-(3-(Alamnyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine
(a) 6-(3 -(Boc-alaniny l-amino)-4-methylpheny l)-3 -(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine. A mixture of 6-(3-amino-4- methylphenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine (4.0 g, 1 1.4 mmol), Boc-Ala (4.31 g, 22.79 mmol), PyBOP (1 1.08 g, 25.07 mmol), ΗOBt (4.62 g, 34.19 mmol) and triethyl amine (2 mL) in acetonitrile (100 mL) was stirred at 5O0C for 15 h. The solvent was removed and the residue was mixed with ethyl acetate (50 mL). The mixture was washed with aqueous sodium bicarbonate (50 mL), 2N citric acid (30 mL), brine, dried and concentrated. The residue was purified by column chromatography to give 4.2 g of crude product as pale white solid, which was determined by 1H NMR to be a mixture of the amide product and amine starting material in a ratio of 52:48.
(b) 6-(3 -( Alaniny l-amino)-4-methylphenyl)-3 -(2-methoxypheny 1)-7H- [l,2,4]triazolo[3 ,4-Z)][1, 3,4]thiadiazine hydrochloride. To a solution of 4.2 g of the above solid in dichloromethane (50 mL) was added 4N hydrochloride in dioxane (7 mL) at room temperature. The mixture was stirred at room temperature for 1 h and become cloudy. It was evaporated and the residue was dissolved in ethyl acetate (50 mL) and the solution was neutralized with aqueous sodium bicarbonate. The organic layer was separated, dried and concentrated. The residue was purified by column chromatography (CΗ2Cl2/Me0Η = 20:1) to give the title compound (1.15 g) as the free base. It was dissolved in methanol (6 mL), acidified with 2N hydrochloride in ether (4 mL), and then added anhydrous ether (20 mL) to precipitate the product. The upper ether was decanted and the solid was filtered and dried to give the 1.18 g (21%) of the title compound. 1H NMR (CD3OD): 8.16 (bs, IH), 7.9-7.8 (m, 2H), 7.68 (t, J = 8.4 Hz, IH), 7.45 (d, J = 8.7 Hz, IH), 7.28 (d, J = 8.7 Hz, IH), 7.25 (t, J = 8.4Hz, IH), 4.47 (s, 2H), 4.22 (m, IH), 3.96 (s, 3H), 2.38 (s, 3H), 1.65 (d, J= 6.9 Hz, 3H).
EXAMPLE 76
6-(3-(Glycinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine hydrochloride
The title compound was prepared in a manner similar to Example 75 in two steps. From 6-(3-amino-4-methylphenyl)-3 -(2-methoxypheny 1)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (420 mg, 1.20 mmol) and Boc-Gly (231 mg, 1.32 mmol) was obtained 200 mg (35%) of the title compound. 1H NMR (CD3OD): 8.27 (s, IH), 7.90 (d, J = 7.8 Hz, IH), 7.81-7.71 (m, 2H), 7.47 (d, J = 8.4 Hz, IH), 7.34 (d, J = 8.4 Hz, IH), 7.27 (d, J = 7.6 Hz, IH), 4.47 (s, 2H), 3.97 (s, 3H), 2.39 (s, 3H).
EXAMPLE 77
6-(3-(Lysinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-b][l ,3,4]thiadiazine hydrochloride
The title compound was prepared in a manner similar to Example 75 in two steps. From 6-(3-amino-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-b][l,3,4]thiadiazine (1.0 g, 2.84 mmol) and Boc- Lys(Boc)OH (1.18 g, 3.42 mmol) was obtained 328 mg (20%) of the title compound. 1H NMR (CD3OD): 8.27 (d, J = 1.8 Hz IH), 7.88-7.85 (d, J = 8.1 Hz, IH), 7.83-7.79 (dd, J1= 8.1 Hz, J2= 2.1 Hz, IH), 7.73 (t, J = 8.4 Hz, IH), 7.47 (d, J = 8.1 Hz, IH), 7.33 (d, J = 8.4 Hz, IH), 7.25 (t, J = 7.2 Hz, IH), 4.47 (s, 2H), 4.24 (t, J = 6.6 Hz, IH), 3.96 (s, 3H), 2.97 (t, J = 7.8 Hz, 2H), 2.20- 1.90 (m, 2H), 1.82-1.73 (m, 2H), 1.67-1.59 (m, 2H).
EXAMPLE 78
6-(4-Methoxy-3-nitrophenyl)-3-(2,5-dimethylfuran-3-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine
The title compound was prepared in a manner similar to Example Ib. From 2-bromo-l-(4-methoxy-3-nitrophenyl)ethanone (438 mg, 1.6 mmol) and 4-amino-5-(2,5-dimethylfuryl)-3-mercapto-(4H)-l,2,4-triazole (336 mg, 1.6 mmol) in isopropanol was obtained 680 mg (98%) of the title compound. 1H NMR (CDCl3): 8.39 (d, J = 2.1 Hz, IH), 8.13 (dd, J = 8.4, 2.1 Hz, IH), 7.25 (dd, J = 8.4, 2.1 Hz, IH), 6.41 (s, IH), 4.07 (s, 3H), 3.97 (s, 2H), 2.62 (s, 3H), 2.32 (s, 3H).
EXAMPLE 79
6-(3-Amino-4-methoxyphenyl)-3-(2,5-dimethylfuran-3-yl)-7H- [1 ,2,4]triazolo[3,4-b][l ,3,4]thiadiazine hydrochloride The title compound was prepared in a manner similar to Example 2. From a mixture of 6-(4-methoxy-3-nitrophenyl)-3-(2,5-dimethylfuran-3-yl)- 7H-[l,2,4]triazolo[3,4-b][l,3,4]thiadiazine (0.45 g, 1.17 mmol) and tin chloride (1.02 g, 4.66 mmol) was obtained 345 mg (80%) of the title compound. 1H NMR (CD3OD): 8.18-8.13 (m, 2H), 7.43 (d, J = 8.7 Hz, IH), 6.53 (s, IH), 4.45 (s, 2H), 4.10 (s, 3H), 2.53 (s, 3H), 2.35 (s, 3H).
EXAMPLE 80
6-(3-(Fmoc-Alaninyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo- [3 ,4-6] [ 1 ,3 ,4]thiadiazine
To an oven-dried round bottom reaction flask charged with a magnetic stir bar at room tmeperature under argon with 6-(3-amino-4-methylphenyl)-3- (2-methoxyphenyl)-7Η-[l,2,4]triazolo[3,4-b][l,3,4]thiadiazine (0.925 g, 2.63 mmol) and deoxygenated CH2Cl2 (65 mL) was added AgCN (0.352 g, 2.63 mmol) and Fmoc-Ala-Cl (0.868 g, 2.63 mmol). The yellow suspension was stirred at room temperature for 25 h and then was diluted with CH2Cl2ZMeOH and filtered through celite (2 in h x 1.5 in w). The filtrate was concentrated by rotary evaporation to give 0.575 g (34%) of the title compound as a gray solid. MS (ESI) m/e for C36H32N6O4S (M+H)+ 645 (100).
EXAMPLE 81
3 -( 1 ,4,5 ,6-Tetrahydrocyclopenta[c]pyrazol-3 -yl)-6-(4-methyl-3 -nitrophenyl)- 7H-[1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine
The title compound was prepared in a manner similar to Example Ib. From 4-amino-5-(l,4,5,6-tetrahydro-3-methylcyclopenta[c]pyrazolyl)-3- mercapto-(4H)-l,2,4-triazole (151 mg, 0.679 mmol) and 2-bromo- 1 -(4- methyl-3-nitrophenyl)ethanone (189 mg, 0.733 mmol) in isopropanol was obtained the title compound as a yellow solid (242 mg, 0.634 mmol, 93%). 1H NMR (DMSO-d6) 8.61 (m, IH), 8.24 (m, IH), 7.73 (d, J = 8.4 Hz, IH), 4.50 (s, 2H), 2.61 (s, 3H), 2.35 - 2.76 (m, 6H).
EXAMPLE 82 3-(l,4,5,6-Tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3-aminophenyl)- 7H-[1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine
The title compound was prepared in a manner similar to Example 21. From 3-(l,4,5,6-tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3- nitrophenyl)-7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (217 mg, 0.569 mmol) was obtained the title compound as a light yellow solid (28 mg, 0.08 mmol, 14%). 1H NMR (DMSO-d6) 7.92 (m, IH), 7.78 (d, J = 7.8 Hz, IH), 7.47 (d, J = 7.8 Hz, IH), 4.20 (s, 2H), 2.70 (m, 2H), 2.55 (s, 3H), 2.51 (m, 2H), 2.41 (m, 2H).
EXAMPLE 83
6-(3-(Alaninyl-amino)-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine
a) 6-(3 -(Boc-alaninyl-amino)-4-methylpheny l)-3 -(2-methoxyphenyl)- 7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine. A mixture of 6-(3-amino-4- methoxyphenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine (1.10 g, 3.0 mmol), Boc-Ala (0.85g, 4.5 mmol), BOP (2.12g, 4.8 mmol), ΗOBt (0.65 g, 4.8 mmol) and triethyl amine (1 mL) in acetonitrile (50 mL) was stirred at 5O0C for 16 h. The solvent was removed and to the residue was added ethyl acetate (50 mL). The mixture was washed with aqueous sodium bicarbonate (50 mL), brine, dried and concentrated. The residue was purified by column chromatography (EtOAc/MeOΗ 40:1) to give 1.34 g (83%) of the title compound. 1H NMR (CDCl3): 8.86 (d, J = 2.4 Hz, IH), 8.62 (bs, IH), 7.61 (m, 2H), 7.48 (m, IH), 7.08 (t, J= 8.7 Hz, IH), 6.92 (d, J= 8.7 Hz, IH), 6.90 (d, J= 8.7 Hz, IH), 5.00 (bs, IH), 4.35 (m, IH), 3.97 (s, 2H), 3.92 (s, 3H), 3.76 (s, 3H), 1.47 (s, 9H), 1.29 (d, J= 6.9 Hz, 3H). b) 6-(3-(Alaninyl-amino)-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine. To a solution of 6-(3-(Boc-alaninyl- amino)-4-methylphenyl)-3 -(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,A- ό][l,3,4]thiadiazine (1.34 g, 2.5 mmol) in dichloromethane (10 mL) was added 4N hydrochloride in dioxane (3 mL) at room temperature and methanol (5 mL). The mixture was stirred at room temperature for 3 h then the solvent was removed by evaporation. The residue was dissolved in methanol (4 mL), mixed with ether (40 mL), and the precipitate was collected by filtration. The solids was dissolved in dichloromethane and the solution was washed with aqueous sodium bicarbonate (20 mL). The organic layer was separated, dried and concentrated. The residue was purified by column chromatography (CH2Cl2/Me0H = 20:1) to give 0.36 g (28% overall yield) of the product as solids. 1H NMR (CDCl3): 10.00 (bs, IH), 8.94 (s, IH), 7.62 (m, 2H), 7.45 (t, J = 8.4 Hz, IH), 7.09 (t, J= 8.4 Hz, IH), 7.02 (d, J = 7.8 Hz, IH), 6.91 (d, J = 7.8 Hz, IH), 3.99 (s, 2H), 3.96 (s, 3H), 3.78 (s, 3H), 3.68 (q, J= 7.2 Hz, IH), 1.45 (d, J= 7.2 Hz, 3H).
EXAMPLE 84
6-(3-(Alaninyl-amino)-4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H- [1 ,2,4]triazolo[3,4-b][l ,3,4]thiadiazine
a) 6-(3-(Boc-alaninyl-amino)-4-methoxyphenyl)-3-(2-methylfuran-3- yl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine The title compound was prepared by a procedure similar to that described for the preparation of compound 83A From 6-(3-amino-4-methoxyphenyl)-3-(2-methylfuran-3-yl)- 7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (0.36 g, 1 mmol) and Boc-Ala (0.28 g, 1.5 mmol) was obtained 0.27 g (50%) of crude product as pale white solid with the ratio of coupling product to S. M. (52% : 48%) by NMR characterization. b) 6-(3 -(Alaninyl-amino)-4-methoxyphenyl)-3 -(2-methy lfuran-3 -yl)- 7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine. To a O0C solution of methanol (20 mL) was added acetyl chloride (5 mL) dropwise. It was then stirred at room temperature for 30 min. To the above solution was added a solution of the crude 6-(3-(Boc-alaninyl-amino)-4-methoxyphenyl)-3-(2-methylfuran-3-yl)- 7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (0.22 g) from step A in methanol (3 mL). The solution was stirred at room temperature for 4 h, and then was evaporated. The residue was dissolved in dichloromethane (30 mL), neutralized with aqueous sodium bicarbonate to pΗ around 8. The organic layer was separated, dried and concentrated to give crude product, which was purified by column chromatography (CΗ2Cl2/Me0Η = 20:1) to give 86 mg (overall yield: 25%) of the title compound. 1H NMR (CD3OD): 8.95 (d, J = 2.4 Hz, IH), 7.76 (q, J= 8.7 Hz5 2.4 Hz, IH), 7.52 (d, J= 2.4 Hz, IH), 7.16 (d, J= 8.7 Hz, IH), 7.02 (d, J= 2.1 Hz, IH), 4.27 (s, 2H), 4.00 (s, 3H), 3.68 (q, J = 6.6 Hz, IH), 2.59 (s, 3H), 1.38 (d, J= 6.6 Hz, 3H).
EXAMPLE 85
6-(3 -(Serinyl-amino)-4-methylphenyl)-3 -(2-methoxyphenyl)-7H- [l,2,4]triazolo-[3,4-δ][l,3,4]thiadiazine
The title compound was prepared in a manner similar to Example 84.
EXAMPLE 86
Identification Of 6-(3-Amino-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H-
[l,2,4]triazolo[3,4-Z>][l,3,4]thiadiazine And Analogs As Caspase Cascade
Activators And Inducers Of Apoptosis In Solid Tumor Cells
[00208] Human breast cancer cell lines T-47D, human colon carcinoma cell line HCTl 16, human hepatocellular carcinoma cell line SNU398, human Burkitt's lymphoma cell line Namalwa were grown according to media component mixtures designated by American Type Culture Collection + 10% FCS (Invitrogen Corporation), in a 5 % CO2 -95 % humidity incubator at 37 0C. T-47D cells were maintained at a cell density between 50 and 80 % confluency at a cell density of 0.1 to 0.6 x 106 cells/mL. Cells were harvested at 600xg and resuspended at 0.65 x 106 cells/mL into appropriate media + 10 % FCS. An aliquot of 22.5 μL of cells was added to a well of a 384-well microtiter plate containing 2.5 μL of a 10 % DMSO in RPMI- 1640 media solution containing 0.16 to 100 μM of 6-(3-amino-4-methoxyphenyl)-3-(2- methoxyphenyl)-7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine or other test compound (0.016 to 10 μM final). An aliquot of 22.5 μL of cells was added to a well of a 384-well microtiter plate containing 2.5 μL of a 10 % DMSO in RPMI- 1640 media solution without test compound as the control sample. The samples were mixed by agitation and then incubated at 37 0C for 48 h in a 5 % CO2-95 % humidity incubator. After incubation, the samples were removed from the incubator and 25 μL of a solution containing 14 μM of N-(Ac- DEVD)-W-ethoxycarbonyl-R110 (SEQ ID No.:l) fluorogenic substrate (Cytovia, Inc.; WO99/18856), 20 % sucrose (Sigma), 20 mM DTT (Sigma), 200 mM NaCl (Sigma), 40 mM Na PIPES buffer pH 7.2 (Sigma), and 500 μg/mL lysolecithin (Calbiochem) was added. The samples were mixed by agitation and incubated at room temperature. Using a fluorescent plate reader (Model SPECTRAfluor Plus, Tecan), an initial reading (T = 0) was made approximately 1-2 min after addition of the substrate solution, employing excitation at 485 nm and emission at 530 nm, to determine the background fluorescence of the control sample. After the 3 h incubation, the samples were read for fluorescence as above (T = 3 h).
[00209] Calculation:
[00210] The Relative Fluorescence Unit values (RFU) were used to calculate the sample readings as follows:
RFU (τ=3h) - Control RFU σ=o) = Net RFU(τ=3h)
[00211] The activity of caspase cascade activation was determined by the ratio of the net RFU value for 6-(3-amino-4-methoxyphenyl)-3-(2-methoxyphenyl)- 7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine (Example 2) or other test compound to that of control samples. The EC50 (nM) was determined by a sigmoidal dose-response calculation (Prism 3.0, GraphPad Software Inc.).
[00212] The caspase activation potency (EC50) are summarized in Table I:
Figure imgf000087_0001
Figure imgf000088_0001
Figure imgf000089_0001
Figure imgf000090_0001
Figure imgf000091_0001
Figure imgf000092_0001
Figure imgf000093_0001
Thus, 6-(3 -amino-4-methoxyphenyl)-3 -(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (Example 2) and analogs are identified as potent caspase cascade activators and inducers of apoptosis in several solid tumor cells.
EXAMPLE 87
Identification Of 6-(3-Amino-4-methoxyphenyl)-3-(2-methoxyphenyl)-
7H-[l,2,4]triazolo[3,4-b][l,3,4]thiadiazine And Analogs As Antineoplastic Compound That Inhibits Cell Proliferation (GI50)
[00213] Human breast cancer cell lines T-47D, human colon carcinoma cell line HCTl 16, human hepatocellular carcinoma cell line SNU398, human Burkitt's lymphoma cell line Namalwa were grown and harvested as in Example 86. An aliquot of 90 μL of cells (4.4 x 104 cells/mL) was added to a well of a 96-well microtiter plate containing 5 μL of a 10 % DMSO in RPMI- 1640 media solution containing 10 nM to 100 μM of 6-(3-amino-4- methoxyphenyl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine (1 nM to 10 μM final). An aliquot of 45 μL of cells was added to a well of a 96-well microtiter plate containing 5 μL of a 10 % DMSO in RPMI- 1640 media solution without compound as the control sample for maximal cell proliferation (LM∞)- The samples were mixed by agitation and then incubated at 37 0C for 48 h in a 5% CO2-95% humidity incubator. After incubation, the samples were removed from the incubator and 25 μL of CellTiter-Glo ™ reagent (Promega) was added. The samples were mixed by agitation and incubated at room temperature for 10-15 min. Plates were then read using a luminescent plate reader (Model SPECTRAfluor Plus, Tecan) to give Ltest values.
[00214] Baseline for GI50 (dose for 50% inhibition of cell proliferation) of initial cell numbers was determined by adding an aliquot of 45 μL of cells or 45 μL of media, respectively, to wells of a 96-well microtiter plate containing 5 μL of a 10% DMSO in RPMI- 1640 media solution. The samples were mixed by agitation and then incubated at 37 0C for 0.5 h in a 5% CO2-95% humidity incubator. After incubation, the samples were removed from the incubator and 25 μL of CellTiter-Glo ™ reagent (Promega) was added. The samples were mixed by agitation and incubated at 37 0C for 10-15 min at room temperature in a 5% CC>2-95% humidity incubator. Fluorescence was read as above, (Lstan) defining luminescence for initial cell number used as baseline in GI5o determinations. Calculation:
[00215] GI50 (dose for 50% inhibition of cell proliferation) is the concentration where [(LTest - LStart) / (LMax- LStart)] = 0.5.
[00216] The GI50 (nM) are summarized in Table II:
Figure imgf000094_0001
[00217] Thus, 6-(3-amino-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H-
[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine (Example 2) and analogs are identified as antineoplastic compound that inhibits cell proliferation in several solid tumor cell lines. Having now fully described this invention, it will be understood by those of ordinary skill in the art that the same can be performed within a wide and equivalent range of conditions, formulations and other parameters without affecting the scope of the invention or any embodiment thereof. All patents, patent applications and publications cited herein are fully incorporated by reference herein in their entirety.

Claims

WHAT IS CLAIMED IS:
1. A compound having the Formula IV:
Figure imgf000096_0001
or a pharmaceutically acceptable salt or prodrug or tautomer thereof, wherein: Rg is halo or H; each of R4, R5, R7, Rg, and R\ \ is independently H, methoxy, ethoxy, trifluoromethoxy, dimethylamino, or methyl; each of R9, Rio> R-12» anc^ R-13 ιs independently H, amino, dimethylamino, or nitro; with the proviso that: at least one of R9, R\Q, R] 2> and Rl 3 is not H; at least one of R4, R5, R7, and Rg is not H; and when R4 or Rg is methoxy or methyl and Ri 1 is methyl, then none of
K\ 0 and Rj 2 is nitro or amino.
2. The compound of claim 1, wherein said compound is selected from the group consisting of
3-(2-Methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(4-Chloro-2-methoxyphenyl)-6-(4-methoxy-3-nitrophenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(4-chloro-2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine ; 6-(3-Dimethylamino-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(3-Dimethylamino-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-Z)][l ,3,4]thiadiazine;
3 -(3 ,5-Dimethoxyphenyl)-6-(4-methyl-3 -nitrophenyl)-7H- [1 ,2,4]triazolo[3,4-Z>][l ,3,4]thiadiazine;
6-(4-Methoxy-3 -nitrophenyl)-3 -(3 ,5-dimethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(3,5-dimethoxyphenyl)-7//- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(3,5-dimethoxyphenyl)-7//- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Dimethylamino-4-methoxyphenyl)-3-(3,5-dimethoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
3-(2-Dimethylaminophenyl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(2-dimethylaminophenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(4-Methyl-3-nitrophenyl)-3-(2-trifluoromethoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine; and
6-(3-Amino-4-methylphenyl)-3-(2-trifluoromethoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-(4-Methyl-3-nitrophenyl)-3-(2-ethoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(2-ethoxyphenyl)-7H- [l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine; and pharmaceutically acceptable salts or prodrugs thereof.
3. A compound selected from the group consisting of: 3-(3,5-Dimethoxyphenyl)-6-(4-nitrophenyl)-7//-[l,2,4]triazolo[3,4- b][l ,3,4]thiadiazine;
6-(4-Aminophenyl)-3-(3,5-dimethoxyphenyl)-7H-[l,2,4]triazolo[3,4- ό][l,3,4]thiadiazine; 6-(4-Methoxy-3 -nitrophenyl)-3 -(2-methylfuran-3 -yl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(4-Bromo-2-methoxyphenyl)-6-(4-methylphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
3-(2-Methoxy-4-methylphenyl)-6-(4-methylphenyl)-7H- [ 1 ,2,4]triazolo[3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Methylphenyl)-3 -(3 -methylthiophen-2-yl)-7H-[ 1 ,2,4]triazolo [3 ,4- b][\ ,3,4]thiadiazine;
3-(4-Bromo-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
3 -(4-Chloro-2-methoxyphenyl)-6-(4-methyl-3 -nitrophenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(4-chloro-2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-ό][l ,3,4]thiadiazine;
6-(3 - Amino-4-methylpheny l)-3 -(4-bromo-2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3,4]thiadiazine;
6-(4-Methylphenyl)-3-(lH-pyrazol-5-yl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methoxyphenyl)-3-(2-methylfuran-3-yl)-7H-[l,2,4]triazolo[3,4- b] [1 ,3,4]thiadiazine;
6-(4-Ηydroxyphenyl)-3-(2-methylfuran-3-yl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine;
3 -(2-Methylfuran-3 -yl)-6-(4-methyl-3 -nitrophenyl)-7H- [ 1 ,2,4]triazolo[3,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(2-methylfuran-3-yl)-7//- [ 1 ,2,4]triazolo [3 ,4-b] [ 1 ,3 ,4]thiadiazine;
6-(4-Methyl-3 -nitrophenyl)-3 -(2-methyl-H-imidazo [ 1 ,2-α]pyridin-3 - yl)-7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(2-methyl-H-imidazo[l,2-α]pyridin-3- yl)-7H-[l,2,4]triazolo[3,4-ό][l,3,4]thiadiazine;
6-(4-Methylphenyl)-3-(l -methyl- lH-pyrazol-5-yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine; 6-(4-Methyl-3-nitrophenyl)-3-(l-methyl-4-nitro-lH-pyrrol-2-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(4-Chloro-2-methylphenyl)-6-(4-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(4-Chloro-2-methylphenyl)-6-(4-hydroxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-Z)] [ 1 ,3 ,4]thiadiazine;
3-(4-Methyl-2-morpholinothiazol-5-yl)-6-(4-methylphenyl)-7//- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(4-Methyl-lH-imidazol-5-yl)-6-(4-methylphenyl)- 7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2,5-Dimethylftiran-3-yl)-6-(4-methylphenyl)-7H-[l,2,4]triazolo[3,4- b] [ 1 ,3 ,4]thiadiazine;
3 -(2-Methoxyphenyl)-6-( 1 -methyl- 1 H-indol-5-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-Acetamido-3-aminophenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(2-methyl-lH-benzo[<i]imidazol-6-yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(Benzofiiran-5-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- b][\ ,3,4]thiadiazine;
6-(Benzo[6]thiophen-5-yl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
3-(4-Fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3 - Amino-4-methylphenyl)-3 -(4-fluoro-2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(4-Methoxy-3-nitrophenyl)-3-(5-methyl-l//-imidazol-4-yl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(5-methyl-lH-imidazol-4-yl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(l-methylsulfonyl-2,3-dihydro-lH-indol-5- yl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2-Methyl-lH-imidazo[l,2-α]pyridin-3-yl)-6-(4-methoxy-3- nitrophenyl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine; 6-(3-Amino-4-methoxyphenyl)-3-(2-methyl-lH-imidazo[l,2- α]pyridin-3-yl)-7H-[ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(2-Methoxy-5-methylphenyl)-6-(4-methylphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(2-Methoxy-5-methylphenyl)-6-(2-oxoindolin-5-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Acetamide-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-( 1 H- 1 - Acetylindazol-6-yl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(4-N-Ηydroxyamidinophenyl)-3-(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(5-Fluoro-2-methoxyphenyl)-6-(4-methylphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(4-(N-methylacetamido)methylphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2-Methoxyphenyl)-6-(4-(N-methylamino)methylphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine; and
6-(4-(N, N-Dimethylamino)methylphenyl)-3 -(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
3-(5-Fluoro-2-methoxyphenyl)-6-(4-methyl-3-nitrophenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(5-fluoro-2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3-(2,5-Dimethylfiαran-3-yl)-6-(4-methyl-3-nitrophenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methylphenyl)-3-(2,5-dimethylfuran-3-yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
3 -(2-Methoxyphenyl)-6-(4-methylphenyl)-7H- [ 1 ,2 ,4]triazolo [3 ,4- 6][1 ,3,4]thiadiazin-7-one;
6-(N- Acetyl-indolin-5 -yl)-3 -(2-methoxyphenyl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(Indolin-5-yl)-3-(2-methoxyphenyl)-7H-[l,2,4]triazolo[3,4- 6][l,3,4]thiadiazine; 6-(3-Fluoro-4-methyl-5-nitrophenyl)-3-(2-methoxyphenyl)-7H- - [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-5-fluoro-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-(4-Methoxy-3-nitrophenyl)-3-(2,5-dimethylfuran-3-yl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-Amino-4-methoxyphenyl)-3-(2,5-dimethylfuran-3-yl)-7H- [1 ,2,4]triazolo[3,4-Z?][l ,3,4]thiadiazine;
3-(l,4,5,6-Tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3- nitrophenyl)-7H- [ 1 ,2,4]triazolo[3 ,4 -b] [ 1 ,3 ,4]thiadiazine;
3-(l,4,5,6-Tetrahydrocyclopenta[c]pyrazol-3-yl)-6-(4-methyl-3- aminophenyl)-7H-[l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(N-Boc-O-'Bu-Serinyl-amino)-4-methylphenyl)-3-(2- methoxyphenyl)-7H-[l ,2,4]triazolo-[3,4-6][l ,3,4]thiadiazine;
6-(3 -(Glycinyl-amino)-4-methoxyphenyl)-3 -(2-methylfuran-3 -yl)-7H- [ 1 ,2,4]triazolo [3 ,4-6] [ 1 ,3 ,4]thiadiazine;
6-(3-(Alaninyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [1 ,2,4]triazolo[3,4-6][l ,3,4]thiadiazine;
6-(3-(Glycinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(Lysinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3 -(Fmoc- Alaninyl-amino)-4-methy lphenyl)-3 -(2-methoxyphenyl)- 7H-[l,2,4]triazolo-[3,4-6][l,3,4]thiadiazine;
6-(3-(Alaninyl-amino)-4-methoxyphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(Alaninyl-amino)-4-methoxyphenyl)-3-(2-methylfuran-3-yl)-7H- [l,2,4]triazolo[3,4-6][l,3,4]thiadiazine;
6-(3-(Serinyl-amino)-4-methylphenyl)-3-(2-methoxyphenyl)-7H- [l,2,4]triazolo-[3,4-6][l,3,4]thiadiazine; and pharmaceutically acceptable salts or prodrugs thereof.
4. A method of treating a disorder responsive to the induction of apoptosis in an animal suffering therefrom, comprising administering to an animal in need of such treatment an effective amount of a compound of claim l or 3.
5. The method of claim 4, wherein said disorder is cancer.
6. The method according to claim 5, wherein said cancer is Hodgkin's disease, non-Hodgkin's lymphomas, acute or chronic lymphocytic leukemia, multiple myeloma, neuroblastoma, breast carcinoma, ovarian carcinoma, lung carcinoma, Wilms1 tumor, cervical carcinoma, testicular carcinoma, soft-tissue sarcoma, chronic lymphocytic leukemia, primary macroglobulinemia, bladder carcinoma, chronic granulocytic leukemia, primary brain carcinoma, malignant melanoma, small-cell lung carcinoma, stomach carcinoma, colon carcinoma, malignant pancreatic insulinoma, malignant carcinoid carcinoma, malignant melanoma, choriocarcinoma, mycosis fungoide, head or neck carcinoma, osteogenic sarcoma, pancreatic carcinoma, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary carcinoma, thyroid carcinoma, esophageal carcinoma, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial carcinoma, polycythemia vera, essential thrombocytosis, adrenal cortex carcinoma, skin cancer, or prostatic carcinoma.
7. The method of claim 6, wherein said cancer is a Myc deregulated cancer.
8. The method of claim 6, wherein said cancer is Burkitt's lymphoma, small-cell lung carcinomas, osteosarcomas, glioblastomas, melanoma, myeloid leukemias, breast, colon or cervical cancer.
9. The method of claim 6, wherein said cancer is drug resistant cancer.
10. The method of claim 6, further comprising administering at least one known cancer chemotherapeutic agent, or a pharmaceutically acceptable salt of said agent.
11. The method according to claim 6, wherein said compound is administered together with at least one compound selected from the group consisting of busulfan, cis-platin, mitomycin C, carboplatin, colchicine, vinblastine, paclitaxel, docetaxel, camptothecin, topotecan, doxorubicin, etoposide, 5-azacytidine, 5-fluorouracil, methotrexate, 5-fluoro-2'-deoxy- uridine, ara-C, hydroxyurea, thioguanine, melphalan, chlorambucil, cyclophosamide, ifosfamide, vincristine, mitoguazone, epirubicin, aclarubicin, bleomycin, mitoxantrone, elliptinium, fludarabine, octreotide, retinoic acid, tamoxifen, Herceptin®, Rituxan®, arsenic trioxide, gemcitabine, doxazosin, terazosin, tamsulosin, CB-64D, CB- 184, haloperidol, lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, cerivastatin, amprenavir, abacavir, CGP- 73547, CGP-61755, DMP-450, indinavir, nelfinavir, tipranavir, ritonavir, saquinavir, ABT-378, AG 1776, BMS-232,632, bexarotene, tretinoin, 13-cis- retinoic acid, 9-cis-retinoic acid, α-difluoromethylornithine, ILX23-7553, fenretinide, N-4-carboxyphenyl retinamide, lactacystin, MG-132, PS-341, Gleevec®, ZD1839 (Iressa), SH268, genistein, CEP2563, SU6668, SUl 1248, EMD121974, Rl 15777, SCH66336, L-778,123, BAL9611, TAN-1813, flavopiridol, UCN-01, roscovitine, olomoucine, celecoxib, valecoxib, rofecoxib and alanosine.
12. The method of claim 6, further comprising treating said animal with radiation-therapy.
13. The method of claim 6, wherein said compound is administered after surgical treatment of said animal for said cancer.
14. The method of claim 4, wherein said disorder is an autoimmune disease.
15. The method of claim 4, wherein said disorder is rheumatoid arthritis.
16. The method of claim 4, wherein said disorder is an inflammatory disease.
17. The method of claim 4, wherein said disorder is a skin disease.
18. The method of claim 4, wherein said disorder is psoriasis.
19. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of claim 1 or 3.
20. The pharmaceutical composition of claim 19, further comprising at least one known cancer chemotherapeutic agent, or a pharmaceutically acceptable salt of said agent.
21. The pharmaceutical composition of claim 19, further comprising at least one compound selected from the group consisting of busulfan, cis-platin, mitomycin C, carboplatin, colchicine, vinblastine, paclitaxel, docetaxel, camptothecin, topotecan, doxorubicin, etoposide, 5-azacytidine, 5-fluorouracil, methotrexate, 5-fluoro-2'-deoxy-uridine, ara-C, hydroxyurea, thioguanine, melphalan, chlorambucil, cyclophosamide, ifosfamide, vincristine, mitoguazone, epirubicin, aclarubicin, bleomycin, mitoxantrone, elliptinium, fludarabine, octreotide, retinoic acid, tamoxifen, Herceptin®, Rituxan®, arsenic trioxide, gemcitabine, doxazosin, terazosin, tamsulosin, CB-64D, CB- 184, haloperidol, lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, cerivastatin, amprenavir, abacavir, CGP-73547, CGP- 61755, DMP-450, indinavir, nelfinavir, tipranavir, ritonavir, saquinavir, ABT- 378, AG 1776, BMS-232,632, bexarotene, tretinoin, 13-cis-retinoic acid, 9- cis-retinoic acid, α-difluoromethylornithine, ILX23-7553, fenretinide, N-4- carboxyphenyl retinamide, lactacystin, MG-132, PS-341, Gleevec®, ZDl 839 (Iressa), SH268, genistein, CEP2563, SU6668, SUl 1248, EMD121974, Rl 15777, SCH66336, L-778,123, BAL9611, TAN-1813, flavopiridol,
UCN-Ol, roscovitine, olomoucine, celecoxib, valecoxib, rofecoxib and alanosine.
PCT/US2009/000471 2008-01-23 2009-01-23 3-aryl-6-aryl-7h-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazines and analogs as activators of caspases and inducers of apoptosis and the use thereof WO2009094205A2 (en)

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