WO2003029241A1 - Inhibiteurs de chk1 kinase - Google Patents

Inhibiteurs de chk1 kinase Download PDF

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Publication number
WO2003029241A1
WO2003029241A1 PCT/US2002/031752 US0231752W WO03029241A1 WO 2003029241 A1 WO2003029241 A1 WO 2003029241A1 US 0231752 W US0231752 W US 0231752W WO 03029241 A1 WO03029241 A1 WO 03029241A1
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WIPO (PCT)
Prior art keywords
ureido
carboxylic acid
thiophene
phenyl
acid amide
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PCT/US2002/031752
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English (en)
Inventor
Cynthia A. Parrish
James F. Callahan
Yue Li
Robert A. Stavenger
Dennis A. Holt
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Smithkline Beecham Corporation
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Publication of WO2003029241A1 publication Critical patent/WO2003029241A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • A61K31/381—Heterocyclic compounds having sulfur as a ring hetero atom having five-membered rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D333/30—Hetero atoms other than halogen
    • C07D333/36—Nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D333/38—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals

Definitions

  • the present invention relates to damage response kinase inhibitors, especially checkpoint kinase ("chkl kinase”) inhibitors, pharmaceutical compositions comprising these compounds and methods of using these compounds to treat various forms of cancer and hyperproliferative disorders.
  • chkl kinase checkpoint kinase
  • pharmaceutical compositions comprising these compounds and methods of using these compounds to treat various forms of cancer and hyperproliferative disorders.
  • chkl kinase checkpoint kinase
  • pharmaceutical compositions comprising these compounds and methods of using these compounds to treat various forms of cancer and hyperproliferative disorders.
  • Chkl was linked to survival responses including checkpoints. Mice lacking CHKl die in early embryogenesis (Liu et al. Gene & Dev. 2000 14: 1448-1459; Takai et al. Gene & Dev. 2000 14: 1439-1447). ES cells expressing a conditional CHKl gene die of p53 -independent apoptosis after loss of CHKl. Prior to their death, these cells become incapable of preventing mitotic entry in response to IR (Liu et al. Gene & Dev.
  • Chkl is required for the G2 DNA damage checkpoint in mammals as previously observed in other organisms. Chkl prevents mitotic entry as follows. Anest in G2 is regulated by the maintenance of inhibitory phosphorylation of Cdc2 (Nurse Cell 1997 91: 865-867). Cdc2 dephosphorylation and activation is catalyzed by the dual specificity phosphatase Cdc25 (Morgan Nature 1995 374: 131-134). Recent evidence indicates that part of the G2/M DNA checkpoint mechanism involves inactivation and translocation of Cdc25C into the cytoplasm.
  • the present invention involves 2-ureidothiophene compounds represented by Formula (I) hereinbelow, pharmaceutical compositions comprising such compounds and methods of inhibiting chkl kinase as well as specific assays to detect inhibition of chkl kinase activity.
  • Rl is selected from the group consisting of H, C, 2 alkyl, XH, XCH 3 , C, 2 alkyl-XH, C, 2 alkyl-XCH 3 , C(O)NH 2 , C(O)NHCH 3 , and C(O)-C, 2 alkyl, with the preferred substitution being H or CH 3 ,
  • X is selected from the group consisting of O, S, and NH
  • R2 is selected from the group consisting of C(O)R 5 , CO 2 R 5 , C(O)NHR 5 ,
  • R 5 and R 6 are, independently, selected from the group consisting of hydrogen, C, ]0 alkyl, C, 10 alkanoyl, C 2 10 alkenyl, C 2 10 alkynyl, C 3 10 cycloalkyl, C 06 alkylaryl, C 06 alkylheterocyclyl, and C 06 alkylheteroaryl, or R 5 and R 6 taken together with the nitrogen to which they are attached may optionally form a ring having 3 to 7 carbon atoms, optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C, 6 alkyl or (CH 2 ) ⁇ 3 aryl, such that any of the foregoing may be optionally substituted by one or more of group A and on any position,
  • R3 is H or halogen, with the prefened substitution being H,
  • R4 is aryl or heteroaryl optionally substituted by one or more of group A and on any position
  • R 7 and R 8 taken together with the nitrogen to which they are attached may optionally form a ring having 3 to 7 carbon atoms optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C, 6 alkyl or (CH 2 ) 03 aryl, wherein any of the foregoing may be optionally substituted by one or more of group E and on any position;
  • alkyl refers to an optionally substituted hydrocarbon group joined together by single carbon-carbon bonds.
  • the alkyl hydrocarbon group may be linear, branched or cyclic, saturated or unsaturated.
  • the group is saturated linear or cyclic.
  • alkanoyl is used herein at all occunences to mean a C(O)alkyl group, wherein the alkyl portion is as defined below, including, but not limited to, acetyl, pivaloyl, and the like.
  • alkenyl is used herein at all occunences to mean a straight or branched chain radical, wherein there is at least one double bond between two of the carbon atoms in the chain, including, but not limited to, ethenyl, 1-propenyl, 2- propenyl, 2-methyl- 1-propenyl, 1-butenyl, 2-butenyl, and the like.
  • alkoxy is used herein at all occurrences to mean a straight or branched chain radical bonded to an oxygen atom, including, but not limited to, methoxy, ethoxy, n- propoxy, isopropoxy, and the like.
  • alkylaryl is used herein at all occunences to mean an aryl group as defined below attached to an alkyl group as defined above, including, but not limited to, benzyl and phenethyl, and the like.
  • alkylheterocyclyl is used herein at all occunences to mean a heterocyclic group as defined below attached to an alkyl group as defined above, including, but not limited to, (tetrahydro-3-furanyl)methyl and 3-(4- morpholinyl)propyl, and the like.
  • alkylheteroaryl is used herein at all occunences to mean a heteroaryl group as defined below attached to an alkyl group as defined above, including, but not limited to, 3-(furanyl)methyl and (2-pyridinyl)propyl, and the like.
  • alkynyl is used herein at all occunences to mean a straight or branched chain radical, wherein there is at least one triple bond between two of the carbon atoms in the chain, including, but not limited to, acetylene, 1- propylene, 2- propylene, and the like.
  • aralkyl is used herein at all occunences to mean an aryl moiety as defined below, which is connected to an alkyl moiety as defined above, including, but not limited to, benzyl or phenethyl, and the like.
  • aryl is used herein at all occunences to mean 6-14-membered substituted or unsubstituted aromatic ring(s) or ring systems which may include bi- or tri-cyclic systems, including, but not limited to phenyl, naphthalenyl, biphenyl, phenanthryl, anthracenyl, and the like.
  • aryloxy is used herein at all occunences to mean an aryl group as defined above linked via an oxy group, including, but not limited to, phenoxy, and the like.
  • cycloalkyl is used herein at all occunences to mean cyclic radicals, which may be mono- or bicyclo- fused ring systems which may additionally include unsaturation, including, but not limited to, cyclopropyl, cyclopentyl, cyclohexyl, 1,2,3,4-tetrahydronaphthalenyl, and the like.
  • halo or halogen are used interchangeably herein at all occurrences to mean radicals derived from the elements chlorine, fluorine, iodine and bromine.
  • heteroaryl is used herein at all occunences to mean a 5-14- membered substituted or unsubstituted aromatic ring(s) or ring systems which may include bi- or tri-cyclic systems, which ring or ring systems contain 1 to 4 heteroatoms selected from nitrogen, which may be optionally substituted with hydrogen or Ci .galkyl, oxygen, and sulfur, including, but not limited to, indolyl, benzofuranyl, thianaphthenyl, quinolyl, isoquinolyl, pynolyl, furanyl, thienyl, pyridyl, and the like.
  • heteroaryloxy is used herein at all occunences to mean an heteroaryl group as defined above linked via an oxy group, including, but not limited to, 2-pyridinyloxy, and the like.
  • heterocyclic is used herein at all occurrences to mean a saturated or wholly or partially unsaturated 5-10-membered ring system (unless the cyclic ring system is otherwise limited) in which one or more rings contain one or more heteroatoms selected from nitrogen, which may be optionally substituted with hydrogen or Ci.galkyl, oxygen, and sulfur, including, but not limited to, pynolidine, piperidine, piperazine, morpholine, imidazolidine, pyrazolidine, 1,2,3,6- tetrahydropyridine, hexahydroazepine, and the like.
  • Preferred compounds useful in the present invention include:
  • More prefened compounds useful in the present invention include: SB-677216: 2-(3-Methyl-ureido)-5-phenyl-thiophene-3-carboxylic acid amide SB-696446: 5-Phenyl-2-ureido-thiophene-3-carboxylic acid amide SB-703954: 5-(4-Fluoro-phenyl)-2-ureido-thiophene-3-carboxylic acid amide SB-710860: 5-Ureido-[2,31bithiophenyl-4-carboxylic acid amide SB-714903: 5-(3-Methyl-ureido)-[2,3 r ]bithiophenyl-4-carboxylic acid amide SB-715135: 5-(4-Fluoro-phenyl)-2-ureido-thiophene-3-carboxylic acid methylamide SB-719113: 5-(3-Fluoro-phenyl)
  • the compounds of the invention can exist in unsolvated as well as solvated forms, including hydrated forms.
  • solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, are equivalent to the unsolvated forms for purposes of this invention.
  • the compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. All of these compounds and diastereomers are contemplated to be within the scope of the present invention.
  • Geometric isomers and tautomers of the present compounds are also within the scope of the present invention.
  • the present compounds can also be formulated as pharmaceutically acceptable salts and complexes thereof.
  • Pharmaceutically acceptable salts are non- toxic salts in the amounts and concentrations at which they are administered.
  • Pharmaceutically acceptable salts include acid addition salts such as those containing sulfate, hydrochloride, fumarate, maleate, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p- toluenesulfonate, cyclohexylsulfamate and quinate.
  • Pharmaceutically acceptable salts can be obtained from acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p- toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.
  • acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p- toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.
  • Pharmaceutically acceptable salts also include basic addition salts such as those containing benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when acidic functional groups, such as carboxylic acid or phenol are present.
  • Thiophenes of formula (I) can be prepared using two main methods from known or commercially available starting materials. Following the procedure of
  • Example 22 Preparation of 5-(4-Amino-phenyl)-2-(3-methyl-ureido)-thiophene-3-carboxylic acid amide Following the procedure described in Example 14(a) with 4- aminophenylboronic acid provided the title compound. ESLMS [M+H] + : 291.2.
  • Example 39 Preparation of 5-(3,4-Dichloro-phenyl)-2-ureido-thiophene-3-carboxylic acid amide Following the procedure described in Example 33 with 3,4-dichloro- phenylboronic acid provided the title compound. ESEVIS [M+H] + : 331.
  • a compound of Formula (I) or a pharmaceutically acceptable salt thereof for the treatment of humans and other mammals, it is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition.
  • Formulations for pharmaceutical use incorporating compounds of the present invention can be prepared in various forms and with numerous excipients. Examples of such formulations are given below:
  • the present ligands can be administered by different routes including intravenous, intraperitoneal, subcutaneous, intramuscular, oral, topical, transdermal, or transmucosal administration.
  • oral administration is prefened.
  • the compounds can be formulated into conventional oral dosage forms such as capsules, tablets and liquid preparations such as syrups, elixirs and concentrated drops.
  • injection parenteral administration
  • the compounds of the invention are formulated in liquid solutions, preferably, in physiologically compatible buffers or solutions, such as saline solution, Hank's solution, or Ringer's solution.
  • the compounds may be formulated in solid form and redissolved or suspended immediately prior to use. Lyophilized forms can also be produced.
  • Systemic administration can also be by transmucosal or transdermal means.
  • penetrants appropriate to the banier to be permeated are used in the formulation.
  • penetrants are generally known in the art, and include, for example, for transmucosal administration, bile salts and fusidic acid derivatives.
  • detergents may be used to facilitate permeation.
  • Transmucosal administration for example, may be through nasal sprays, rectal suppositories, or vaginal suppositories.
  • the compounds of the invention can be formulated into ointments, salves, gels, or creams, as is generally known in the art.
  • the amounts of various compounds to be administered can be determined by standard procedures taking into account factors such as the compound IC 50 , EC J0 , the biological half-life of the compound, the age, size and weight of the patient, and the disease or disorder associated with the patient. The importance of these and other factors to be considered are known to those of ordinary skill in the art.
  • Amounts administered also depend on the routes of administration and the degree of oral bioavailability. For example, for compounds with low oral bioavailability, relatively higher doses will have to be administered.
  • the composition is in unit dosage form.
  • a tablet, or capsule may be administered, for nasal application, a metered aerosol dose may be administered, for transdermal application, a topical formulation or patch may be administered and for transmucosal delivery, a buccal patch may be administered. In each case, dosing is such that the patient may administer a single dose.
  • Each dosage unit for oral administration contains suitably from 0.01 to 500 mg/Kg, and preferably from 0.1 to 50 mg/Kg, of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, calculated as the free base.
  • the daily dosage for parenteral, nasal, oral inhalation, transmucosal or transdermal routes contains suitably from 0.01 mg to 100 mg/Kg, of a compound of Formula (I).
  • a topical formulation contains suitably 0.01 to 5.0% of a compound of Formula (I).
  • the active ingredient may be administered from 1 to 6 times per day, preferably once, sufficient to exhibit the desired activity, as is readily apparent to one skilled in the art.
  • treatment includes, but is not limited to prevention, retardation and prophylaxis of the disease.
  • diseases treatable using the present compounds include, but are not limited to leukemias, solid tumor cancers and metastases, lymphomas, soft tissue cancers, brain cancer, esophageal cancer, stomach cancer, pancreatic cancer, liver cancer, lung cancer, bladder cancer, bone cancer, prostate cancer, ovarian cancer, cervical cancer, uterine cancer, testicular cancer, kidney cancer, head cancer and neck cancer, chronic inflammatory proliferative diseases such as psoriasis and rheumatoid arthritis; proliferative cardiovascular diseases such as restenosis; prolifertive ocular disorders such as diabetic retinopathy; and benign hyperproliferative diseases such as hemangiomas.
  • Composition of Formula (I) and their pharmaceutically acceptable salts which are active when given orally can be formulated as syrups, tablets, capsules and lozenges.
  • a syrup formulation will generally consist of a suspension or solution of the compound or salt in a liquid canier for example, ethanol, peanut oil, olive oil, glycerine or water with a flavoring or coloring agent.
  • a liquid canier for example, ethanol, peanut oil, olive oil, glycerine or water with a flavoring or coloring agent.
  • any pharmaceutical canier routinely used for preparing solid formulations may be used. Examples of such caniers include magnesium stearate, tena alba, talc, gelatin, acacia, stearic acid, starch, lactose and sucrose.
  • composition is in the form of a capsule
  • any routine encapsulation is suitable, for example using the aforementioned caniers in a hard gelatin capsule shell.
  • composition is in the form of a soft gelatin shell capsule
  • any pharmaceutical canier routinely used for preparing dispersions or suspensions may be considered, for example aqueous gums, celluloses, silicates or oils, and are incorporated in a soft gelatin capsule shell.
  • Typical parenteral compositions consist of a solution or suspension of a compound or salt in a sterile aqueous or non-aqueous canier optionally containing a parenterally acceptable oil, for example polyethylene glycol, polyvinylpynolidone, lecithin, arachis oil or sesame oil.
  • a parenterally acceptable oil for example polyethylene glycol, polyvinylpynolidone, lecithin, arachis oil or sesame oil.
  • compositions for inhalation are in the form of a solution, suspension or emulsion that may be administered as a dry powder or in the form of an aerosol using a conventional propellant such as dichlorodifluoromethane or trichlorofluoromethane.
  • a typical suppository formulation comprises a compound of Formula (I) or a pharmaceutically acceptable salt thereof which is active when administered in this way, with a binding and/or lubricating agent, for example polymeric glycols, gelatins, cocoa-butter or other low melting vegetable waxes or fats or their synthetic analogs.
  • Typical dermal and transdermal formulations comprise a conventional aqueous or non-aqueous vehicle, for example a cream, ointment, lotion or paste or are in the form of a medicated plaster, patch or membrane.
  • the composition is in unit dosage form, for example a tablet, capsule or metered aerosol dose, so that the patient may administer a single dose.
  • Streptavidin coated SPA beads, ATP and 33 P-ATP were obtained from Amersham Pharmacia Biotech, Biotin labeled peptide
  • KVSRSGLYRSPSMPENLNRK(Biotin-xx)NH 2 was obtained from Affiniti Research Products Ltd, assay buffer reagents were obtained from Sigma-Aldrich Co. Ltd. fff84 well assay plates were obtained from Corning Inc.
  • Assay buffer 50 mM HEPES, 50 mM KC1, 5% Glycerol, 1 mM EGTA, 0.001% Tween-20; enzyme/peptide mix: 25 nM Chkl, 2.5 ⁇ M biotin peptide, 7.5 mM 2- mercaptoethanol in assay buffer; ATP mix: 20 ⁇ M ATP at 650kBq/mL, 5mM MgCl, in assay buffer.
  • Inhibitors of decreasing concentration, from lO ⁇ M were incubated at room temperature for 1 hour together with 5 ⁇ L enzyme/peptide mix and 5 ⁇ L ATP mix. The reaction was stopped with 5 ⁇ L of 0.5M EDTA followed by a further addition of
  • a GST-Chkl expression construct was constructed which has the glutathione-S-transferase gene fused to the amino terminus of Chkl kinase via a linker containing a thrombin cleavage site. This construct was cloned into the Baculovirus expression vector, pFASTBAC, and this was used to make the viral stock for the subsequent infection. Spodoptera frugiperda cells (Sf9) were infected with the virus expressing the GST-Chkl and the cells were grown for 3 days, then harvested and frozen down.
  • GST-Chkl protein was purified as follows: An Sf9 cell pellet expressing
  • GST-Chkl was resuspended on ice in lysis buffer (50mM Tris-Cl, pH 7.5, 250mM NaCl2, lmM dithiothreitol (DTT), 0.1%Brij, 5% (v/v) protease inhibitor cocktail, ImM sodium orthovanadate), cells were lysed by sonication and centrifuged at 100,000xg for 30min The supernatant was added to Glutathione Sepharose 4B, beads, equilibrated in wash buffer (20mM Tris-Cl, pH 7.0, lOmM MgCl2, lOOmM NaCl2, ImM DTT, 0.5%(v/v) protease inhibitor cocktail, ImM sodium orthovanadate).
  • wash buffer 20mM Tris-Cl, pH 7.0, lOmM MgCl2, lOOmM NaCl2, ImM DTT, 0.5%(v/v) protease inhibitor cocktail, ImM sodium orthovanadate
  • the mixture was rocked for 30min
  • the resin with the bound GST- Chkl was spun down at 500xg for 5min and washed with 14mls of wash buffer.
  • the beads were spun as above and resuspended in another 14mls of wash buffer.
  • the suspension was transfened into a column and allowed to pack, then the wash buffer was allowed to flow through by gravity.
  • the GST-Chkl was eluted from the column with lOmM Glutathione in 50mM Tris-Cl, pH 8.0 in 500ul fractions. Protein concentrations were determined on the fractions using Bio-Rad's Protein assay kit as per instructions.
  • Cell Cvcle Studies Drug studies considering cellular effects were performed in the Hela S3 adherent cell line. Cells were plated at a concentration sufficiently low such that 24 hours later they were at 10-20% confluence (typically 2xl ⁇ 5 cells/15cm e3). Cells were then synchronized in S phase by a repeated thymidine block. Briefly, cells were treated with 2mM thymidine for l ⁇ hours, released for 8 hours by 3 washes, and then treated again with thymidine. Following the second release from thymidine, 95% of cells were in S phase.
  • Synchronized cells were then returned to complete media containing a DNA-damaging drug such as 50nM topotecan (a dosage we have found to be sufficient to anest cells in early G2 phase without inducing apoptosis) alone and in combination with test compounds for up to 18 hours.
  • Cell cycle profiles were then performed cytometrically using a procedure for propidium iodide staining of nuclei. (Vindelov et al, Cytometry Vol.3, No.5, 1983, 323-327)
  • CHKl inhibitors would be expected to reverse the G2 anest caused by the DNA damaging agent. Typical concentration ranges for such activity would be 0.001 to 10 uM.
  • Proliferation/Apoptosis Studies were performed in a variety of adherent and non- adherent cell lines including Hela S3, HT29, and Jurkat.
  • the proliferation assay utilized a colorimetric change resulting from reduction of the tetrazolium reagent XTT into a formazan product by metabolically active cells (Scudiero et al. Cancer Research, 48, 1981, 4827-4833) Cells were seeded in lOOul in 96 well plates to roughly 10% confluence (cell concentration varied with cell lines) and grown for 24 hours. Compounds were then added with or without sufficient vehicle- containing media to raise the cells to a 200ul final volume containing chemical reagents in 0.2% DMSO.
  • DNA-damaging anti-proliferative drugs such as topotecan, test compounds, and combination treatment at 37°C 5% CO2. 72 hours later, 50 uls of an XTT/ phenazine methosulfate mixture were added to each well and cells
  • IKK- ⁇ was expressed as a GST-tagged protein, and its activity was assessed in a 96- well scintillation proximity assay (SPA). Briefly, IKK- ⁇ was diluted in assay buffer (20 mM Hepes, pH 7.7, 2 mM MgCl2, 1 mM MnCl2, 10 mM ⁇ - glycerophosphate, 10 mM NaF, 10 mM PNPP, 0.3 mM Na ⁇ VO/ , 1 mM benzamidine, 2 ⁇ M PMSF, 10 ⁇ g/ml aprotinin, 1 ug/mL leupeptin, 1 ug/mL pepstatin, ImM DTT; 20 nM final), with various concentrations of compound or DMSO vehicle, 240 nM ATP and 200 nCi [ « - 33 P]-ATP (10 mCi/mL, 2000 Ci/mmol; NEN Life Science Products, Boston, MA).
  • assay buffer 20 m
  • the reaction was started with the addition of a biotinylated peptide comprising amino acids 15 - 46 of I ⁇ B- ⁇ (American Peptide) to a final concentration of 2.4 ⁇ M, in a total volume of 50 uL.
  • the sample incubated for one hour a 30 °C, followed by the addition of 150 uL of stop buffer (PBS w/o Ca 2 +, Mg 2 + 0.1% Triton X-100 (v/v), 10 mM EDTA) containing 0.2 mg streptavidin-coated SPA PVT beads (Amersham Pharmacia Biotech, Piscataway, NJ).
  • stop buffer PBS w/o Ca 2 +, Mg 2 + 0.1% Triton X-100 (v/v), 10 mM EDTA

Abstract

L'invention porte sur de nouveaux composés utiles dans l'inhibition des kinases en réaction à un dommage.
PCT/US2002/031752 2001-10-04 2002-10-04 Inhibiteurs de chk1 kinase WO2003029241A1 (fr)

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WO2005016909A1 (fr) * 2003-08-15 2005-02-24 Astrazeneca Ab Thiophenes substitues et leurs utilisations
WO2005027907A1 (fr) * 2003-09-17 2005-03-31 Icos Corporation Utilisation d'inhibiteurs de la chk1 pour lutter contre la proliferation cellulaire
WO2005066163A3 (fr) * 2004-01-05 2005-09-01 Astrazeneca Ab Heterocycles substitues et leurs utilisations
JP2006510676A (ja) * 2002-12-06 2006-03-30 スミスクライン・ビーチャム・コーポレイション ＮＦ−κＢ阻害剤
US7125896B2 (en) * 2001-07-25 2006-10-24 Astrazeneca Ab Thiophene carboxamide compounds as inhibitors of enzyme IKK-2
US7553868B2 (en) 2003-01-15 2009-06-30 Astrazeneca Ab Thiophene carboxamides as inhibitors of the enzyme IKK-2
WO2009096198A1 (fr) * 2008-02-01 2009-08-06 Pharma Ip Limited Liability Intermediary Corporations Nouveau derive de biaryle
US7572826B2 (en) 2003-01-15 2009-08-11 Astrazeneca Ab Thiophene-carboxamide derivatives and their use as inhibitors of the enzyme IKK-2
CN101967141A (zh) * 2010-10-13 2011-02-09 信实生物医药（上海）有限公司 一种Chk蛋白激酶拮抗剂AZD-7762的制备方法
EP2330105A1 (fr) 2005-03-29 2011-06-08 ICOS Corporation Dérivés d'uree d'heteroaryle utilises pour inhiber CHK1
JP2011522890A (ja) * 2008-06-11 2011-08-04 ジェネンテック， インコーポレイテッド 置換されたピロール及び使用方法
WO2012046793A1 (fr) * 2010-10-07 2012-04-12 参天製薬株式会社 Nouvel inhibiteur de jak3 contenant, à titre de principe actif, un dérivé de thiophène portant un groupe uréido et un groupe aminocarbonyle à titre de substituants
US8426355B2 (en) 2006-03-15 2013-04-23 Theralogics, Inc. Methods of treating muscular wasting diseases using NF-κB activation inhibitors
EP2826780A1 (fr) 2009-09-18 2015-01-21 Zhanggui Wu Dérivés de thieno[2,3-d]pyridazine et leur utilisation thérapeutique pour l'inhibition de la protéine kinase
CN109438416A (zh) * 2018-12-06 2019-03-08 河南师范大学 一种合成噻吩类抑制剂tpca-1的方法
EP3461480A1 (fr) 2017-09-27 2019-04-03 Onxeo Combinaison d'inhibiteurs de point de contrôle du cycle cellulaire de réponse à un dommage à l'adn et de belinostat pour traiter le cancer
WO2023031246A1 (fr) * 2021-09-01 2023-03-09 Rijksuniversiteit Groningen Composés thiophènes substitués utilisés en tant qu'inhibiteurs de d-dopachrome tautomérase

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6344476B1 (en) * 1997-05-23 2002-02-05 Bayer Corporation Inhibition of p38 kinase activity by aryl ureas
US6414013B1 (en) * 2000-06-19 2002-07-02 Pharmacia & Upjohn S.P.A. Thiophene compounds, process for preparing the same, and pharmaceutical compositions containing the same background of the invention
US6476023B1 (en) * 1999-03-12 2002-11-05 Boehringen Ingelheim Pharmaceuticals, Inc. Aromatic heterocyclic compounds as anti-inflammatory agents

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6344476B1 (en) * 1997-05-23 2002-02-05 Bayer Corporation Inhibition of p38 kinase activity by aryl ureas
US6476023B1 (en) * 1999-03-12 2002-11-05 Boehringen Ingelheim Pharmaceuticals, Inc. Aromatic heterocyclic compounds as anti-inflammatory agents
US6414013B1 (en) * 2000-06-19 2002-07-02 Pharmacia & Upjohn S.P.A. Thiophene compounds, process for preparing the same, and pharmaceutical compositions containing the same background of the invention

Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7956084B2 (en) 2001-07-25 2011-06-07 Astrazeneca Ab Phenyl thiophene carboxamide compounds as inhibitors of the enzyme IKK-2
US7125896B2 (en) * 2001-07-25 2006-10-24 Astrazeneca Ab Thiophene carboxamide compounds as inhibitors of enzyme IKK-2
JP2006510676A (ja) * 2002-12-06 2006-03-30 スミスクライン・ビーチャム・コーポレイション ＮＦ−κＢ阻害剤
US7572826B2 (en) 2003-01-15 2009-08-11 Astrazeneca Ab Thiophene-carboxamide derivatives and their use as inhibitors of the enzyme IKK-2
US7553868B2 (en) 2003-01-15 2009-06-30 Astrazeneca Ab Thiophene carboxamides as inhibitors of the enzyme IKK-2
JP2007502308A (ja) * 2003-08-15 2007-02-08 アストラゼネカ アクチボラグ 置換チオフェンとその使用
WO2005016909A1 (fr) * 2003-08-15 2005-02-24 Astrazeneca Ab Thiophenes substitues et leurs utilisations
US7423061B2 (en) 2003-08-15 2008-09-09 Astrazeneca Ab Substitute thiophenes and uses thereof
WO2005027907A1 (fr) * 2003-09-17 2005-03-31 Icos Corporation Utilisation d'inhibiteurs de la chk1 pour lutter contre la proliferation cellulaire
EP2305671A1 (fr) 2004-01-05 2011-04-06 AstraZeneca AB Dérivés de thiophène et thiazole comme inhibiteurs de CHK 1
WO2005066163A3 (fr) * 2004-01-05 2005-09-01 Astrazeneca Ab Heterocycles substitues et leurs utilisations
JP2007517843A (ja) * 2004-01-05 2007-07-05 アストラゼネカ アクチボラグ Ｃｈｋ１阻害剤であるチオフェン誘導体
EP2330105A1 (fr) 2005-03-29 2011-06-08 ICOS Corporation Dérivés d'uree d'heteroaryle utilises pour inhiber CHK1
US9173920B2 (en) 2006-03-15 2015-11-03 Theralogics, Inc. Methods of treating muscular wasting diseases using NF-KB activation inhibitors
US8426355B2 (en) 2006-03-15 2013-04-23 Theralogics, Inc. Methods of treating muscular wasting diseases using NF-κB activation inhibitors
WO2009096198A1 (fr) * 2008-02-01 2009-08-06 Pharma Ip Limited Liability Intermediary Corporations Nouveau derive de biaryle
JP2011522890A (ja) * 2008-06-11 2011-08-04 ジェネンテック， インコーポレイテッド 置換されたピロール及び使用方法
EP2826780A1 (fr) 2009-09-18 2015-01-21 Zhanggui Wu Dérivés de thieno[2,3-d]pyridazine et leur utilisation thérapeutique pour l'inhibition de la protéine kinase
WO2012046793A1 (fr) * 2010-10-07 2012-04-12 参天製薬株式会社 Nouvel inhibiteur de jak3 contenant, à titre de principe actif, un dérivé de thiophène portant un groupe uréido et un groupe aminocarbonyle à titre de substituants
CN101967141A (zh) * 2010-10-13 2011-02-09 信实生物医药（上海）有限公司 一种Chk蛋白激酶拮抗剂AZD-7762的制备方法
EP3461480A1 (fr) 2017-09-27 2019-04-03 Onxeo Combinaison d'inhibiteurs de point de contrôle du cycle cellulaire de réponse à un dommage à l'adn et de belinostat pour traiter le cancer
CN109438416A (zh) * 2018-12-06 2019-03-08 河南师范大学 一种合成噻吩类抑制剂tpca-1的方法
CN109438416B (zh) * 2018-12-06 2020-04-10 河南师范大学 一种合成噻吩类抑制剂tpca-1的方法
WO2023031246A1 (fr) * 2021-09-01 2023-03-09 Rijksuniversiteit Groningen Composés thiophènes substitués utilisés en tant qu'inhibiteurs de d-dopachrome tautomérase
NL2029098B1 (en) * 2021-09-01 2023-03-17 Univ Groningen Substituted thiophene compounds as D-dopachrome tautomerase inhibitors

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