EP1150950A2 - Anthranilic acid derivatives - Google Patents

Anthranilic acid derivatives

Info

Publication number
EP1150950A2
EP1150950A2 EP99968160A EP99968160A EP1150950A2 EP 1150950 A2 EP1150950 A2 EP 1150950A2 EP 99968160 A EP99968160 A EP 99968160A EP 99968160 A EP99968160 A EP 99968160A EP 1150950 A2 EP1150950 A2 EP 1150950A2
Authority
EP
European Patent Office
Prior art keywords
phenylamino
methyl
benzamide
chloro
difluoro
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP99968160A
Other languages
German (de)
French (fr)
Inventor
Haile Tecle
Stephen Douglas Barrett
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Warner Lambert Co LLC
Original Assignee
Warner Lambert Co LLC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Warner Lambert Co LLC filed Critical Warner Lambert Co LLC
Publication of EP1150950A2 publication Critical patent/EP1150950A2/en
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D285/00—Heterocyclic compounds containing rings having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by groups C07D275/00 - C07D283/00
    • C07D285/01—Five-membered rings
    • C07D285/02—Thiadiazoles; Hydrogenated thiadiazoles
    • C07D285/04—Thiadiazoles; Hydrogenated thiadiazoles not condensed with other rings
    • C07D285/10—1,2,5-Thiadiazoles; Hydrogenated 1,2,5-thiadiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/08—Drugs for disorders of the urinary system of the prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/52—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton
    • C07C229/54—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton with amino and carboxyl groups bound to carbon atoms of the same non-condensed six-membered aromatic ring
    • C07C229/56—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton with amino and carboxyl groups bound to carbon atoms of the same non-condensed six-membered aromatic ring with amino and carboxyl groups bound in ortho-position
    • C07C229/58—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton with amino and carboxyl groups bound to carbon atoms of the same non-condensed six-membered aromatic ring with amino and carboxyl groups bound in ortho-position having the nitrogen atom of at least one of the amino groups further bound to a carbon atom of a six-membered aromatic ring, e.g. N-phenyl-anthranilic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C317/00—Sulfones; Sulfoxides
    • C07C317/26—Sulfones; Sulfoxides having sulfone or sulfoxide groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton
    • C07C317/32—Sulfones; Sulfoxides having sulfone or sulfoxide groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton with sulfone or sulfoxide groups bound to carbon atoms of six-membered aromatic rings of the carbon skeleton
    • C07C317/34—Sulfones; Sulfoxides having sulfone or sulfoxide groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton with sulfone or sulfoxide groups bound to carbon atoms of six-membered aromatic rings of the carbon skeleton having sulfone or sulfoxide groups and amino groups bound to carbon atoms of six-membered aromatic rings being part of the same non-condensed ring or of a condensed ring system containing that ring
    • C07C317/36—Sulfones; Sulfoxides having sulfone or sulfoxide groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton with sulfone or sulfoxide groups bound to carbon atoms of six-membered aromatic rings of the carbon skeleton having sulfone or sulfoxide groups and amino groups bound to carbon atoms of six-membered aromatic rings being part of the same non-condensed ring or of a condensed ring system containing that ring with the nitrogen atoms of the amino groups bound to hydrogen atoms or to carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/23—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton
    • C07C323/31—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton having the sulfur atom of at least one of the thio groups bound to a carbon atom of a six-membered aromatic ring of the carbon skeleton
    • C07C323/33—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton having the sulfur atom of at least one of the thio groups bound to a carbon atom of a six-membered aromatic ring of the carbon skeleton having at least one of the nitrogen atoms bound to a carbon atom of the same non-condensed six-membered aromatic ring
    • C07C323/35—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton having the sulfur atom of at least one of the thio groups bound to a carbon atom of a six-membered aromatic ring of the carbon skeleton having at least one of the nitrogen atoms bound to a carbon atom of the same non-condensed six-membered aromatic ring the thio group being a sulfide group
    • C07C323/36—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton having the sulfur atom of at least one of the thio groups bound to a carbon atom of a six-membered aromatic ring of the carbon skeleton having at least one of the nitrogen atoms bound to a carbon atom of the same non-condensed six-membered aromatic ring the thio group being a sulfide group the sulfur atom of the sulfide group being further bound to an acyclic carbon atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
    • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
    • C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles

Definitions

  • This invention relates to 4' heteroaryl diarylamines having pharmaceutical activity.
  • MEK enzymes are dual specificity kinases involved in, for example, immunomodulation, inflammation, and proliferative diseases such as cancer and restenosis.
  • Proliferative diseases are caused by a defect in the intracellular signaling system, or the signal transduction mechanism of certain proteins.
  • Defects include a change either in the intrinsic activity or in the cellular concentration of one or more signaling proteins in the signaling cascade .
  • the cell may produce a growth factor that binds to its own receptors, resulting in an autocrine loop, which continually stimulates proliferation. Mutations or overexpression of intracellular signaling proteins can lead to spurious mitogenic signals within the cell. Some of the most common mutations occur in genes encoding the protein known as Ras, a G-protein that is activated when bound to GTP, and inactivated when bound to GDP.
  • Ras leads in turn to the activation of a cascade of serine/threonine kinases.
  • One of the groups of kinases known to require an active Ras-GTP for its own activation is the Raf family. These in turn activate MEK (e.g., MEK ⁇
  • MEK e.g., MEK ⁇
  • MAP kinase e.g., MEK ⁇
  • Activation of MAP kinase by mitogens appears to be essential for proliferation; constitutive activation of this kinase is sufficient to induce cellular transformation.
  • Blockade of downstream Ras signaling for example by use of a dominant negative Raf-1 protein, can completely inhibit mitogenesis, whether induced from cell surface receptors or from oncogenic Ras mutants.
  • Ras is not itself a protein kinase, it participates in the activation of Raf and other kinases, most likely through a phosphorylation mechanism.
  • Raf and other kinases phosphorylate MEK on two closely adjacent serine residues, s218 and S ⁇ 22 j n he case of MEK-1 , which are the prerequisite for activation of MEK as a kinase.
  • MEK in turn phosphorylates MAP kinase on both a tyrosine, Y 1 8 5 , and a threonine residue, T ' '83 j separated by a single amino acid. This double phosphorylation activates MAP kinase at least 100-fold.
  • MAP kinase can then catalyze the phosphorylation of a large number of proteins, including several transcription factors and other kinases. Many of these MAP kinase phosphorylations are mitogenically activating for the target protein, such as a kinase, a transcription factor, or another cellular protein.
  • target protein such as a kinase, a transcription factor, or another cellular protein.
  • other kinases activate MEK, and MEK itself appears to be a signal integrating kinase. Current understanding is that MEK is highly specific for the phosphorylation of MAP kinase.
  • MEK does not phosphorylate peptides based on the MAP kinase phosphorylation sequence, or even phosphorylate denatured MAP kinase.
  • MEK also appears to associate strongly with MAP kinase prior to phosphorylating it, suggesting that phosphorylation of MAP kinase by MEK may require a prior strong interaction between the two proteins.
  • the invention features a compound having the formula (I) below:
  • W is OR1, NR20R!, NR A R B , NR 2 NR A RB, 0(CH 2 ) 1 - 4 NR A R B , or NR 2 (CH 2 ) ⁇ -4 NR A RB- RI is H, C ⁇ alkyl, C 3 - ⁇ alkenyl, C 3 . 8 alkynyl,
  • C 3-8 cycloalkyl phenyl, (phenyl)C ⁇ . 4 alkyl, (phenyl)C 3 - 4 alkenyl, (phenyl)- C 3-4 alkynyl, (C 3 . 8 cycloalkyl)C 1-4 alkyl, (C 3- 8 cycloalkyl)C 3 -4 alkenyl, (C 3-8 cycloalkyl)C 3- alkynyl, C 3-8 heterocyclic radical, (C 3 -s heterocyclic radical)C 1-4 alkyl, (C 3-8 heterocyclic radical)C 3 -4 alkenyl, or (C 3-8 heterocyclic radical)C 3-4 alkynyl.
  • Each of R 2 and R 3 is independently H, phenyl, C 1.4 alkyl, C 3-8 alkenyl, C 3 . 8 alkynyl, C 3 . 8 cycloalkyl, or (C 3 - 8 cycloalkyl)C 1- 4 alkyl.
  • Each of R 4 , R5 and R 6 is independently H, F, Br, Cl, or NO 2 .
  • RA is H, C ⁇ - 6 alkyl, C 3 . 8 alkenyl, C 3 - 8 alkynyl, C 3 - 8 cycloalkyl, phenyl, (C 3 - 8 cycloalkyl)C 1. 4 alkyl, (C 3 .
  • J is SR C , OR c , SO 2 R c , SOR c , SO 2 NR D RE, C 1 - ⁇ alkyl, C 3 -s alkenyl, C 3 . 8 alkynyl, C 3 - ⁇ cycloalkyl, C 5 - ⁇ cycloalkenyl, phenyl, (C 3 - 8 cycloalkyl)C 1 - 4 alkyl, (C 3 . 8 cycloalkyl)C 3 .
  • M' is O, SO, SO 2 , NR E , (CO)NR E , NR E (CO), SO 2 NR E , NR E S0 2 , or CH 2 .
  • E' is absent (in other words, a covalent bond), (CH 2 ) ⁇ -4 or (CH 2 )m O(CH 2 ) p where 1 ⁇ (each of m and p independently) ⁇ 3 and 2 ⁇ (m + p) ⁇ 4.
  • G' is OR 3 , SORc, SO 2 R C
  • or NR F R G ; provided that where p 1 , then G' is H.
  • R c , RD > RE, RF and RG is independently selected from H, C 1-6 alkyl, C 3 -4 alkenyl, C 3 - 4 alkynyl, C 3 - 6 cycloalkyl, C 3 .
  • NRFRG and NRDR E can each also independently be selected from morpholinyl, pyrazinyl, piperazinyl, pyrrolidinyl, or piperadinyl.
  • R 10 is H, C 1- 4 alkyl, halo, NO 2) or S0 2 NR H R
  • Rn is H, halo, or NO 2 .
  • Each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C 1-4 alkyl, C 3 - 6 cycloalkyl, C 3 - 4 alkenyl, C 3 . 4 alkynyl, phenyl, hydroxy, amino, (amino)sulfonyl, and NO 2 , wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 3 substituents independently selected from halo, C 1 -2 alkyl, hydroxy, amino, and NO 2 .
  • the invention also encompasses a pharmaceutically acceptable salt or C 1 . 7 ester of a compound of formula (I).
  • the invention also relates to a pharmaceutical composition including (a) a compound of formula (I) and (b) a pharmaceutically-acceptable carrier.
  • the invention further relates to a method for treating proliferative diseases, such as cancer, restenosis, psoriasis, autoimmune disease, and atherosclerosis.
  • Other aspects of the invention include methods for treating MEK-related (including ras-related) cancers, whether solid or hematopoietic.
  • cancers include colorectal, cervical, breast, ovarian, brain, acute leukemia, gastric, non-small cell lung, pancreatic, prostatic, and renal cancers.
  • Further aspects of the invention include methods for treating or reducing the symptoms of xenograft (cell(s), llimb, skin, organ or bone marrow transplant) rejection, osteoarthritis, rheumatoid arthritis, cystic fibrosis, complications of diabetes (including diabetic retinopathy and diabetic nephropathy), hepatomegaly, cardiomegaly, stroke (such as acute focal ischemic stroke and global cerebral ischemia), heart failure, septic shock, asthma, and Alzheimer's disease.
  • xenograft cell(s), llimb, skin, organ or bone marrow transplant
  • osteoarthritis rheumatoid arthritis
  • cystic fibrosis cystic fibrosis
  • complications of diabetes including diabetic retinopathy and diabetic nephropathy
  • hepatomegaly such as acute focal ischemic stroke and global cerebral ischemia
  • stroke such as acute focal ischemic stroke and global cerebral ischemia
  • heart failure such as acute focal ischemic stroke
  • Compounds of the invention are also useful as antiviral agents for treating viral infections such as HIV, hepatitis (B) virus (HBV), human papilloma virus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV).
  • B hepatitis virus
  • HPV human papilloma virus
  • CMV cytomegalovirus
  • EBV Epstein-Barr virus
  • These methods include the step of administering to a patient in need of such treatment, or suffering from such a disease or condition, a pharmaceutically-effective amount of a disclosed compound or pharmaceutical composition thereof.
  • the invention also features methods of combination therapy, such as a method for treating cancer, wherein the method further includes providing radiation therapy or chemotherapy, for example, with mitotic inhibitors such as a taxane or a vinca alkaloid.
  • mitotic inhibitors include paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine, and vinflunine
  • Other therapeutic combinations include a MEK inhibitor of the invention and an anticancer agent such as cisplatin, 5-fluoro-2-4(1 H,3H)-pyrimidinedione (5FU), flutamide, and gemcitabine.
  • the chemotherapy or radiation therapy may be administered before, concurrently, or after the administration of a disclosed compound according to the needs of the patient.
  • the invention further includes synthetic intermediates and methods disclosed herein.
  • the invention features 4-heteroaryl diarylamine compounds, pharmaceutical compositions thereof, and methods of using such compounds and compositions.
  • the compounds are MEK inhibitors.
  • MEK inhibition assays include the in vitro MEK/MAP described at column 6, line 36 to column 7, line 4 of U.S. Patent Number 5,525,625 and the in vitro MEK assay described at column 7, lines 4-27 of the same patent, the entire disclosure of which is incorporated by reference (see also Examples 13 et seq.). A whole cell assay is described in Example 16.
  • Alkyl groups include aliphatic (i.e., hydrocarbyl or hydrocarbon radical structures containing hydrogen and carbon atoms) with a free valence. Alkyl groups are understood to include straight chain and branched structures.
  • Examples include methyl, ethyl, propyl, isopropyl, butyl, n-butyl, isobutyl, t-butyl, pentyl, isopentyl, 2,3-dimethylpropyl, hexyl, 2,3-dimethylhexyl, 1 ,1-dimethylpentyl, heptyl, and octyl.
  • Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl.
  • Alkyl groups can be substituted with 1 , 2, 3 or more substituents which are independently selected from halo (fluoro, chloro, bromo, or iodo), hydroxy, amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy.
  • substituents are independently selected from halo (fluoro, chloro, bromo, or iodo), hydroxy, amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy.
  • Specific examples include fluoromethyl, hydroxyethyl, 2, 3-d i hydroxyethyl, (2- or 3-furanyl)methyl, cyclopropylmethyl, benzyloxyethyl, (3-pyridinyl)methyl, (2- or 3-furanyl)methyl, (2- thienyl)ethyl, hydroxypropyl, aminocyclohexyl, 2-dimethylaminobutyl, methoxymethyl, /V-pyridinylethyl, diethylaminoethyl, and cyclobutylmethyl.
  • Alkenyl groups are analogous to alkyl groups, but have at least one double bond (two adjacent sp 2 carbon atoms). Depending on the placement of a double bond and substituents, if any, the geometry of the double bond may be Mais (E), or sixteen (Z), cis, or trans. Similarly, alkynyl groups have at least one triple bond (two adjacent sp carbon atoms). Unsaturated alkenyl or alkynyl groups may have one or more double or triple bonds, respectively, or a mixture thereof; like alkyl groups, unsaturated groups may be straight chain or branched, and they may be substituted as described both above for alkyl groups and throughout the disclosure by example.
  • alkenyls, alkynyls, and substituted forms include cis-2-butenyl, trans-2-butenyl, 3-butynyl, 3-phenyl-2- propynyl, 3-(2'-fluorophenyl)-2-propynyl, 3-methyl(5-phenyl)-4-pentynyl, 2- hydroxy-2-propynyI, 2-methyl-2-propynyl, 2-propenyl, 4-hydroxy-3-butynyl, 3-(3- fluorophenyl)-2-propynyl, and 2-methyl-2-propenyl.
  • alkenyls and alkynyls can be C 2 - ⁇ or C 2 .
  • substituted hydrocarbon radicals include hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl, hydroxyaryl, and corresponding forms for the prefixes amino-, halo- (e.g., fluoro-, chloro-, or bromo-), nitro-, alkyl-, phenyl-, cycloalkyl- and so on, or combinations of substituents.
  • halo- e.g., fluoro-, chloro-, or bromo-
  • substituted alkyls include hydroxyalkyl, aminoalkyl, nitroalkyl, haloalkyl, alkylalkyl (branched alkyls, such as methylpentyl), (cycloalkyl)alkyl, phenylalkyl, alkoxy, alkylaminoalkyl, dialkylaminoalkyl, arylalkyl, aryloxyalkyl, arylalkyloxyalkyl, (heterocyclic radical)alkyl, and (heterocyclic radical)oxyalkyl.
  • Ri thus includes hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl, hydroxyaryl, aminoalkyl, aminoalkenyl, aminoalkynyl, aminocycloalkyl, aminoaryl, alkylalkenyl,
  • R A includes hydroxyalkyl and aminoaryl
  • RB includes hydroxyalkyl, aminoalkyl, and hydroxyalkyl(heterocyclic radical)alkyl.
  • Heterocyclic radicals which include but are not limited to heteroaryls, include: furyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, pyrrolyl, imidazolyl, 1 ,3,4- triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, indolyl, 1 ,2,5-thiadiazolyl and their nonaromatic counterparts.
  • heterocyclic radicals include piperidyl, quinolyl, isothiazolyl, piperidinyl, morpholinyl, piperazinyl, tetrahydrofuryl, tetrahydropyrrolyl, pyrrolidinyl, octahydroindolyl, thiadiazolyl, octahydrobenzothiofuranyl, and octahydrobenzofuranyl.
  • Selective MEK 1 or MEK 2 inhibitors are those compounds which inhibit the MEK 1 or MEK 2 enzymes, respectively, without substantially inhibiting other enzymes such as MKK3, PKC, Cdk2A, phosphorylase kinase, EGF, and PDGF receptor kinases, and C-src.
  • a selective MEK 1 or MEK 2 inhibitor has an IC 50 for MEK 1 or MEK 2 that is at least one-fiftieth (1/50) that of its IC 50 for one of the above-named other enzymes.
  • a selective inhibitor has an IC 50 that is at least 1/100, more preferably 1/500, and even more preferably 1/1000, 1/5000, or less than that of its IC 5 o or one or more of the above-named enzymes.
  • Embodiments of the invention include compounds wherein: (a) R c is C ⁇ . 2 alkyl; (b) W is OH, or W is NHOR-i (c) Rio is methyl or chloro; (d) Rn is fluoro; (e) Rn is H; (f) J is trihalomethyl or methylthio; (g) J is SO 2 CH 3 ; (h) J is SOCH 3 ; (i) J is C 3 - ⁇ alkynyl where the triple bond is between the carbon atoms alpha and beta to the phenyl group; (j) Ri has at least one hydroxy substituent; (k) Ri is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 3 - 5 alkenyl, C 3 - 5 alkynyl, C
  • R 2 is H, methyl, C 3 - 4 alkynyl, C 3 .
  • R A is H, methyl, ethyl, isobutyl, hydroxyethyl, hydroxypropyl, cyclopropylmethyl, cyclobutylmethyl, C 3 - 4 alkynyl, phenyl, 2- piperjdin-1 -yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2-hydroxyethyl)-piperazin-1-yl]- propyl, 2-pyrrolidin-1 -yl-ethyl, or 2-diethylamino-ethyl; and RB is H; or where R B is methyl and R A is phenyl; (0) each of R 4 and R ⁇ is H, and R 5 is F; (p) each of R 4 , R 5 , and R 6 is F; (q) R 5 is F; (r) each R 5 and R 6 is F and R 6 is
  • Ri, R 2 , R A , RB, Rc, RD, RE, RF, and RG is an alkenyl or alkynyl group, its double or triple bond, respectively, is not adjacent the point of attachment.
  • W is NR 2 OR ⁇
  • R 2 is preferably prop-2-ynyl, or but-2 or 3-enyl, and less preferably prop-1-ynyl or but- 1-enyl.
  • Examples of compounds of formula (I) include: 4-fluoro-2-(2-methyl-4- methylsulfanyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2-(2-methyl-4- methylsulfanyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4-methanesulfinyl-2- methyl-phenylamino)-benzoic acid; 2-(4-methanesulfinyl-2-methyl-phenylamino)- 4-nitro-benzoic acid; 3,4,5-trifluoro-2-(4-methanesulfonyl-2-methyl-phenylamino)- benzoic acid; 3,4-difluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 2-(2-methyl-4-methylsulfanyl-phenylamino)-4-nitro-benzoic acid; 3,4, 5-
  • Preferred examples of compounds of formula (I) are : 4-Fluoro-2-(4- methanesulfanyl-phenylamino)-benzoic acid (1); 4-Fluoro-2-(4-methanesulfinyl- phenylamino)-benzoic acid (2); 4-Fluoro-2-(4-methanesulfonyl-phenylamino)- benzoic acid (3); 4-Fluoro-2-(2-methyl-4-trimethylsilanylethynyl-phenylamino)- benzoic acid (6); 4-Fluoro-2-(2-methyl-4-ethynyl-phenylamino)-benzoic acid (7).
  • Additional preferred compounds include the following: (a) 5-Bromo-2-(4- ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy- 2-(4-ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzamide; 2-(4-Ethynyl-2- methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4- ethynyl-2-methyl-phenylamino)-3,4,5-trifluoro-benzamide; 2-(4-Ethynyl-2-methyl- phenylamino)-3,4,5-trifluoro-benzoic acid; 5-
  • Further preferred compounds include: (a) 5-bromo-2-(2-chloro-4- methylsulfanyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-chloro-4- methanesulfinyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-chloro-4- methanesulfonyl-phenylamino)-3,4,5-trifluoro-benzoic acid; 2-(2-chloro- methylsulfanyl-phenylamino)-3,4-difluoro-benzoic acid; 5-bromo-2-(2-chloro-4- methanesulfonyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-Chloro-4- methanesulfonyl-phenylamino)-3,4-difluoro-benzoic acid; (b) 5-bromo-2
  • the disclosed compounds can be synthesized according to the following five schemes, or variants thereof.
  • PyBOP is (benzotriazolyl- oxy)-tripyrrolidino phosphonium hexafluorophosphate.
  • compositions are useful as both prophylactic and therapeutic treatments for diseases or conditions as provided in the Summary section, as well as diseases or conditions modulated by the MEK cascade.
  • diseases or conditions modulated by the MEK cascade include stroke, heart failure, osteoarthritis, rheumatoid arthritis, organ transplant rejection, and a variety of tumors such as ovarian, non-small cell lung, pancreatic, brain, prostatic, renal, and colon.
  • an effective amount will be between 0.1 and 1000 mg/kg per day, preferably between 1 and 300 mg/kg body weight, and daily dosages will be between 10 and 5000 mg for an adult subject of normal weight.
  • Capsules, tablets or other formulations may be of between 5 and 200 mg, such as 10, 15, 25, 35, 50 mg, 60 mg, and 100 mg and can be administered according to the disclosed methods. 2.
  • Dosage unit forms include tablets, capsules, pills, powders, granules, aqueous and nonaqueous oral solutions and suspensions, and parenteral solutions packaged in containers adapted for subdivision into individual doses. Dosage unit forms can also be adapted for various methods of administration, including controlled release formulations, such as subcutaneous implants. Administration methods include oral, rectal, parenteral (intravenous, intramuscular, subcutaneous), intracistemal, intravaginal, intraperitoneal, intravesical, local (drops, powders, ointments, gels, or cream), and by inhalation (a buccal or nasal spray).
  • Parenteral formulations include pharmaceutically acceptable aqueous or nonaqueous solutions, dispersion, suspensions, emulsions, and sterile powders for the preparation thereof.
  • carriers include water, ethanol, polyols (propylene glycol, polyethylene glycol), vegetable oils, and injectable organic esters such as ethyl oleate. Fluidity can be maintained by the use of a coating such as lecithin, a surfactant, or maintaining appropriate particle size.
  • Carriers for solid dosage forms include (a) fillers or extenders, (b) binders, (c) humectants, (d) disintegrating agents, (e) solution retarders, (f) absorption acccelerators, (g) adsorbants, (h) lubricants, (i) buffering agents, and (j) propellants.
  • Compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispensing agents; antimicrobial agents such as parabens, chlorobutanol, phenol, and sorbic acid; isotonic agents such as a sugar or sodium chloride; absorption-prolonging agents such as aluminum monostearate and gelatin; and absorption-enhancing agents.
  • adjuvants such as preserving, wetting, emulsifying, and dispensing agents
  • antimicrobial agents such as parabens, chlorobutanol, phenol, and sorbic acid
  • isotonic agents such as a sugar or sodium chloride
  • absorption-prolonging agents such as aluminum monostearate and gelatin
  • absorption-enhancing agents such as aluminum monostearate and gelatin.
  • the invention provides the disclosed compounds and closely related, pharmaceutically acceptable forms of the disclosed compounds, such as salts, esters, amides, hydrates or solvated forms thereof; masked or protected forms; and racemic mixtures, or enantiomerically or optically pure forms.
  • Pharmaceutically acceptable salts, esters, and amides include carboxylate salts (e.g., C ⁇ . 8 alkyl, cycloalkyl, aryl, heteroaryl, or non-aromatic heterocyclic), amino acid addition salts, esters, and amides which are within a reasonable benefit/risk ratio, pharmacologically effective, and suitable for contact with the tissues of patients without undue toxicity, irritation, or allergic response.
  • carboxylate salts e.g., C ⁇ . 8 alkyl, cycloalkyl, aryl, heteroaryl, or non-aromatic heterocyclic
  • amino acid addition salts esters, and amides which are within a reasonable benefit/risk ratio, pharmacologically effective, and suitable for contact with the tissues of patients without undue toxicity, irritation, or allergic response.
  • Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, iaurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactiobionate, and laurylsulfonate.
  • alkali metal and alkali earth cations such as sodium, potassium, calcium, and magnesium, as well as non-toxic ammonium, quaternary ammonium, and amine cations such as tetramethyl ammonium, methylamine, trimethylamine, and ethylamine.
  • alkali metal and alkali earth cations such as sodium, potassium, calcium, and magnesium
  • non-toxic ammonium, quaternary ammonium, and amine cations such as tetramethyl ammonium, methylamine, trimethylamine, and ethylamine.
  • amine cations such as tetramethyl ammonium, methylamine, trimethylamine, and ethylamine.
  • Representative pharmaceutically acceptable amides of the invention include those derived from ammonia, primary C -
  • Secondary amines include 5- or 6-membered heterocyclic or heteroaromatic ring moieties containing at least one nitrogen atom and optionally between 1 and 2 additional heteroatoms.
  • Preferred amides are derived from ammonia, C ⁇ 3 alkyl primary amines, and di (C ⁇ - 2 alkyl)amines.
  • Representative pharmaceutically acceptable esters of the invention include C 1. 7 alkyl, C 5 - 7 cycloalkyl, phenyl, and phenyl(C ⁇ - 6 )alkyl esters.
  • Preferred esters include methyl esters.
  • the invention also includes disclosed compounds having one or more functional groups (e.g., hydroxyl, amino, or carboxyl) masked by a protecting group. Some of these masked or protected compounds are pharmaceutically acceptable; others will be useful as intermediates. Synthetic intermediates and processes disclosed herein, and minor modifications thereof, are also within the scope of the invention.
  • one or more functional groups e.g., hydroxyl, amino, or carboxyl
  • HYDROXYL PROTECTING GROUPS Hydroxyl protecting groups include: ethers, esters, and protection for 1 ,2- and 1 ,3-diols.
  • the ether protecting groups include: methyl, substituted methyl ethers, substituted ethyl ethers, substituted benzyl ethers, silyl ethers and conversion of silyl ethers to other functional groups.
  • Substituted Methyl Ethers include: methoxymethyl, methylthiomethyl, t- butylthiomethyl, (phenyldimethylsilyl) methoxymethyl, benzyloxymethyl, p-ethoxybenzyloxymethyl, (4-methoxyphenoxy) methyl, guaiacolmethyl, f-butoxymethyl, 4-pentenyloxymethyl, siloxymethyl, 2-methoxyethoxymethyl, 2,2,2-trichloroethoxymethyl, bis(2-chloro- ethoxy)methyl, 2-(trimethylsilyl)- ethoxymethyl, tetrahydropyranyl, 3-bromotetrahydro-pyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl, 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl, 4-methoxytetra
  • Substituted ethyl ethers include: 1-ethoxyethyl, 1-(2,chloroethoxy)ethyl, 1 -methyl- 1-methoxyethyl, 1-methyl-1 -benzyloxyethyl, 1 -methy 1-1 -benzyloxy-2- fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilyethyl, 2-(phenylselenyl)ethyl, f-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, and benzyl.
  • Substituted Benzyl Ethers include: 1-ethoxyethyl, 1-(2,chloroethoxy)ethyl, 1 -methyl- 1-methoxyethyl, 1-methyl-1 -benzyloxyethyl, 1 -methy 1-1-1
  • Substituted benzyl ethers include: p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2- and 4-picolyl, 3-methyl-2-picolyl ⁇ /-oxido, diphenylmethyl, p, p -dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, ⁇ -naphthyl- diphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)- phenylmethyl, tri-(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxy)phenyl- diphenylmethyl, 4,4',4"-ths(4,5-dichlorophthalimid
  • Silyl Ethers Silyl ethers include: trimethylsilyl, triethylsilyl, triisopropylsilyl, dimethylisopropylsilyl, diethylisopropylsilyl, dimethylthexylsilyl, f-butyldimethylsilyl, f-butyldiphenylsilyl, tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl, and f-butylmethoxyphenylsilyl.
  • Esters protecting groups include: esters, carbonates, assisted cleavage, miscellaneous esters, and sulfonates.
  • protective esters include: formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, p-P- phenylacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4- (ethylenedithio) pentanoate, pivaloate, adamantoate,crotonate,4- methoxycrotonate, benzoate, p-phenylbenzoate, and 2,4,6-trimethylbenzoate (mesitoate).
  • Carbonates include: methyl, 9-fluorenylmethyl, ethyl, 2,2,2-thchloroethyl, 2-(trimethylsilyl) ethyl, 2-(phenylsulfonyl) ethyl, 2-(triphenylphosphonio) ethyl, isobutyl, vinyl, allyl, p-nitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl, and methyl dithiocarbonate.
  • assisted Cleavage protecting groups include: 2-iodobenzoate, 4- azido-butyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl) benzoate, 2- formylbenzene-sulfonate, 2-(methylthiomethoxy) ethyl carbonate, 4- (methylthiomethoxymethyl) benzoate, and 2-(methylthiomethoxymethyl) benzoate.
  • 2-iodobenzoate 4- azido-butyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl) benzoate, 2- formylbenzene-sulfonate, 2-(methylthiomethoxy) ethyl carbonate, 4- (methylthiomethoxymethyl) benzoate, and 2-(methylthiomethoxymethyl) benzoate.
  • miscellaneous esters include: 2,6-dichloro-4- methylphenoxyacetate, 2,6-dichloro-4-(1 ,1 ,3,3-tetramethylbutyl) phenoxyacetate, 2,4-bis(1 ,1-dimethylpropyl) phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (£)-2-methyl-2-butenoate (tigloate), o-(methoxycarbonyl) benzoate, p-P-benzoate, ⁇ -naphthoate, nitrate, alkyl N,N,N ',N '- tetramethylphosphorodiamidate, ⁇ /-phenylcarbamate, borate, dimethylphosphinothioyl, and 2,4-dinitrophenylsulfenate.
  • Sulfonates Protective sulfates includes: sulfate, methan
  • the protection for 1 ,2 and 1 ,3-diols group includes: cyclic acetals and ketals, cyclic ortho esters, and silyl derivatives.
  • Cyclic Acetals and Ketals include: methylene, ethylidene, 1-f-butylethylidene, 1-phenylethylidene, (4-methoxyphenyl) ethylidene, 2,2,2-trichloroethylidene, acetonide (isopropylidene), cyclopentylidene, cyclohexylidene, cycloheptylidene, benzylidene, p-methoxybenzylidene, 2,4-dimethoxybenzylidene, 3,4- dimethoxybenzylidene, and 2-nitrobenzylidene.
  • Cyclic ortho esters include: methoxymethylene, ethoxymethylene, dimethoxy- methylene, 1-methoxyethylidene, 1-ethoxyethylidine, 1 ,2-dimethoxyethylidene, ⁇ -methoxybenzylidene, 1-( ⁇ /, ⁇ /-dimethylamino)ethylidene derivative, ⁇ -( ⁇ /, ⁇ /- dimethylamino) benzylidene derivative, and 2-oxacyclopentylidene.
  • Ester protecting groups include: esters, substituted methyl esters, 2-substituted ethyl esters, substituted benzyl esters, silyl esters, activated esters, miscellaneous derivatives, and stannyl esters.
  • Substituted Methyl Esters include: 9-fluorenylmethyl, methoxymethyl, methylthiomethyl, tetrahydropyranyl, tetrahydrofuranyl, methoxyethoxymethyl, 2- (t methylsilyl)ethoxy-methyl, benzyloxymethyl, phenacyl, p-bromophenacyl, ⁇ - methylphenacyl, p-methoxyphenacyl, carboxamidomethyl, and N- phthalimidomethyl.
  • 2-Substituted Ethyl Esters include: 2,2,2-trichloroethyl, 2-haloethyl, l-chloroalkyl, 2-(trimethylsily)ethyl, 2-methylthioethyl, 1 ,3-dithianyl-2-methyl, 2(p- nitrophenylsulfenyl)-ethyl, 2-(p-toluenesulfonyl)ethyl, 2-(2'-pyridyl)ethyl, 2-
  • Substituted Benzyl esters include: triphenylmethyl, diphenylmethyl, bis(o-nitrophenyl)methyl, 9-anthrylmethyl, 2-(9,10-dioxo)anthrylmethyl, 5-dibenzo- suberyl, 1 -pyrenylmethyl, 2-(trifluoromethyl)-6-chromylmethyl, 2,4,6- thmethylbenzyl, p-bromobenzyl, o-nitrobenzyl, p-nitrobenzyl, p-methoxybenzyl, 2,6-dimethoxybenzyl, 4-(methylsulfinyl)benzyl, 4-sulfobenzyl, piperonyl, and 4-P- benzyl.
  • Silyl esters include: trimethylsilyl, t ethylsilyl, f-butyldimethylsilyl, /- propyldimethylsilyl, phenyldimethylsilyl, and di- f-butylmethylsilyl.
  • Miscellaneous Derivatives Miscellaneous derivatives includes: oxazoles, 2-alkyl-1 ,3-oxazolines, 4-alkyl-5- oxo-1 , 3-oxazolidines, 5-alkyl-4-oxo-1 ,3-dioxolanes, ortho esters, phenyl group, and pentaaminocobalt(lll) complex.
  • Stannyl Esters examples include: triethylstannyl and tri-n-butylstannyl. AMIDES AND HYDRAZIDES
  • Amides include: N,N -dimethyl, pyrrolidinyl, piperidinyl, 5,6-dihydrophen- anthridinyl, o-nitroanilides, ⁇ /-7-nitroindolyl, /V-8-nitro-1,2,3,4-tetrahydroquinolyl, and p-P-benzenesulfonamides.
  • Hydrazides include: /V-phenyl, ⁇ /./V '-diisopropyl and other dialkyl hydrazides.
  • CARBAMATES Carbamates include: carbamates, substituted ethyl, assisted cleavage, photolytic cleavage, urea-type derivatives, and miscellaneous carbamates.
  • Carbamates include: methyl and ethyl, 9-fluorenylmethyl, 9-(2- sulfo)fluorenylmethyl, 9-(2,7-dibromo)fluorenylmethyl, 2,7-di- -butyl-[9-(10,10- dioxo-10, 10, 10, 10-tetrahydro- thioxanthyl)]methyl, and 4-methoxyphenacyl.
  • Substituted Ethyl protective groups include: 2,2,2-trichloroethyl, 2- trimethylsilylethyl, 2-phenylethyl, 1-(1-adamantyl)-1-methylethyl, 1 ,1-dimethyl-2- haloethyl, 1 ,1dimethyl-2,2-dibromoethyl, 1 ,1-dimethyl-2,2,2-trichloroethyl, 1- methyl-1-(4-biphenylyl)ethyl, 1-(3,5-di-f-butylphenyl)-1-methylethyl, 2-(2'-and 4'- pyridyl)ethyl, 2-(/V, ⁇ /-icyclohexylcarboxamido)- ethyl, f-butyl, 1-adamantyl, vinyl, allyl, 1-isopropylallyl, connam
  • Assisted Cleavage Protection via assisted cleavage includes: 2-methylthioethyl, 2- methylsulfonylethyl, 2-(p-toluenesulfonyl)ethyl, [2-(1 ,3-dithianyl)]methyl, 4- methylthiophenyl, 2,4-dimethyl-thiophenyl, 2-phosphonioethyl, 2- triphenylphosphonioisopropyl, 1 ,1-dimethyl-2-cyanoethyl, m-chloro-p- acyloxy benzyl, p-(dihydroxyboryl)benzyl, 5-benzisoxazolyl-methyl, and 2- (trifluoromethyl)-6-chromonyimethyl.
  • Photolytic Cleavage uses groups such as: m-nitrophenyl, 3,5- dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl, and phenyl(o- nitrophenyl)methyl.
  • groups such as: m-nitrophenyl, 3,5- dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl, and phenyl(o- nitrophenyl)methyl.
  • urea-type derivatives include: phenothiazinyl-(10)-carbonyl derivative, N '-p-toluenesulfonylaminocarbonyl, and N '-phenylaminothiocarbonyl.
  • miscellaneous carbamates include: f-amyl, S-benzyl thiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclopropylmethyl, p-decyloxy-benzyl, diisopropylmethyl, 2,2- dimethoxycarbonylvinyl, o-( ⁇ /, ⁇ /-dimethyl-carboxamido)-benzyl, 1 ,1-dimethyl- 3( ⁇ /, ⁇ /-dimethylcarboxamido)propyl, 1 ,1-dimethyl-propynyl, di(2-pyridyl)methyl, 2- furanylmethyl, 2-iodoethyl, isobornyl, isobutyl, isonicotinyl, p(p - methoxyphenylazo)benzyl, 1-methylcyclobuty
  • Amides includes: ⁇ /-formyl, ⁇ /-acetyl, ⁇ /-chloroacetyl, /V-trichloroacetyl, ⁇ /-trifluoroacetyl, /V-phenylacetyl, /V-3-phenylpropionyl, /V-picolinoyl, ⁇ /-3-pyridyl- carboxamide, ⁇ /-benzoylphenylalanyl derivative, ⁇ /-benzoyl, and ⁇ /-p- phenylbenzoyl.
  • Assisted Cleavage Assisted cleavage groups include: /V-o-nitrophenylacetyl, N-o- nitrophenoxyacetyl, ⁇ /-acetoacetyl, ( ⁇ / -dithiobenzyloxycarbonylamino)acetyl, ⁇ /-3- (p-hydroxphenyl) propionyl, ⁇ /-3-(o-nitrophenyl)propionyl, ⁇ /-2-methyl-2-(o- nitrophenoxy)propionyl, ⁇ /-2-methyl-2-(o-phenylazophenoxy)propionyl, ⁇ /-4- chlorobutyryl, ⁇ /-3-methyl-3-nitrobutyryl, ⁇ /-o-nitrocinnamoyl, ⁇ /-acetylmethionine derivative, ⁇ /-o-nitrobenzoyl, ⁇ /-o-(benzoyloxymethyl)benzoyl, and 4,5-diphenyl-3- o
  • Cyclic imide derivatives include: ⁇ /-phthalimide, ⁇ /-dithiasuccinoyl, ⁇ /-2,3-diphenyl-maleoyl, ⁇ /-2,5-dimethylpyrrolyl, ⁇ /-1 ,1 ,4,4-tetramethyldisilylazacyclopentane adduct, 5-substituted 1 ,3-dimethyl-1 ,3,5-triazacyclohexan-2-one, 5-substituted 1 ,3-dibenzyl- 1 ,3,5-triazacyclohexan-2-one, and 1 -substituted 3,5-dinitro-4-pyridonyl.
  • Protective groups for - NH include: ⁇ /-alkyl and ⁇ /-aryl amines, imine derivatives, enamine derivatives, and ⁇ /-hetero atom derivatives (such as /V-metal, N-N, N-P, N-Si, and N-S), /V-sulfenyl, and ⁇ /-sulfonyl.
  • ⁇ /-Alkyl and ⁇ /-Aryl Amines ⁇ /-alkyl and ⁇ /-aryl amines include: ⁇ /-methyl, ⁇ /-allyl, ⁇ /-[2-(trimethylsilyl)ethoxyl]- methyl, ⁇ /-3-acetoxypropyl, ⁇ /-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl), quaternary ammonium salts, /V-benzyl, ⁇ /-di(4-methoxyphenyl)methyl, ⁇ /-5-dibenzosuberyl, /V-triphenylmethyl, ⁇ /-(4-methoxyphenyl)diphenylmethyl, ⁇ /-9-phenylfluorenyl, ⁇ /-2,7-dichloro-9-fluorenylmethylene, ⁇ /-ferrocenylmethyl, and ⁇ /-2-picolylamine N '-oxid
  • Imine derivatives include: ⁇ /-1 ,1-dimethylthiomethylene, ⁇ /-benzylidene, ⁇ /-p-methoxybenzylidene, ⁇ /-diphenylmethylene, ⁇ /-[(2-pyridyl)mesityl]methylene, N-(N ',N '-dimethylaminomethylene), N,N '-isopropylidene, ⁇ /-p-nitrobenzylidene, ⁇ /-salicylidene, ⁇ /-5-chlorosalicylidene, ⁇ /-(5-chloro-2-hydroxyphenyl)phenyl- methylene, and ⁇ /-cyclohexylidene.
  • Enamine Derivatives include: ⁇ /-1 ,1-dimethylthiomethylene, ⁇ /-benzylidene, ⁇ /-p-methoxybenzylidene, ⁇ /-diphenylmethylene, ⁇ /-
  • An example of an enamine derivative is ⁇ /-(5,5-dimethyl-3-oxo-1-cyclohexenyl). ⁇ /-Hetero Atom Derivatives
  • /V-metal derivatives include: ⁇ /-borane derivatives, ⁇ /-diphenylborinic acid derivative, ⁇ /-[phenyl(pentacarbonylchromium- or -tungsten)]carbenyl, and
  • N-N derivatives include: ⁇ /-nitro,
  • tV-P derivatives include:
  • ⁇ /V-dialkyl phosphoryl examples include: ⁇ /-benzenesulfenyl,
  • ⁇ /-sulfonyl derivatives include: /V-p-toluenesulfonyl, /V-benzenesulfonyl, N- 2,3,6-trimethyl- 4-methoxybenzenesulfonyl, ⁇ /-2,4,6-trimethoxybenzenesulfonyl, N-
  • Disclosed compounds which are masked or protected may be prodrugs, compounds metabolized or otherwise transformed in vivo to yield a disclosed compound, e.g., transiently during metabolism.
  • This transformation may be a hydrolysis or oxidation which results from contact with a bodily fluid such as blood, or the action of acids, or liver, gastrointestinal, or other enzymes.
  • Example 4 to a solution of 2-methyl-4-trimethyl- silanylethynyl aniline (1.50 g, 0.008 mole) in THF (10 ml) at " 78°C, LDA (2 M in THF, 6 ml, 0.012 mole) was added and the mixture was stirred at " 78°C for 30 minutes. Solid 2,4-difluoro-benzoic acid (0.633 g, 0.004 mole) was added and the stirred for 16 hours while it warmed up to room temperature. The solvents were removed and water (30 ml) and Et 2 0 (50 ml) added to the oil residue.
  • EXAMPLE 6 4-Fluoro-2-(2-methyl-4-ethvnyl-phenylamino)-benzoic acid (7). To a solution of 6 in CH 3 OH (30 ml), aqueous 1 N KOH (10 ml) was added.
  • Incorporation of 32 P into myelin basic protein is assayed in the presence of a glutathione S-transferase fusion protein containing p44MAP kinase (GST-MAPK) and a glutathione S-transferase fusion protein containing p45MEK (GST-MEK).
  • the assay solution contains 20 mM HEPES, pH 7.4, 10 mM MgCI 2 , 1 mM MnCI 2 , 1 mM EGTA, 50, ⁇ M [ ⁇ - 32 P]ATP, 10 ⁇ g GST-MEK, 0.5 ⁇ g GST-MAPK and 40 ⁇ g MBP in a final volume of 100 ⁇ L.
  • Reactions are stopped after 20 minutes by addition of trichloroacetic acid and filtered through a GF/C filter mat. 32 P retained on the filter mat is determined using a 120S Betaplate. Compounds are assessed at 10 ⁇ M for ability to inhibit incorporation of 32 P.
  • two additional protocols are employed. In the first protocol, compounds are added to tubes containing GST-MEK, followed by addition of GST-MAPK, MBP and [ ⁇ - 32 P]ATP. In the second protocol, compounds are added to tubes containing both GST-MEK and GST-MAPK, followed by MBP and [ ⁇ - 32 P]ATP. Compounds that show activity in both protocols are scored as MAPK inhibitors, while compounds showing activity in only the first protocol are scored as MEK inhibitors.
  • Inhibitory activity can be confirmed in direct assays.
  • MAP kinase 1 ⁇ g GST-MAPK is incubated with 40 ⁇ g MBP for 15 minutes at 30°C in a final volume of 50 ⁇ L containing 50 mM Tris (pH 7.5), 10 ⁇ M MgC1 2 , 2 ⁇ M EGTA, and 10 ⁇ M [ ⁇ - 32 P]ATP.
  • the reaction is stopped by addition of Laemmli SDS sample buffer and phosphorylated MBP resolved by electrophoresis on a 10% polyacrylamide gel. Radioactivity incorporated into MBP is determined by both autoradiography, and scintillation counting of excised bands.
  • GST-MEKi For evaluation of direct MEK activity, 10 ⁇ g GST-MEKi is incubated with 5 ⁇ g of a glutathione S-transferase fusion protein containing p44MAP kinase with a lysine to alanine mutation at position 71 (GST-MAPK-KA). This mutation eliminates kinase activity of MAPK, so only kinase activity attributed to the added MEK remains. Incubations are 15 minutes at 30°C in a final volume of 50 ⁇ L containing 50 mM Tris (pH 7.5), 10 ⁇ M MgCI 2 , 2 , ⁇ M EGTA, and 10 ⁇ M [ ⁇ - 32 P]ATP.
  • the reaction is stopped by addition of Laemmli SDS sample buffer.
  • Phosphorylated GST-MAPK-KA is resolved by electrophoresis on a 10% polyacrylamide gel. Radioactivity incorporated into GST-MAPK-KA is determined by autoradiography, and subsequent scintillation counting of excised bands.
  • an artificially activated MEK containing serine to glutamate mutations at positions 218 and 222 (GST-MEK-2E) is used. When these two sites are phosphorylated, MEK activity is increased. Phosphorylation of these sites can be mimicked by mutation of the serine residues to glutamate.
  • 5 ⁇ g GST-MEK-2E is incubated with 5 ⁇ g GST-MAPK-KA for 15 minutes at 30°C in the same reaction buffer as described above. Reactions are terminated and analyzed as above.
  • Lysates are clarified by centrifugation at 13,000 x g for 10 minutes. Five to fifteen micrograms of protein from the resulting supematants are subjected to SDS/PAGE and Western blotting for phosphorylated MAP kinase levels.
  • EXAMPLE 17 Monolaver growth Cells are plated into multi-well plates at 10 to 20,000 cells/mL. Forty-eight hours after seeding, test compounds are added to the cell growth medium and incubation is continued for 2 additional days. Cells are then removed from the wells by incubation with trypsin and enumerated with a Coulter counter. EXAMPLE 18 Growth in soft-agar
  • Cells are seeded into 35-mm dishes at 5 to 10,000 cells/dish using growth medium containing 0.3% agar. After chilling to solidify the agar, cells are transferred to a 37°C incubator. After 7 to 10 days' growth, visible colonies are manually enumerated with the aid of a dissecting microscope.
  • Type II collagen-induced arthritis in mice is an experimental model of arthritis that has a number of pathologic, immunologic, and genetic features in common with rheumatoid arthritis.
  • the disease is induced by immunization of DBA/1 mice with 100 ⁇ g type II collagen, which is a major component of joint cartilage, delivered intradermally in Freund's complete adjuvant.
  • the disease susceptibility is regulated by the class II MHC gene locus, which is analogous to the association of rheumatoid arthritis with HLA-DR4.
  • a progressive and inflammatory arthritis develops in the majority of mice immunized, characterized by paw width increases of up to 100%.
  • a test compound is administered to mice in a range of amounts, such as 20, 60, 100, and 200 mg/kg body weight/day. The duration of the test can be several weeks to a few months, such as 40, 60, or 80 days.
  • a clinical scoring index is used to assess disease progression from erythema and edema (stage 1), joint distortion (stage 2), to joint ankylosis (stage 3). The disease is variable in that it can affect one or all paws in an animal, resulting in a total possible score of 12 for each mouse. Histopathology of an arthritic joint reveals synovitis, pannus formation, and cartilage and bone erosions. All mouse strains that are susceptible to CIA are high antibody responders to type II collagen, and there is a marked cellular response to CH.
  • EXAMPLE 20 SCW-induced monoarticular arthritis
  • Rats receive 6 ⁇ g sonicated SCW [in 10 ⁇ l Dulbecco's PBS (DPBS)] by an intraarticular injection into the right tibiotalar joint on day 0. On day 21 , the DTH is initiated with 100 ⁇ g of SCW (250 ⁇ l) administered i.v.
  • SCW 250 ⁇ l
  • SCW twice daily (10 ml/kg volume) beginning 1 hr prior to reactivation with SCW.
  • Compounds are administered in amounts between 10 and 500 mg/kg body weight/day, such as 20, 30, 60, 100, 200, and 300 mg/kg/day. Edema measurements are obtained by determining the baseline volumes of the sensitized hindpaw before reactivation on day 21 , and comparing them with volumes at subsequent time points such as day 22, 23, 24, and 25. Paw volume is determined by mercury plethysmography.
  • Each animal is anesthetized and an incision is made at the base of the recipient ear, cutting only the dorsal epidermis and dermis.
  • the incision is spread open and down to the cartilage parallel to the head, and sufficiently wide to accommodate the appropriate tunneling for a rat or insertion tool for a mouse.
  • a neonatal mouse or rat pup less than 60 hours old is anesthetized and cervically dislocated.
  • the heart is removed from the chest, rinsed with saline, bisected longitudinally with a scalpel, and rinsed with sterile saline.
  • the donor heart fragment is placed into the preformed tunnel with the insertion tool and air or residual fluid is gently expressed from the tunnel with light pressure. No suturing, adhesive bonding, bandaging, or treatment with antibiotics is required.
  • Implants are examined at 10-20-fold magnification with a stereoscopic dissecting microscope without anesthesia. Recipients whose grafts are not visibly beating may be anesthetized and evaluated for the presence of electrical activity using Grass E-2 platinum subdermal pin microelectodes placed either in the pinna or directly into the graft and a tachograph. Implants can be examined 1-4 times a day for 10, 20, 30 or more days. The ability of a test compound to ameliorate symptoms of transplant rejection can be compared with a control compound such as cyciosporine, tacrolimus, or orally-administered lefluonomide.
  • a control compound such as cyciosporine, tacrolimus, or orally-administered lefluonomide.
  • mice Female C57BLJ6 mice are obtained from the Jackson Laboratory (Bar Harbor, ME). All animals are given food and water ad libitum. Mice are sensitized with a single i.p. injection of OVA (grade V, Sigma Chemical Company, St. Louis, MO) adsorbed to alum, (10 ⁇ g OVA + 9 mg alum in 200 ⁇ l saline) or vehicle control, (9 mg alum in 200 ⁇ l saline) on day 0. On day 14, the mice are challenged with a 12-minute inhalation of an aerosol consisting of 1.5% OVA (weight/volume) in saline produced by a nebulizer (small particle generator, model SPAG-2; ICN Pharmaceuticals, Costa Mesa, CA).
  • OVA grade V
  • vehicle control 9 mg alum in 200 ⁇ l saline
  • mice are dosed with oral vehicle (0.5% hydroxypropylmethylcellulose / 0.25% TWEEN- 80), or a test compound at 10, 30, or 100 mg/kg in oral vehicle, 200 ⁇ l per mouse p.o. Dosing is performed once per day starting on day 7 or day 13, and extending through day 16. For determination of pulmonary eosinophilia, three days after the first
  • mice are anesthetized with an i.p. injection of anesthetic (Ketamine/Acepromazine/Xylazine), and the tracheae is exposed and cannulated.
  • anesthetic Ketamine/Acepromazine/Xylazine
  • the lungs and upper airways are lavaged twice with 0.5 ml of cold PBS.
  • a portion (200 ⁇ l) of the bronchoalveolar lavage (BAL) fluid is enumerated using a Coulter counter Model ZB1 (Coulter Electronics, Hialeah, FL).
  • the remaining BAL fluid is then centrifuged at 300 x g for five minutes, and the cells are resuspended in 1 ml of HBSS (Gibco BRL) containing 0.5% fetal calf serum (HyClone) and 10 mM HEPES (Gibco BRL).
  • the cell suspension is centrifuged in a cytospin (Shandon Southern Instruments, Sewickley, PA) and stained by Diff Quick (American Scientific Products, McGraw Park, IL) to differentiate BAL leukocytes into neutrophil, eosinophil, monocyte or lymphocyte subsets.
  • the number of eosinophils in the BAL fluid is determined by multiplying the percentage of eosinophils by the total cell count.
  • Transepithelial flux measurements were made by mounting the cell monolayers in a side-by-side diffusion chamber system (Precision Instrument Design, Tahoe City, CA). Temperature was maintained at 37°C with a circulating water jacket. The solutions were mixed with gas-lift circulation with 95% oxygen-5% carbon dioxide. Donor solutions with PD compounds, [ 14 C] mannitol (leakage marker) and [ 3 H] metoprolol (reference compound) were added to the apical chamber. Donor and receiver samples were collected at selected time intervals for up to 3 hours. Radiolabelled mannitol and metoprolol were analyzed using scintillation counting (TopCount, Packard Instruments, Downers Grove, IL).
  • Step a Preparation of ⁇ /-Cvclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide
  • a stirring suspension comprised of O-cyclopropylmethyl-hydroxylamine hydrochloride (5.40 g, 43.7 x 10 "3 mol) in dichloromethane (20 ml) at ambient temperature under a nitrogen atmosphere was added directly diisopropylethylamine (10.8 ml, 0.062 mol).
  • a solution comprised of 2,3,4- trifluorobenzenesulfonyl chloride (1.00 g, 4.34 x 10 "3 mol) in dichloromethane (120 ml) was added as a slow steady stream over twelve minutes to the first solution.
  • Step b Preparation of 2-(2-Chloro-4-iodo-phenylamino)- ⁇ /-cvclopropylmetho ⁇ y- 3,4-difluoro-benzenesulfonamide
  • 2-chloro-4-iodoaniline (0.80 g, 3.2 x 10 "3 mol) in tetrahydrofuran (10 ml) at -78 °C under a nitrogen atmosphere was added 1.0 molar lithium b/ ' strimethylsilylamide solution in tetrahydrofuran (6.2 ml, 6.2 x 10 "3 mol) to form a green suspension.
  • the suspension was stirred for five minutes before a stirring suspension comprised of lithiated N- cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide [prepared by adding 3.0 ml of the 1.0 molar lithium D/ ' strimethylsilylamide solution to a stirring solution comprised of 0.83 g (2.95 x 10 "3 mol) of ⁇ /-cyclopropylmethoxy-2,3,4-trifluoro- benzenesulfonamide in 10 ml of tetrahydrofuran at -78 °C] was added. The cold bath was removed and the reaction mixture was stirred for one hour.
  • lithiated N- cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide prepared by adding 3.0 ml of the 1.0 molar lithium D/ ' strimethylsilylamide solution to a stirring solution comprised of 0.83 g (2.95
  • Aqueous (10 %) hydrochloric acid 50 ml was added to the reaction mixture, and the biphasic mixture was concentrated in vacuo to an aqueous suspension that was extracted with diethyl ether (200 ml). The ether phase was dried (MgSO 4 ) and concentrated in vacuo to afford 2 g of a tan oil.
  • the crude product was purified by flash silica chromatography.

Abstract

The invention features a compound having the formula (I) below. The invention also relates to a pharmaceutical composition including (a) a compound of formula (I) and (b) a pharmaceutically-acceptable carrier. The invention further relates to a method for treating proliferative diseases such as cancer, restenosis, psoriasis, autoimmune disease, and atherosclerosis. Other aspects of the invention include methods for treating MEK-related (including ras-related) cancers, whether solid or hematopoietic.

Description

4' HETEROARYL D1ARYLAMINES
This invention relates to 4' heteroaryl diarylamines having pharmaceutical activity.
BACKGROUND
MEK enzymes are dual specificity kinases involved in, for example, immunomodulation, inflammation, and proliferative diseases such as cancer and restenosis.
Proliferative diseases are caused by a defect in the intracellular signaling system, or the signal transduction mechanism of certain proteins. Defects include a change either in the intrinsic activity or in the cellular concentration of one or more signaling proteins in the signaling cascade . The cell may produce a growth factor that binds to its own receptors, resulting in an autocrine loop, which continually stimulates proliferation. Mutations or overexpression of intracellular signaling proteins can lead to spurious mitogenic signals within the cell. Some of the most common mutations occur in genes encoding the protein known as Ras, a G-protein that is activated when bound to GTP, and inactivated when bound to GDP. The above-mentioned growth factor receptors, and many other mitogenic receptors, when activated, lead to Ras being converted from the GDP-bound state to the GTP-bound state. This signal is an absolute prerequisite for proliferation in most cell types. Defects in this signaling system, especially in the deactivation of the Ras-GTP complex, are common in cancers, and lead to the signaling cascade below Ras being chronically activated.
Activated Ras leads in turn to the activation of a cascade of serine/threonine kinases. One of the groups of kinases known to require an active Ras-GTP for its own activation is the Raf family. These in turn activate MEK (e.g., MEK<| and MEK2) which then activates MAP kinase, ERK (ERKi and ERK2). Activation of MAP kinase by mitogens appears to be essential for proliferation; constitutive activation of this kinase is sufficient to induce cellular transformation. Blockade of downstream Ras signaling, for example by use of a dominant negative Raf-1 protein, can completely inhibit mitogenesis, whether induced from cell surface receptors or from oncogenic Ras mutants. Although Ras is not itself a protein kinase, it participates in the activation of Raf and other kinases, most likely through a phosphorylation mechanism. Once activated, Raf and other kinases phosphorylate MEK on two closely adjacent serine residues, s218 and S^22 jn he case of MEK-1 , which are the prerequisite for activation of MEK as a kinase. MEK in turn phosphorylates MAP kinase on both a tyrosine, Y185, and a threonine residue, T''83j separated by a single amino acid. This double phosphorylation activates MAP kinase at least 100-fold. Activated MAP kinase can then catalyze the phosphorylation of a large number of proteins, including several transcription factors and other kinases. Many of these MAP kinase phosphorylations are mitogenically activating for the target protein, such as a kinase, a transcription factor, or another cellular protein. In addition to Raf-1 and MEKK, other kinases activate MEK, and MEK itself appears to be a signal integrating kinase. Current understanding is that MEK is highly specific for the phosphorylation of MAP kinase. In fact, no substrate for MEK other than the MAP kinase , ERK, has been demonstrated to date and MEK does not phosphorylate peptides based on the MAP kinase phosphorylation sequence, or even phosphorylate denatured MAP kinase. MEK also appears to associate strongly with MAP kinase prior to phosphorylating it, suggesting that phosphorylation of MAP kinase by MEK may require a prior strong interaction between the two proteins. Both this requirement and the unusual specificity of MEK are suggestive that it may have enough difference in its mechanism of action to other protein kinases that selective inhibitors of MEK, possibly operating through allosteric mechanisms rather than through the usual blockade of the ATP binding site, may be found. SUMMARY OF THE INVENTION
The invention features a compound having the formula (I) below:
(I)
In formula (I), W is OR1, NR20R!, NRARB, NR2NRARB, 0(CH2)1-4NRARB, or NR2(CH2)ι-4 NRARB- RI is H, C ^ alkyl, C 3-β alkenyl, C 3.8 alkynyl,
C 3-8 cycloalkyl, phenyl, (phenyl)C ι.4 alkyl, (phenyl)C 3-4 alkenyl, (phenyl)- C 3-4 alkynyl, (C 3.8 cycloalkyl)C 1-4 alkyl, (C 3-8 cycloalkyl)C 3-4 alkenyl, (C 3-8 cycloalkyl)C 3- alkynyl, C 3-8 heterocyclic radical, (C 3-s heterocyclic radical)C 1-4 alkyl, (C 3-8 heterocyclic radical)C 3-4 alkenyl, or (C 3-8 heterocyclic radical)C 3-4 alkynyl. Each of R2 and R3 is independently H, phenyl, C 1.4 alkyl, C 3-8 alkenyl, C 3.8 alkynyl, C 3.8 cycloalkyl, or (C 3-8 cycloalkyl)C 1-4 alkyl. Each of R4, R5 and R6 is independently H, F, Br, Cl, or NO2. RA is H, C ι-6 alkyl, C 3.8 alkenyl, C 3-8 alkynyl, C 3-8 cycloalkyl, phenyl, (C 3-8 cycloalkyl)C 1.4 alkyl, (C 3.8 cycloalkyl)C 3-4 alkenyl, (C 3-8 cycloalkyl)C 3-4 alkynyl, C 3.8 heterocyclic radical, (C 3-8 heterocyclic radical)C 1-4 alkyl, (aminosulfonyl)phenyl, (aminosulfonyl)- phenyl]C 1-4 alkyl, (aminosulfonyl)C 1-6 alkyl, (aminosulfonyl)C 3.6 cycloalkyl, or [(aminosulfonyl)C 3.6 cycloalkyljC alkyl. RB is H, C ι.8 alkyl, C 3.8 alkenyl, C 3.8 alkynyl, C 3-8 cycloalkyl, or phenyl. J is SRC, ORc, SO2Rc, SORc, SO2NRDRE, C 1-β alkyl, C 3-s alkenyl, C 3.8 alkynyl, C 3-β cycloalkyl, C 5-β cycloalkenyl, phenyl, (C 3-8 cycloalkyl)C 1-4 alkyl, (C 3.8 cycloalkyl)C 3.4 alkenyl, (C 3-8 cycloalkyl)- C 3-4 alkynyl, C 3.8 heterocyclic radical (e.g., 1 ,2,5-thiadiazol-3-yl), (C 3-8 heterocyclic radical) C 1-4 alkyl, -M'E'G', (heterocyclic radical)-M'-E'-G', or (cycloalkyl)-M'-E'-G'. M' is O, SO, SO2, NRE, (CO)NRE, NRE (CO), SO2NRE, NRES02, or CH2. E' is absent (in other words, a covalent bond), (CH2)ι-4 or (CH2)m O(CH2)p where 1 < (each of m and p independently) < 3 and 2 < (m + p) < 4. G' is OR3, SORc, SO2RC| or NRFRG; provided that where p = 1 , then G' is H. Each of Rc, RD> RE, RF and RG is independently selected from H, C 1-6 alkyl, C 3-4 alkenyl, C 3-4 alkynyl, C 3-6 cycloalkyl, C 3.6 heterocyclic radical, and phenyl; NRFRG and NRDRE can each also independently be selected from morpholinyl, pyrazinyl, piperazinyl, pyrrolidinyl, or piperadinyl. R10 is H, C 1-4 alkyl, halo, NO2) or S02NRHR|. Rn is H, halo, or NO2.
Each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C 1-4 alkyl, C 3-6 cycloalkyl, C 3-4 alkenyl, C 3.4 alkynyl, phenyl, hydroxy, amino, (amino)sulfonyl, and NO2, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 3 substituents independently selected from halo, C 1-2 alkyl, hydroxy, amino, and NO2. The invention also encompasses a pharmaceutically acceptable salt or C 1.7 ester of a compound of formula (I).
The invention also relates to a pharmaceutical composition including (a) a compound of formula (I) and (b) a pharmaceutically-acceptable carrier.
The invention further relates to a method for treating proliferative diseases, such as cancer, restenosis, psoriasis, autoimmune disease, and atherosclerosis. Other aspects of the invention include methods for treating MEK-related (including ras-related) cancers, whether solid or hematopoietic. Examples of cancers include colorectal, cervical, breast, ovarian, brain, acute leukemia, gastric, non-small cell lung, pancreatic, prostatic, and renal cancers. Further aspects of the invention include methods for treating or reducing the symptoms of xenograft (cell(s), llimb, skin, organ or bone marrow transplant) rejection, osteoarthritis, rheumatoid arthritis, cystic fibrosis, complications of diabetes (including diabetic retinopathy and diabetic nephropathy), hepatomegaly, cardiomegaly, stroke (such as acute focal ischemic stroke and global cerebral ischemia), heart failure, septic shock, asthma, and Alzheimer's disease. Compounds of the invention are also useful as antiviral agents for treating viral infections such as HIV, hepatitis (B) virus (HBV), human papilloma virus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV). These methods include the step of administering to a patient in need of such treatment, or suffering from such a disease or condition, a pharmaceutically-effective amount of a disclosed compound or pharmaceutical composition thereof. The invention also features methods of combination therapy, such as a method for treating cancer, wherein the method further includes providing radiation therapy or chemotherapy, for example, with mitotic inhibitors such as a taxane or a vinca alkaloid. Examples of mitotic inhibitors include paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine, and vinflunine Other therapeutic combinations include a MEK inhibitor of the invention and an anticancer agent such as cisplatin, 5-fluoro-2-4(1 H,3H)-pyrimidinedione (5FU), flutamide, and gemcitabine.
The chemotherapy or radiation therapy may be administered before, concurrently, or after the administration of a disclosed compound according to the needs of the patient.
The invention further includes synthetic intermediates and methods disclosed herein.
Other aspects of the invention are provided in the description, examples, and claims below.
DETAILED DESCRIPTION The invention features 4-heteroaryl diarylamine compounds, pharmaceutical compositions thereof, and methods of using such compounds and compositions. According to one aspect of the invention, the compounds are MEK inhibitors. MEK inhibition assays include the in vitro MEK/MAP described at column 6, line 36 to column 7, line 4 of U.S. Patent Number 5,525,625 and the in vitro MEK assay described at column 7, lines 4-27 of the same patent, the entire disclosure of which is incorporated by reference (see also Examples 13 et seq.). A whole cell assay is described in Example 16.
A. Terms
Certain terms are defined below and by their usage throughout this disclosure.
Alkyl groups include aliphatic (i.e., hydrocarbyl or hydrocarbon radical structures containing hydrogen and carbon atoms) with a free valence. Alkyl groups are understood to include straight chain and branched structures.
Examples include methyl, ethyl, propyl, isopropyl, butyl, n-butyl, isobutyl, t-butyl, pentyl, isopentyl, 2,3-dimethylpropyl, hexyl, 2,3-dimethylhexyl, 1 ,1-dimethylpentyl, heptyl, and octyl. Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Alkyl groups can be substituted with 1 , 2, 3 or more substituents which are independently selected from halo (fluoro, chloro, bromo, or iodo), hydroxy, amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy. Specific examples include fluoromethyl, hydroxyethyl, 2, 3-d i hydroxyethyl, (2- or 3-furanyl)methyl, cyclopropylmethyl, benzyloxyethyl, (3-pyridinyl)methyl, (2- or 3-furanyl)methyl, (2- thienyl)ethyl, hydroxypropyl, aminocyclohexyl, 2-dimethylaminobutyl, methoxymethyl, /V-pyridinylethyl, diethylaminoethyl, and cyclobutylmethyl.
Alkenyl groups are analogous to alkyl groups, but have at least one double bond (two adjacent sp2 carbon atoms). Depending on the placement of a double bond and substituents, if any, the geometry of the double bond may be entgegen (E), or zusammen (Z), cis, or trans. Similarly, alkynyl groups have at least one triple bond (two adjacent sp carbon atoms). Unsaturated alkenyl or alkynyl groups may have one or more double or triple bonds, respectively, or a mixture thereof; like alkyl groups, unsaturated groups may be straight chain or branched, and they may be substituted as described both above for alkyl groups and throughout the disclosure by example. Examples of alkenyls, alkynyls, and substituted forms include cis-2-butenyl, trans-2-butenyl, 3-butynyl, 3-phenyl-2- propynyl, 3-(2'-fluorophenyl)-2-propynyl, 3-methyl(5-phenyl)-4-pentynyl, 2- hydroxy-2-propynyI, 2-methyl-2-propynyl, 2-propenyl, 4-hydroxy-3-butynyl, 3-(3- fluorophenyl)-2-propynyl, and 2-methyl-2-propenyl. In formula (I), alkenyls and alkynyls can be C 2-Λ or C 2.8, for example, and are preferably C 3-4 or C 3-8- More general forms of substituted hydrocarbon radicals include hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl, hydroxyaryl, and corresponding forms for the prefixes amino-, halo- (e.g., fluoro-, chloro-, or bromo-), nitro-, alkyl-, phenyl-, cycloalkyl- and so on, or combinations of substituents. According to formula (I), therefore, substituted alkyls include hydroxyalkyl, aminoalkyl, nitroalkyl, haloalkyl, alkylalkyl (branched alkyls, such as methylpentyl), (cycloalkyl)alkyl, phenylalkyl, alkoxy, alkylaminoalkyl, dialkylaminoalkyl, arylalkyl, aryloxyalkyl, arylalkyloxyalkyl, (heterocyclic radical)alkyl, and (heterocyclic radical)oxyalkyl. Ri thus includes hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl, hydroxyaryl, aminoalkyl, aminoalkenyl, aminoalkynyl, aminocycloalkyl, aminoaryl, alkylalkenyl,
(alkylaryl)alkyl, (haloaryl)alkyl, (hydroxyaryl)alkynyl, and so forth. Similarly, RA includes hydroxyalkyl and aminoaryl, and RB includes hydroxyalkyl, aminoalkyl, and hydroxyalkyl(heterocyclic radical)alkyl.
Heterocyclic radicals, which include but are not limited to heteroaryls, include: furyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, pyrrolyl, imidazolyl, 1 ,3,4- triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, indolyl, 1 ,2,5-thiadiazolyl and their nonaromatic counterparts. Further examples of heterocyclic radicals include piperidyl, quinolyl, isothiazolyl, piperidinyl, morpholinyl, piperazinyl, tetrahydrofuryl, tetrahydropyrrolyl, pyrrolidinyl, octahydroindolyl, thiadiazolyl, octahydrobenzothiofuranyl, and octahydrobenzofuranyl.
Selective MEK 1 or MEK 2 inhibitors are those compounds which inhibit the MEK 1 or MEK 2 enzymes, respectively, without substantially inhibiting other enzymes such as MKK3, PKC, Cdk2A, phosphorylase kinase, EGF, and PDGF receptor kinases, and C-src. In general, a selective MEK 1 or MEK 2 inhibitor has an IC50 for MEK 1 or MEK 2 that is at least one-fiftieth (1/50) that of its IC50 for one of the above-named other enzymes. Preferably, a selective inhibitor has an IC50 that is at least 1/100, more preferably 1/500, and even more preferably 1/1000, 1/5000, or less than that of its IC5o or one or more of the above-named enzymes.
B. Compounds One aspect of the invention features disclosed compounds shown in formula (I) in the Summary section. Embodiments of the invention include compounds wherein: (a) Rc is C ι.2 alkyl; (b) W is OH, or W is NHOR-i (c) Rio is methyl or chloro; (d) Rn is fluoro; (e) Rn is H; (f) J is trihalomethyl or methylthio; (g) J is SO2CH3; (h) J is SOCH3; (i) J is C 3-β alkynyl where the triple bond is between the carbon atoms alpha and beta to the phenyl group; (j) Ri has at least one hydroxy substituent; (k) Ri is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 3-5 alkenyl, C 3-5 alkynyl, C 3-6 cycloalkyl, (C 3-5 cycloalkyl)C ι.2 alkyl, or (C 3-5 heterocyclic radical)C ι.2 alkyl; (I) Ri is H or (C 3.4 cycloalkyl)C ι.2 alkyl; (m) R2 is H, methyl, C 3-4 alkynyl, C 3.5 cycloalkyl, or (C 3-5 cycloalkyl)methyl; (n) RA is H, methyl, ethyl, isobutyl, hydroxyethyl, hydroxypropyl, cyclopropylmethyl, cyclobutylmethyl, C 3-4 alkynyl, phenyl, 2- piperjdin-1 -yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2-hydroxyethyl)-piperazin-1-yl]- propyl, 2-pyrrolidin-1 -yl-ethyl, or 2-diethylamino-ethyl; and RB is H; or where RB is methyl and RA is phenyl; (0) each of R4 and RΘ is H, and R5 is F; (p) each of R4, R5, and R6 is F; (q) R5 is F; (r) each R5 and R6 is F and R6 is Br; (s) each R5 and RΘ is F and R$ is H; (t) J is 1 ,2,5-thiadiazol-3-yl; or a combination thereof.
Preferably, where one of Ri, R2, RA, RB, Rc, RD, RE, RF, and RG, for example, is an alkenyl or alkynyl group, its double or triple bond, respectively, is not adjacent the point of attachment. For example, where W is NR2ORι, R2 is preferably prop-2-ynyl, or but-2 or 3-enyl, and less preferably prop-1-ynyl or but- 1-enyl.
Examples of compounds of formula (I) include: 4-fluoro-2-(2-methyl-4- methylsulfanyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2-(2-methyl-4- methylsulfanyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4-methanesulfinyl-2- methyl-phenylamino)-benzoic acid; 2-(4-methanesulfinyl-2-methyl-phenylamino)- 4-nitro-benzoic acid; 3,4,5-trifluoro-2-(4-methanesulfonyl-2-methyl-phenylamino)- benzoic acid; 3,4-difluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 2-(2-methyl-4-methylsulfanyl-phenylamino)-4-nitro-benzoic acid; 3,4, 5-trifluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 4-fluoro- 2-(4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro- 2-(4-methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 3,4,5-trifluoro-2- (2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 4-fluoro-2-(4-methane- sulfinyl-2-methyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2- (4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 3,4-difluoro-2- (4-methanesulfonyl-2-methyl-phenylamino)-benzoic acid; and 2-(4-methanesulfonyl-2-methyl-phenylamino)-4-nitro-benzoic acid; and the corresponding hydroxamic acid or cyclopropylhydroxamic acid of each.
Preferred examples of compounds of formula (I) are : 4-Fluoro-2-(4- methanesulfanyl-phenylamino)-benzoic acid (1); 4-Fluoro-2-(4-methanesulfinyl- phenylamino)-benzoic acid (2); 4-Fluoro-2-(4-methanesulfonyl-phenylamino)- benzoic acid (3); 4-Fluoro-2-(2-methyl-4-trimethylsilanylethynyl-phenylamino)- benzoic acid (6); 4-Fluoro-2-(2-methyl-4-ethynyl-phenylamino)-benzoic acid (7). Biological data on these seven compounds is given on page 17; full characterization of the compounds - MP, NMR, MS, IR and CHN- is given on pages 28-31. Additional preferred compounds include the following: (a) 5-Bromo-2-(4- ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy- 2-(4-ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzamide; 2-(4-Ethynyl-2- methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4- ethynyl-2-methyl-phenylamino)-3,4,5-trifluoro-benzamide; 2-(4-Ethynyl-2-methyl- phenylamino)-3,4,5-trifluoro-benzoic acid; 5-Bromo-N-cyclopropylmethoxy-2-(4- ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzamide; (b) 5-Bromo-2-(4-ethynyl- CI-methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4- ethynyl-CI-methyl-phenylamino)-3,4-difiuoro-benzamide; 2-(4-Ethynyl-CI-methyl- phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4-ethynyl-CI- methyl-phenylamino)-3,4,5-trifluoro-benzamide; 2-(4-Ethynyl-CI-methyl- phenylamino)-3,4,5-trifluoro-benzoic acid; 5-Bromo-N-cyclopropylmethoxy-2-(4- ethynyl-CI-methyl-phenylamino)-3,4-difluoro-benzamide; (c) 5-bromo-3,4-difluoro- 2-(2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4- methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 3,4,5-trifluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(2-methyl-4- methylsulfanyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzoic acid; (d) 5-bromo-N- cyclopropylmethoxy-3,4-difluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)- benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-methanesulfinyl-2-methyl- phenylamino)-benzamide; N-cyclopropylmethoxy-3,4, 5-trifluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4- difluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzamide; N- cyclopropylmethoxy-3,4,5-trifluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)- benzamide; 5-bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(4-methanesulfonyl-2- methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4, 5-trifiuoro-2-(2- methyl-4-methylsulfanyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4- difluoro-2-(4-methanesulfonyl-2-methyl-phenylamino)-benzamide; (e) N- cyclopropylmethoxy-3,4-difluoro-2-(4-imidazol-1-yl-2-methyl-phenylamino)- benzamide; (f) N-cyclopropylmethoxy-3,4, 5-trifluoro-2-(2-methyl-4- [1 ,2,5]thiadiazol-3-yl-phenylamino)-benzamide; 2-[4-(4-chloro-[1 ,2,5]thiadiazol-3- yl)-2-methyl-phenylamino]-3,4,5-trifluoro-benzoic acid; 2-[4-(4-chloro-
[1 ,2,5]thiadiazol-3-yl)-2-methyl-phenylamino]-N-cyclopropylmethoxy-3,4,5- trifluoro-benzamide; (g) 2-{4-[4-(2-dimethylamino-ethoxy)-[1 ,2,5]thiadiazol-3-yl]-2- methyl-phenylamino}-3,4,5-trifluoro-benzoic acid; (h) N-cyclopropylmethoxy- 3,4, 5-trifluoro-2-{2-methyl-4-[4-(2-piperidin-1-yl-ethoxy)-[1 , 2, 5]thiadiazol-3-yl]- phenylaminoj-benzamide.
Further preferred compounds include: (a) 5-bromo-2-(2-chloro-4- methylsulfanyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-chloro-4- methanesulfinyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-chloro-4- methanesulfonyl-phenylamino)-3,4,5-trifluoro-benzoic acid; 2-(2-chloro- methylsulfanyl-phenylamino)-3,4-difluoro-benzoic acid; 5-bromo-2-(2-chloro-4- methanesulfonyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-Chloro-4- methanesulfonyl-phenylamino)-3,4-difluoro-benzoic acid; (b) 5-bromo-2-(2- chloro-4-methylsulfanyl-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzamide; 2-(2-chloro-4-methanesuIfinyl-phenylamino)-N-cyclopropylmethoxy- 3,4-difluoro-benzamide; 2-(2-chloro-4-methanesulfonyl-phenylamino)- N- cyclopropylmethoxy-3,4,5-trifluoro-benzamide; 2-(2-chloro-4-methylsulfanyl- phenylamino)- N-cyclopropylmethoxy-3,4-difluoro-benzamide;
2-(2-chloro-4-methanesulfinyl-phenylamino)- N-cyclopropylmethoxy-3,4, 5- trifluoro-benzamide; 5-bromo-2-(2-chloro-4-methanesulfonyl-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-methylsulfanyl- phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-benzamide; 2-(2-chloro-4- methanesulfonyl-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide; and (c) 2-[2-chloro 4-(3H-imidazol-1-yl)-phenylamino]-N-cyclopropylmethoxy-3,4- difluoro-benzamide; 2-(2-chloro-4-[1 ,2,5]thiadiazol-3-yl-phenylamino)-N- cyclopropylmethoxy-3,4,5-trifluoro-benzamide; 2-[4-(2-chloro-4-chioro- [1 ,2,5]thiadiazol-3-yl)-phenylamino]-3,4,5-trifluoro-benzoic acid; 2-[2-chloro-4-(4- chloro-[1 ,2,5]thiadiazol-3-yl)-phenylamino]-N-cyclopropylmethoxy-3,4,5-trifiuoro- benzamide; 2-{4-[4-(2-dimethylamino-ethoxy)-[1 ,2,5]thiadiazol-3-yl]-2-methyl- phenylamino}-3,4,5-thfluoro-benzoic acid; 2-{2-chloro-4-[4-(2-piperidin-1-yl- ethoxy)-[1 ,2,5]thiadiazol-3-yl]-phenylamino}-N-cyclopropylmethoxy-3,4,5-trifluoro- benzamide; Additional preferred compounds include: (a) 2-(2-Chloro-4-ethynyl- phenylamino)-4-fluoro-benzoic acid; 5-Bromo-2-(2-chloro-4-ethynyl- phenylamino)-3,4-difluoro-benzoic acid; 2-(2-Chloro-4-ethynyl-phenylamino)- N- cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)- N-cyclopropylmethoxy-4-nitro-benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)- N-hydroxy-3,4,5-trifluoro- benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)-3,4- difluoro-benzoic acid; 2-(4-Ethynyl-2-chloro-phenylamino)-4-nitro-benzoic acid; 2- (2-Chloro-4-ethynyl-phenylamino)- N-Cyclopropylmethoxy-3,4,5-trifluoro- benzamide; 2-(2-chloro-4-methanesulfinyl-phenylamino)- 4-fluoro-N-hydroxy- benzamide; 5-Bromo-2-(4-ethynyl-2-chloro-phenylamino)-3,4-difluoro-N-hydroxy- benzamide; (b) 2-(2-Chloro-4-ethynyl-phenylamino)-3,4,5-trifluoro-benzoic acid;
2-(2-Chloro-4-ethynyl-phenylamino)- N-cyclopropylmethoxy-4-fluoro-benzamide;
5-Bromo-2-(2-chloro-4-ethynyl-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro- benzamide; 2-(4-Ethynyl-2-chloro-phenylamino)-3,4-difluoro-N-hydroxy- benzamide; 2-(4-Ethynyl-2-chloro-phenylamino)-N-hydroxy-4-nitro-benzamide; and (c) 2-(2-Chloro-4-ethynyl-phenylamino)-4-fluoro-benzoic acid; 2-(2-Chloro-4-ethynyl-phenylamino)- N-cyclopropylmethoxy-4-fluoro-benzamide; 2-(2-Chloro-4-methanesulfinyl-phenylamino)- 4-fluoro-N-hydroxy-benzamide; 2-(2-chloro-4-imidazol-1-yl-phenylamino)- 3,4-Difluoro-benzoic acid.
C. Synthesis
The disclosed compounds can be synthesized according to the following five schemes, or variants thereof. The abbreviation PyBOP is (benzotriazolyl- oxy)-tripyrrolidino phosphonium hexafluorophosphate. These synthetic strategies are further exemplified in Examples 1-12 below:
Scheme 1
PhS0 W2 HPh
RTONHR2 PyBOP
Scheme 2
Scheme 3
Scheme 4 3-Aryl-1 ,2,5-thiadiazols
Y = solubilizing side chain
Scheme 5
18 19 D. Uses
The disclosed compositions are useful as both prophylactic and therapeutic treatments for diseases or conditions as provided in the Summary section, as well as diseases or conditions modulated by the MEK cascade. Examples include stroke, heart failure, osteoarthritis, rheumatoid arthritis, organ transplant rejection, and a variety of tumors such as ovarian, non-small cell lung, pancreatic, brain, prostatic, renal, and colon.
MEK INHIBITION
1. Dosages
Those skilled in the art will be able to determine, according to known methods, the appropriate dosage for a patient, taking into account factors such as age, weight, general health, the type of symptoms requiring treatment, and the presence of other medications. In general, an effective amount will be between 0.1 and 1000 mg/kg per day, preferably between 1 and 300 mg/kg body weight, and daily dosages will be between 10 and 5000 mg for an adult subject of normal weight. Capsules, tablets or other formulations (such as liquids and film-coated tablets) may be of between 5 and 200 mg, such as 10, 15, 25, 35, 50 mg, 60 mg, and 100 mg and can be administered according to the disclosed methods. 2. Formulations
Dosage unit forms include tablets, capsules, pills, powders, granules, aqueous and nonaqueous oral solutions and suspensions, and parenteral solutions packaged in containers adapted for subdivision into individual doses. Dosage unit forms can also be adapted for various methods of administration, including controlled release formulations, such as subcutaneous implants. Administration methods include oral, rectal, parenteral (intravenous, intramuscular, subcutaneous), intracistemal, intravaginal, intraperitoneal, intravesical, local (drops, powders, ointments, gels, or cream), and by inhalation (a buccal or nasal spray).
Parenteral formulations include pharmaceutically acceptable aqueous or nonaqueous solutions, dispersion, suspensions, emulsions, and sterile powders for the preparation thereof. Examples of carriers include water, ethanol, polyols (propylene glycol, polyethylene glycol), vegetable oils, and injectable organic esters such as ethyl oleate. Fluidity can be maintained by the use of a coating such as lecithin, a surfactant, or maintaining appropriate particle size. Carriers for solid dosage forms include (a) fillers or extenders, (b) binders, (c) humectants, (d) disintegrating agents, (e) solution retarders, (f) absorption acccelerators, (g) adsorbants, (h) lubricants, (i) buffering agents, and (j) propellants.
Compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispensing agents; antimicrobial agents such as parabens, chlorobutanol, phenol, and sorbic acid; isotonic agents such as a sugar or sodium chloride; absorption-prolonging agents such as aluminum monostearate and gelatin; and absorption-enhancing agents.
3. Related compounds
The invention provides the disclosed compounds and closely related, pharmaceutically acceptable forms of the disclosed compounds, such as salts, esters, amides, hydrates or solvated forms thereof; masked or protected forms; and racemic mixtures, or enantiomerically or optically pure forms.
Pharmaceutically acceptable salts, esters, and amides include carboxylate salts (e.g., C ι.8 alkyl, cycloalkyl, aryl, heteroaryl, or non-aromatic heterocyclic), amino acid addition salts, esters, and amides which are within a reasonable benefit/risk ratio, pharmacologically effective, and suitable for contact with the tissues of patients without undue toxicity, irritation, or allergic response. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, iaurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactiobionate, and laurylsulfonate. These may include alkali metal and alkali earth cations such as sodium, potassium, calcium, and magnesium, as well as non-toxic ammonium, quaternary ammonium, and amine cations such as tetramethyl ammonium, methylamine, trimethylamine, and ethylamine. See, for example, S.M. Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977, 66:1-19 which is incorporated herein by reference. Representative pharmaceutically acceptable amides of the invention include those derived from ammonia, primary C -|.6 alkyl amines and secondary di (C ι_6 alkyl) amines. Secondary amines include 5- or 6-membered heterocyclic or heteroaromatic ring moieties containing at least one nitrogen atom and optionally between 1 and 2 additional heteroatoms. Preferred amides are derived from ammonia, C ^3 alkyl primary amines, and di (C ι-2 alkyl)amines. Representative pharmaceutically acceptable esters of the invention include C 1.7 alkyl, C 5-7 cycloalkyl, phenyl, and phenyl(C ι-6)alkyl esters. Preferred esters include methyl esters.
The invention also includes disclosed compounds having one or more functional groups (e.g., hydroxyl, amino, or carboxyl) masked by a protecting group. Some of these masked or protected compounds are pharmaceutically acceptable; others will be useful as intermediates. Synthetic intermediates and processes disclosed herein, and minor modifications thereof, are also within the scope of the invention.
HYDROXYL PROTECTING GROUPS Hydroxyl protecting groups include: ethers, esters, and protection for 1 ,2- and 1 ,3-diols. The ether protecting groups include: methyl, substituted methyl ethers, substituted ethyl ethers, substituted benzyl ethers, silyl ethers and conversion of silyl ethers to other functional groups.
Substituted Methyl Ethers Substituted methyl ethers include: methoxymethyl, methylthiomethyl, t- butylthiomethyl, (phenyldimethylsilyl) methoxymethyl, benzyloxymethyl, p-ethoxybenzyloxymethyl, (4-methoxyphenoxy) methyl, guaiacolmethyl, f-butoxymethyl, 4-pentenyloxymethyl, siloxymethyl, 2-methoxyethoxymethyl, 2,2,2-trichloroethoxymethyl, bis(2-chloro- ethoxy)methyl, 2-(trimethylsilyl)- ethoxymethyl, tetrahydropyranyl, 3-bromotetrahydro-pyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl, 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxido, 1 -[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl, 1 ,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, and 2,3,3a,4,5,6,7,7a-octahydro-7,8,8- trimethyl-4,7-ethanobenzofuran-2-yl. Substituted Ethyl Ethers
Substituted ethyl ethers include: 1-ethoxyethyl, 1-(2,chloroethoxy)ethyl, 1 -methyl- 1-methoxyethyl, 1-methyl-1 -benzyloxyethyl, 1 -methy 1-1 -benzyloxy-2- fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilyethyl, 2-(phenylselenyl)ethyl, f-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, and benzyl. Substituted Benzyl Ethers
Substituted benzyl ethers include: p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2- and 4-picolyl, 3-methyl-2-picolyl Λ/-oxido, diphenylmethyl, p, p -dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyl- diphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)- phenylmethyl, tri-(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxy)phenyl- diphenylmethyl, 4,4',4"-ths(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"- tris(levulinoyloxyphenyl) methyl, 4,4',4"tris(benzoyloxyphenyl)methyl, 3-(imidazol- 1-ylmethyl)bis(4',4"-dimethoxyphenyl)-methyl, 1 ,1-bis(4-methoxyphenyl)-1'- pyrenylmethyl, 9-anthryl, 9-(9-phenyl) xanthenyl, 9-(9-phenyl-10-oxo) anthryl, 1 ,3- benzodithiolan-2-yl, and benzisothiazolyl S,S-dioxido. Silyl Ethers Silyl ethers include: trimethylsilyl, triethylsilyl, triisopropylsilyl, dimethylisopropylsilyl, diethylisopropylsilyl, dimethylthexylsilyl, f-butyldimethylsilyl, f-butyldiphenylsilyl, tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl, and f-butylmethoxyphenylsilyl.
ESTERS
Esters protecting groups include: esters, carbonates, assisted cleavage, miscellaneous esters, and sulfonates. Esters
Examples of protective esters include: formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, p-P- phenylacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4- (ethylenedithio) pentanoate, pivaloate, adamantoate,crotonate,4- methoxycrotonate, benzoate, p-phenylbenzoate, and 2,4,6-trimethylbenzoate (mesitoate).
Carbonates Carbonates include: methyl, 9-fluorenylmethyl, ethyl, 2,2,2-thchloroethyl, 2-(trimethylsilyl) ethyl, 2-(phenylsulfonyl) ethyl, 2-(triphenylphosphonio) ethyl, isobutyl, vinyl, allyl, p-nitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl, and methyl dithiocarbonate. Assisted Cleavage Examples of assisted cleavage protecting groups include: 2-iodobenzoate, 4- azido-butyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl) benzoate, 2- formylbenzene-sulfonate, 2-(methylthiomethoxy) ethyl carbonate, 4- (methylthiomethoxymethyl) benzoate, and 2-(methylthiomethoxymethyl) benzoate. Miscellaneous Esters
In addition to the above classes, miscellaneous esters include: 2,6-dichloro-4- methylphenoxyacetate, 2,6-dichloro-4-(1 ,1 ,3,3-tetramethylbutyl) phenoxyacetate, 2,4-bis(1 ,1-dimethylpropyl) phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (£)-2-methyl-2-butenoate (tigloate), o-(methoxycarbonyl) benzoate, p-P-benzoate, α-naphthoate, nitrate, alkyl N,N,N ',N '- tetramethylphosphorodiamidate, Λ/-phenylcarbamate, borate, dimethylphosphinothioyl, and 2,4-dinitrophenylsulfenate. Sulfonates Protective sulfates includes: sulfate, methanesulfonate(mesylate), benzylsulfonate, and tosylate.
PROTECTION FOR 1 ,2- AND 1.3-DIOLS
The protection for 1 ,2 and 1 ,3-diols group includes: cyclic acetals and ketals, cyclic ortho esters, and silyl derivatives.
Cyclic Acetals and Ketals Cyclic acetals and ketals include: methylene, ethylidene, 1-f-butylethylidene, 1-phenylethylidene, (4-methoxyphenyl) ethylidene, 2,2,2-trichloroethylidene, acetonide (isopropylidene), cyclopentylidene, cyclohexylidene, cycloheptylidene, benzylidene, p-methoxybenzylidene, 2,4-dimethoxybenzylidene, 3,4- dimethoxybenzylidene, and 2-nitrobenzylidene. Cyclic Ortho Esters Cyclic ortho esters include: methoxymethylene, ethoxymethylene, dimethoxy- methylene, 1-methoxyethylidene, 1-ethoxyethylidine, 1 ,2-dimethoxyethylidene, α-methoxybenzylidene, 1-(Λ/,Λ/-dimethylamino)ethylidene derivative, α-(Λ/,Λ/- dimethylamino) benzylidene derivative, and 2-oxacyclopentylidene.
PROTECTION FOR THE CARBOXYL GROUP ESTERS Ester protecting groups include: esters, substituted methyl esters, 2-substituted ethyl esters, substituted benzyl esters, silyl esters, activated esters, miscellaneous derivatives, and stannyl esters. Substituted Methyl Esters Substituted methyl esters include: 9-fluorenylmethyl, methoxymethyl, methylthiomethyl, tetrahydropyranyl, tetrahydrofuranyl, methoxyethoxymethyl, 2- (t methylsilyl)ethoxy-methyl, benzyloxymethyl, phenacyl, p-bromophenacyl, α- methylphenacyl, p-methoxyphenacyl, carboxamidomethyl, and N- phthalimidomethyl.
2-Substituted Ethyl Esters 2-Substituted ethyl esters include: 2,2,2-trichloroethyl, 2-haloethyl, l-chloroalkyl, 2-(trimethylsily)ethyl, 2-methylthioethyl, 1 ,3-dithianyl-2-methyl, 2(p- nitrophenylsulfenyl)-ethyl, 2-(p-toluenesulfonyl)ethyl, 2-(2'-pyridyl)ethyl, 2-
(diphenylphosphino)ethyl, 1-methyl-1-phenylethyl, .--butyl, cyclopentyl, cyclohexyl, allyl, 3-buten-1-yl, 4-(trimethylsily)-2-buten-1-yl, cinnamyl, α-methylcinnamyl, phenyl, p-(methylmercapto)-phenyl, and benzyl. Substituted Benzyl Esters Substituted benzyl esters include: triphenylmethyl, diphenylmethyl, bis(o-nitrophenyl)methyl, 9-anthrylmethyl, 2-(9,10-dioxo)anthrylmethyl, 5-dibenzo- suberyl, 1 -pyrenylmethyl, 2-(trifluoromethyl)-6-chromylmethyl, 2,4,6- thmethylbenzyl, p-bromobenzyl, o-nitrobenzyl, p-nitrobenzyl, p-methoxybenzyl, 2,6-dimethoxybenzyl, 4-(methylsulfinyl)benzyl, 4-sulfobenzyl, piperonyl, and 4-P- benzyl.
Silyl Esters Silyl esters include: trimethylsilyl, t ethylsilyl, f-butyldimethylsilyl, /- propyldimethylsilyl, phenyldimethylsilyl, and di- f-butylmethylsilyl. Miscellaneous Derivatives Miscellaneous derivatives includes: oxazoles, 2-alkyl-1 ,3-oxazolines, 4-alkyl-5- oxo-1 , 3-oxazolidines, 5-alkyl-4-oxo-1 ,3-dioxolanes, ortho esters, phenyl group, and pentaaminocobalt(lll) complex.
Stannyl Esters Examples of stannyl esters include: triethylstannyl and tri-n-butylstannyl. AMIDES AND HYDRAZIDES
Amides include: N,N -dimethyl, pyrrolidinyl, piperidinyl, 5,6-dihydrophen- anthridinyl, o-nitroanilides, Λ/-7-nitroindolyl, /V-8-nitro-1,2,3,4-tetrahydroquinolyl, and p-P-benzenesulfonamides. Hydrazides include: /V-phenyl, Λ/./V '-diisopropyl and other dialkyl hydrazides.
PROTECTION FOR THE AMINO GROUP
CARBAMATES Carbamates include: carbamates, substituted ethyl, assisted cleavage, photolytic cleavage, urea-type derivatives, and miscellaneous carbamates.
Carbamates Carbamates include: methyl and ethyl, 9-fluorenylmethyl, 9-(2- sulfo)fluorenylmethyl, 9-(2,7-dibromo)fluorenylmethyl, 2,7-di- -butyl-[9-(10,10- dioxo-10, 10, 10, 10-tetrahydro- thioxanthyl)]methyl, and 4-methoxyphenacyl. Substituted Ethyl Substituted ethyl protective groups include: 2,2,2-trichloroethyl, 2- trimethylsilylethyl, 2-phenylethyl, 1-(1-adamantyl)-1-methylethyl, 1 ,1-dimethyl-2- haloethyl, 1 ,1dimethyl-2,2-dibromoethyl, 1 ,1-dimethyl-2,2,2-trichloroethyl, 1- methyl-1-(4-biphenylyl)ethyl, 1-(3,5-di-f-butylphenyl)-1-methylethyl, 2-(2'-and 4'- pyridyl)ethyl, 2-(/V,Λ/-icyclohexylcarboxamido)- ethyl, f-butyl, 1-adamantyl, vinyl, allyl, 1-isopropylallyl, connamyl, 4-nitrocinnamyl, quinolyl, V-hydroxypiperidinyl, alkyldithio, benzyl, p-methoxybenzyl, p-nitrobenzyl, p-bromobenzyl, p- chlorobenzyl, 2,4-dichlorobenzyl, 4-methylsulfinylbenzyl, 9-anthrylmethyl, and diphenylmethyl.
Assisted Cleavage Protection via assisted cleavage includes: 2-methylthioethyl, 2- methylsulfonylethyl, 2-(p-toluenesulfonyl)ethyl, [2-(1 ,3-dithianyl)]methyl, 4- methylthiophenyl, 2,4-dimethyl-thiophenyl, 2-phosphonioethyl, 2- triphenylphosphonioisopropyl, 1 ,1-dimethyl-2-cyanoethyl, m-chloro-p- acyloxy benzyl, p-(dihydroxyboryl)benzyl, 5-benzisoxazolyl-methyl, and 2- (trifluoromethyl)-6-chromonyimethyl. Photolytic Cleavage Photolytic cleavage methods use groups such as: m-nitrophenyl, 3,5- dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl, and phenyl(o- nitrophenyl)methyl. Urea-Type Derivatives
Examples of urea-type derivatives include: phenothiazinyl-(10)-carbonyl derivative, N '-p-toluenesulfonylaminocarbonyl, and N '-phenylaminothiocarbonyl.
Miscellaneous Carbamates In addition to the above, miscellaneous carbamates include: f-amyl, S-benzyl thiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclopropylmethyl, p-decyloxy-benzyl, diisopropylmethyl, 2,2- dimethoxycarbonylvinyl, o-(Λ/,Λ/-dimethyl-carboxamido)-benzyl, 1 ,1-dimethyl- 3(Λ/,Λ/-dimethylcarboxamido)propyl, 1 ,1-dimethyl-propynyl, di(2-pyridyl)methyl, 2- furanylmethyl, 2-iodoethyl, isobornyl, isobutyl, isonicotinyl, p(p - methoxyphenylazo)benzyl, 1-methylcyclobutyl, 1-methylcyclohexyl, 1 -methyl-1 -cyclopropylmethyl, 1 -methyl-(3,5-dimethoxyphenyl)ethyl, 1 -methyl- 1 (p-henylazophenyl)- ethyl, 1 -methyl-1 -phenylethyl, 1 -methyl-1 -(4-pyridyl)ethyl, phenyl, p-(phenylazo)benzyl, 2,4,6-tri-f-butylphenyl, 4-trimethylammonium)- benzyl, and 2,4,6-trimethylbenzyl.
AMIDES
Amides Amides includes: Λ/-formyl, Λ/-acetyl, Λ/-chloroacetyl, /V-trichloroacetyl, Λ/-trifluoroacetyl, /V-phenylacetyl, /V-3-phenylpropionyl, /V-picolinoyl, Λ/-3-pyridyl- carboxamide, Λ/-benzoylphenylalanyl derivative, Λ/-benzoyl, and Λ/-p- phenylbenzoyl.
Assisted Cleavage Assisted cleavage groups include: /V-o-nitrophenylacetyl, N-o- nitrophenoxyacetyl, Λ/-acetoacetyl, (Λ/ -dithiobenzyloxycarbonylamino)acetyl, Λ/-3- (p-hydroxphenyl) propionyl, Λ/-3-(o-nitrophenyl)propionyl, Λ/-2-methyl-2-(o- nitrophenoxy)propionyl, Λ/-2-methyl-2-(o-phenylazophenoxy)propionyl, Λ/-4- chlorobutyryl, Λ/-3-methyl-3-nitrobutyryl, Λ/-o-nitrocinnamoyl, Λ/-acetylmethionine derivative, Λ/-o-nitrobenzoyl, Λ/-o-(benzoyloxymethyl)benzoyl, and 4,5-diphenyl-3- oxazolin-2-one.
Cyclic Imide Derivatives Cyclic imide derivatives include: Λ/-phthalimide, Λ/-dithiasuccinoyl, Λ/-2,3-diphenyl-maleoyl, Λ/-2,5-dimethylpyrrolyl, Λ/-1 ,1 ,4,4-tetramethyldisilylazacyclopentane adduct, 5-substituted 1 ,3-dimethyl-1 ,3,5-triazacyclohexan-2-one, 5-substituted 1 ,3-dibenzyl- 1 ,3,5-triazacyclohexan-2-one, and 1 -substituted 3,5-dinitro-4-pyridonyl.
SPECIAL -NH PROTECTIVE GROUPS
Protective groups for - NH include: Λ/-alkyl and Λ/-aryl amines, imine derivatives, enamine derivatives, and Λ/-hetero atom derivatives (such as /V-metal, N-N, N-P, N-Si, and N-S), /V-sulfenyl, and Λ/-sulfonyl.
Λ/-Alkyl and Λ/-Aryl Amines Λ/-alkyl and Λ/-aryl amines include: Λ/-methyl, Λ/-allyl, Λ/-[2-(trimethylsilyl)ethoxyl]- methyl, Λ/-3-acetoxypropyl, Λ/-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl), quaternary ammonium salts, /V-benzyl, Λ/-di(4-methoxyphenyl)methyl, Λ/-5-dibenzosuberyl, /V-triphenylmethyl, Λ/-(4-methoxyphenyl)diphenylmethyl, Λ/-9-phenylfluorenyl, Λ/-2,7-dichloro-9-fluorenylmethylene, Λ/-ferrocenylmethyl, and Λ/-2-picolylamine N '-oxide.
Imine Derivatives Imine derivatives include: Λ/-1 ,1-dimethylthiomethylene, Λ/-benzylidene, Λ/-p-methoxybenzylidene, Λ/-diphenylmethylene, Λ/-[(2-pyridyl)mesityl]methylene, N-(N ',N '-dimethylaminomethylene), N,N '-isopropylidene, Λ/-p-nitrobenzylidene, Λ/-salicylidene, Λ/-5-chlorosalicylidene, Λ/-(5-chloro-2-hydroxyphenyl)phenyl- methylene, and Λ/-cyclohexylidene. Enamine Derivative
An example of an enamine derivative is Λ/-(5,5-dimethyl-3-oxo-1-cyclohexenyl). Λ/-Hetero Atom Derivatives
/V-metal derivatives include: Λ/-borane derivatives, Λ/-diphenylborinic acid derivative, Λ/-[phenyl(pentacarbonylchromium- or -tungsten)]carbenyl, and
Λ/-copper or N-zinc chelate. Examples of N-N derivatives include: Λ/-nitro,
Λ/-nitroso, and /V-oxide. Examples of tV-P derivatives include:
Λ/-diphenylphosphinyl, Λ/-dimethylthiophosphinyl, Λ/-diphenylthiophosphinyl,
/V-dialkyl phosphoryl, Λ/-dibenzyl phosphoryl, and Λ/-diphenyl phosphoryl. Examples of Λ/-sulfenyl derivatives include: Λ/-benzenesulfenyl,
Λ/-o-nitrobenzenesulfenyl, Λ/-2,4-dinitrobenzenesulfenyl,
Λ/-pentachlorobenzenesulfenyl, Λ/-2-nitro-4-methoxy-benzenesulfenyl, N- triphenylmethylsulfenyl, and Λ/-3-nitropyridinesulfenyl.
Λ/-sulfonyl derivatives include: /V-p-toluenesulfonyl, /V-benzenesulfonyl, N- 2,3,6-trimethyl- 4-methoxybenzenesulfonyl, Λ/-2,4,6-trimethoxybenzenesulfonyl, N-
2,6-dimethyl-4-methoxy-benzenesulfonyl, Λ/-pentamethylbenzenesulfonyl, N-
2,3,5,6-tetramethyl-4-methoxybenzene- sulfonyl, Λ/-4-methoxybenzenesulfonyl, N-
2,4,6-trimethylbenzenesulfonyl, Λ/-2,6-dimethoxy- 4-methylbenzenesulfonyl, N-
2,2,5,7,8-pentamethylchroman-6-sulfonyl, Λ/-methanesulfonyl, Λ/- ?-trimethylsilylethanesulfonyl, Λ/-9-anthracenesulfonyl, N-
4-(4',8'-dimethoxynaphthylmethyl)-benzenesulfonyl, Λ/-benzylsulfonyl, N- trifluoromethylsulfonyl, and Λ/-phenacylsulfonyl.
Disclosed compounds which are masked or protected may be prodrugs, compounds metabolized or otherwise transformed in vivo to yield a disclosed compound, e.g., transiently during metabolism. This transformation may be a hydrolysis or oxidation which results from contact with a bodily fluid such as blood, or the action of acids, or liver, gastrointestinal, or other enzymes.
Features of the invention are further descπbed in the examples below. E. EXAMPLES
EXAMPLE 1
4-Fluoro-2-(4-methanesulfanyl-phenylamino)-benzoic acid (1). To a solution of 4-(methylmercapto)aniline (3.1622 g, 0.02 mole) in THF at
"78°C, a solution of LDA in THF (2M, 30 ml, 0.06 mole) was added and the reaction mixture stirred for 30 minutes at ~78°C (Scheme 1) . Solid 2,4-diflluoro benzoic acid (3.1622 g, 0.02 mole) was added and the reaction stirred for 16 hours while it wormed up to room temperature. The reaction mixture was pour in to ether saturated with HCl gas. HCl gas was bubbled into until precipitation of salts ceased. The precipitated salts were separated by filtration and discarded. The ether layer was concentrated to give 1 as a white solid. Yield 5.63 g (100%); mp 173-179 °C (DEC); 1H-NMR (400 MHz; CDCI3) ™ 9.39 (s, 1 H), 8.04 (dd, 1H, J=9.2, 6.8 Hz), 7.32-7.17 (AB quartet, 4H), 6.74 (dd, 1 H, J=12.1 , 2.4 Hz), 6.46- 6.41 (m, 1 H), 2.51 (s, 3H); 13C-NMR (100 MHz; CDCI3) ™ 172.79, 167.57 (d, JC- F=253.4 Hz), 151.55 (d, JC-F=12.2 HZ), 136.83, 135.40 (d, JC-F=12.2 HZ), 134.72, 128.31 , 124.60, 106.51 , 105.12 (d, JC-F=22.9 HZ), 99.79 (d, JC-F=26.7 HZ), 16.51 ; 19F-NMR (376 MHz; CDCI3) ™ -101.39 to -101.46 (m); MS (APCI+) 278 (M+1 , 100); IR (KBr) 3319, 1664, 1589, 1258 cm"1; Anal, calcd/found for: Cι4H12FN02S C, 60.64/60.99; H, 4.36/4.63; N, 5.05/4.80; S, 11.56/10.97.
EXAMPLE 2 4-Fluoro-2-(4-methanesulfinyl-phenylamino)-benzoic acid (2).
A mixture of 1 (Scheme 1) (0.286 g, 0.001031 mole) and oxaziridine (0.235 g, 0.0009 mole) in CHCI3 (30 ml) at room temperature for 2 hours. The solvent was removed and the resulting brown oil chromatographed on silica column. Elution with CH2CI2 removed fast moving byproduct. Further elution with CH2CI2:CH3OH (9.5:05), R = 0.27, gave pure 2 as a light brown solid. Yield 132.8 mg (50%); mp 191-192 °C; 1H-NMR (400 MHz; CDCI3) δ 9.77 (s, 1 H), 8.08 (dd, 1 H, J=8.9, 6.7 Hz), 7.70-7.39 (AB quartet, 4H), 6.98 (dd, 1 H, J=11.6, 2.4 Hz), 6.57-6.52 (m, 1H), 2.80 (s, 3H); 13C-NMR (100 MHz; CDCI3) ™ 170.76, 167.18 (d, JC-F=253.3 Hz), 149.33 (d, JC-F=12.2 Hz), 143.02, 139.50, 135.37 (d, JC-F=12.2 Hz), 125.47, 122.32, 108.22, 106.35 (d, JC-F=22.8 Hz), 100.69, (d, JC-F=25.9 Hz), 43.75; MS (APCI+) 294 (M+1 , 100); IR (KBr) 1673, 1592, 1228 cm"1; Anal, calcd/found for: Cι4H12FNO3S C, 57.33/57.48; H, 4.12/4.27; N, 4.78/4.67.
EXAMPLE 3
4-Fluoro-2-(4-methanesulfonyl-phenylamino)-benzoic acid (3).
A solution of 1 (Scheme 1) (0.4458 g, 0.00152 mole) and tetrabutylammonium oxon (1.1 g, 0.0030 mole) in CH2CI2 (20 ml) was stirred at room temperature for 16 hours. TLC showed the presence of starting material; so additional 1.1 g (0.0030 mole) of the tetrabutylammonium oxon was added and reaction mixture stirred for 16 more hours. The reaction mixture was loaded on to a silica column and eluted with CH2Cl2:CH3OH (9.75:0.25) and the fast moving fraction collected and concentrated to give 3 as a white solid. Yield, 0.3856 g (82%); mp 200-202 °C; 1H-NMR (400 MHz; CDCI3) δ 9.78 (s, 1H), 8.13 (dd, 1H, J=8.9, 6.5 Hz), 7.94-7.38 (AB quartet, 4H), 7.10 (dd, 1 H, J=11.3, 2.4 Hz), 6.66- 6.61 (m, 1 H), 3.09 (s, 3H); 13C-NMR (100 MHz; CDCI3) ™171.52, 167.28 (d, JC-F=254.9 Hz), 148.32, 145.21 , 135.59 (d, Jc-F=11.5 Hz), 134.50, 129.39, 120.62, 108.74, 107.46 (d, JC-F=22.8 HZ), 101.61 (d, JC-F=26.7 HZ), 44.78; 19F- NMR (376 MHz; CDCI3) ™ -100.29 to -100.45 (m); MS (APCI+) 310 (M+1 , 100); (APCI-) 308 (M-1 , 100); Anal, calcd/found for: C14H12FNO4SO.75 H2O C, 52.08/52.36; H, 4.22/3.88; N.4.34/4.26.
EXAMPLE 4 2-methyl-4-trimethylsilanylethvnyl-aniline (5) To a solution of 4-iodo-2-methyl-aniline (2.33g, 10 mmol), bis(triphenylphosphine)palladium(ll)chloride (1.4g, 0.2 mmol), Cul (0.19 g, 0.1 mmol) in EtsN (40 ml) at ice-bath temperature, (trimethylsilyl)acetylene (1.18 g, 12 mmol) was added dropwise (Scheme 2). After an hour stirring, the ice-bath was removed and the reaction mixture heated at 40°C (oil-bath temperature) for one hour; cooled to room temperature and the solvent removed. The residue was partitioned between H20 and Et20. The Et20 layer was separated, dried
(MgS04) and concentrated to give an oil. The oil was purified by silica column, eluting with CH2CI2. The fraction with Rf = 0.37 was collected and concentrated to give 2-methyl-4-trimethylsilanylethynyl-aniline as a dark brown oil. Yield 1.50 g (83%).
EXAMPLE 5
4-Fluoro-2-(2-methyl-4-trimethylsilanylethynyl-phenylamino)-benzoic acid (6)
Continuing after Example 4, to a solution of 2-methyl-4-trimethyl- silanylethynyl aniline (1.50 g, 0.008 mole) in THF (10 ml) at"78°C, LDA (2 M in THF, 6 ml, 0.012 mole) was added and the mixture was stirred at "78°C for 30 minutes. Solid 2,4-difluoro-benzoic acid (0.633 g, 0.004 mole) was added and the stirred for 16 hours while it warmed up to room temperature. The solvents were removed and water (30 ml) and Et20 (50 ml) added to the oil residue. The mixture was stirred vigorously and the Et2O layer separated, dried (MgSO4) and concentrated to give a brown solid. The solid was purified on silica column, eluted with CH2CI2. The fraction with Rf = 0.37 was collected and concentrated to give a light brown solid. The solid was added to pentane; some insoluble brown particulate was separated by filtration and discarded. The pentane layer was concentrated to give 6 as a light yellow solid. Yield 0.65 g (47%); mp 170-171°C; 1H-NMR (400 MHz; CDCI3) δ 9.33 (s, 1 H), 8.05 (dd, 1 H, J=8.9, 6.8 Hz), 7.43 (d, 1 H, J=1.2 Hz), 7.35 (dd, 1 H, J=8.2, 1.7 Hz), 7.25 (d, 1 H, J=8.2 Hz), 6.53 (dd, 1 H, J=11.8, 2.4 Hz), 6.47-6.42 (m, 1 H), 2.25 (s, 3H), 0.26 (s, 9H); 13C-NMR (100 MHz; CDCI3) D 172.86, 167.61 (d, JC-F=253.3), 151.24 (d, JC-F=12.3 HZ), 138.28, 135.38 (d, Jc-F=11.4 Hz), 134.85, 132.82, 130.63, 123.81 , 119.91 , 106.63, 105.23 (d, Jc-F=22.8 Hz), 104.77, 99.98 (d, JC-F=26.7 HZ), 94.05, 17.78, 0.00; MS (APCI+) 342 (M+1 , 100); IR (KBr) 2151 , 1661 , 1249 cm'1; Anal, calcd/found for: Ci9H2oFN02Si C, 66.83/67.02; H, 5.90/6.00; N, 4.10/4.09; F, 5.56/5.45.
EXAMPLE 6 4-Fluoro-2-(2-methyl-4-ethvnyl-phenylamino)-benzoic acid (7). To a solution of 6 in CH3OH (30 ml), aqueous 1 N KOH (10 ml) was added.
After stirring at room temperature for 16 hours, the CH3OH was removed and the aqueous layer was acidified with 6N HCl (Scheme 2). The resulting white precipitation was extracted in to Et2O, the Et2O layer was dried (MgS04) and concentrated to give 7 as tan colored solid. Yield 0.4274 g (91%); mp 177-178 ° C; 1H-NMR (400 MHz; CDCI3) δ 9.35 (s, 1 H), 8.08-8.04 (m, 1 H), 7.44 (s, 1 H), 7.38-7.25 (m, 2H), 6.57 (d, 1 H, J=11.8 Hz), 6.48-6.44 (m, 1 H), 3.08 (s, 1 H), 2.27 (s, 3H); 13C-NMR (100 MHz; CDCI3) ™ 172.84, 167.61 (d, JC-F=253.3), 151.15 (d, JC-F=12.3 Hz), 138.63, 135.40 (d, JC-F=12.3 HZ), 135.00, 132.87, 130.81 , 123.76, 118.79, 106.75, 105.33 (d, JC-F=22.8 HZ), 100.03 (d, JC-F=26.0 HZ), 83.37, 17.83, 0.00; 19F-NMR (376 MHz; CDCI3) ™ -101.24 to -101.31 (m); MS (APCI+) 270 (M+1 , 100); IR (KBr) 3315, 1672, 1594, 1253 cm"1; Anal, calcd/found for: Cι6H12FN02 C, 71.37/71.08; H, 4.49/4.82; N, 5.20/5.09.
EXAMPLE 7 1-(4-nitro-phenyl)-1 H-pyrrole (9a) To a gently refluxing mixture of 4-nitroaniline (6.906 g, 0.05 mole), and sodium acetate (23 g, 0.28 mole) in acetic acid (100 ml) was added 2,5- dimethoxytetrahydrofuran (7.26 g, 7.12 ml, 0.055 mole) dropwise (Scheme 3). After refluxing for 3 hours, the reaction mixture was poured on to crushed ice (~250 ml), basified with 10 % sodium hydroxide (250 ml) and extracted with CH2CI2. The CH2CI2 layer was dried (K2C03) to afford the product as a dark brown oil. Yield 9.40 g (100 %).
EXAMPLE 8 1-(4-nitro-phenyl)-1 H-pyrazole (9b) A mixture of pyrrazole (6.808 g, 0.1 mole) tetrabutylammonium bromide
(3.22 g, 0.01 mole) and KOH (11.22 g, 0.2 mole) were ground together and sonicated for 16 hours. To this 1-fluoro-4-nitrobenzene (15.521 g, 11.67 ml, 0.11 mole) was added and the mixture sonicated for 24 hours. The reaction mixture was extracted with CH2CI2. The CH2CI2 layer was dried (MgS0 ) and concentrated to give dark brown solid. This was purified by silica column chromatography. Elution with CH2CI2 (Rf = 0.44) gave the product as a light brown solid. Yield 8.80 g (47 %); mp 171-172 °C; Anal, calcd/found for: C9H7N302 C, 57.14/56.52; H, 3.73/3.62; N, 22.21/21.95.
EXAMPLE 9 3,5-dimethyl-1-(4-nitro-phenyl)-1 H-pyrazole (9c)
To a solution of 4-nitro-phenyl-hydrazine (15.3 g, 0.1 mole) and 2,4- pentanedione (10.01 g, 10.27 ml, 0.1 mole) in EtOH (200 ml) were added 5 drops of concentrated HCl. The mixture was refluxed for 15 minutes; and the solvent removed to give a gummy product. This was purified by silica column chromatography. Elution with CH2CI gave the desired product (Rf = 0.10) as a brown solid. Yield 7.22 g (33 %).
EXAMPLE 10 4-Pyrrol-1-yl-phenylamine (10a) Catalytic reduction (H2/RaNi (5 g) /THF) of 1 -(4-nitro-phenyl)- 1 H-pyrrole
(9.69 g, 0.05149 mole) at 51 psi gave crude product as an oil (Scheme 3). The product was purified by silica column chromatography. Elution with CH2CI2 (Rf = 0.13) gave the pure product as white solid. Yield 8.06 g (99 %); mp 80-81 °C.
EXAMPLE 11 In a manner similar to the preparation of 4-pyrrol-1-yl-phenylamine, the following were prepared:
4-1 H-Pyrazol-1-yl-phenylamine (10b). Dark brown oil, yield 6.26 g (100 %).
Benzenamine, 4-(3,5-dimethyl-1 H-pyrazzol-1-yl) (10c). Dark brown oil. Yield 6.45 g (100 %).
EXAMPLE 12
4-Fluoro-2-(4-pyrrol-1-yl-phenylamino)-benzoic acid (11a)
To a solution of 4-pyrrol-1-yl-phenylamine (3.16 g, 0.02 mole) in THF (30 ml) at "78°C, a solution of LDA (2M, 15 ml, 0.03 mole) was added and the mixture stirred for 30 minutes. Solid 2,4-difluorobenzoic acid was added and the reaction mixture stirred for 16 hours as it warmed up to room temperature. The solvent was removed and ether (100 ml) added to the dark oily residue. This was stirred vigorously and the insoluble gummy precipitate separated by filtration. The gamy residue was dissolved in H2O, acidified to pH 1 with 10% HCl, and extracted with Et2O. The Et20 layer was dried (MgSO4) and concentrated to give the target compound as a brown solid. Yield 2.74 g (93 %); mp 223-225 °C (DEC); 19F-NMR (376 MHz; CDCI3) δ -101.44 (s); MS (APCI+) 297 (M+1 , 100); IR (KBr) 1658, 1526, 1254 cm"1.
In a manner similar to the preparation of 4-Fluoro-2-(4-pyrrol-1-yl- phenylamino)-benzoic acid, the following were prepared:
4-Fluoro-2-(4-pyrazol-1-yl-phenylamino)-benzoic acid (11b). Light brown solid, mp 212-213 °C.
2-r4-(3,5-Dimethyl-pyrazol-1-yl)-phenylamino1- 4-Fluoro benzoic acid (11c). Tan powder, mp 198 -200 °C.
EXAMPLE 13 Cascade assay for inhibitors of the MAP kinase pathway
Incorporation of 32P into myelin basic protein (MBP) is assayed in the presence of a glutathione S-transferase fusion protein containing p44MAP kinase (GST-MAPK) and a glutathione S-transferase fusion protein containing p45MEK (GST-MEK). The assay solution contains 20 mM HEPES, pH 7.4, 10 mM MgCI2, 1 mM MnCI2, 1 mM EGTA, 50,μM [γ-32P]ATP, 10 μg GST-MEK, 0.5 μg GST-MAPK and 40 μg MBP in a final volume of 100 μL. Reactions are stopped after 20 minutes by addition of trichloroacetic acid and filtered through a GF/C filter mat. 32P retained on the filter mat is determined using a 120S Betaplate. Compounds are assessed at 10 μM for ability to inhibit incorporation of 32P. To ascertain whether compounds are inhibiting GST-MEK or GST MAPK, two additional protocols are employed. In the first protocol, compounds are added to tubes containing GST-MEK, followed by addition of GST-MAPK, MBP and [γ-32P]ATP. In the second protocol, compounds are added to tubes containing both GST-MEK and GST-MAPK, followed by MBP and [γ-32P]ATP. Compounds that show activity in both protocols are scored as MAPK inhibitors, while compounds showing activity in only the first protocol are scored as MEK inhibitors.
EXAMPLE 14
In vitro MAP kinase assay
Inhibitory activity can be confirmed in direct assays. For MAP kinase, 1 μg GST-MAPK is incubated with 40 μg MBP for 15 minutes at 30°C in a final volume of 50 μL containing 50 mM Tris (pH 7.5), 10 μM MgC12, 2 μM EGTA, and 10 μM [γ-32P]ATP. The reaction is stopped by addition of Laemmli SDS sample buffer and phosphorylated MBP resolved by electrophoresis on a 10% polyacrylamide gel. Radioactivity incorporated into MBP is determined by both autoradiography, and scintillation counting of excised bands.
EXAMPLE 15
In vitro MEK assay
For evaluation of direct MEK activity, 10 μg GST-MEKi is incubated with 5 μg of a glutathione S-transferase fusion protein containing p44MAP kinase with a lysine to alanine mutation at position 71 (GST-MAPK-KA). This mutation eliminates kinase activity of MAPK, so only kinase activity attributed to the added MEK remains. Incubations are 15 minutes at 30°C in a final volume of 50 μL containing 50 mM Tris (pH 7.5), 10 μM MgCI2, 2 , μM EGTA, and 10 μM [γ-32P]ATP. The reaction is stopped by addition of Laemmli SDS sample buffer. Phosphorylated GST-MAPK-KA is resolved by electrophoresis on a 10% polyacrylamide gel. Radioactivity incorporated into GST-MAPK-KA is determined by autoradiography, and subsequent scintillation counting of excised bands. Additionally, an artificially activated MEK containing serine to glutamate mutations at positions 218 and 222 (GST-MEK-2E) is used. When these two sites are phosphorylated, MEK activity is increased. Phosphorylation of these sites can be mimicked by mutation of the serine residues to glutamate. For this assay, 5 μg GST-MEK-2E is incubated with 5 μg GST-MAPK-KA for 15 minutes at 30°C in the same reaction buffer as described above. Reactions are terminated and analyzed as above.
EXAMPLE 16 Whole cell MAP kinase assay
To determine if compounds block activation of MAP kinase in whole cells, the following protocol is used. Cells are plated in multi-well plates and grown to confluence. Cells are serum-deprived overnight. Cells are exposed to the desired concentrations of compound or vehicle (DMSO) for 30 minutes, followed by addition of a growth factor, for example, PDGF (100 ng/mL). After a 5-minute treatment with the growth factor, cells are washed with PBS, and lysed in a buffer consisting of 70 mM NaCI, 10 mM HEPES (pH 7.4), 50 mM glycerol phosphate, and 1 % Triton X-100. Lysates are clarified by centrifugation at 13,000 x g for 10 minutes. Five to fifteen micrograms of protein from the resulting supematants are subjected to SDS/PAGE and Western blotting for phosphorylated MAP kinase levels.
EXAMPLE 17 Monolaver growth Cells are plated into multi-well plates at 10 to 20,000 cells/mL. Forty-eight hours after seeding, test compounds are added to the cell growth medium and incubation is continued for 2 additional days. Cells are then removed from the wells by incubation with trypsin and enumerated with a Coulter counter. EXAMPLE 18 Growth in soft-agar
Cells are seeded into 35-mm dishes at 5 to 10,000 cells/dish using growth medium containing 0.3% agar. After chilling to solidify the agar, cells are transferred to a 37°C incubator. After 7 to 10 days' growth, visible colonies are manually enumerated with the aid of a dissecting microscope.
EXAMPLE 19 Collagen-Induced Arthritis in Mice Type II collagen-induced arthritis (CIA) in mice is an experimental model of arthritis that has a number of pathologic, immunologic, and genetic features in common with rheumatoid arthritis. The disease is induced by immunization of DBA/1 mice with 100 μg type II collagen, which is a major component of joint cartilage, delivered intradermally in Freund's complete adjuvant. The disease susceptibility is regulated by the class II MHC gene locus, which is analogous to the association of rheumatoid arthritis with HLA-DR4.
A progressive and inflammatory arthritis develops in the majority of mice immunized, characterized by paw width increases of up to 100%. A test compound is administered to mice in a range of amounts, such as 20, 60, 100, and 200 mg/kg body weight/day. The duration of the test can be several weeks to a few months, such as 40, 60, or 80 days. A clinical scoring index is used to assess disease progression from erythema and edema (stage 1), joint distortion (stage 2), to joint ankylosis (stage 3). The disease is variable in that it can affect one or all paws in an animal, resulting in a total possible score of 12 for each mouse. Histopathology of an arthritic joint reveals synovitis, pannus formation, and cartilage and bone erosions. All mouse strains that are susceptible to CIA are high antibody responders to type II collagen, and there is a marked cellular response to CH. EXAMPLE 20 SCW-induced monoarticular arthritis
Arthritis is induced as described by Schwab, et al., Infection and Immunity, 59:4436-4442 (1991) with minor modifications. Rats receive 6 μg sonicated SCW [in 10 μl Dulbecco's PBS (DPBS)] by an intraarticular injection into the right tibiotalar joint on day 0. On day 21 , the DTH is initiated with 100 μg of SCW (250 μl) administered i.v. For oral compound studies, compounds are suspended in vehicle (0.5% hydroxypropyi-methylcellulose/0.2% Tween 80), sonicated, and administered twice daily (10 ml/kg volume) beginning 1 hr prior to reactivation with SCW. Compounds are administered in amounts between 10 and 500 mg/kg body weight/day, such as 20, 30, 60, 100, 200, and 300 mg/kg/day. Edema measurements are obtained by determining the baseline volumes of the sensitized hindpaw before reactivation on day 21 , and comparing them with volumes at subsequent time points such as day 22, 23, 24, and 25. Paw volume is determined by mercury plethysmography.
EXAMPLE 21 Mouse ear-heart transplant model
Fey, T.A. et al. describe methods for transplanting split-heart neonatal cardiac grafts into the ear pinna of mice and rats (J. Pharm. and Toxic. Meth. 39:9-17 (1998)). Compounds are dissolved in solutions containing combinations of absolute ethanol, 0.2% hydroxypropyl methylcellulose in water, propylene glycol, cremophor, and dextrose, or other solvent or suspending vehicle. Mice are dosed orally or intraperitoneally once, twice or three times daily from the day of transplant (day 0) through day 13 or until grafts have been rejected. Rats are dosed once, twice, or three times daily from day 0 through day 13. Each animal is anesthetized and an incision is made at the base of the recipient ear, cutting only the dorsal epidermis and dermis. The incision is spread open and down to the cartilage parallel to the head, and sufficiently wide to accommodate the appropriate tunneling for a rat or insertion tool for a mouse. A neonatal mouse or rat pup less than 60 hours old is anesthetized and cervically dislocated. The heart is removed from the chest, rinsed with saline, bisected longitudinally with a scalpel, and rinsed with sterile saline. The donor heart fragment is placed into the preformed tunnel with the insertion tool and air or residual fluid is gently expressed from the tunnel with light pressure. No suturing, adhesive bonding, bandaging, or treatment with antibiotics is required.
Implants are examined at 10-20-fold magnification with a stereoscopic dissecting microscope without anesthesia. Recipients whose grafts are not visibly beating may be anesthetized and evaluated for the presence of electrical activity using Grass E-2 platinum subdermal pin microelectodes placed either in the pinna or directly into the graft and a tachograph. Implants can be examined 1-4 times a day for 10, 20, 30 or more days. The ability of a test compound to ameliorate symptoms of transplant rejection can be compared with a control compound such as cyciosporine, tacrolimus, or orally-administered lefluonomide.
EXAMPLE 22
Murine ovalbumin-induced eosinophilia
Female C57BLJ6 mice are obtained from the Jackson Laboratory (Bar Harbor, ME). All animals are given food and water ad libitum. Mice are sensitized with a single i.p. injection of OVA (grade V, Sigma Chemical Company, St. Louis, MO) adsorbed to alum, (10 μg OVA + 9 mg alum in 200 μl saline) or vehicle control, (9 mg alum in 200 μl saline) on day 0. On day 14, the mice are challenged with a 12-minute inhalation of an aerosol consisting of 1.5% OVA (weight/volume) in saline produced by a nebulizer (small particle generator, model SPAG-2; ICN Pharmaceuticals, Costa Mesa, CA). Groups of eight mice are dosed with oral vehicle (0.5% hydroxypropylmethylcellulose / 0.25% TWEEN- 80), or a test compound at 10, 30, or 100 mg/kg in oral vehicle, 200 μl per mouse p.o. Dosing is performed once per day starting on day 7 or day 13, and extending through day 16. For determination of pulmonary eosinophilia, three days after the first
OVA aerosol challenge (day 17), the mice are anesthetized with an i.p. injection of anesthetic (Ketamine/Acepromazine/Xylazine), and the tracheae is exposed and cannulated. The lungs and upper airways are lavaged twice with 0.5 ml of cold PBS. A portion (200 μl) of the bronchoalveolar lavage (BAL) fluid is enumerated using a Coulter counter Model ZB1 (Coulter Electronics, Hialeah, FL). The remaining BAL fluid is then centrifuged at 300 x g for five minutes, and the cells are resuspended in 1 ml of HBSS (Gibco BRL) containing 0.5% fetal calf serum (HyClone) and 10 mM HEPES (Gibco BRL). The cell suspension is centrifuged in a cytospin (Shandon Southern Instruments, Sewickley, PA) and stained by Diff Quick (American Scientific Products, McGraw Park, IL) to differentiate BAL leukocytes into neutrophil, eosinophil, monocyte or lymphocyte subsets. The number of eosinophils in the BAL fluid is determined by multiplying the percentage of eosinophils by the total cell count.
EXAMPLE 23
Caco-2 cell studies
Cell transport studies were conducted with Caco-2 cells grown on
Snapwells between 22 to 28 days postseeding. Typically, 10 mM MES buffer (pH
6.5) with 5 mM KCI, 135 mM NaCI and 1.8 mM CaCI2 was used for the apical side and 10 mM MOPS (pH 7.4) with 5 mM KCI, 132.5 mM NaCI and 1.8 mM CaCI2 with 5 mM D-Glucose was used for the basolateral side. After washing the monolayers, appropriate buffers were pipetted into the respective chambers and the cells were pre-equilibrated at 37°C for at least 15 min. On the day of the experiment the growth media was aspirated and the cell monolayers were preequilibrated with appropriate buffers at 37°C for at least 15 min. Thereafter, TEER measurements were performed to confirm the integrity of the monolayers. Transepithelial flux measurements were made by mounting the cell monolayers in a side-by-side diffusion chamber system (Precision Instrument Design, Tahoe City, CA). Temperature was maintained at 37°C with a circulating water jacket. The solutions were mixed with gas-lift circulation with 95% oxygen-5% carbon dioxide. Donor solutions with PD compounds, [14C] mannitol (leakage marker) and [3H] metoprolol (reference compound) were added to the apical chamber. Donor and receiver samples were collected at selected time intervals for up to 3 hours. Radiolabelled mannitol and metoprolol were analyzed using scintillation counting (TopCount, Packard Instruments, Downers Grove, IL). PD compounds were analyzed using a LC/MS/MS method. Apparent permeability coefficients were calculated using the following equation: Papp = (V* dC) / (A.Co. dt) where V= volume of the receiver solution in ml, A = surface area in cm2, C0 = initial donor concentration in mM and dC/dt = change in the drug concentration in the receiver chamber over time.
EXAMPLE 24
Metabolic Stability in Human and Rat Liver Microsomes
Compounds are individually incubated (5 μM, dissolved in DMSO) with human and rat liver microsomes (0.5 mg/mL protein) in 50 mM KHPO4 buffer at 37°C in the presence of 1.0 mM NADPH. At 0, 10, 20 and 40 minutes, 100 μL aliquots are removed and added to 300 μL of acetonitrile. Standard curves are run in a similar manner with each compound at concentrations: 7.5 μM, 3.75 μM, 2.5 μM, 1.25 μM. The samples are analyzed for parent concentration by LC/MS/MS. The in vitro metabolic half-life determinations are determined from the concentration vs. time plots using WinNonlin. These in vitro data represent the rate of oxidative and hydrolytic metabolism.
EXAMPLE 25
Preparation of 2-(2-Chloro-4-iodo-phenylamino)-Λ/-cvclopropylmethoxy-3,4- difluoro-benzenesulfonamide (PD 297447)
Step a: Preparation of Λ/-Cvclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide To a stirring suspension comprised of O-cyclopropylmethyl-hydroxylamine hydrochloride (5.40 g, 43.7 x 10"3 mol) in dichloromethane (20 ml) at ambient temperature under a nitrogen atmosphere was added directly diisopropylethylamine (10.8 ml, 0.062 mol). A solution comprised of 2,3,4- trifluorobenzenesulfonyl chloride (1.00 g, 4.34 x 10"3 mol) in dichloromethane (120 ml) was added as a slow steady stream over twelve minutes to the first solution. After twelve minutes of stirring the combined reaction mixture, a 10 % aqueous hydrochloric acid solution (140 ml) was added. The biphasic mixture was stirred vigorously and the layers were separated. The organic phase was dried (MgSO4) and concentrated to 6 ml volume. The concentrate was applied to a flash silica column (90 g of silica). Elution with dichloromethane afforded 0.83 g of the white amorphous solid product; 68 % yield; 1H-NMR (400 MHz; CDCI3) δ 7.50 (m, 1 H), 7.10 (s, 1 H), 6.95 (m, 1 H), 3.59 (d, 2H, J=7.2 Hz), 0.80 (m, 1 H), 0.31 (m, 2H), 0.02 (m, 2H); 19F-NMR (376 MHz; CDCI3) δ -122.65 (m, 1F), -
129.37 (m, 1 F), -156.20 (m, 1 F); MS (APCI-) 280 (M-1 , 100), 210 (55), 195 (45).
Step b: Preparation of 2-(2-Chloro-4-iodo-phenylamino)-Λ/-cvclopropylmethoχy- 3,4-difluoro-benzenesulfonamide To a stirring solution comprised of 2-chloro-4-iodoaniline (0.80 g, 3.2 x 10"3 mol) in tetrahydrofuran (10 ml) at -78 °C under a nitrogen atmosphere was added 1.0 molar lithium b/'strimethylsilylamide solution in tetrahydrofuran (6.2 ml, 6.2 x 10"3 mol) to form a green suspension. The suspension was stirred for five minutes before a stirring suspension comprised of lithiated N- cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide [prepared by adding 3.0 ml of the 1.0 molar lithium D/'strimethylsilylamide solution to a stirring solution comprised of 0.83 g (2.95 x 10"3 mol) of Λ/-cyclopropylmethoxy-2,3,4-trifluoro- benzenesulfonamide in 10 ml of tetrahydrofuran at -78 °C] was added. The cold bath was removed and the reaction mixture was stirred for one hour. Aqueous (10 %) hydrochloric acid (50 ml) was added to the reaction mixture, and the biphasic mixture was concentrated in vacuo to an aqueous suspension that was extracted with diethyl ether (200 ml). The ether phase was dried (MgSO4) and concentrated in vacuo to afford 2 g of a tan oil. The crude product was purified by flash silica chromatography. Elution with a gradient [99:1 hexanes-ethyl acetate -» (2 min) 9:1 -> (25 min) 3:1] afforded 1.10 g of a clear amorphous foam; 73 % yield; 1H-NMR (400 MHz; DMSO) δ 7.69 (m, 1 H), 7.59 (d, 1 H, J=1.9 Hz), 7.34 (dd, 1H, J=8.7, 1.9 Hz), 7.27 (s, 1 H), 7.00 (s, 1H), 6.95 (m, 1H), 6.43 (dd, 1 H, J=8.7, 5.8 Hz), 3.52 (d, 2H, J=7.5 Hz), 0.74 (m, 1 H), 0.34 (m, 2H), 0.02 (m, 2H); 19F-NMR (376 MHz; CDCI3) δ -124.76 (m, 1 F), -136.69 (d, 1 F, J=18.3 Hz); MS (APCI+) 515 (M+1 , 100); (APCI-) 513 (M-1 , 50), 443 (73), 428 (100); IR (KBr) 1491 cm"1; Anal. Calcd/found for C16H14CIF2lN2O3S C, 37.34/36.54; H, 2.74/2.71 ; N, 5.44/5.15; F, 7.38/7.57.
F. OTHER EMBODIMENTS
From the above disclosure and examples, and from the claims below, the essential features of the invention are readily apparent. The scope of the invention also encompasses various modifications and adaptations within the knowledge of a person of ordinary skill. Examples include a disclosed compound modified by addition or removal of a protecting group, or an ester, pharmaceutical salt, hydrate, acid, or amide of a disclosed compound. Publications cited herein are hereby incorporated by reference in their entirety.
What is claimed is:

Claims

A compound of formula (I):
(I)
wherein
W is ORi, NR2OR!, NRARB, NR2NRARB, O(CH2)ι^NRARB, or NR2(CH2)ι-4 NRARB; 0(CH2)i-4ORi, or NR2(CH2)ι-4 θRι;
Ri is H, C 1-8 alkyl, C 3-e alkenyl, C 3-8 alkynyl, C 3.8 cycloalkyl, phenyl, (phenyl)C 1-4 alkyl, (phenyl)C 3-4 alkenyl, (phenyl)C 3- alkynyl, (C 3.8 cycloalkyl)- C 1-4 alkyl, (C 3.8 cycloalkyl)C 3-4 alkenyl, (C 3.8 cycloalkyl)C 3-4 alkynyl, C 3-8 heterocyclic radical, (C 3.8 heterocyclic radical)C 1-4 alkyl, (C 3-8 heterocyclic radical)C 3-4 alkenyl, or (C 3.8 heterocyclic radical)C 3.4 alkynyl;
each of R2 and R3 is independently H, phenyl, C 1-4 alkyl, C 3.8 alkynyl, C 3-8 cycloalkyl, or (C 3-8 cycloalkyl)C 1-4 alkyl;
each of R4, R5 and R6 is independently H, CI, F, or Br; RA is H, C 1-6 alkyl, C 3.8 alkenyl, C 3-β alkynyl, C 3.8 cycloalkyl, phenyl, (C 3-β cycloalkyl)C 1-4 alkyl, (C 3.8 cycloalkyl)C 3.4 alkenyl, (C 3-8 cycloalkyl)C 3-4 alkynyl, C 3-8 heterocyclic radical, (C 3-8 heterocyclic radical)C 1-4 alkyl, (aminosulfonyl)phenyl, [(aminosulfonyl)phenyl]C 1-4 alkyl, (aminosulfonyl)C 1-6 alkyl, (aminosulfonyl)C 3^ cycloalkyl, or [(aminosulfonyl)C 3.6 cycloalkyljC 1-4 alkyl;
RB is H, C 1-8 alkyl, C 3.8 alkenyl, C 3.8 alkynyl, C 3.8 cycloalkyl, or phenyl;
J is SRc, ORc, SO2Rc, SORc, SO2NRDRE, C ι.8 alkyl, C 3-8 alkenyl, C 3-8 alkynyl, C 3-8 cycloalkyl, C 5.8 cycloalkenyl, phenyl, (C 3.3 cycloalkyl)C 1. alkyl,
(C 3-8 cycloalkyl)C 3- alkenyl, (C 3-8 cycloalkyl)C 3.4 alkynyl, C 3-8 heterocyclic radical, (C 3.8 heterocyclic radical)C 1.4 alkyl, -M'E'G', (heterocyclic radical)-M'-E'- G', or (cycloalkyl)-M'-E'-G';
M' is O, SO, SO2, NRE, (CO)NRE, NRE (CO), SO2NRE, NRESO , or CH2;
E' is absent (a covalent bond), (CH2)ι-4 or (CH2)m O(CH )p where 1 < (each of m and p independently) < 3 and 2 < (m + p) < 4;
G' is OR3, S02Rc, or NRFRG; provided that where p = 1 , then G' is H;
each of Rc, RD, RE, RF and RG is independently selected from H, C 1.6 alkyl, C 3-4 alkenyl, C 3.4 alkynyl, C 3-6 cycloalkyl, C 3-6 heterocyclic radical, and phenyl; NR RG and NRDRE can each also independently be selected from morpholinyl, pyrazinyl, piperazinyl, pyrrolidinyl, or piperadinyl;
R10 is H, C - alkyl, halo, NO2, or SO2NRHRι; and
R11 is H, halo, or NO2; wherein each hydrocarbon radical or heterocyclic radical above is optionally substituted with between 1 and 3 substituents independently selected from halo, C ι_ alkyl, C 3-6 cycloalkyl, C 2-A alkenyl, C 2-4 alkynyl, phenyl, hydroxy, amino, (amino)sulfonyl, and NO2, wherein each substituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substituted with between 1 and 3 substituents independently selected from halo, C ι-2 alkyl, hydroxy, amino, and NO2;
or a pharmaceutically acceptable salt or C 1-7 ester thereof.
2. A compound of claim 1 , wherein Rc is C 1-2 alkyl.
3. A compound of claim 1 , wherein W is OH.
4. A compound of claim 1 , wherein W is NHOH.
5. A compound of claim 1 , wherein W is NHO(cyclopropylmethyl).
6. A compound of claim 1 , wherein Rio is methyl or chloro.
7. A compound of claim 1 , where Rn is fluoro.
8. A compound of claim 1 , where Rn is H.
9. A compound of claim 1 , wherein J is trihalomethyl or methylthio.
10. A compound of claim 1 , wherein J is 1 ,2,5-thiadiazol-3-yl.
11. A compound of claim 1 , wherein J is S02CH3
12. A compound of claim 1 , wherein J is SOCH3
13. A compound of claim 1 , wherein J is C 2-B alkynyl where the triple bond is between the carbon atoms alpha and beta to the phenyl group.
14. A compound of claim 1 , wherein Ri has at least one hydroxy substituent.
15. A compound of claim 1 , wherein Ri is H, methyl, ethyl, propyl, isopropyl, isobutyl, benzyl, phenethyl, allyl, C 3.5 alkenyl, C 3-5 alkynyl,
C 3-6 cycloalkyl, (C 3.5 cycloalkyl)C 1-2 alkyl, or (C 3-5 heterocyclic radical)- C 1-2 alkyl.
16. A compound of claim 15, wherein Ri is H or (C 3-4 cycloalkyl)- C 1-2 alkyl.
17. A compound of claim 1 , wherein R2 is H, methyl, C 3.4 alkynyl, C 3-5 cycloalkyl, or (C 3.5 cycloalkyl)methyl.
18. A compound of claim 1 , wherein RA is H, methyl, ethyl, isobutyl, hydroxyethyl, hydroxypropyl, cyclopropylmethyl, cyclobutylmethyl, C 2-4 alkynyl, phenyl, 2-piperidin-1 -yl-ethyl, 2,3-dihydroxy-propyl, 3-[4-(2-hydroxyethyl)- piperazin-1-yl]-propyl, 2-pyrrolidin-1 -yl-ethyl, or 2-diethylamino-ethyl; and RB is H; or where RB is methyl and RA is phenyl.
19. A compound of claim 1 , wherein each of R and R6 is H, and R5 is F.
20. A compound of claim 1 , wherein each of R4, R5, and Re is F.
21. A compound of claim 1 , wherein each of R4 and R5 is F and R6 is Br.
22. A compound of claim 1 , wherein R5 is F.
23. A compound of claim 1, having the structure: 4-fluoro-2-(2-methyl- 4-methylsulfanyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2-(2-methyl-4- methylsulfanyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4-methanesulfinyl-2- methyl-phenylamino)-benzoic acid; 2-(4-methanesulfinyl-2-methyl-phenylamino)- 4-nitro-benzoic acid; 3,4,5-trifluoro-2-(4-methanesulfonyl-2-methyl-phenylamino)- benzoic acid; 3,4-difluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 2-(2-methyl-4-methylsulfanyl-phenylamino)-4-nitro-benzoic acid; 3,4,5-trifluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 4-fluoro- 2-(4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro- 2-(4-methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 3,4,5-trifluoro-2- (2-methyl-4-methylsulfanyl-phenylamino)-benzoic acid; 4-fluoro-2-(4-methane- sulfinyl-2-methyl-phenylamino)-benzoic acid; 5-bromo-3,4-difluoro-2- (4-methanesulfinyl-2-methyl-phenylamino)-benzoic acid; 3,4-difluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzoic acid; 2-(4-methanesulfonyl-2- methyl-phenylamino)-4-nitro-benzoic acid; N-cyclopropylmethoxy~4-fluoro-2-(2- methyl-4-methylsulfanyl-phenylamino)-benzamide; 5-bromo-N- cyclopropylmethoxy-3,4-difluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)- benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4-methanesulfinyl-2-methyl- phenylamino)-benzamide; N-cyclopropylmethoxy-2-(4-methanesulfinyl-2-methyl- phenylamino)-4-nitro-benzamide; N-cyclopropylmethoxy-3,4, 5-trifluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4- difluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzamide; N- cyclopropylmethoxy-2-(2-methyl-4-methylsulfanyl-phenylamino)-4-nitro- benzamide; N-cyclopropylmethoxy-3,4, 5-trifluoro-2-(4-methanesulfinyl-2-methyl- phenylamino)-benzamide; N-cyclopropylmethoxy-4-fluoro-2-(4-methanesulfιnyl-2- methyl-phenylamino)-benzamide; 5-bromo-N-cyclopropylmethoxy-3,4-difluoro-2- (4-methanesulfonyl-2-methyl-phenylamino)-benzamide; N-cyclopropylmethoxy- 3,4, 5-trifluoro-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzamide; N- cyclopropylmethoxy-4-fluoro-2-(4-methanesulfinyl-2-methyl-phenylamino)- benzamide; 5-bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(4-methanesulfinyl-2- methyl-phenylamino)-benzamide; N-cyclopropylmethoxy-3,4-difluoro-2-(4- methanesulfonyl-2-methyl-phenylamino)-benzamide; or N-cyclopropylmethoxy-2- (4-methanesulfonyl-2-methyl-phenylamino)-4-nitro-benzamide.
24. A compound of claim 1 , having the structure: 4-fluoro-N-hydroxy-2- (2-methyl-4-methylsulfanyl-phenylamino)-benzamide; 5-bromo-3,4-difluoro-N- hydroxy-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzamide; 3,4-difluoro-N- hydroxy-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzamide; N-hydroxy-2- (4-methanesulfinyl-2-methyl-phenylamino)-4-nitro-benzamide; 3,4,5-trifluoro-N- hydroxy-2-(4-methanesulfonyl-2-methyl-phenylamino)-benzamide; 3,4-difluoro-N- hydroxy-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzamide; N-hydroxy-2-(2- methyl-4-methylsulfanyl-phenylamino)-4-nitro-benzamide; 8: 3,4,5-trifluoro-N- hydroxy-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzamide; 4-fluoro-N- hydroxy-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzamide; 5-bromo-3,4- difluoro-N-hydroxy-2-(4-methanesulfonyl-2-methyl-phenylamino)-benzamide; 3,4,5-trifluoro-N-hydroxy-2-(2-methyl-4-methylsulfanyl-phenylamino)-benzamide; 4-fluoro-N-hydroxy-2-(4-methanesulfinyl-2-methyl-phenylamino)-benzamide; 5- bromo-3,4-difluoro-N-hydroxy-2-(4-methanesulfinyl-2-methyl-phenylamino)- benzamide; 3,4-difluoro-N-hydroxy-2-(4-methanesulfonyl-2-methyl-phenylamino)- benzamide; or N-hydroxy-2-(4-methanesulfonyl-2-methyl-phenylamino)-4-nitro- benzamide.
25. A compound of claim 1 , having the structure: 3,4-difluoro-2-(4- imidazol-1-yl-2-methyl-phenylamino)-benzoic acid; N-cyclopropylmethoxy-3,4- difluoro-2-(4-imidazol-1-yl-2-methyl-phenylamino)-benzamide; 3,4-difluoro-N- hydroxy-2-(4-imidazol-1-yl-2-methyl-phenylamino)-benzamide; 3,4,5-trifluoro-2- (2-methyl-4-[1 ,2,5]thiadiazol-3-yl-phenylamino)-benzoic acid; N- cyclopropylmethoxy-3,4,5-trifluoro-2-(2-methyl-4-[1 ,2,5]thiadiazol-3-yl- phenylamino)-benzamide; 3,4,5-trifluoro-N-hydroxy-2-(2-methyl-4- [1 ,2,5]thiadiazol-3-yl-phenylamino)-benzamide; 2-[4-(4-chloro-[1 ,2,5]thiadiazol-3- yl)-2-methyl-phenylamino]-3,4,5-trifluoro-benzoic acid; 2-[4-(4-chloro-
[1 ,2,5]thiadiazol-3-yl)-2-methyl-phenylamino]-N-cyclopropylmethoxy-3,4,5- trifluoro-benzamide; 2-[4-(4-chloro-[1,2,5]thiadiazol-3-yl)-2-methyl-phenylamino]- 3,4,5-trifluoro-N-hydroxy-benzamide; 2-{4-[4-(2-dimethylamino-ethoxy)- [1 ,2,5]thiadiazol-3-yl]-2-methyl-phenylamino}-3,4,5-trifluoro-benzoic acid; N- cyclopropylmethoxy-3,4,5-trifiuoro-2-{2-methyl-4-[4-(2-piperidin-1-yl-ethoxy)- [1 ,2,5]thiadiazol-3-yl]-phenylamino}-benzamide; or 3,4,5-trifluoro-N-hydroxy-2-{2- methyl-4-[4-(2-morpholin-4-yl-ethoxy)-[1 ,2,5]thiadiazol-3-yl]-phenylamino}- benzamide.
26. The compound of claim 1 , having a structure selected from: 5-bromo-2-(2-chloro-4-methylsulfanyl-phenylamino)-3,4-difluoro-benzoic acid; 2- (2-chloro-4-methanesulfinyl-phenylamino)-3,4-difluoro-benzoic acid;
2-(2-chloro-4-methanesulfonyl-phenylamino)-3,4,5-trifluoro-benzoic acid; 2-(2-chloro-methylsulfanyl-phenylamino)-3,4-difluoro-benzoic acid; 5-bromo-2-(2-chloro-4-methanesulfonyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-Chloro-4-methanesulfonyl-phenylamino)-3,4-difluoro-benzoic acid; 5-bromo-2-(2-chloro-4-methylsulfanyl-phenylamino)-N-cyclopropylmethoxy-3,4- difluoro-benzamide; 2-(2-chloro-4-methanesulfinyl-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-methanesulfonyl- phenylamino)- N-cyclopropylmethoxy-3,4, 5-trifluoro-benzamide; 2-(2-chloro-4- methylsulfanyl-phenylamino)- N-cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-methanesulfinyl-phenylamino)- N-cyclopropylmethoxy-3,4,5- trifluoro-benzamide; 5-bromo-2-(2-chloro-4-methanesulfonyl-phenylamino)-N- cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-chloro-4-methylsulfanyl- phenylamino)-N-cyclopropylmethoxy-3,4, 5-trifluoro-benzamide; 2-(2-chloro-4- methanesulfonyl-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide; 2-[2-chloro 4-(3H-imidazol-1-yl)-phenylamino]-N-cyclopropylmethoxy-3,4-difluoro- benzamide; 2-(2-chloro-4-[1 ,2,5]thiadiazol-3-yl-phenylamino)-N- cyclopropylmethoxy-3,4, 5-trifluoro-benzamide; 2-[4-(2-chloro-4-chloro- [1 ,2,5]thiadiazol-3-yl)-phenylamino]-3,4,5-trifluoro-benzoic acid; 2-[2-chloro-4-(4- chloro-[1 ,2,5]thiadiazol-3-yl)-phenylamino]-N-cyclopropylmethoxy-3,4,5-trifluoro- benzamide; 2-{4-[4-(2-dimethylamino-ethoxy)-[1 ,2,5]thiadiazol-3-yl]-2-methyl- phenylamino}-3,4,5-trifluoro-benzoic acid; 2-{2-chloro-4-[4-(2-piperidin-1-yl- ethoxy)-[1 ,2,5]thiadiazol-3-yl]-phenylamino}-N-cyclopropylmethoxy-3,4,5-trifluoro- benzamide.
27. The compound of claim 1 , having a structure selected from: 2-(4-Ethynyl-2-methyl-phenylamino)-4-fluoro-benzoic acid; 5-Bromo-2-(4-ethynyl- 2-methyl-phenylamino)-3,4-difluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4- ethynyl-2-methyl-phenylamino)-3,4-difluoro-benzamide; N-Cyclopropylmethoxy-2- (4-ethynyl-2-methyl-phenylamino)-4-nitro-Benzamide; 2-(4-Ethynyl-2-methyl- phenylamino)-3,4,5-trifluoro-N-hydroxy-benzamide; 2-(4-Ethynyl-2-methyl- phenylamino)-3,4-difluoro-benzoic acid; 2-(4-Ethynyl-2-methyl-phenylamino)-4- nitro-benzoic acid; N-Cyclopropylmethoxy-2-(4-ethynyl-2-methyl-phenylamino)- 3,4, 5-trifluoro-benzamide; 4-Fluoro-N-hydroxy-2-(4-methanesulfinyl-2-methyl- phenylamino)-benzamide; 5-Bromo-2-(4-ethynyl-2-methyl-phenylamino)-3,4- difluoro-N-hydroxy-benzamide; 2-(4-Ethynyl-2-methyl-phenylamino)-3,4,5- trifluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4-ethynyl-2-methyl- phenylamino)-4-fluoro-benzamide; 5-Bromo-N-cyclopropylmethoxy-2-(4-ethynyl- 2-methyl-phenylamino)-3,4-difluoro-benzamide; 2-(4-Ethynyl-2-methyl- phenylamino)-3,4-difluoro-N-hydroxy-benzamide; 2-(4-Ethynyl-2-methyl- phenylamino)-N-hydroxy-4-nitro-benzamide; 2-(4-Ethynyl-2-methyl-phenylamino)- 4-fluoro-benzoic acid; N-Cyclopropylmethoxy-2-(4-ethynyl-2-methyl- phenylamino)-4-fluoro-benzamide; and 4-Fluoro-N-hydroxy-2-(4-methanesulfιnyl- 2-methyl-phenylamino)-benzamide.
28. The compound of claim 1 , having a structure selected from: 2-(2-Chloro-4-ethynyl-phenylamino)-4-fluoro-benzoic acid; 5-Bromo-2-(2-chloro- 4-ethynyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(2-Chloro-4-ethynyl- phenylamino)- N-cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(2-Chloro-4- ethynyl-phenylamino)- N-cyclopropylmethoxy-4-nitro-benzamide; 2-(2-Chloro-4- ethynyl-phenylamino)- N-hydroxy-3,4,5-trifluoro- benzamide; 2-(2-Chloro-4- ethynyl-phenylamino)-3,4-difluoro-benzoic acid; 2-(4-Ethynyl-2-chloro- phenylamino)-4-nitro-benzoic acid; 2-(2-Chloro-4-ethynyl-phenylamino)- N- Cyclopropylmethoxy-3,4, 5-trifluoro-benzamide; 2-(2-chloro-4-methanesulfinyl- phenylamino)- 4-fluoro-N-hydroxy-benzamide; 5-Bromo-2-(4-ethynyl-2-chloro- phenylamino)-3,4-difiuoro-N-hydroxy-benzamide; 2-(2-Chloro-4-ethynyl- phenylamino)-3,4,5-trifluoro-benzoic acid; 2-(2-Chloro-4-ethynyl-phenylamino)- N- cyclopropylmethoxy-4-fluoro-benzamide; 5-Bromo-2-(2-chloro-4-ethynyl- phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide; 2-(4-Ethynyl-2- chloro-phenylamino)-3,4-difluoro-N-hydroxy-benzamide; 2-(4-Ethynyl-2-chloro- phenylamino)-N-hydroxy-4-nitro-benzamide; 2-(2-Chloro-4-ethynyl-phenylamino)- 4-fluoro-benzoic acid; 2-(2-Chloro-4-ethynyI-phenylamino)- N- cyclopropylmethoxy-4-fluoro-benzamide; 2-(2-Chloro-4-methanesulfinyl- phenylamino)- 4-fluoro-N-hydroxy-benzamide; and 2-(2-chloro-4-imidazol-1-yl- phenylamino)- 3,4-Difluoro-benzoic acid.
29. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically-acceptable carrier.
30. A method for treating a proliferative disease, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
31. A method of claim 30, wherein said proliferative disease is selected from psoriasis, restenosis, autoimmune disease, and atherosclerosis.
32. A method for treating cancer, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
33. A method of claim 32, wherein said cancer is MEK-related.
34. A method of claim 32, wherein said cancer is colorectal, cervical, breast, ovarian, brain, acute leukemia, gastric, non-small cell lung, pancreatic, prostatic, or renal.
35. A method for treating, or ameliorating the sequelae of, a stroke, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
36. A method for treating, or ameliorating the sequelae of, heart failure, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
37. A method for treating or reducing the symptoms of xenograft rejection, said method comprising administering to a cell transplant, limb transplant, skin transplant, an organ transplant or bone marrow transplant patient a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
38. A method for treating osteoarthritis, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
39. A method for treating rheumatoid arthritis, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
40. A method for treating cystic fibrosis, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
41. A method for treating hepatomegaly, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
42. A method for treating cardiomegaly, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
43. A method for treating Alzheimer's disease, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
44. A method for treating a complication of diabetes, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
45. A method for treating septic shock, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
46. A method for treating a viral infection, said method comprising administering to a patient in need of such treatment a pharmaceutically-effective amount of a composition comprising a compound of claim 1.
47. A method of claim 46, wherein said viral infection is a HIV infection.
48. A method for treating cancer, said method comprising
(a) administering to a patient in need of such treatment, a pharmaceutically- effective amount of a composition comprising a compound of claim 1 ; and
(b) administering a therapy selected from radiation therapy and chemotherapy.
49. A method of claim 48, wherein said chemotherapy comprises a mitotic inhibitor.
50. A method of claim 49, wherein said mitotic inhibitor is selected from paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine, and vinflunine.
EP99968160A 1999-01-13 1999-12-21 Anthranilic acid derivatives Withdrawn EP1150950A2 (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US11587699P 1999-01-13 1999-01-13
US115876P 1999-01-13
US12258399P 1999-03-02 1999-03-02
US122583P 1999-03-02
PCT/US1999/030491 WO2000041505A2 (en) 1999-01-13 1999-12-21 Anthranilic acid derivatives

Publications (1)

Publication Number Publication Date
EP1150950A2 true EP1150950A2 (en) 2001-11-07

Family

ID=26813668

Family Applications (1)

Application Number Title Priority Date Filing Date
EP99968160A Withdrawn EP1150950A2 (en) 1999-01-13 1999-12-21 Anthranilic acid derivatives

Country Status (6)

Country Link
EP (1) EP1150950A2 (en)
JP (1) JP2002534446A (en)
AU (1) AU2483400A (en)
BR (1) BR9916857A (en)
CA (1) CA2349180A1 (en)
WO (1) WO2000041505A2 (en)

Families Citing this family (52)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001005393A2 (en) * 1999-07-16 2001-01-25 Warner-Lambert Company Method for treating chronic pain using mek inhibitors
DOP2003000556A (en) 2002-01-23 2003-10-31 Warner Lambert Co ESTERES HYDROXAMATE ACID N- (4-PHENYL-REPLACED) -ANTRANILICO.
BR0307060A (en) 2002-01-23 2004-10-26 Warner Lambert Co n- (phenyl-4-substituted) -anthanilic acid hydroxamate esters
US7235537B2 (en) 2002-03-13 2007-06-26 Array Biopharma, Inc. N3 alkylated benzimidazole derivatives as MEK inhibitors
PT3000810T (en) 2002-03-13 2017-10-25 Array Biopharma Inc N3 alkylated benzimidazole derivative as mek inhibitor
US7144907B2 (en) 2003-09-03 2006-12-05 Array Biopharma Inc. Heterocyclic inhibitors of MEK and methods of use thereof
US7538120B2 (en) 2003-09-03 2009-05-26 Array Biopharma Inc. Method of treating inflammatory diseases
MXPA06004363A (en) 2003-10-21 2006-06-14 Warner Lambert Co Polymorphic form of n-[(r)-2, 3-dihydroxy -propoxy]-3, 4-difluoro-2 -(2-fluoro-4 -iodophenylamino) -benzamide.
US7772234B2 (en) 2003-11-19 2010-08-10 Array Biopharma Inc. Bicyclic inhibitors of MEK and methods of use thereof
US7517994B2 (en) 2003-11-19 2009-04-14 Array Biopharma Inc. Heterocyclic inhibitors of MEK and methods of use thereof
US7732616B2 (en) 2003-11-19 2010-06-08 Array Biopharma Inc. Dihydropyridine and dihydropyridazine derivatives as inhibitors of MEK and methods of use thereof
NZ590160A (en) 2003-11-21 2012-07-27 Array Biopharma Inc AKT protein kinase inhibitors
UA89035C2 (en) 2003-12-03 2009-12-25 Лео Фарма А/С Hydroxamic acid esters and pharmaceutical use thereof
DK1802579T3 (en) 2004-10-20 2014-01-20 Merck Serono Sa Derivatives of 3-arylaminopyridine
DK1922307T3 (en) 2005-05-18 2012-04-02 Array Biopharma Inc Heterocyclic Inhibitors of MEK and Methods for Using Them
US7547782B2 (en) * 2005-09-30 2009-06-16 Bristol-Myers Squibb Company Met kinase inhibitors
JP5129143B2 (en) * 2005-10-07 2013-01-23 エグゼリクシス， インコーポレイテッド MEK inhibitor and method of use thereof
GB0601962D0 (en) 2006-01-31 2006-03-15 Ucb Sa Therapeutic agents
EP2025347A4 (en) 2006-05-15 2010-08-11 Takeda Pharmaceutical Prophylactic and therapeutic agent for cancer
EP2049546B1 (en) 2006-07-06 2010-12-29 Array Biopharma, Inc. Dihydrofuro pyrimidines as akt protein kinase inhibitors
WO2008006039A1 (en) 2006-07-06 2008-01-10 Array Biopharma Inc. Dihydrothieno pyrimidines as akt protein kinase inhibitors
ATE523499T1 (en) 2006-07-06 2011-09-15 Array Biopharma Inc CYCLOPENTA [D]-PYRIMIDINE AS AN AKT PROTEIN KINASE INHIBITOR
US8063050B2 (en) 2006-07-06 2011-11-22 Array Biopharma Inc. Hydroxylated and methoxylated pyrimidyl cyclopentanes as AKT protein kinase inhibitors
WO2008140553A2 (en) 2006-10-23 2008-11-20 Takeda Pharmaceutical Company Limited Mapk/erk kinase inhibitors
PL2101759T3 (en) 2006-12-14 2019-05-31 Exelixis Inc Methods of using mek inhibitors
AU2008272832B2 (en) 2007-07-05 2014-02-20 Array Biopharma Inc. Pyrimidyl cyclopentanes as AKT protein kinase inhibitors
US8846683B2 (en) 2007-07-05 2014-09-30 Array Biopharma, Inc. Pyrimidyl cyclopentanes as Akt protein kinase inhibitors
AU2008272830B8 (en) 2007-07-05 2013-12-12 Array Biopharma Inc. Pyrimidyl cyclopentanes as AKT protein kinase inhibitors
US9409886B2 (en) 2007-07-05 2016-08-09 Array Biopharma Inc. Pyrimidyl cyclopentanes as AKT protein kinase inhibitors
US8022057B2 (en) 2007-11-12 2011-09-20 Takeda Pharmaceutical Company Limited MAPK/ERK kinase inhibitors
US8853216B2 (en) 2008-01-09 2014-10-07 Array Biopharma, Inc. Hydroxylated pyrimidyl cyclopentane as AKT protein kinase inhibitor
JP5346345B2 (en) 2008-01-09 2013-11-20 アレイ バイオファーマ、インコーポレイテッド Hydroxylated pyrimidylcyclopentanes as AKT protein kinase inhibitors
PL2307376T3 (en) 2008-08-04 2016-04-29 Merck Patent Gmbh Novel phenylamino isonicotinamide compounds
ES2399384T3 (en) 2008-11-10 2013-04-01 Bayer Schering Pharma Ag Sulfonamido substituted phenoxybenzamides
JP2013508318A (en) 2009-10-21 2013-03-07 バイエル・ファルマ・アクチェンゲゼルシャフト Substituted benzosulfonamide derivatives
US20120263714A1 (en) 2009-10-21 2012-10-18 Bayer Intellectual Property Gmbh Substituted halophenoxybenzamide derivatives
WO2011047795A1 (en) 2009-10-21 2011-04-28 Bayer Schering Pharma Aktiengesellschaft Substituted benzosulphonamides
JP5860045B2 (en) 2010-07-09 2016-02-16 ファイザー・リミテッドＰｆｉｚｅｒ Ｌｉｍｉｔｅｄ Compound
JP2013542214A (en) 2010-10-29 2013-11-21 バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツング Substituted phenoxypyridines
CA2831935A1 (en) 2011-04-01 2012-10-04 Genentech, Inc. Combinations of akt inhibitor compounds and chemotherapeutic agents, and methods of use
PL2694073T3 (en) 2011-04-01 2019-06-28 Genentech, Inc. Combinations of akt and mek inhibitors for treating cancer
EP2714037B1 (en) 2011-05-25 2016-07-13 Université Paris Descartes Erk inhibitors for use in treating spinal muscular atrophy
CN103204827B (en) 2012-01-17 2014-12-03 上海科州药物研发有限公司 Benzothiadiazole compounds as protein kinase inhibitors, and preparation method and application thereof
CN104136021B (en) * 2012-03-14 2016-04-20 中国中化股份有限公司 Substituted diphenylamine aminated compounds is as the application preparing antitumor drug
TR201807861T4 (en) 2012-10-12 2018-06-21 Exelixis Inc New process for making compounds for use in the treatment of cancer.
EP3043790B1 (en) * 2013-09-11 2021-05-26 The Administrators of the Tulane Educational Fund Novel anthranilic amides and the use thereof
EP3043822A1 (en) 2013-09-11 2016-07-20 The J. David Gladstone Institutes, A Testamentary Trust Established under The Will of J. David Gladstone Compositions for preparing cardiomyocytes
EP3204516B1 (en) 2014-10-06 2023-04-26 Dana-Farber Cancer Institute, Inc. Angiopoietin-2 biomarkers predictive of anti-immune checkpoint response
MA41866A (en) 2015-03-31 2018-02-06 Massachusetts Gen Hospital SELF-ASSEMBLING MOLECULES FOR TARGETED DRUG DELIVERY
WO2018157843A1 (en) * 2017-03-02 2018-09-07 中国科学院上海药物研究所 2-(substituted benzene matrix) aromatic formate fto inhibitor, preparation method therefor, and applications thereof
CN114907267A (en) * 2021-02-08 2022-08-16 中国科学院上海药物研究所 Pharmaceutical composition for resisting tumor
TW202342018A (en) 2022-03-04 2023-11-01 美商奇奈特生物製藥公司 Inhibitors of mek kinase

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2082171A (en) * 1933-05-24 1937-06-01 Winthrop Chem Co Inc Basically substituted amino acridine derivatives
NL62219C (en) * 1945-05-07
US2553914A (en) * 1948-03-22 1951-05-22 Ward Blenkinsop & Co Ltd Substituted diphenylamine 2-monocarboxylic acids and the production thereof
US3781358A (en) * 1968-05-29 1973-12-25 E Anderson Intermediates for preparing acridines
CH541608A (en) * 1970-11-06 1973-09-15 Ciba Geigy Ag Process for the preparation of nitrodiphenylamines and their use
US4510139A (en) * 1984-01-06 1985-04-09 Sterling Drug Inc. Substituted aminobenzamides and their use as agents which inhibit lipoxygenase activity
DE3608726A1 (en) * 1986-03-15 1987-09-17 Hoechst Ag USE OF AMINO-SUBSTITUTED BENZOESAURS AS A MEDICINE AGAINST DIARRHOE AND MEDICINAL PRODUCTS BASED ON THESE COMPOUNDS
US5155110A (en) * 1987-10-27 1992-10-13 Warner-Lambert Company Fenamic acid hydroxamate derivatives having cyclooxygenase and 5-lipoxygenase inhibition
US6251943B1 (en) * 1997-02-28 2001-06-26 Warner-Lambert Company Method of treating or preventing septic shock by administering a MEK inhibitor
ES2195397T3 (en) * 1997-10-28 2003-12-01 Warner Lambert Co QUINAZOLIN-2,4-DIONAS REPLACED IN POSITION 7 USEFUL AS ANTIBACTERIAL AGENTS.

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0041505A2 *

Also Published As

Publication number Publication date
AU2483400A (en) 2000-08-01
CA2349180A1 (en) 2000-07-20
JP2002534446A (en) 2002-10-15
WO2000041505A3 (en) 2000-10-19
WO2000041505A2 (en) 2000-07-20
BR9916857A (en) 2001-12-04

Similar Documents

Publication Publication Date Title
EP1144372B1 (en) Sulphohydroxamic acids and sulphohydroxamates and their use as mek inhibitors
WO2000041505A2 (en) Anthranilic acid derivatives
EP1144371B1 (en) Benzenesulphonamide derivatives and their use as mek inhibitors
EP1144385B1 (en) Benzoheterocycles and their use as mek inhibitors
EP1144394B1 (en) 1-heterocycle substituted diarylamines
WO2000041994A1 (en) 4-arylamino, 4-aryloxy, and 4-arylthio diarylamines and derivatives thereof as selective mek inhibitors
US6506798B1 (en) 4-Arylamino, 4-aryloxy, and 4-arylthio diarylamines and derivatives thereof as selective MEK inhibitors
US7001905B2 (en) Substituted diarylamines as MEK inhibitors
CA2403017A1 (en) 5-amide substituted diarylamines as mex inhibitors
EP1202724A2 (en) Method for treating chronic pain using mek inhibitors
JP2000212141A (en) Diarylamine
JP2001055376A (en) Diaryl amine
JP2000212157A (en) Diarylamine
MXPA01004180A (en) 4-arylamino, 4-aryloxy, and 4-arylthio diarylamines and derivatives thereof as selective mek inhibitors
MXPA01006568A (en) Benzoheterocycles and their use as mek inhibitors

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20010813

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

17Q First examination report despatched

Effective date: 20030512

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: WARNER-LAMBERT COMPANY LLC

18D Application deemed to be withdrawn

Effective date: 20041228