WO2005065686A1 - Agents de modulation de la differenciation et utilisations associees - Google Patents

Agents de modulation de la differenciation et utilisations associees Download PDF

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WO2005065686A1
WO2005065686A1 PCT/AU2005/000008 AU2005000008W WO2005065686A1 WO 2005065686 A1 WO2005065686 A1 WO 2005065686A1 AU 2005000008 W AU2005000008 W AU 2005000008W WO 2005065686 A1 WO2005065686 A1 WO 2005065686A1
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Johannes Bernhard Prins
Louise Joyce Hutley
Ross Peter Mcgeary
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Adipogen Pharmaceuticals Pty Limited
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/731—Carrageenans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4184—1,3-Diazoles condensed with carbocyclic rings, e.g. benzimidazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/42—Oxazoles
    • A61K31/423—Oxazoles condensed with carbocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/737—Sulfated polysaccharides, e.g. chondroitin sulfate, dermatan sulfate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
    • A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
    • 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
    • A61P3/04—Anorexiants; Antiobesity agents

Definitions

  • FIELD OF THE INVENTION relates generally to methods and agents for antagonizing the differentiation and or proliferation of preadipocytes. More particularly, the present invention relates to fibroblast growth factor (FGF) signalling, especially FGF-1 and FGF-2 signalling, which causes the proliferation of preadipocytes and which potentiates preadipocytes to differentiate into adipocytes. Even more particularly, the invention relates to molecules that reduce, impair or abrogate FGF signalling, including antagonist molecules that are specific for Fgf or Fgfr polynucleotides or their expression products, and to the use of these molecules for the down- regulation of adipogenesis, including down-regulating the differentiation potential and/or proliferation of preadipocytes.
  • FGF fibroblast growth factor
  • the present invention extends to methods of screening for agents that are useful for antagonizing FGF signalling, including modulating the expression of a gene selected from a Fgf gene or a Fgfr gene or a gene belonging to the same biosynthetic or regulatory pathway as the Fgf gene or the Fgfr gene or for modulating the level or functional activity of an expression product of that gene.
  • the invention relates to the use of such antagonizing agents in methods for treating or preventing adiposity-related conditions including, but not limited to, obesity, lipoma and lipomatosis.
  • OBESITY Obesity represents a major health problem worldwide which is no longer confined to traditional 'Westernised' communities, as the high-fat diet and sedentary lifestyle of the traditional 'Western' countries is adopted in preference to traditional ethnic lifestyles (Doll et al. Int J Obes Relat Metab Disord 26(1): 48-57 2002) (Fall. Br Med Bull 60: 33-50 2001).
  • the incidence of obesity and, in particular, obesity in children, is increasing at a faster rate than almost any other medical condition.
  • Around 22 million children under the age of five years are overweight worldwide (Deckelbaum et al.
  • Obes Res 9 Suppl 4: 239S-243S 2001 and over 7% of adults worldwide are obese with around a further 21% of adults being classified as overweight (Seidell Acta Paediatr Suppl 88(428): 46-50 1999).
  • the World Health Organisation describes the high worldwide incidence of obesity in adults as a 'global pandemic'.
  • the association of obesity with serious co-morbidities such as cardiovascular diseases and type II diabetes (Fall 2001 supra) is the cause of its classification as a serious medical condition (James et al Obes Res 9 Suppl 4: 228S-233S 2001).
  • the consequential and significant financial impact of obesity on healthcare budgets has made obesity management and prevention a major priority for health promotion strategies.
  • New adipocytes are formed by the proliferation and diffeientiation of preadipocytes, a process known as adipogenesis.
  • Preadipocytes are f ⁇ broblast-hke cells found m the stromo-vascular compartment of adipose tissue.
  • Therapeutic interventions which inhibit adipogenesis would have profound chnical applications m the management of severely overweight patients
  • Research has, thus far, discovered several protein, neuropeptide and transcriptional regulators of the cellular and molecular events underlying changes in adipose cell size or number.
  • adipocyte number and size are altered in a complex interplay involving hormonal and nutritional cues, which trigger downstream signalling via molecules which act in a cell-cell or cell-matrix manner (Gregoire Exp Biol Med (Maywood) 226(H)- 997-1002 2001).
  • the full repertoire of these molecules has yet to be established, as well as the way in which they interact and exert their effects on adipocytes.
  • Much has been learned about adipogenesis through development of techniques allowing the isolation and in vitro replication and differentiation of animal and human preadipocytes. Further insight has been gamed by the study of murme preadipocyte cell lines (e.g., 3T3-L1) that differentiate in vitro to an adipocyte-hke cell.
  • preadipocytes are isolated from adipose tissue using collagenase digestion and plated m serum-contammg medium. Upon reaching confluence (with or without previous subculture) the cells are differentiated in a serum-free chemically and hormonally modified medium. This process is relatively inefficient, both in time and m the low percentage of cells that acquire a mature adipocyte phenotype.
  • the replication phase is enhanced by mitogens and insulin, and requires serum.
  • the differentiation phase is completely inhibited by serum, and enhanced by insulin, corticosteroids, thyroid hormone and growth hormone.
  • thiazolidinedione (TZD) class of drugs stimulate differentiation via binding to PPAR ⁇ , a ligand-dependant transcription factor central to adipogenesis.
  • PPAR ⁇ a ligand-dependant transcription factor central to adipogenesis.
  • adipogenesis is preceded by the establishment of a fine vascular network (Hutley et al. Am J Physiol Endocrinol Metab 281(5): El 037-44 2001) and a paracrine interaction between preadipocytes and the endothelial cells of the micro vasculature had been proposed (Hutley et al. supra) (Varzaneh et al.
  • FGF acidic fibroblast growth factor
  • MVEC microvascular endothelial cells
  • FGF-1 acidic fibroblast growth factor
  • basic FGF also known as FGF-2
  • the fibroblast growth factor family of structurally related polypeptide growth factors comprises over 20 members with protean recognised actions. There is limited direct coding sequence homology across the family. The name is misleading as stimulation of growth is not universal among family members but, as a family, the FGFs have critical roles in growth and development, cell replication and angiogenesis, cell survival and apoptosis, tumour development and morphogenesis.
  • the FGFs belong to the larger Heparin-Binding Growth Factor family which comprises a large number of growth factors, some with similar or complementary actions to the FGFs. FGFs are encoded by a number of different genes and have similar intron-exon organisation, with three coding regions in FGF- 1-6.
  • FGFs vary in the presence of signal peptides or localisation sequences and in glycosylation sites and post-translational modification. Many of the FGFs show diversity with alternative promoter usage (e.g., FGF-1), alternative splicing (e.g., FGF-1 and -2) and the use of alternative polyadenylation sites (e g , FGF-1 and -2)
  • alternative promoter usage e.g., FGF-1
  • alternative splicing e.g., FGF-1 and -2
  • alternative polyadenylation sites e.g , FGF-1 and -2
  • tissue-specific promoter usage e.g., FGF-1
  • FGFs can be released from cells but some also accumulate m the nucleus or cytoplasm of producing and target cells, hi addition, secreted FGFs are stored in the extracellular mat ⁇ x and their further release is under protease control. FGFs are "released" from the extracellular mat ⁇ x by one of two mechanisms. First, enzymatic cleavage of extracellular mat ⁇ x components by proteases or hepa ⁇ nases results in release of FGF Second, FGF can bind to a carrier protein (FGF-BP) that can m turn deliver FGF to its receptor It is accepted that hepa ⁇ n or heparan-hke glycosammoglycans are essential for efficient FGF signalling.
  • FGF-BP carrier protein
  • FGF Tissue-specificity and/or differentiation stage-specificity of expression of some FGFs has been reported
  • the association of the FGF family with components of the extracellular matrix is thought to serve two purposes: a) protection of FGFs from circulating protease degradation; and b) creation of a local reservoir of growth factor(s)
  • the latter feature allows for strict spatial regulation of FGF signalling, as only cells m contact with the extracellular matrix are recipient to the FGF signal.
  • the FGF receptors comprise a gene family encoding five (at least) structurally related proteins They are members of the tyrosme-kmase class of receptors and are widely expressed. Ammo acid sequence homology of the five receptors is 60-95% with the best-conserved areas involved m signal transduction. FGFs have differing specificity m their binding to the receptors and this, along with cell-specific expression of the receptors and their splice variants, provides further diversity in signalling options.
  • FGFRs are also expressed withm the nuclear envelope and matrix Signal transduction m response to FGFs occurs through receptor dime ⁇ sahon and complex formation with heparan sulfate proteoglycans (HSPGs).
  • HSPGs heparan sulfate proteoglycans
  • FGF signalling diversity is provided by cell specific expression of receptor combinations, cell specific expression of receptor isoform combinations, va ⁇ ous hetero-dimer combinations and different repertoires of FGFs.
  • HSPGS HSPGs are sulfated glycosammoglycans covalently bound to a core protein that act to facilitate FGF-FGFR interaction This may be due either to the HSPG inducing conformational changes in FGF and FGFR allowing each to dime ⁇ se and bind or due to the HSPG forming part of an active signalling complex with the FGF and FGFR. Expe ⁇ mental evidence to support both models exists, and it is highly conceivable that both mechanisms exist. Some FGF early responses may be elicited m the absence of HSPG but the latter appears essential for sustained signalling. HSPG also acts to protect FGFs from degradation in the extracellular mat ⁇ x.
  • HSPGs implicated to date m FGF signalling include the syndecans (cell-associated transmembrane proteoglycans), the glypicans (proteoglycans anchored to the plasma membrane by a glycosylphosphatidylmositol group) and perlecan (an extracellular, basal lammaproteoglycan).
  • syndecans cell-associated transmembrane proteoglycans
  • glypicans proteoglycans anchored to the plasma membrane by a glycosylphosphatidylmositol group
  • perlecan an extracellular, basal lammaproteoglycan
  • Cysteine- ⁇ ch FGFR is an integral membrane sialoglycoprotem that lacks heparan sulfate chains and binds FGFs FGF binding to CFR and FGFR is mutually exclusive. CFR appears to have a role in FGF targeting to intracellular sites and in regulation of intracellular FGF concentrations
  • FGFR-Dependent Intracellular Signalling As outlined above, and with reference to the schematic representation of the FGF signalling pathway shown in Figure 1, ligand binding induces receptor dimerisation and auto- phosphorylation. Mutational analysis indicates that dimerisation alone is sufficient for signal transduction FGFRs have a number of intracellular phosphorylation sites (seven in the case of FGFR-1) and phosphorylation site mutated, kinase dead, receptors are unable to transduce many biological signals of FGFs. However, some effects are retained, indicating that non receptor- mediated signalling pathways are an important consideration.
  • the signalling pathways known to be utilized by FGF/FGFR are (1) the SHC/FRS2- RAF MAPKKK-MAPKK-MAPK pathway, and (2) the PLCy, PKC, Ca 2+ pathway.
  • SHC/FRS2-RAF/MAPKKK-MAPKK-MAPK pathway Subsequent to receptor phosphorylation src homology (SH-2) domain-containing and phosphotyrosine-bmdmg (PTB) domain proteins bind to specific intracellular FGFR phosphotyrosmes
  • SH-2 receptor phosphorylation src homology
  • PTB phosphotyrosine-bmdmg
  • PLC ⁇ PKC
  • Ca ⁇ pathway PLC ⁇ is a SH-2 domain protein that binds to a specific phosphotyrosme m FGFRs (Y766 m FGFR-1) and subsequently hydrolyses phosphomositol to mositol 1,4,5 t ⁇ phosphate (IP 3 ) and diacyglycerol (DAG) IP 3 induces Ca 2+ release from intracellular stores, whereas DAG activates PKC, a serme/threomne-specific kmase
  • DAG diacyglycerol
  • FGF Target Genes FGF treatment alters expression of many genes, and can do so via non FGFR-mediated mechanisms. This is presumed to be a dnect effect, and many FGFs have nuclear targeting motifs and are found m the nucleus, the nucleolus and in association with chromatm. The effect of FGFs on gene transcription is cell-type specific. Further, FGFs have been demonstrated to maintain the expression of genes whose initial induction is dependent on other factors, hi addition to transcriptional regulation, FGFs also influence mRNA stability and translation and post- translational modification of proteins. 3. Interaction with other Growth Factor Signalling Pathways FGFs can antagonize or synergize with many other growth factors.
  • FGF co-operativity with transforming growth factor (TGF), insulin-like growth factor- 1 (IGF-1) and WNT signalling is common.
  • TGF transforming growth factor
  • IGF-1 insulin-like growth factor- 1
  • WNT signalling is common.
  • molecules of a FGF signalling pathway especially of the FGF-1 or FGF-2 signalling pathway, can be used to provide both drug targets and regulators to inhibit adipogenesis in wter alia adiposity- related conditions and also to provide diagnostic markers for predisposition to obesity, as described hereinafter.
  • the present invention provides methods for inhibiting adipogenesis, which are useful inter alia in the treatment or prevention of adiposity-related conditions. These methods generally comprise contacting a cell with an agent for a time and under conditions sufficient to inhibit a FGF signalling pathway, hi some embodiments, the FGF signalling pathway is selected from the FGF-1 signalling pathway and the FGF-2 signalling pathway.
  • Representative members of these pathways include, but are not limited to, FGFRs, HSPGs, members of the SHC/FRS2-RAF/MAPKKK-MAPKK-MAPK pathway, members of the PLC ⁇ -PKC-Ca 2+ pathway, members of the FGF-1 nuclear translocation pathway and intracellular binding partners such as P34 and FIF (FGF-interacting factor).
  • suitable agents include small molecules, such as nucleic acids, peptides, polypeptides, peptidomimetics, carbohydrates, lipids or other organic (carbon containing) or inorganic molecules, as further described herein.
  • the cell is contacted with an agent that inhibits the expression of a gene or the level or functional activity of an expression product of the gene, wherein the gene is selected from a Fgf gene (e.g., Fgf-1 or Fgf-2) and a gene belonging to the same regulatory or biosynthetic pathway as the Fgf gene (e.g., P34 and FIF).
  • the cell is suitably a microvascular endothelial cell, or precursor thereof.
  • the cell is contacted with an agent that inhibits the expression of a gene or the level or functional activity of an expression product of the gene, wherein the gene is selected from a Fgfr gene (e.g., Fgfr-1, Fgfr-2, Fgfr- 3, Fgfr-4, Fgfr-5, especially Fgfr-1, Fgfr-2, Fgfr-3, Fgfr-4), a gene belonging to the same regulatory or biosynthetic pathway as the Fgfr gene (e.g., a gene involved in signalling via the Ras-Raf-MAPkinase pathway and or via the phospholipase C pathway), a gene whose expression is inhibited directly or indirectly by an expression product of the Fgf gene (e.g, PPAR ⁇ , IGFBP-3, IGFBP-6, IGF-2, IRS-2, PI3 kinase, PKC ⁇ ), or that antagonizes the function of a FGFR with which a Fgfr
  • the cell is suitably a preadipocyte or precursor thereof.
  • the agent reduces the expression of a gene (e.g, Fgfr-1, Fgfr-2, Ppar ⁇ , C/Ebpa, Plc ⁇ 2, Igfbp-3, Igfbp-6) or the level or functional activity of an expression product of that gene (e.g, FGFR-1, FGFR-2, PPAR ⁇ , C/EBP ⁇ , PLC ⁇ 2, IGFBP-3, IGFBP-6).
  • a gene e.g, Fgfr-1, Fgfr-2, Ppar ⁇ , C/Ebpa, Plc ⁇ 2, Igfbp-3, Igfbp-6
  • an expression product of that gene e.g, FGFR-1, FGFR-2, PPAR ⁇ , C/EBP ⁇ , PLC ⁇ 2, IGFBP-3, IGFBP-6.
  • the agent increases the expression of a gene (e.g, Fgf-1, Fgfr-3, Igf-2, Irs-2, Pi3 kinase, Pkc ⁇ ) or the level or functional activity of an expression product of that gene (e.g, FGF-1, FGF-3, IGF-2, IRS-2, PI3 kinase, PKC ⁇ ).
  • the agent antagonizes the function of a FGFR, including reducing or abrogating the interaction between a FGFR and a FGF.
  • the agents antagonize a FGF signalling pathway and are therefore useful for directly or indirectly reducing or abrogating the differentiation potential and or proliferation of a preadipocyte.
  • the agent increases or reduces the expression of the gene or the level or functional activity of an expression product of that gene by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%), 90% relative to the expression, level or functional activity in the absence of the agent.
  • the invention provides methods for identifying agents that inhibit a FGF signalling pathway.
  • These methods typically comprise contacting a preparation with a test agent, wherein the preparation comprises (i) a polypeptide comprising an amino acid sequence corresponding to at least a biologically active fragment of a polypeptide component of the FGF signalling pathway, or to a variant or derivative thereof; or (ii) a polynucleotide comprising at least a portion of a genetic sequence that regulates the component, which is operably linked to a reporter gene.
  • Still another aspect of the present invention provides methods for identifying agents that inhibit a FGF signalling pathway. These methods generally comprise contacting a first sample of cells expressing a FGFR with a FGF and measuring a marker; contacting a second sample of cells expressing the FGFR with an agent and the FGF, and measuring the marker; and comparing the marker of the first sample of cells with the marker of the second sample of cells. In various embodiments, these methods measure the levels of various markers (e.g, glycerol 3-phosphate dehydrogenase; G3PDH, and intracellular components of the FGF pathway), or combinations of markers, associated with the proliferation and/or differentiation of preadipocytes.
  • markers e.g, glycerol 3-phosphate dehydrogenase; G3PDH, and intracellular components of the FGF pathway
  • the agents broadly described above are useful for inhibiting adipogenesis in adiposity-related conditions.
  • the adiposity-related conditions include, but are not restricted to, obesity, lipoma or lipomatosis.
  • another aspect of the present invention contemplates the use of an agent, which is optionally formulated with a pharmaceutically acceptable carrier or diluent, for inhibiting or decreasing adipogenesis, or for controlling adipogenesis in obesity or in conditions of localized, abnormal increases in adipogenesis, wherein the agent antagonizes a FGF signalling pathway as broadly described above.
  • the agent used in the above methods is characterized in that it binds to an expression product of a gene as broadly described above or to a genetic sequence (e.g, a transcriptional element) that inhibits the expression of the gene, as determined by: contacting a preparation comprising at least a portion of an expression product of a gene as broadly described above, or a variant or derivative of the expression product, or a genetic sequence that modulates the expression of the gene, with the agent; and detecting a decrease in the level or functional activity of the at least a portion of the expression product, or the variant or derivative, or of a product expressed from the genetic sequence.
  • a genetic sequence e.g, a transcriptional element
  • an agent which inhibits or otherwise decreases adipogenesis binds to a FGF or FGFR or to a genetic sequence (e.g, a transcriptional element) that modulates the expression of a Fgfo ⁇ Fgfr gene, as determined by: contacting a preparation comprising a FGF or FGFR polypeptide or biologically active fragment thereof, or variant or derivative of these, or a genetic sequence that modulates the expression of a Fgf ox Fgfr gene; and detecting a decrease in the level or functional activity of the FGF or FGFR polypeptide or biologically active fragment thereof, or variant or derivative, or of a product expressed from the genetic sequence.
  • a genetic sequence e.g, a transcriptional element
  • an agent which inhibits or otherwise decreases adipogenesis antagonizes a FGF signalling pathway, as determined by: contacting a FGFR and a FGF with the agent and measuring the binding of the FGFR with the FGF.
  • agents can bind to FGF or FGFR and test positive when they reduce or abrogate the binding of the FGFR with the FGF.
  • the agents can be small molecules or antigen-binding molecules specific for the FGF or the FGFR.
  • an agent which inhibits or otherwise decreases adipogenesis antagonizes a FGF signalling pathway, as determined by: contacting a FGFR and an HSPG with the agent and measuring the binding of the FGFR with the HSPG.
  • agents can bind to FGF or HSPG and test positive when they reduce or abrogate the binding of the HSPG with the FGFR.
  • the compounds can be small molecules or antigen-binding molecules specific for the FGFR or the HSPG.
  • an agent which inhibits or otherwise decreases adipogenesis antagonizes a FGF signalling pathway, as determined by: contacting a FGF and a CFR with the agent and measuring the binding of the FGF with the CFR.
  • agents can bind to FGF or CFR and test positive when they reduce or abrogate the binding of the FGF with the CFR.
  • the compounds can be small molecules or antigen-binding molecules specific for the FGF or the CFR.
  • an agent which inhibits or otherwise decreases adipogenesis antagonizes a FGF signalling pathway, as determined by: contacting a first sample of cells selected from preadipocytes or their precursors with a FGF and measuring differentiation and/or proliferation of the cells; contacting a second sample of cells selected from preadipocytes or their precursors with an agent and the FGF, and measuring differentiation and/or proliferation of the cells; comparing the differentiation and/or proliferation of the first sample of cells with the differentiation and/or proliferation of the second sample of cells.
  • the agents antagonize the FGF signalling pathway by interfering with the association of the FGF and a FGFR, by interfering with the phosphorylation of a FGFR, by interfering with components of the signalling pathway upstream or downstream of the FGF/FGFR interaction, by interfering with the association of a FGFR with an HSPG, by interfering with the association of the FGF and CFR, or by interfering with the dimerisation of a FGFR.
  • agents that antagonize the FGF signalling pathway interfere with a signalling pathway selected from the TGF, IGF-1 and WNT signalling pathways.
  • an agent which inhibits or otherwise decreases adipogenesis antagonizes a FGF signalling pathway, as determined by: administering to an animal model, or a human, an agent that antagonizes the signalling pathway, and measuring the animal's responsiveness to the agent.
  • the method can be practiced with agents as described above and animals can be examined for inhibition or reduction of adipogenesis in obesity or in conditions of localized, abnormal increases in adipogenesis.
  • Still another aspect of the present invention provides methods of producing an agent for inhibiting adipogenesis in adiposity-related conditions.
  • These methods generally comprise: testing an agent suspected of inhibiting a FGF signalling pathway as broadly described above; and synthesizing the agent on the basis that it tests positive for the inhibition.
  • the method further comprises derivatizing the agent, and optionally formulating the derivatized agent with a pharmaceutically acceptable carrier and/or diluent, to improve the efficacy of the agent for treating or preventing the adiposity-related condition(s).
  • Another aspect of the present invention contemplates methods for inhibiting or reducing adipogenesis in obesity or m conditions of localized, abnormal increases in adipogenesis.
  • These methods generally comprise administering to a patient in need of such treatment an adipogenesis- lnhibitmg effective amount of an agent which impairs or interferes with a FGF signalling pathway as broadly described above, and optionally a pharmaceutically acceptable carrier or diluent
  • an agent as broadly described above in the preparation of a medicament for treating or preventing an adiposity-related condition.
  • Figure 1 is a schematic representation of the FGF signalling pathway.
  • Figure 2 is a schematic representation of the method for isolation and separation of microvascular endothelial cells (MVEC) and preadipocytes (PA) from human adipose tissue DPBS deiomsed phosphate buffered saline, RT room temperature; HBSS: Hank's balanced salt solution; FCS. fetal calf serum; EC: endothelial cells; PECAM-1 : platelet-endothelial cell adhesion molecule 1.
  • Figure 3 is a photographic representation illustrating the morphology of adipose tissue- derived MVEC A.
  • phase-contrast photomicrograph of MVEC isolated from human adipose tissue Note the typical cobblestone morphology and the prominent, centrally located nuclei.
  • FIG 4 is a photographic representation of a Western blot analysis, showing strong expression of FGF-1 m adipose-derived MVEC and also in 3T3-L1 adipocytes (expression was also shown m 3T3-L1 fibroblasts).
  • FGF-1 protein is undetectable in both human preadipocytes (+/- exposure to FGF-1) and adipocytes.
  • RT-PCR analysis co ⁇ oborated these expression patterns.
  • Figure 5 is a graphical representation showing a marked increase in proliferation of human preadipocytes (PAs) m response to both FGF-1 and FGF-2 (with FGF-1 effects on proliferation greater than FGF-2).
  • PAs human preadipocytes
  • Figure 6 is a graphical representation showing a marked increase in differentiation of human preadipocytes (PAs) m response to both FGF-1 and FGF-2 (with FGF-1 effects on differentiation greater than FGF-2).
  • Figure 7 is a graphical representation showing the effects of combination treatments of
  • FIG. 8 is a photographic representation showing the differentiation of human preadipocytes (PAs) using a 3T3-L1 differentiation protocol that utilizes serum-containing medium (SCM) (+ insulin and, for the first 3 days, dexamethasone and rosiglitazone).
  • Panel (A) shows PAs that have not been exposed to FGF-1 during proliferation prior to differentiation.
  • Panels (B) and (C) show subcutaneous & omental PAs, respectively, that have been proliferated for six weeks in the presence of FGF-1 and subsequently differentiated in SCM. This is the first report of human PAs differentiating in the presence of serum.
  • Figure 9 is a tabular representation showing the results of two separate gene array experiments which compared gene expression in human PAs grown to confluence in serum- containing medium in the presence and absence of FGF-1. Gene expression was considered to be influenced by FGF-1 if expression was consistently (CV ⁇ 5%) increased or reduced by at least
  • Figure 11 is a graphical representation showing that inhibition of PLC markedly reduces FGF-1 induced differentiation of preadipocytes.
  • Figure 12 is a graphical representation showing that neutralising anti-FGF-1 antibody abrogates FGF-1 -induced human preadipocyte replication, (Black bars, subcutaneous preadipocytes and white bars, omental preadipocytes).
  • Figure 13 is a graphical representation showing that inhibition of post FGFR signal transduction pathways has marked effects on FGF-1 -mediated human adipogenesis.
  • Figure 14 is a graphical representation showing that inhibition of post FGFR signal transduction pathways with anti-FGF-1 antibody has a marked effect on FGF-1 mediated human adipogenesis.
  • Figure 15 is a photographic representation showing the inhibition of FGF-1 induced differentiation of preadipocytes with anti-FGF-1 antibody.
  • Figure 16 is a graphical representation showing inhibition of FGF-1 mediated subcutaneous preadipocyte proliferation by Methyl l-oxo-3-phenyl-17J-indene-2-carboxylate.
  • Figure 17 is a graphical representation showing inhibition of FGF-1 mediated omental preadipocyte proliferation by Methyl l-oxo-3 -phenyl- lH-indene-2-carboxylate.
  • Figure 18 is a graphical representation showing inhibition of FGF-1 mediated subcutaneous preadipocyte proliferation by l-Deoxy-l- ⁇ [(2,4-dichlorophenoxy)acetyl]amino ⁇ - ⁇ - D-glucopyranuronic acid.
  • Figure 19 is a graphical representation showing inhibition of FGF-1 mediated omental preadipocyte proliferation by l-Deoxy-l- ⁇ [(2,4-dichlorophenoxy)acetyl]amino ⁇ - ⁇ -D- glucopyranuronic acid.
  • Figure 20 is a graphical representation showing inhibition of FGF-1 mediated subcutaneous preadipocyte proliferation by 3-[(3-(2-carboxyethyl)-4-methylpyrrol-2- yl)methylene]-2-indolinone.
  • Figure 21 is a graphical representation showing inhibition of FGF-1 mediated subcutaneous preadipocyte proliferation by phosphopeptide ThrAsnGluLeuTyr(0- P0 3 H 2 )MefMetMetArg.
  • Figure 22 is a graphical representation showing inhibition of FGF-1 mediated subcutaneous preadipocyte differentiation by ThrAsnGluLeuTyr(0-P0 3 H 2 )MefMetMetArg.
  • Figure 23 is a graphical representation showing effects of anti-FGF Receptor 1, 2 and 3 antibodies on FGF-1 mediated subcutaneous preadipocyte proliferation.
  • Figure 24 is a graphical representation showing effects of anti-FGF Receptor 1, 2 and 3 antibodies on FGF-1 mediated omental preadipocyte proliferation.
  • a gene belonging to the same regulatoiy or biosynthetic pathway is meant a gene whose expression product can modulate or otherwise influence FGF or FGFR protein levels and or Fgf or Fgfr transcription levels.
  • a gene belonging to the same regulatory pathway as g may encode an upstream regulator of Fgf/FGF, or a downstream regulatory target of Fgf/FGF, instead of Fgf/FGF.
  • a gene belonging to the same regulatory or biosynthetic pathway as a Fgfr gene includes genes which directly or indirectly modulate the expression of a Fgfr gene as well as genes which act as signal transducers for FGFR activation.
  • Such signalling molecules are involved in communicating and/or mediating the effects of FGFR activation and are commonly known in the art. They include inter alia molecules involved in the phospholipase C (PLC)- ⁇ , Crk, SNT-1/FRS2 and/or Src signalling pathways.
  • PLC phospholipase C
  • the term "aberrant polynucleotide” as used herein refers to a polynucleotide which is distinguished from a "normal" reference polynucleotide by the substitution, deletion or addition of at least one nucleotide and which correlates with the presence or risk of adipogenic defects including an elevated rate of adipogenesis compared to a non-obese, reference value.
  • Aberrant polypeptide refers to a polypeptide which is distinguished from a "normal” reference polypeptide by the substitution, deletion or addition of at least one amino acid residue and which correlates with the presence or risk of adipogenic defects including an elevated rate of adipogenesis compared to a non-obese, reference value.
  • Amplification product refers to a nucleic acid product generated by a nucleic acid amplification technique.
  • antigen-binding molecule is meant a molecule that has binding affinity for a target antigen. It will be understood that this term extends to immunoglobulins, immunoglobulin fragments and non-immunoglobulin derived protein frameworks that exhibit antigen-binding activity.
  • Antigenic or immunogenic activity refers to the ability of a polypeptide, fragment, variant or derivative according to the invention to produce an antigenic or immunogenic response in an animal, suitably a mammal, to which it is administered, wherein the response includes the production of elements which specifically bind the polypeptide or fragment thereof.
  • biologically active fragment is meant a fragment of a full-length parent polypeptide which fragment retains an activity of the parent polypeptide.
  • biologically active fragment includes deletion variants and small peptides, for example of at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50 contiguous amino acid residues, which comprise an activity of the parent polypeptide.
  • Peptides of this type may be obtained through the application of standard recombinant nucleic acid techniques or synthesized using conventional liquid or solid phase synthesis techniques. For example, reference may be made to solution synthesis or solid phase synthesis as described, for example, in Chapter 9 entitled “Peptide Synthesis” by Atherton and Shephard which is included in a publication entitled “Synthetic Vaccines” edited by Nicholson and published by Blackwell Scientific Publications.
  • peptides can be produced by digestion of a polypeptide of the invention with proteinases such as endoLys-C, endoArg-C, endoGlu-C and staphylococcus V8-protease.
  • the digested fragments can be purified by, for example, high performance liquid chromatographic (HPLC) techniques.
  • HPLC high performance liquid chromatographic
  • biological sample refers to a sample that may extracted, untreated, treated, diluted or concentrated from a patient.
  • the biological sample is a tissue biopsy, more preferably from subcutaneous or omental tissue biopsy.
  • condition of localized, abnormal increases in adipogenesis includes pathologies characterized by and/or associated with anatomically localized disregulated adipogenesis that lead to circumscribed depositions of fat tissue. Such conditions include but are not limited to lipoma and lipomatosis.
  • derivative is meant a polypeptide that has been derived from the basic sequence by modification, for example by conjugation or complexing with other chemical moieties or by post- translational modification techniques as would be understood in the art.
  • the term “derivative” also includes within its scope alterations that have been made to a parent sequence including additions or deletions that provide for functional equivalent molecules.
  • the term “differentiation potential” as used herein means the capacity of a preadipocyte to respond, or the magnitude of the response, to a signal which promotes its functional maturation into an adipocyte.
  • an "increase in differentiation potential" may be seen to be conferred by a test molecule wherein, for example, a co-culture of preadipocytes with the test molecule for a sufficient time and under appropriate conditions results in an increase in the response of the preadipocytes to a differentiation-inducing agent, which may be observed inter alia as a rise in the number of preadipocytes undergoing differentiation or an increase in the rate at which the preadipocytes undergo differentiation.
  • an effective amount in the context of modulating an activity or of treating or preventing a condition is meant the administration of that amount of active ingredient to an individual in need of such modulation, treatment or prophylaxis, either in a single dose or as part of a series, that is effective for modulation of that effect or for treatment or prophylaxis or improvement of that condition.
  • improvements in an individual suffering conditions of localized, abnormal increases in adipogenesis include reduced fat deposits, and an improvement in the symptoms relating to cardiovascular disease and diabetes.
  • the effective amount will vary depending upon the health and physical condition of the individual to be treated, the taxonomic group of individual to be treated, the formulation of the composition, the assessment of the medical situation, and other relevant factors.
  • the term “function” refers to a biological, enzymatic, or therapeutic function.
  • functional Fgf polynucleotide or “functional FGF polypeptide” is meant an Fgf polynucleotide or an FGF polypeptide having no structural or functional defects and which do not correlate with the presence or risk of adipogenic defects including elevated or impaired adipogenesis.
  • gene refers to any and all discrete coding regions of the cell's genome, as well as associated non-coding and regulatory regions.
  • the gene is also intended to mean the open reading frame encoding specific polypeptides, introns, and adjacent 5' and 3' non- coding nucleotide sequences involved in the regulation of expression.
  • the gene may further comprise control signals such as promoters, enhancers, termination and/or polyadenylation signals that are naturally associated with a given gene, or heterologous control signals.
  • the DNA sequences may be cDNA or genomic DNA or a fragment thereof.
  • the gene may be introduced into an appropriate vector for extrachromosomal maintenance or for integration into the host. "Hybridization” is used herein to denote the pairing of complementary nucleotide sequences to produce a DNA-DNA hybrid or a DNA-RNA hybrid.
  • Complementary base sequences are those sequences that are related by the base-pairing rules.
  • the terms "match” and “mismatch” as used herein refer to the hybridisation potential of paired nucleotides in complementary nucleic acid strands. Matched nucleotides hybridize efficiently, such as the classical A-T and G-C base pair mentioned above. Mismatches are other combinations of nucleotides that do not hybridize efficiently.
  • Reference herein to "immuno-interactive” includes reference to any interaction, reaction, or other form of association between molecules and in particular where one of the molecules is, or mimics, a component of the immune system.
  • isolated is meant material that is substantially or essentially free from components that normally accompany it in its native state.
  • modulating is meant increasing or decreasing, either directly or indirectly, the level or functional activity of a target molecule.
  • an agent may indirectly modulate the level/activity by interacting with a molecule other than the target molecule.
  • indirect modulation of a gene encoding a target polypeptide includes within its scope modulation of the expression of a first nucleic acid molecule, wherein an expression product of the first nucleic acid molecule modulates the expression of a nucleic acid molecule encoding the target polypeptide.
  • obesity includes conditions where there is an increase in body fat beyond the physical requirement as a result of excess accumulation of adipose tissue in the body.
  • the term obesity includes but is not limited to the following conditions: adult-onset obesity; alimentary obesity; endogenous or metabolic obesity; endocrine obesity; familial obesity; hyperinsulinar obesity; hyperplastic-hypertrophic obesity; hypogonadal obesity; hypothyroid obesity; lifelong obesity; morbid obesity and exogenous obesity.
  • obtained from is meant that a sample such as, for example, a polynucleotide extract or polypeptide extract is isolated from, or derived from, a particular source of the host.
  • the extract can be obtained from a tissue or a biological fluid isolated directly from the host.
  • oligonucleotide refers to a polymer composed of a multiplicity of nucleotide residues (deoxyribonucleotides or ribonucleotides, or related structural variants or synthetic analogues thereof) linked via phosphodiester bonds (or related structural variants or synthetic analogues thereof).
  • oligonucleotide typically refers to a nucleotide polymer in which the nucleotide residues and linkages between them are naturally occurring, it will be understood that the term also includes within its scope various analogues including, but not restricted to, peptide nucleic acids (PNAs), phosphoramidates, phosphorothioates, methyl phosphonates, 2-O-methyl ribonucleic acids, and the like. The exact size of the molecule can vary depending on the particular application.
  • PNAs peptide nucleic acids
  • phosphoramidates phosphoramidates
  • phosphorothioates phosphorothioates
  • methyl phosphonates 2-O-methyl ribonucleic acids
  • oligonucleotide is typically rather short in length, generally from about 10 to 30 nucleotide residues, but the term can refer to molecules of any length, although the te ⁇ n "polynucleotide” or “nucleic acid” is typically used for large oligonucleotides.
  • operably linked is meant that transcriptional and translational regulatory polynucleotides are positioned relative to a polypeptide-encoding polynucleotide in such a manner that the polynucleotide is transcribed and the polypeptide is translated.
  • patient refers to patients of human or other animal origin and includes any individual it is desired to examine or treat using the methods of the invention.
  • Suitable animals that fall within the scope of the invention include, but are not restricted to, primates, livestock animals (e.g, sheep, cows, horses, donkeys, pigs), laboratory test animals (e.g, rabbits, mice, rats, guinea pigs, hamsters), companion animals (e.g, cats, dogs) and captive wild animals (e.g, foxes, deer, dingoes, avians, reptiles).
  • pharmaceutically acceptable carrier is meant a solid or liquid filler, diluent or encapsulating substance that can be safely used in topical or systemic administration to a mammal.
  • polynucleotide or “nucleic acid” as used herein designates mRNA, RNA, cRNA, cDNA or DNA. The term typically refers to oligonucleotides greater than 30 nucleotide residues in length.
  • polynucleotide variant and “variant” refer to polynucleotides displaying substantial sequence identity with a reference polynucleotide sequence or polynucleotides that hybridize with a reference sequence under stringent conditions as known in the art (see for example Sambrook et al, Molecular Cloning. A Laboratory Manual", Cold Spring Harbor Press, 1989).
  • polynucleotides in which one or more nucleotides have been added or deleted, or replaced with different nucleotides.
  • certain alterations inclusive of mutations, additions, deletions and substitutions can be made to a reference polynucleotide whereby the altered polynucleotide retains a biological function or activity of the reference polynucleotide.
  • polynucleotide variant and “variant” also include naturally-occurring allelic variants.
  • Polypeptide “peptide” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues and to variants and synthetic analogues of the same.
  • polypeptide variant refers to polypeptides in which one or more amino acids have been replaced by different amino acids. It is well understood in the art that some amino acids may be changed to others with broadly similar properties without changing the nature of the activity of the polypeptide (conservative substitutions) as described hereinafter. These terms also encompass polypeptides in which one or more amino acids have been added or deleted, or replaced with different amino acids.
  • primer an oligonucleotide which, when paired with a strand of DNA, is capable of initiating the synthesis of a primer extension product in the presence of a suitable polymerizing agent.
  • the primer is preferably single-stranded for maximum efficiency in amplification but can alternatively be double-stranded.
  • a primer must be sufficiently long to prime the synthesis of extension products in the presence of the polymerization agent. The length of the primer depends on many factors, including application, temperature to be employed, template reaction conditions, other reagents, and source of primers. For example, depending on the complexity of the target sequence, the oligonucleotide primer typically contains 15 to 35 or more nucleotide residues, although it can contain fewer nucleotide residues.
  • Primers can be large polynucleotides, such as from about 200 nucleotide residues to several kilobases or more. Primers can be selected to be “substantially complementary” to the sequence on the template to which it is designed to hybridize and serve as a site for the initiation of synthesis. By “substantially complementary”, it is meant that the primer is sufficiently complementary to hybridize with a target polynucleotide. Preferably, the primer contains no mismatches with the template to which it is designed to hybridize but this is not essential. For example, non-complementary nucleotide residues can be attached to the 5' end of the primer, with the remainder of the primer sequence being complementary to the template.
  • non-complementary nucleotide residues or a stretch of non-complementary nucleotide residues can be interspersed into a primer, provided that the primer sequence has sufficient complementarity with the sequence of the template to hybridize therewith and thereby form a template for synthesis of the extension product of the primer.
  • Probe refers to a molecule that binds to a specific sequence or sub-sequence or other moiety of another molecule. Unless otherwise indicated, the term “probe” typically refers to a polynucleotide probe that binds to another polynucleotide, often called the "target polynucleotide", through complementary base pairing.
  • Probes can bind target polynucleotides lacking complete sequence complementarity with the probe, depending on the stringency of the hybridization conditions. Probes can be labelled directly or indirectly.
  • the term "recombinant polynucleotide" as used herein refers to a polynucleotide formed in vitro by the manipulation of a polynucleotide into a fonn not normally found in nature.
  • the recombinant polynucleotide can be in the form of an expression vector.
  • expression vectors include transcriptional and translational regulatory polynucleotide operably linked to the polynucleotide.
  • reporter molecule as used in the present specification is meant a molecule that, by its chemical nature, provides an analytically identifiable signal that allows the detection of a complex comprising an antigen-binding molecule and its target antigen.
  • reporter molecule also extends to use of cell agglutination or inhibition of agglutination such as red blood cells on latex beads, and the like.
  • vector is meant a polynucleotide molecule, preferably a DNA molecule derived, for example, from a plasmid, bacteriophage, yeast or virus, into which a polynucleotide can be inserted or cloned.
  • a vector preferably contains one or more unique restriction sites and can be capable of autonomous replication in a defined host cell including a target cell or tissue or a progenitor cell or tissue thereof, or be integrable with the genome of the defined host such that the cloned sequence is reproducible.
  • the vector can be an autonomously replicating vector, i.e., a vector that exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g, a linear or closed circular plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome.
  • the vector can contain any means for assuring self- replication.
  • the vector can be one which, when introduced into the host cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated.
  • a vector system can comprise a single vector or plasmid, two or more vectors or plasmids, which together contain the total DNA to be introduced into the genome of the host cell, or a transposon.
  • the choice of the vector will typically depend on the compatibility of the vector with the host cell into which the vector is to be introduced, hi the present case, the vector is preferably a viral or viral-derived vector, which is operably functional in animal and preferably mammalian cells.
  • Such vector may be derived from a poxvirus, an adenovirus or yeast.
  • the vector can also include a selection marker such as an antibiotic resistance gene that can be used for selection of suitable transformants.
  • resistance genes examples include the nptll gene that confers resistance to the antibiotics kanamycin and G418 (Geneticin®) and the hph gene which confers resistance to the antibiotic hygromycin B.
  • wild-type and normal are used interchangeably to refer to the phenotype that is characteristic of most of the members of the species occurring naturally and contrast for example with the phenotype of a mutant.
  • underscoring or italicizing the name of a gene shall indicate the gene, in contrast to its protein product, which is indicated by the name of the gene in the absence of any underscoring or italicizing.
  • Fgf-1 shall mean the Fgf-1 gene
  • FGF-1 shall indicate the protein product or products generated from transcription and translation and alternative splicing of the "Fgf-1” gene.
  • alkyl is intended to include both branched and straight-chain saturated aliphatic hydrocarbon group and may have a specified number of carbon atoms.
  • - Cio as in “C Ci 0 alkyl” is defined to include groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbons in linear or branched arrangement.
  • C ⁇ -C ⁇ 0 alkyl specifically includes, but is not limited to, methyl, ethyl, «-propyl, t-propyl, n-butyl, t-butyl, -butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl.
  • cycloalkyl or "aliphatic ring” means a monocyclic saturated aliphatic hydrocarbon group and may have a specified number of carbon atoms.
  • cycloallcyl includes, but is not limited to, cyclopropyl, methyl-cyclopropyl, 2,2-dimethyl-cyclobutyl, 2-ethyl- cyclopentyl, cyclohexyl.
  • cycloalkenyl means a monocyclic unsaturated hydrocarbon group and may have a specified number of carbon atoms.
  • cycloalkenyl includes but is not limited to, cyclobutenyl, cyclopentenyl, 1-methylcyclopentenyl, cyclohexenyl and cyclohexadienyl.
  • alkoxy represents either a cyclic or non-cyclic allcyl group attached through an oxygen bridge. “Alkoxy” therefore encompasses the definitions of allcyl and cycloallcyl above.
  • alkoxy groups include but are not limited to methoxy, oxy ethoxy, n-propyloxy, i- propyloxy, cyclopentyloxy and cyclohexyloxy. If no number of carbon atoms is specified, the term “alkenyl” refers to a non-aromatic hydrocarbon radical, straight, branched or cyclic, containing from 2 to 10 carbon atoms and at least one carbon to carbon double bond.
  • C 2 -C 6 alkenyl means an alkenyl radical having from 2 to 6 carbon atoms.
  • Alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, 2-methylbutenyl and cyclohexenyl. The straight, branched or cyclic portion of the alkenyl group may contain double bonds and may be substituted if a substituted alkenyl group is indicated.
  • alkynyl refers to a hydrocarbon radical straight, branched or cyclic, containing from 2 to 10 carbon atoms and at least one carbon to carbon triple bond. Up to three carbon-carbon triple bonds may be present.
  • C 2 -C 6 alkynyl means an alkynyl radical having from 2 to 6 carbon atoms.
  • Alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, 3- methylbutynyl and so on.
  • the straight, branched or cyclic portion of the alkynyl group may contain triple bonds and may be substituted if a substituted alkynyl group is indicated.
  • substituents may be defined with a range of carbons that includes zero, such as (Co-C 6 )allcylene-aryl. If aryl is taken to be phenyl, this definition would include phenyl itself as well as, for example, -CH 2 Ph, -CH 2 CH 2 Ph, CH(CH 3 )CH 2 CH(CH 3 )Ph.
  • aromatic or aryl is intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 atoms in each ring, wherein at least one ring is aromatic.
  • aryl elements include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl.
  • halo or halogen as used herein is intended to include chloro, fluoro, bromo and iodo.
  • heterocycle , “heteroaliphatic” or “heterocyclyl” as used herein is intended to mean a 5- to 10-membered nonaromatic heterocycle containing from 1 to 4 heteroatoms selected from the group consisting of O, N and S, and includes bicyclic groups.
  • heteroaryl or “heteroaromatic” , as used herein, represents a stable monocyclic or bicyclic ring of up to 7 atoms in each ring, wherein at least one ring is aromatic and contains from 1 to 4 heteroatoms selected from the group consisting of O, N and S.
  • Heteroaryl groups within the scope of this definition include but are not limited to: acridinyl, carbazolyl, cirmolinyl, quinoxalinyl, pyrrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, bezofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinoline.
  • heteroaryl is also understood to include the N-oxide derivative of any nitrogen-containing heteroaryl.
  • alkylene refers to a straight, branched or cyclic, preferably straight or branched, bivalent aliphatic hydrocarbon group, preferably having from 1 to about 20 carbon atoms, more preferably 1 to 12 carbons, even more preferably lower allcylene.
  • the alkylene group is optionally substituted with one or more "allcyl group substituents.” There may be optionally inserted along the allcylene group one or more oxygen, sulphur or substituted or unsubstituted nitrogen atoms, where the nitrogen substituent is allcyl as previously described.
  • Exemplary allcylene groups include methylene (-CH 2 -), ethylene (-CH 2 CH 2 -), propylene (-(CH 2 ) 3 -), cyclohexylene (- C 6 H ⁇ 0 -), methylenedioxy (-0-CH 2 -0-) and ethylenedioxy (-0-(CH 2 ) 2 -0-).
  • the term "lower allcylene” refers to allcylene groups having 1 to 6 carbons.
  • Preferred alkylene groups are lower allcylene, with allcylene of 1 to 3 carbon atoms being particularly preferred.
  • alkenylene refers to a straight, branched or cyclic, preferably straight or branched, bivalent aliphatic hydrocarbon group, preferably having from 2 to about 20 carbon atoms and at least one double bond, more preferably 2 to 12 carbons, even more preferably lower alkenylene.
  • the alkenylene group is optionally substituted with one or more "allcyl group substituents.” There may be optionally inserted along the alkenylene group one or more oxygen, sulphur or substituted or unsubstituted nifrogen atoms, where the nifrogen substituent is allcyl as previously described.
  • lower alkenylene refers to alkenylene groups having 2 to 6 carbons. Preferred alkenylene groups are lower alkenylene, with alkenylene of 3 to 4 carbon atoms being particularly preferred.
  • alkynylene refers to a straight, branched or cyclic, preferably straight or branched, bivalent aliphatic hydrocarbon group, preferably having from 2 to about 20 carbon atoms and at least one triple bond, more preferably 2 to 12 carbons, even more preferably lower allcynylene.
  • the alkynylene group is optionally substituted with one or more "allcyl group substituents.” There may be optionally inserted along the allcynylene group one or more oxygen, sulphur or substituted or unsubstituted nitrogen atoms, where the nitrogen substituent is allcyl as previously described.
  • the term "lower allcynylene” refers to allcynylene groups having 2 to 6 carbons. Preferred allcynylene groups are lower allcynylene, with allcynylene of 3 to 4 carbon atoms being particularly preferred.
  • arylene refers to a monocyclic or polycyclic, preferably monocyclic, bivalent aromatic group, preferably having from 3 to about 20 carbon atoms and at least one aromatic ring, more preferably 3 to 12 carbons, even more preferably lower arylene.
  • the arylene group is optionally substituted with one or more "allcyl group substituents.” There may be optionally inserted around the arylene group one or more oxygen, sulphur or substituted or unsubstituted nitrogen atoms, where the nitrogen substituent is allcyl as previously described.
  • Exemplary arylene groups include 1,2-, 1,3- and 1,4- phenylene.
  • the term "lower arylene” refers to arylene groups having 5 or 6 carbons.
  • Preferred arylene groups are lower arylene.
  • Preferred acyl includes C(0)-R, wherein R is hydrogen or an allcyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclyl residue, preferably a C
  • acyl examples include formyl; straight chain or branched alkanoyl such as, acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl, 2,2-dimethylpropanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, tridecanoyl, tefradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl and icosanoyl; cycloallcylcarbonyl such as cyclopropylcarbonyl, cyclobutylcarbonyl, cyclopentylcarbonyl and cyclohexylcarbonyl; aroyl such as benzoyl,
  • phenylacetyl phenylpropanoyl, phenylbutanoyl, phenylisobutanoyl, phenylpentanoyl and phenylhexanoyl
  • naphthylalkanoyl e.g. naphthylacetyl, naphthylpropanoyl and naphthylbutanoyl
  • aralkenoyl such as phenylalkenoyl (e.g.
  • phenylpropenoyl e.g., phenylbutenoyl, phenylmethacryloyl, phenylpentenoyl and phenylhexenoyl and naphthylalkenoyl (e.g.
  • aryloxyalkanoyl such as phenoxyacetyl and phenoxypropionyl
  • arylthiocarbamoyl such as phenylthiocarbamoyl
  • arylglyoxyloyl such as phenylglyoxyloyl and naphthylglyoxyloyl
  • arylsulfonyl such as phenylsulfonyl and napthylsulfonyl
  • heterocycliccarbonyl heterocyclicalkanoyl such as thienylacetyl, thienylpropanoyl, thienylbutanoyl, thienylpentanoyl, thienylhexanoyl, thiazolylacetyl, thiadiazolylacetyl and tetrazolylacetyl
  • heteroarylene refers to a bivalent monocyclic or multicyclic ring system, preferably of about 3 to about 15 members where one or more, more preferably 1 to 3 of the atoms in the ring system is a heteroatom, that is, an element other than carbon, for example, nifrogen, oxygen and sulfur atoms.
  • the heteroarylene group may be optionally substituted with one or more, preferably 1 to 3, aryl group substituents.
  • Exemplary heteroarylene groups include, for example, 1 ,4-imidazolylene.
  • arylallcylidene refers to an alkylidene group in which either R' or R" is and aryl group.
  • diarylalkylidene refers to an alkylidene group in which R' and R" are both aryl groups.
  • Diheteroarylalkylidene refers to an alkylidene group in which R' and R" are both heteroaryl groups.
  • arylidene refers to an unsaturated cyclic bivalent group where both points of attachment are on the same atom of the ring.
  • exemplary arylidene groups include, but are not limited to, quinone methide moieties that have the formula:
  • Heteroarylidene groups are arylidene groups where one or two, preferably two, of the atoms in the ring are heteroatoms, such as, but not limited to, O, S and N.
  • “Aralkyl” means allcyl as defined above which is substituted with an aryl group as defined above, e.g, -CH 2 phenyl, -(CH 2 ) 2 phenyl, -(CH 2 ) 3 phenyl, -H 2 CH(CH 3 )CH 2 phenyl, and the like and derivatives thereof.
  • Phenylallcyl means allcyl as defined above which is substituted with phenyl, e.g, - CH 2 phenyl also known as benzyl, -(CH 2 ) 2 phenyl, -(CH 2 ) 3 phenyl, CH 3 CH(CH 3 )CH 2 ⁇ henyl, and the like and derivatives thereof. Phenylallcyl is a subset of the aralkyl group.
  • Heteroaralkyl means allcyl as defined above which is substituted with a heteroaryl group, e.g, -CH 2 pyridinyl, -(CH 2 ) 2 pyrimidinyl, -(CH 2 ) 3 imidazolyl, and the likes and derivatives thereof.
  • Heterocyclylallcyl means allcyl as defined above which is substituted with a heterocycle group, e.g, -CH 2 pyrrolidin-l-yl, -(CH 2 ) 2 piperidin-l-yl, and the like, and derivatives thereof.
  • hydrocarbyl as used herein includes any radical containing carbon and hydrogen including saturated, unsaturated, aromatic, straight or branched chain or cyclic including polycyclic groups. Hydrocarbyl includes but is not limited to C ⁇ -C 8 alkyl, C 2 -C 8 alkenyl, C 2 - Cgalkynyl, C 3 -C ⁇ 0 cycloallcyl, aryl such as phenyl and naphthyl, Ar (C 1 -C 8 )alkyl such as benzyl, any of which may be optionally substituted.
  • pseudohalides are groups that behave substantially similar to halides.
  • Such groups can be used in the same manner and treated in the same manner as halides (X, in which X is a halogen, such as CI or Br).
  • Pseudohalides include, but are not limited to cyanide, cyanate, thiocyanate, selenocyanate, trifluoromethyl and azide. It will also be recognised that the compounds described herein may possess asymmetric centres and are therefore capable of existing in more than one stereoisomeric form.
  • the invention thus also relates to compounds in substantially pure isomeric form at one or more asymmetric centres e.g, greater than about 90% ee, such as about 95% or 97% ee or greater than 99% ee, as well as mixtures, including racemic mixtures, thereof.
  • Such isomers may be naturally occurring or may be prepared by asymmetric synthesis, for example using chiral intermediates, or by chiral resolution.
  • the present invention relates to a method of inhibiting or reducing adipogenesis in obesity or conditions of localized, abnormal increases in adipogenesis comprising administering to a patient in need of such treatment an adipogenesis inhibiting amount of an agent which impairs or interferes with a FGF signalling pathway, and optionally a pharmaceutically acceptable carrier or diluent.
  • the agent may be selected from small organic molecules, peptides, polypeptides, proteoglycans, proteins, sugars, oligosaccharides and carbohydrates as defined below.
  • Suitable small organic molecules that impair or interfere with a FGF signalling pathway include:
  • X is CH or N
  • B is halo, hydroxy, or NR 3 R ;
  • Ri, R 2 , R 3 and R 4 independently are hydrogen, C C 8 allcyl, C 2 -C 8 alkenyl, C 2 -C 8 allcynyl, Ar 1 , amino, C ⁇ -C 8 allcylamino or di-Ci-Cg alkylamino; and wherein the allcyl, alkenyl, and alkynyl groups may be substituted by NR 5 R 6 , where R 5 and R 6 are independently hydrogen, C C 8 alkyl,
  • any of the foregoing alkyl, alkenyl, and allcynyl groups may be substituted with hydroxy or a 5- or 6-membered carbocyclic or heterocyclic ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen, and sulfur, and R 9 , Rjo, Rn and R[ 2 independently are hydrogen, nitro, trifluoromethyl, phenyl, substituted phenyl, -C ⁇ N, -COOR 8 , -COR 8 ,
  • n 0, 1, 2 or 3; and wherein R 5 and R 6 together with the nitrogen to which they are attached can complete a ring having 3 to 6 carbon atoms and optionally containing a heteroatom selected from nitrogen, oxygen, and sulfur;
  • R ! and R 2 together with the nitrogen to which they are attached, and R 3 and R 4 together with the nitrogen to which they are attached, can also be
  • R can complete a ring having 3 to 6 carbon atoms and optionally containing 1 or 2 heteroatoms selected from nifrogen, oxygen, and sulfur, and R] and R additionally can be an acyl analog selected from
  • R 8 is hydrogen, -C 8 allcyl, C 2 -C 8 alkenyl, C 2 -C 8 allcynyl, C 3 -C ⁇ 0 cycloallcyl optionally containing an oxygen, nitrogen, or sulfur atom,
  • R allcyl, alkenyl, and allcynyl groups can be substituted by NR 5 R 6 ;
  • Ar and Ar are unsubstituted or substituted aromatic or heteroaromatic groups selected from phenyl, imidazolyl, pyrrolyl, pyridyl, pyrimidyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, pyrazinyl, thiazolyl, oxazolyl, isoxazolyl, furnanayl, thienyl, naphthyl, wherein the substituents are R 9 , Rio, Ri 1 and R 12 as defined above; and the pharmaceutically acceptable acid and base addition salts thereof; provided that when X is N, B is NHCONHtbutyl and Ar is 2,6- dichlorophenyl, Ri and R 2 cannot be hydrogen and 4-diethylaminobutyl. Particularly
  • R 3 , P . R S , Re, R 9 and R, 0 are defined in formula (I) above.
  • Exemplary compounds include: l-tert-Butyl-3-[6-(2,6-dichlorophenyl)-2-(3-diethylamino-propylamino)-pyrido[2,3-d]-pyrimidin- 7-yl]-urea; l-tert-Butyl-3-[6-(2,6-dichlorophenyl)-2-(3-dimethylamino-propylamino)-pyrido[2,3-d]pyrimidin-
  • Ri and R 2 are at each occurrence independently selected from halogen, nitro, cyano, trifluoromethyl, hydrocarbyl, ORt, SR 4 , SOR 5 , S0 2 R 5 , COOH, COR 6 , SONR 7 R 8 , S0 2 NR 7 R 8 and
  • R 3 is selected from H or R and is absent when X is O;
  • R 9 and R 10 are independently selected from H and R ⁇ R 4 is selected from H, hydrocarbyl, COR 6 , and CONR 7 R 8 ;
  • R 5 is hydrocarbyl
  • R 6 is selected from H, hydrocarbyl, OR 5 and NR 7 R 8 ;
  • R 7 and R 8 are each independently selected from H or hydrocarbyl, or one of R 7 and R 8 is H or hydrocarbyl and the other is COR 5 , COOR 5 , or CONR 7 R 8 , or R 7 and R 8 together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring optionally containing 1-2 further heteroatoms selected from oxygen, nifrogen and sulfur; m is 0 to 3 and n is 0 to 5.
  • Exemplary compounds are those wherein
  • X is N, m is 1, R, at position 5 is a radical NHCOCH 3 , R 9 and R 10 are H, R 3 is CH 2 -CH 2 -COOH, CH 2 -CH 2 -COOR 5 , or CH 2 -CH 2 -CONR 7 R 8 , wherein R 5 is C C g allcyl, preferably methyl, and R 7 and
  • R 8 are each independently selected from H or hydrocarbyl or R 7 and R 8 together with the nifrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring optionally containing 1-2 further heteroatoms selected from oxygen, nifrogen and sulfur, n is 2 and R 2 is C ⁇ -C 8 alkoxy, preferably methoxy, most preferably at positions 3 and 5 of the phenyl radical.
  • a preferred compound is 3-(5-acetylamino-4-carbamoyl-2-(3,5-dimethoxyphenyl)-benzimidazol-lyl)-propionic acid.
  • R, and R 2 are at each occurrence are independently selected from halogen, nitro, cyano, frifluoromethyl, hydrocarbyl, OR,, SR 4 , SOR 5 , S0 2 R 5 , COOH, COR 6 , SONR 7 R 8 , S0 2 NR 7 R 8 and
  • R 4 is selected from H, hydrocarbyl, COR 6) and CONR 7 R 8 ;
  • R 5 is hydrocarbyl
  • R 6 is selected from H, hydrocarbyl, OR 5 and NR 7 R 8 ;
  • R 7 and R 8 are each independently selected from H or hydrocarbyl, or one of R 7 and R 8 is H or hydrocarbyl and the other is COR 5 , COOR 5 , or CONR 7 R 8 , or R 7 and R 8 together with the nifrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring optionally containing 1-2 further heteroatoms selected from oxygen, nitrogen and sulfur; m and n independently are an integer from 0 to 4.
  • Exemplary compounds are those wherein R 3 is H, m is 1, Ri is selected from OH or dimethyl carboxamoyl, n is 2, R 2 is selected from N0 2 or NH 2 , especially 3 -hydroxy 9-nifro-5H- benzofuro[3,2-c] quinoline-6-one and 3-methylcarbamoyloxy-9-amino-5H-benzofuro[3,2-c] quinoline-6-one.
  • These compounds, methods for their preparation and their biological activity are disclosed in WO 03/020698. The disclosed compounds are described as having an inhibitory effect on tyrosine kinase activity associated with an FGFR.
  • R la is independently selected from H, unsubstituted or substituted C ⁇ -C 10 allcyl, OR 8 , and N(R 8 ) 2 ;
  • R 1 is independently selected from H, unsubstituted or substituted C]-C ⁇ o allcyl, unsubstituted or substituted C 3 -C 10 cycloallcyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, halo, CF 3 , -(CH 2 ) t R 9 C(0)R 8 , -C(0)R 9 , -(CH 2 ) t OR 8 , unsubstituted or substituted C 2 -C 6 alkenyl, unsubstituted or substituted C 2 -C 6 alkynyl, CN, -(CH 2 ) t NR 7 R 8 , -(CH 2 ) t C(0)NR 7 R 8 , -C(0)OR 8 , and
  • R 7 and R 8 when attached to the same nitrogen atom may be joined to form a 5-7 membered heterocycle containing, in addition to the nitrogen, one or two more heteroatoms selected from N, O, or S, said heterocycle being optionally substituted with one to three R 2 substituents;
  • R 9 is independently selected from unsubstituted or substituted C C ⁇ o allcyl, unsubstituted or substituted heterocycle, and unsubstituted or substituted aryl;
  • W is selected from aryl, and heterocycle;
  • m is 0, 1 or 2;
  • n is independently 0, 1, 2, 3, 4, 5 or 6;
  • p is 0, 1, 2, 3 or 4;
  • q is independently 0, 1 or 2;
  • t is independently 0, 1, 2, 3, 4, 5 or 6; or a pharmaceutically acceptable salt, hydrate or stereoisomer thereof.
  • heterocyclyl encompasses saturated, unsaturated and heteroaromatic groups.
  • Exemplary compounds include:
  • R 1 is independently selected from H, unsubstituted or substituted Ci-Cio allcyl, halo, unsubstituted or substituted aryl, unsubstituted or substituted heterocycle, CF 3 , -(CH 2 ) t R 9 C(0)R 8 , -C(0)R 9 , and
  • R 2 is independently selected from H, unsubstituted or substituted C ⁇ -C 10 alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocycle, halo, OR 8 , N(R 8 ) 2 , and CN;
  • R 3 is independently selected from H, unsubstituted or substituted C ⁇ -C ⁇ 0 allcyl, and unsubstituted or substituted aralkyl;
  • R 8 is independently selected from H, unsubstituted or substituted CpCio alkyl, and unsubstituted or substituted aryl;
  • R 9 is independently selected from unsubstituted or substituted aryl, and unsubstituted or substituted heterocycle; m is 0, 1 or 2; n is O, 1, 2, 3, 4, 5 or 6; p is O, 1, 2, 3 or 4; t is independently 0, 1, 2, 3, 4, 5 or 6; or a pharmaceutically acceptable salt, hydrate or stereoisomer thereof.
  • Preferred compounds include: 4-(2-amino-5-bromo-l,3-thiazol-4-yl)-N-(3,5-dimethylphenyl)pyrimidin-2-amine; 4-(2-amino-l,3-thiazol-4-yl)-N-(3,5-dimethylphenyl)pyrimidin-2-amine; 4-(2-amino-5-phenyl-l,3-thiazol-4-yl)-N-(3,5-dimethylphenyl)pyrimidin-2-amine; 2-amino-4- ⁇ 2-[(3 ,5 -dimethylphenyl)amino]pyrimidin-4-yl ⁇ - 1 ,3 -thiazole-5 -carbonitrile; 4- ⁇ 2-[(3,5-dimethylphenyl)amino]pyrimidin-4-yl ⁇ -l,3-thiazole-5-carbonitrile; or a pharmaceutically acceptable salt or hydrate thereof.
  • W is selected from:
  • X and Y are independently selected from C or N, provided that when X is N, then Y is C and when
  • X is C, then Y is N;
  • V is C orN;
  • R 1 is selected from unsubstituted and substituted aryl or unsubstituted or substituted heterocycle, where the substituted group may have from 1 to 3 substituents selected from unsubstituted or substituted C ⁇ -C 6 allcyl, unsubstituted or substituted C 3 -C 10 cycloallcyl, unsubstituted or substituted aryl, unsubstituted or substituted aralkyl, CF 3 , OR 4 , halo, CN, -(CH 2 ) t R 9 C(0)R 4 , -(CH 2 ) t OR 4 ,
  • R 4 and R 7 may be taken together with the nitrogen to which they are attached to form a 5-7 membered heterocycle containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said heterocycle being optionally substituted with one to three substituents selected from R 2 ; and -C(0)R 4 ;
  • R 2 is selected from H, halo, unsubstituted or substituted C ⁇ -C 6 alkyl, unsubstituted or substituted aryl, unsubstituted or substituted C 2 -C 6 alkenyl, unsubstituted or substituted C 2 -C 6 alkynyl, OR 4 , CN and N(R 4 ) 2 ;
  • R 3 is independently selected from H, unsubstituted or substituted C]-C 6 allcyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, CN, halo, OR 4 , and N(R 4 ) 2 ;
  • R 4 is selected from H, unsubstituted or substituted C x -C 6 allcyl, unsubstituted or substituted aryl, unsubstituted or substituted aralkyl, and unsubstituted or substituted heterocyclyl;
  • R 7 is selected from H, unsubstituted or substituted C ⁇ -C 6 allcyl, unsubstituted or substituted aryl, unsubstituted or substituted aralkyl, and unsubstituted or substituted heterocycle;
  • R 9 is selected from unsubstituted or substituted heterocycle; m is 0, 1 or 2; n is 0, 1, 2, 3, 4 or 5; and t is 0, 1, 2, 3, 4 or 5; or a pharmaceutically acceptable salt, hydrate or stereoisomer thereof.
  • heterocyclyl or “heterocycle” included saturated, unsaturated and heteroaromatic groups.
  • Especially desirable compounds have the following formula:
  • X and Y are independently selected from C or N, provided that when X is N, then Y is C and when X is C, then Y is N;
  • R 2 is selected from H, halo, unsubstituted or substituted C C 6 allcyl, and OR 4 ;
  • R 3 is independently selected from H, unsubstituted or substituted C]-C 6 allcyl, unsubstituted or substituted aryl, and unsubstituted or substituted heterocyclyl.
  • R 4 is selected from H, unsubstituted or substituted C ⁇ -C 6 allcyl, unsubstituted or substituted aryl, unsubstituted or substituted aralkyl, and unsubstituted or substituted heterocyclyl;
  • R 5 is independently selected from unsubstituted or substituted C ⁇ -C 6 allcyl, OR 4 , halo, and CN;
  • R 7 is selected from H, unsubstituted or substituted -C ⁇ allcyl, unsubstituted or substituted aryl, unsubstituted or substituted aralkyl, and unsubstituted or substituted heterocyclyl;
  • R 9 is selected from unsubstituted or substituted heterocyclyl; m is 0, 1 or 2; n is 0, 1, 2, 3, 4 or 5; and q is O, 1, 2, 3 or 4; or a pharmaceutically acceptable salt, hydrate or stereoisomer thereof.
  • exemplary compounds include: (4-Indol-l -yl-pyrimidin-2-yl)-phenyl-amine;
  • X is selected from CH or ⁇ ;
  • Ri is selected from H, C 6 allcyl, C 2 - 6 alkenyl and C ⁇ - 6 alkyl ⁇ (R 4 ) 2 ;
  • R 2 is selected from H, halogen, C 6 alkyl, hydroxy, C ⁇ - 6 allcoxy, -OCOC 6 alkoxy, frifluoromethyl, cyano, nitro, NH 2 , NHCp ⁇ aikyl and (C ⁇ - 6 alkyl) 2 ;
  • R 3 is selected from COR 5 , C r6 alkyl, phenyl, SO z R 5 and cyano;
  • Each R is independently selected from H and C ⁇ - 6 allcyl
  • R 5 is selected from [C(R 6 ) 2 ] m N(R 7 ) 2) [C(R 6 ) 2 ] m C0 2 R, [C(Rs) 2 ] m phenyl, Ci-ealkyl or heterocyclyl;
  • Each R 6 is independently selected from H, C 3 alkyl, hydroxy, C ⁇ al oxy frifluoromethyl, cyano, nitro and halo;
  • Each R 7 is independently selected from hydrogen, C ⁇ - 3 alkyl, [C(R 5 ) 2 ] m phenyl, [C(R 6 ) 2 ] m N(R 8 ) 2 ,
  • Each R 8 is independently selected from H and d- 3 alkyl; and m is 0 or an integer from 1 to 3; and wherein each phenyl group is optionally substituted with R 2 , C0 2 H or C0 2 C ⁇ -3alkyl.
  • Preferred compounds are those in which X is CH or (CR 2 );
  • R is hydrogen or methyl
  • R 3 is CH 2 CH 2 C0 2 H, CH 2 CH 2 C0 2 CH 3 , CH 2 CH(NH 2 )CONHPh or
  • CONHPh and R 2 is selected from H, 4-chloro, 4-methyl, 4-methoxy, 4-acetyloxy, 5-fluoro, 5- chloro, 5-bromo, 5-methyl, 5-methoxy, 5-acetyloxy, 5-hydroxy, 5-frifluoromethyl, 5-cyano, 5- nitro, 6-chloro, 6-methyl, 6-methoxy, 6-acetyloxy, 6-hydroxy, 7-chloro, 7-methyl, 7-methoxy, 7- acetyloxy, 7-hydroxy, or when X is N, R, is methyl, R 2 is hydrogen and R 3 is CONHPh, or those in which X is CH, R, is hydrogen, methyl or (CH 2 ) 3 N(CH 3 ) 2 , R 2 is H and R 3 is CH 3 , phenyl, CONH 2 ,
  • CONHCH 3 CON(CH 3 ) 2 , CONHPh, CONHCH 2 Ph, CONHCH 2 C0 2 H, CONH(CH 2 ) 2 N(CH 3 ), CONHCH 2 CH(OH)CH 2 OH, CONHCH 2 Ph(4-C0 2 H), CONHCH 2 Ph(4-C0 2 CH 3 ), CON(CH 3 )Ph,
  • the disclosed compounds are described as having an inhibitory effect on tyrosine kinase activity of growth factor receptors.
  • Ri is selected from halo, hydroxy, C ⁇ -3 allcoxy, SH, SC ⁇ -3 allcyl, C ⁇ -3 allcyl, C 2-3 alkenyl, C 2-3 allcynyl or cyano;
  • R 2 is selected from H, OC ⁇ _ 3 allcyl, OC 2-3 alkenyI, OC 2 . 3 alkynyl or OC 1 . 3 allcylOC ⁇ -3 allcyl; and R 3 is selected from Cj. 6 allcyl, C 2 . 6 alkenyl, C 2 _ 6 alkynyl, C ⁇ . 3 allcylO-C ⁇ . 3 allcyl, C ⁇ . 3 allcylS-C ⁇ -3 allcyl, heterocycle, heterocycleC ⁇ . 6 allcyl-, heterocycleC 2-6 alkenyl, heteroaryl, heteroarylC].
  • exemplary compounds of formula (VII) include those where m is 1 to 3, R] is selected from F, CI, Br, I, OH, CH 3 and cyano, R 2 is selected from H, methoxy or 0(CH 2 ) 2 OCH 3 and R 3 is selected from H, CH 3 , (CH 2 ) 2 0CH 3 , heterocyclyl, heterocyclylC 1 . 4 allcyl-, heterocyclylC 2-4 alkenyl-, heteroaryl, or heteroarylC 2 . 4 aIkenyl-.
  • Preferred compounds of formula (VII) are those in which m is 1 to 3, Ri is selected from F,
  • Ri is selected from halo, hydroxy, C ]-3 alkoxy, SH, SC ⁇ . 3 allcyl, C ⁇ . 3 alkyl, C 2-3 alkenyl, C 2-3 alkynyl or cyano;
  • R 2 is selected from H, OC ]-3 allcyl, OC 2 . 3 alkenyl, OC 2-3 alkynyl or and
  • R is selected from C ⁇ . 6 allcyl, C 2-6 alkenyl, C 2-6 alkynyl, C ⁇ -3 allcylO-C ⁇ . 3 allcyl-, C ⁇ . 3 a ⁇ lcyl8-C ⁇ . 3 alkyl-, heterocycle, heterocycleC ⁇ allcyl-, heterocycleC 2-6 alkenyl-, heteroaryl, heteroarylCi- ⁇ al yl-, heteroarylC 2-6 alkenyl-; and m is 0 or an integer from 1 to 4.
  • Exemplary compounds are those in which X is CH or N where m is 1 to 3, Ri is selected from F, CI, Br, I, OH, CH 3 and cyano, R 2 is selected from H, methoxy or 0(CH 2 ) 2 OCH 3 and R 3 is selected from H, CH 3 , -(CH 2 ) 2 OCH 3 , heterocyclyl, heterocyclylC ⁇ - allcyl-, heterocyclylC 2 . 4 alkenyl-, heteroaryl, heteroarylC ⁇ . 4 allcyl- or heteroarylC 2-4 alkenyl-.
  • Preferred compounds are those in which X is CH or N, m is 1 to 3, Ri is F, CI or OH, R 2 is
  • OCH 3 and R 3 is (CH 2 ) 2 OCH 3 .
  • the disclosed compounds are described as having an inhibitory effect on tyrosine kinase activity of growth factor receptors.
  • X is CH, C(R ⁇ ) or N; m is 0 or an integer from 1 to 2; each R] and R 2 is independently selected from H, C]. 3 allcyl, halo, N0 2 , CN, OH, OC ⁇ -3 allcyl, NH 2 ,
  • R 3 is selected from C h alky!, unsubstituted or substituted phenyl or R 3 and R 2 together may be -
  • R 4 is hydrogen or when R 3 is allcyl or forms a ring with R 2 , R 4 together with the first carbon atom of R 3 may form a double bond;
  • R 5 is selected from OH, OC ⁇ alkyl, NH 2 , NH(C 1-3 allcyl), N(C 1-3 alkyl) 2 , NH(CH 2 ) n N(R 8 ) 2 ;
  • R 6 is hydroxy
  • Each R 8 is independently selected from hydrogen and C ⁇ -3 allcyl; is a single or double bond; n is an integer from 1 to 3, and the phenyl in R 3 may be substituted one or more times with a group selected from C,. 3 allcyl, frifluoromethyl, halo, hydroxy, OC 1-3 alkyl, N0 2 , CN, NH 2 , NH(C ⁇ -3 alkyl) and N(C,. 3 allcyl) 2 .
  • Exemplary compounds include those in which:
  • X is CH or C(R,); m is 0 or 1 ; each R, is selected from CH 3 , CI, N0 2 and OCH 3 ;
  • R 2 is H or OCH 3 ;
  • R 3 is C 1-3 allcyl, unsubstituted phenyl or phenyl substituted with one or more substituents selected from methyl or halo; or R 3 and R 2 together are -CH 2 -CH 2 -CH 2 - or -CH 2 -CH 2 -CH 2 -CH 2 -;
  • R 4 is hydrogen or together with the first carbon atom of R 3 forms a double bond
  • R 5 is selected from OH, OCH 3 , NH 2 , NHCH 3 , N(CH 3 ) 2 or NH(CH 2 ) 2 N(CH 2 CH 3 ) 2 ;
  • Preferred compounds include:
  • R is selected from cycloallcyl, cycloalkenyl, heterocyclyl, aryl or heteroaryl;
  • Each R 2 is selected from hydrogen or C ⁇ -6 alkyl;
  • R 3 is selected from H, OH, C 1-6 alkoxy, halo, substituted C ⁇ -6 allcyl, halo, CN, N0 2 , cycloallcyl, C0 2 H, C0 2 C ⁇ _ 6 alkyl, halosubstituted C ⁇ . 6 alkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, NR 5 R 6 , CONR 5 R 6 or -C 1-6 alkylene CONR 5 R 6 ;
  • R 4 is selected from R 3 or
  • R 5 is selected from hydrogen or C ⁇ . 6 alkyl; and Re is selected from aryl, heteroaryl, heterocyclyl, aminoallcyl, alkylaminoalkyl, dialkylaminoallcyl, hydroxyalkyl, acetylalkyl, cyanoalkyl, carboxyalkyl, alkoxycarbonylalkyl, heteroaralkyl, aralkyl, or heterocyclylalkyl wherein the allcyl chain in aminoallcyl, alkylaminoalkyl, dialkylaminoallcyl, aralkyl, heteroaralkyl, or heterocyclylalkyl is optionally substituted with one or two hydroxy or R 5 and R 6 together with the nitrogen atom to which they are attached combine to form saturated or unsaturated heterocyclylamino; wherein each cycloalkyl, cycloalkenyl, heterocyclyl, aryl or heteroaryl in Ri may be optionally substituted with
  • each allcyl, aryl, heteroaryl, heterocyclyl and alkylene may be optionally substituted with C 1-3 alkyl, C 1-3 alkoxy, halo, CN, N0 2 , C0 2 H, COH, C0 2 C ⁇ -3 alkyl, COC,. 3 allcyl, COC,. 3 allcyl, NH 2 , NH(C,. 3 alkyl) or N(C 1-3 allcyl) 2 .
  • Exemplary compounds of formula (X) include those in which any one of the following definitions apply: is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl or optionally substituted heterocyclyl;
  • Each R 2 is hydrogen, or C h alky!;
  • R 3 is hydrogen, C,. 3 alkyl, OH, C,. 3 alkoxy, halo, CN, N0 2 , C0 2 H, C0 2 C 1-3 allcyl, NH 2 , NH(d. 3 alkyl) or N(C,. 3 allcyl) 2 ;
  • R 4 is H or
  • R 5 is H or d. 3 alkyl
  • R ⁇ is selected from aminoallcyl, alkylaminoalkyl, dialkylaminoallcyl, hydroxyalkyl, acetylallcyl, cyanoallcyl, carboxyallcyl, alkoxycarbonylalkyl, heteroarallcyl, or heterocyclylalkyl wherein the allcyl chain in aminoallcyl, heteroarallcyl, heteroarallcyl, or heterocyclylalkyl is optionally substituted with one or two hydroxy group(s); or R 5 and R 6 together with the nitrogen atom to which they are attached form saturated or unsaturated heterocycloamino; preferably saturated 5 or 6 membered heterocycloamino containing one or two nitrogen atoms, the remaining ring atoms being carbon.
  • One of the ring carbons may be optionally replaced by carbonyl or oxygen and the ring may be optionally substituted with one or two substituents independently selected group the group consisting of allcyl, hydroxy, dialkylamino, hydroxyalkyl, alkoxyalkyl, and optionally substituted heterocyclylalkyl wherein said heterocyclyl ring is 5 or 6 membered and contains one or two nitrogen atoms, the rest of the ring atoms being carbon.
  • R 5 and R 6 together with the nitrogen atom to which they are attached form 4-methylpiperazin-l-yl, 3,5-dimethylpiperazin-l-yl, piperidin-1-yl, morpholin-4-yl, 4-(pyrrolidin-l-yl)-piperidin-l-yl, 2- (pyrrolidin-l-ylmethyl)pyrrolidin-l-yl (wherein the stereochemistry at the C-2 carbon atom of the pyrrolidin-1-yl is RS, R or S), 4-hydroxypiperidin-l-yl, 4-aminopiperidin-l-yl, 3- diethylaminopyrrolidin-1-yl (wherein the stereochemistry at the C-3 carbon atom of the pyrrolidin- 1-yl is RS, R or S), 4-(pyrrolidin-l-yl)-piperidin-l-yl (stereochemistry at the C-4 carbon atom of the pyrrolidin-1-yl is RS
  • R 5 and R 6 together with the nitrogen atom to which they are attached form 2- (pyrrolidin-l-ylmethyl)pyrrolidin-l-yl (wherein the stereochemistry at the C-2 carbon atom of the pyrrolidin-1-yl is RS, R or S), preferably (R).
  • Rj is optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted phenyl, optionally substituted pyrrole, optionally substituted pyridine, optionally substituted furan or optionally substituted pyrimidine.
  • Ri may be
  • X is CH 2 , O or NH, especially NH and R 7 is hydrogen, alkyl, cycloalkyl, hydroxyalkyl, aminoallcyl, allcylaminoallcyl, diallcylaminoallcyl, carboxyallcyl, heterocyclylalkyl, aryl, heteroaryl, carboxy, alkoxycarbonyl, heterocyclycarbonyl, aminoalkylcarbonyl, alkylaminoallcylcarbonyl, diallcylaminoallcylcarbonyl, -CONR 5 R 6 , or -(alkylene)-CONR 5 R 6 .
  • R s and R 9 are independently hydrogen, allcyl, cycloallcyl, heterocyclylalkyl, -CORio, -(alkylene)- CORio where io is alkoxy, hydroxy, or heterocycle, alkylamino, dialkylamino), -S0 2 R ⁇ , - CONR ⁇ 2 R ⁇ , or -(allcylene) -CONR ⁇ 2 R n (where R ⁇ 2 is hydrogen or allcyl, and R n is aminoalkyl, allcylaminoallcyl, diallcylaminoallcyl, hydroxyalkyl, acetylalkyl, cyanoallcyl, carboxyallcyl, alkoxycarbonyallcyl, heteroallcyl, or heterocyclylalkyl wherein the allcyl chain is aminoallcyl, heteroarallcyl, heteroarallcyl, or heterocyclylalkyl is optionally
  • substituents include C ⁇ . 3 alkyl, especially methyl, halo and C,. 3 alkyleneCO 2 H.
  • R is optionally substituted phenyl, particularly 4-substituted phenyl.
  • Prefe ⁇ ed substituents include C ⁇ . 3 alkyl, halo, trifluoromethyl, cycloallcyl especially cyclohexyl, and heterocyclyl especially where R' can be H, C 1-3 allcyl, C0 2 H, C0 2 C 1-3 allcyl, C(O)H or C(0)C 1 . 3 alkyl.
  • Particularly prefened compounds include:
  • Ar 1 is a monocyclic or fused bicyclic, tricyclic or tetracyclic aromatic or heteroaromatic group, where the heteroaromatic group contains one or two, preferably two, heteroatoms selected from O, S and N
  • Ar 2 is a monocyclic or fused bicyclic, tricyclic or tetracyclic arylidene or heteroarylidene group, where the heteroarylidene group contains one or two, preferably two, heteroatoms selected from O, S, and N
  • the aryl, heteroaryl, arylidene and heteroarylidene moieties of the compounds of formula (XI) are unsubstituted or are substituted with one or more substituents each independently selected from Z, which, as defined herein, is halogen, hydroxy, nitrile, nitro, formyl, mercapto, carboxy, hydroxysulfonyl, hydroxyphosphoryl, allcyl, haloalkyl, polyhaloallcyl, aminoallcyl, diaminoallcyl, alkenyl containing 1 to 2 double bonds, alkynyl containing 1 to 2 triple bonds, cycloallcyl, cycloallcylallcyl, aryl, heteroaryl, arylalkyl, heteroarylallcyl, allcylidene, arylallcylidene, alkylcarbonyl, arylcarbonyl, heteroarylcarbonyl, alkoxycarbon
  • R' and R 5 are each independently selected from hydrogen, alkyl, arallcyl, heteroarallcyl, aryl, heteroaryl, C0 2 R 20 , S0 3 R 20 and PO 3 (R 20 ) 2 , or, together with R 13 , form oxy;
  • R 2 and R 4 are each independently hydrogen, hahde, pseudohahde, allcyl, arallcyl, heteroarallcyl, aryl or heteroaryl, or, together with R 3 , form allcylenylammo,
  • R 3 is hydrogen, hydroxy, thioxy, alkoxy, aryloxy, SR 40 or NR 40 R 41 , or, together with R 2 or R 4 , fonr s allcylenylammo,
  • R 6 and R 10 are each independently selected from hydrogen, hahde, pseudohahde, C0 2 R 20 , S0 3 R 20 and PO 3 (R 20 ) 2 ;
  • R 7 and R 9 are each independently hydrogen, hahde, pseudohahde, alkyl, aralkyl, heteroarallcyl, aryl or heteroaryl,
  • R 8 is hydrogen, hahde, pseudohahde, hydroxy, alkoxy, aralkoxy, heteroarallcoxy, aryloxy, heteroaryloxy, NR 40 R 41 , C0 2 R °, PO 3 (R 20 ) 2 or SO n R 20 where n is 0-3;
  • R n is selected from hydrogen, halide and pseudohahde, or, together with X, forms alkylenylammonium;
  • R 12 is hydrogen, halide, pseudohahde, alkyl, arallcyl, heteroarallcyl, aryl or heteroaryl, or, together with X, forms allcylenylammonium;
  • R 13 is hydrogen, or, together with R 1 or R 5 , forms oxy
  • R 14 is selected from hydrogen, alkyl, aralkyl, heteroarallcyl, aryl and heteroaryl;
  • X is oxy, thio, NR 40 or NbR ⁇ R 41 , or, together with R ⁇ and or R 12 , forms alkylenylammonium;
  • R 15 is C0 2 R 20 , S0 3 R 20 or PO 3 (R 20 ) 2 ;
  • R 16 is selected from hydrogen, alkoxy, aralkoxy, heteroaralkoxy, aryloxy and heteroaryloxy;
  • R 17 and R 18 are each independently hydrogen, halide or pseudohahde
  • R 20 is selected from hydrogen, allcyl, arallcyl, heteroarallcyl, aryl, heteroaryl and Na;
  • R 40 and R 4i are each independently hydrogen, allcyl, arallcyl, heteroarallcyl, aryl or heteroaryl, or together form allcylene or alkenylene.
  • the compounds are of fonnulae (XIa) where RbR 14 and X are selected as above.
  • the compounds are diphenylmethylidene quinone methides, diphenylmethylidene thiaquinone methides, and imminium derivatives thereof.
  • Exemplary compounds include:
  • R 51 is selected from hydrogen, allcyl, alkenyl, hydroxycarbonylalkyl, hydroxyalkyl, aralkyl, heteroarallcyl, aryl and heteroaryl; n is 0 or 1;
  • R 40 and R 41 are each independently hydrogen, alkyl, arallcyl, heteroaralkyl, aryl or heteroaryl, or together form allcylene or alkenylene;
  • R 52 is selected from aryl, heteroaryl and NR 60 R 61 ;
  • R 55 is allcyl, arallcyl, heteroarallcyl, aryl, heteroaryl, thioalkyl, thioaralkyl, thioheteroarallcyl, thioaryl or thioheteroaryl;
  • R 60 and R 6 ' are each independently hydrogen, aryl, heteroaryl or S(0) m -aryl or -heteroaryl, where m is 1 or 2, or together form allcylidene or cycloallcylidene;
  • R 70 is allcyl, arallcyl, heteroarallcyl, aryl or heteroaryl;
  • R 80 , R 81 , R 82 and R 83 are selected as in (i) or (ii) as follows:
  • R 80 , R 81 , R 82 and R 83 are selected from Z, preferably from hydrogen, allcyl, alkoxy, halide, haloallcyl and pseudohahde; or
  • R 80 and R 81 , or R 81 and R 82 , or R 82 and R 83 form 1,3-butadienylene, l-aza-1,3- butadienylene or 2-aza- 1,3 -butadienylene which are unsubstituted or substituted with 1,3- butadienylene, l-aza-l,3-butadienylene or 2-aza- 1,3 -butadienylene, and the others are selected as in (i); lc is 0-6; and s is 0-2.
  • Preferred compounds include: N-ethyl-2-(2-(4-dimethylaminophenyl)ethenyl)naphtho[2,l-d]thiazolium iodide, 3,3'- dioctadecyloxacarbocyanine perchlorate, N-ethyl-2-(2-ethyl-3-(N-ethylnaphtho[l,2-d]thiazolidin- 2-ylidene)propenyl)naphtho[l,2-d]thiazolium bromide, N,N'-dioctadecyloxacarbocyanine para- toluenesulfonate, 2-(2-acetanilinovinyl)-3-ethylbenzothiazolium iodide, 3-methyl-2-((3-methyl-2- benzothiazolinylidene)aminoazo)benzothiazolium tefr
  • R designates a hydrogen atom, a hydroxyl group in position 2', 3', or 4', a methoxy group in position 2', 3' or 4' or an ethoxy group in position 3' or 4'
  • R 2 designates a hydrogen atom, a hydroxyl group in position 3', 4', 5' or 6', a methoxy group in position 3' or 4' or an ethoxy group in position 5'
  • R 3 designates a hydrogen atom, a hydroxyl group in position 4', 5' or 6' or a methoxy group in position 4', 5' or 6',
  • R 4 designates a hydrogen atom or a hydroxyl group.
  • the compound of formula (XII) is 8-prenymaringenin in which Ri, R 2 and R 4 are hydrogen and R 3 is a 4 '-hydroxyl group.
  • Ri, R 2 and R 4 are hydrogen and R 3 is a 4 '-hydroxyl group.
  • B is an aromatic heterocycle having 1 to 4 N, O and/or S atoms, bonded via N or C, which can be unsubstituted or mono-, di- or tri-substituted by Hal, A and or OA, and can also be fused to a benzene or pyridine ring,
  • Q is absent or is allcylene having 1 -6 C atoms
  • X is CH 2 , S or O
  • R 1 and R 2 in each case independently of one another are H or A
  • R 3 and R 4 in each case independently of one another are -OH, OR 5 , -SR 5 , -SOR 5 , -S0 2 R 5 ,
  • R 5 Hal, methylenedioxy, -N0 2 , -NH 2 , -NHR 5 OR -NR 5 R 6 ,
  • R 5 and R 6 in each case independent of one another are A, cycloallcyl having 3-7 C atoms, mefhylenecycloalkyl having 4-8 C atoms or alkenyl having 2-8 C atoms,
  • A is allcyl having 1 to 10 C atoms, which can be substituted by 1 to 5 F and/or CI atoms, and Hal is F, CI, Br or l and their stereoisomers and physiologically acceptable, salts and solvates;
  • B is a phenyl ring which is unsubstituted or mono- or polysubstituted by R 3 , Q is absent or is allcylene having 1-4 C atoms, R 1 and R 2 each independently of one another are -OR 4 , -SR 4 , -SOR 4 , -S0 2 R 4 or Hal, or R 1 and R 2 together may form -0-CH 2 -0-,
  • R 3 is R 4 , Hal, OH, OR 4 , OPh, N0 2 , NHR 4 , N(R 4 ) 2 , NHCOR 4 , NHS0 2 R 4 or NHCOOR 4 ,
  • R 4 is A, cycloallcyl having 3-7 C atoms, allcylenecycloallcyl having 5-10 C atoms or alkenyl having 2-8 C atoms, A is allcyl having 1 to 10 C atoms, which can be substituted by 1 to 5 F and/or CI atoms, and Hal is F, CI, Br or I and their physiologically acceptable, salts and solvates;
  • R' and R' in each case independently of one another are -OR, OR 5 , -S-Rp -SO-R ⁇ -S0 2 -R 5 or Hal, or
  • R 1 and R 2 together may form -0-CH 2 -0-,
  • R 3 is NH 2 , NHA, NAA' or a saturated heterocycle having 1 to 4 N, O and/or S atoms which can be unsubstituted or mono-, di- or tri-substituted by Hal,
  • a and/or OA Q is absent or is branched or unbranched allcylene having 1-10 C atoms,
  • R 5 is A, cycloallcyl having 3-7 C atoms, allcylenecycloallcyl having 4-8 C atoms or alkenyl having
  • a and A' in each case independently of one another are alkyl which has 1 to 10 C atoms and which can be substituted by 1 to 5 F and/or CI atoms, and Hal is F, CI, Br or I, and the physiologically acceptable salts and solvates thereof;
  • B is A, OA, NH 2 , NHA, NAA' or an unsaturated heterocycle which has 1 to 4 N, O and/or S atoms and which can be unsubstituted or mono- di- or tri-substituted by Hal, A and/or OA,
  • Q is absent or is allcylene having 1-6 C atoms
  • R' and R 2 in each case independently of one another are -OH, OR 5 , -SR 5 , -SOR 5 , -S0 2 R 5 , Hal, - N0 2 , -NH 2 , -NHR 5 or -NR 5 R 6 , or R 1 and R 2 together are also -0-CH 2 -0-,
  • R 3 and R 4 in each case independently of one another are H or A
  • R 5 and R ⁇ in each case independently of one another are A, cycloallcyl having 3-7 C atoms, methylenecycloalkyl having 4-8 C atoms or alkenyl having 2-8 C atoms,
  • a and A 1 in each case independently of one another are alkyl which has 1 to 10 C atoms and which can be substituted by 1 to 5 F and/or CI atoms, and
  • Hal is F, CI, Br or I, and the stereoisomers and physiologically acceptable salts and solvates thereof;
  • R and R in each case independently of one another are H or A
  • R 3 and R 4 in each case independently of one another are -OH, OA, -SA, -SOA, -S0 2 A, Hal, methylenedioxy, -N0 2 , -NH 2 , -NHA or -NAA',
  • a and A' in each case independently of one another are allcyl having 1 to 10 C-atoms, and which can be substituted by 1 to 5 F and/or CI atoms, cycloallcyl having 3-7 C atoms or methylenecycloalkyl having 4-8 atoms,
  • B is -Y-R 5 .
  • Q is absent or is allcylene having 1-4 C atoms
  • Y is absent or is allcylene having 1-10 C atoms
  • X is CH 2 or S
  • R 5 is NH 2 , NHA, NAA' or is a saturated 3-8 membered heterocycle having at least one N atom, and wherein other CH 2 groups optionally may be replaced by NH, NA, S or O, which can be unsubstituted or monosubstituted by A or OH,
  • Hal is F, CI, Br or I
  • R 1 and R 2 in each case independently of one another are H, OH, OA, SA, SOA, S0 2 A, F, CI or
  • A' 2 N-(CH 2 ) n -0-, R 1 and R 2 may also form -0-CH 2 -0-, R 3 and R 4 in each case independently of one another are H, A, Hal, OH, OA, N0 2 , NHA, NA 2 , CN,
  • R 5 and R 6 in each case independently of one another are H or allcyl having 1 to 6 C atoms,
  • A is allcyl having 1 to 10 C atoms, which can be substituted by 1 to 5 F and/or CI atoms, is cycloallcyl having 3-7 C atoms, allcylenecycloallcyl having 5-10 C atoms or alkenyl having 2-8 C atoms,
  • A' is allcyl having 1, 2, 3, 4, 5 or 6 C atoms, n is 1, 2, 3 or 4,
  • Hal is F, CI, Br or I, and their physiologically acceptable salts and solvates;
  • R 1 and R 2 in each case independently of one another are H or A,
  • R 3 and R 4 in each case independently of one another are -OH, -OR 10 , -SR 10 , -SOR 10 , -S0 2 R 10 , Hal, methylenedioxy, -N0 2 , -NH 2 , -NHR 10 or -NR 10 R U ,
  • R 5 is a phenyl radical which is unsubstituted or mono- or disubstituted by R 6 and or R 7 ,
  • Q is absent or is allcylene having 1-6 C atoms
  • R 6 and R 7 in each case independently of one another are -NH 2 , -NR 8 R 9 , -NHR 10 , -NR 10 R ⁇ , -N0 2 ,
  • R 8 and R 9 in each case independently of one another are H, acyl having 1-8 C atoms which can be substituted by 1-5 F and/or CI atoms, -COOA, -S-A, -SO-A, -S0 2 A, -CONH,, -CONHA, -CONA 2 ,
  • A is allcyl having 1 to 6 C atoms which can be substituted by 1-5 F and or CI atoms,
  • R'° and R" in each case independently of one another are A, cycloallcyl having 3-7 C atoms, mefhylenecycloalkyl having 4-8 C atoms or alkenyl having 2-8 C-atoms, and
  • Hal is F, Ch Br or l, and their physiologically acceptable salts and solvates;
  • R 1 and R 2 in each case independently of one another are H or A
  • R 3 and R 4 in each case independently of one another are -OH, -OR 10 , -SR 10 , -S0 2 R 10 , Hal, methylenedioxy, -N0 2 , -NH 2 , -NHR 10 or -NR 10 R n ,
  • R 5 is a phenyl radical which is unsubstituted or mono- or disubstituted by R 6 and/or R 7 ,
  • Q is absent or is allcylene having 1-6 C atoms
  • R 6 and R 7 in each case independently of one another are -NH 2 , -NR 8 R 9 , -NHR 10 , -NR 10 R n , -N0 2 ,
  • R 8 and R 9 in each case independently of one another are H, acyl having 1-8 C atoms which can be substituted by 1-5 F and/or CI atoms, -COOA, -SO-A, -S0 2 A, -CONH 2 , -CONHA, -CONA 2 , -CO-
  • A is allcyl having 1 to 6 C atoms which can be substituted by 1-5 F and/or CI atoms,
  • R 10 and R 11 in each case independently of one another are A, cycloallcyl having 3-7 C atoms, methylenecycloalkyl having 4-8 C atoms or alkenyl having 2-8 C-atoms, and
  • Hal is F, CI, Br or I, and their physiologically acceptable salts and solvates;
  • R 1 and R 2 in each case independently of one another are H or A
  • R 3 and R 4 in each case independently of one another are OH, OA, SA, SOA, -S0 2 A, Hal, methylenedioxy, cycloallcyloxy with 3-7 C-atoms or 0-C m H 2m+ ⁇ - k F k ,
  • R 5 is -NR 6 R 7 or -N (CH 2 ) n ,
  • R 6 and R 7 in each case independently of one another are H or A
  • Q is allcylene with 1-6 C-atoms
  • A is allcyl with 1-6 C-atoms, Hal is F, CI, Br or I, m is 1, 2, 3, 4, 5 or 6, n is 3, 4, 5 or 6, lc is 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13, and their physiologically acceptable salts and solvates;
  • R 1 and R 2 in each case independently of one another are H or A
  • R 3 is H, OA or 0-C m H 2m+1 .
  • n X, rule X is F or CI
  • A is allcyl with 1-6 C-atoms, m is 1, 2, 3, 4, 5 or 6 and n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 and their physiologically acceptable salts and solvates.
  • R 1 and R 2 in each case independently of one another are H, OH, OR 5 , -SR 5 , -SOR 5 , -S0 2 R 5 or Hal, or
  • R 1 and R 2 together may form -OCH 2 0- or -OCH 2 CH 2 0-,
  • R 3 and R 3' in each case independently of one another are H, R 5 , OH, OR 5 , NH 2 , NHR 5 , NAA'
  • NHCOR 5 NHCOOR 5 , Hal, COOH, COOR 5 , CONH 2 , CONHR 5 orCONR 5 A',
  • R 5 is A or cycloallcyl with 3 to 6 C-atoms, which can be substituted by 1 to 5 F and/or CI atoms, or -(CH 2 ) consult-Ar, A and A' in each case independently of one another are alkyl with 1 to 10 C-atoms or are alkenyl with 2 to 8 C-atoms, which can be substituted by 1 to 5 F and/or CI atoms, or
  • a and A' together are also cycloallcyl or cycloallcylene with 3 to 7 C-atoms, wherein one CH 2 group can be replaced by O, NH, NA, NCOA or NCOOA, Ar is phenyl, n is 0, 1 or 2,
  • Hal is F, CI, Br or I and their pharmaceutically useable derivatives, solvates and stereoisomers, including mixtures thereof in all ratios. Also compounds:
  • G is selected from the group consisting of 0(CH 2 ) n C 3 _ 6 cycloalkyl, 0(CH 2 ) n phenyl, 0(CH 2 ) classroomheterocyclyl, 0(CH 2 ) n heteroaryl, NHC(0)(CH 2 ) n C 3-6 cycloalkyl, NHC(0)(CH 2 ) n phenyl, NHC(0)(CH 2 ) n heterocyclyl, NHC(0)(CH 2 ) n heteroaryl, NHC(0)(CH 2 ) m OC 3-6 cycloallcyl,
  • each cycloallcyl and heterocyclyl may also be optionally substituted with one or more carbonyl groups.
  • heteroaryl and heterocyclyl are 5 or 6 membered heteroaryl or heterocyclyl groups.
  • Prefe ⁇ ed compounds are those in which G is 0(CH 2 ) n heterocyclyl, n is 1 or 2 and the heterocyclyl group optionally substituted with one or two carbonyl groups; or G is NHC(O)(CH 2 ) friendshipOphenyl or NHC(0)(CH 2 ) n Oheteroaryl wherein n is 0 or 1 and each phenyl or heteroaryl is optionally substituted with one or more halo or d -3 alkoxy.
  • Especially preferred compounds are those m which G is 0(CH 2 ) 2 N-Succ ⁇ mm ⁇ de, NHC(0)CH 2 0[2,4-d ⁇ chlorophenyl], NHC(0)[3,4-d ⁇ fluorophenyl], NHC(0)[2-th ⁇ ophene], NHC(0)[4-py ⁇ dme], NHC(0)CH 2 0[3,4,5- t ⁇ methoxyphenyl] .
  • a prefe ⁇ ed compound of formula (XTV) is l-Deoxy-l- ⁇ [(2,4- d ⁇ chlorophenoxy)acetyl]am ⁇ no ⁇ - ⁇ -D-glucopyranuron ⁇ c acid.
  • R 7 and R 8 can be taken together with the nitrogen to which they are attached to form a monocyclic or bicyclic heterocycle with 5-7 members in each ring and optionally, in addition to containing nifrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic or bicyclic heterocycle optionally substituted with one or more substituents selected from R 6a ;
  • R a is (C
  • heterocyclyl encompasses all of saturated, unsaturated and aromatic (heteroaryl groups) heterocyclic groups.
  • Preferred compounds are those in which at least one of the following applies.
  • R 2 and R 3 are H and R s is H or F;
  • R) is H; or
  • R ta or ⁇ is H and the other is C ⁇ -C 6 allcyleneNR 7 R 8 , said allcylene optionally substituted with oxo.
  • Prefe ⁇ ed compounds include:
  • the disclosed compounds are described as inhibitors of the tyrosine kinase activity of transmembrane receptors such as growth factor receptors.
  • Other quinolinone compounds that also display tyrosine kinase inhibitory activity are disclosed in WO 03/020699. These compounds include:
  • W is ⁇ or C
  • R 7 and R 8 can be taken together with the nitrogen to which they are attached to form a monocyclic or bicyclic heterocycle with 5-7 members in each ring and optionally, in addition to containing nitrogen, one or two additional heteroatoms selected from N, O and S, said monocyclic or bicyclic heterocycle optionally substituted with one or more substituents selected from R 6a ;
  • R a is (C,-C6)allcyl, (C 3 -C 6 )cycloalkyl, aryl or heterocyclyl;
  • heterocyclyl encompasses saturated, unsaturated and aromatic (heteroaryl) heterocyclic groups. Prefe ⁇ ed compounds are those in which at least one of the following applies; Z is
  • Exemplary compounds include: 6-Chloro-3-(lH -indol-2-yl)-lH-indazole, 3-(lH-Indol-2-yl)-lH-indazole, 3-(lH-Indol-2-yl)-lH-indazol-5-ylamine, 3-(lH-Indol-2-yl)-6-methyl-lH-indazole, 3-(lH-Indol-2-yl)-4-chloro-lH-indazole, 3-(lH-Indol-2-yl)-7-chloro-l/J-indazole, 3-(lH-Indol-2-yl)-4-fluoro-lH-indazole, 3-(l/ -Indol-2-yl)-5- fluoro-lH-indazole, 3-(lH-Indol-2-yl)-5-methyl-lH-indazole, 3-(l
  • R represents OH, (C r C 5 )alkoxy, carboxyl, (C 2 -C 6 )alkoxycarbonyl, NR 5 R 6 , NH-S0 2 -Allc, NH- S0 2 -Phenyl, NH-CO-Ph, N(Alk)-CO-Ph, NH-CO-NHPh, NH-CO-Alk, NH-C0 2 -Alk, 0-(CH 2 ) n - cAllc, 0-Allc-C0 2 R 7 , 0-Allc-OR 8 , O-Allc-OH, 0-Alk-C(NH 2 ):NOH, O-Alk-NRsRe, O-Alk-CN, O- (CH 2 ) n -Ph, 0-Alk-CO-NR 5 R 6 , CO-NH-(CH 2 ) m -C0 2 R 7 , CO-NH-Alk,
  • R 2 represents H, (C C 5 )allcyl, (C ⁇ -C 5 )allcylhalide, (C 3 -C 6 )cycloallcyl or phenyl optionally substituted with one or more halogen, d-dalkoxy, carboxy or alkoxycarbonyl having 2 to 6 carbon atoms;
  • A represents -CO-, -SO- or S0 2 -;
  • R 3 and ) are identical or different and each represent H, (C ⁇ -C 5 )alkoxy, amino, carboxy, (C 2 - C 6 )alkoxycarbonyl, OH, N0 2 , hydroxyamino, -Alk-C0 2 R 7 , NR 5 R 6 , NH-Alk-C0 2 R 7 , NH-C0 2 -Alk, N(Rn)-S0 2 -Alk-NR 9 R ⁇ o, N(R n )-S0 2 -Al
  • Prefe ⁇ ed compounds include
  • Sulfonamide-contammg heterocyclic compounds having FGF inhibiting activity are disclosed m WO 03/074045
  • the disclosed compounds are also encompassed in some embodiments of the present invention.
  • An exemplary compound of this type is:
  • tncychc-based mdolmone compounds pyrazolylamide-based compounds, imidazolyl 2- mdolmone derivatives and phenyl 2-mdolmone derivatives have also been described as modulators of protein kinases.
  • ring A and ring B share one common bond
  • ring B and ring C share one common bond
  • ring A, Ring B and ring R are independently selected from the group consisting of an aromatic ring, a heteroaromatic ring, an aliphatic ring, a heteroaliphatic ring, and a fused aromatic or aliphatic ring system, where the heteroaromatic ring and heteroaliphatic ring each independently contain 0, 1, 2 or 3 heteroatoms independently selected from the group consisting of nifrogen, oxygen and sulfur
  • ring A, ring B, ring Q and ring R are each independently and optionally substituted with one, two or three substituents independently selected from the group consisting of allcyl, an aromatic or heteroaromatic ring, an aliphatic or heteroaliphatic ring, an amine, a nitro group, a halogen or trihalomethyl group, a lcetone, a carboxylic acid or ester, an
  • ring A and ring B of formula (XVIII) and (XIX) are each independently selected from the group consisting of a 5-membered ring, a 6-membered ring, a 7-membered ring, 8-membered ring and a bicyclic or tricyclic fused ring system having preferably 8-13 atoms in the ring backbone.
  • R is a 6-membered ring or a bicyclic or tricyclic fused ring system.
  • Ri and R 2 are independently selected from the group consisting of hydrogen, allcyl, an aromatic or heteroaromatic ring, an aliphatic or heteroaliphatic ring, an amine, a nitro group, a halogen, a lcetone, a carboxylic acid or ester, an alcohol or an alkoxyalkyl group, an amide, a sulfonamide, an alkoxyalkoxy group and a sulfone;
  • R 4 and R 5 are each independently selected from the group consisting of hydrogen, allcyl, an aromatic or heteroaromatic ring, an aliphatic or heteroaliphatic ring, an amine, a nitro group, a halogen, a lcetone, a carboxylic acid or ester, an alcohol or an alkoxyalkyl group, an amide, a sulfonamide, an alkoxyalkoxy group and a sulfone;
  • R 3 are independently selected from the group consisting of hydrogen
  • is selected from the group consisting of hydrogen, alkyl, especially methyl, ra-propyl and t-butyl, an aromatic or heteroaromatic ring, an aliphatic or heteroaliphatic ring.
  • R 2 is selected from hydrogen, allcyl and halogen, especially hydrogen and bromine.
  • R 4 and R 5 are selected from hydrogen, allcyl, an aromatic or heteroaromatic ring, an aliphatic or heteroaliphatic ring, preferably hydrogen or an aromatic or heteroaromatic ring optionally substituted with 1 to 3 substituents selected from allcyl, frihalomethyl and alkoxy moieties.
  • Exemplary compounds include: 2-benzyl-5 -tert-butyl -2H-pyrazole-3 -carboxylic acid (4-frifluoromethylphenyl)amide, 2-benzyl-5-tert-butyl-2H-pyrazole-3-carboxylic acid quinolin-3-ylamide, 2-benzyl-5 -tert-butyl -2H-pyrazole-3 -carboxylic acid (2,6-dimethoxypyridin-3-yl)amide, 2-benzyl-5 -tert-butyl -2H-pyrazole-3 -carboxylic acid (2,3 ,5 ,6-tetrafluoropyridin-4-yl)amide, 2-benzyl-5-tert-butyl-2H-pyrazole-3-carboxylic acid (3-methylquinolin-4-yl)amide, 2-benzyl-5-t ⁇ 7"t-butyl-2/J-pyrazole-3 -carboxylic acid (4,6-
  • R] R 2 and R 3 are independently selected from the group consisting of hydrogen, allcyl, an aromatic or heteroaromatic ring, an aliphatic or heteroaliphatic ring, an amine, a nitro group, a halogen or frihalomethyl group, a lcetone, a carboxylic acid or ester, an alcohol or an alkoxyalkyl group, an amide, a sulfonamide, an aldehyde, a sulfone or a thiol or thioether;
  • A, B, D and E are selected from the group consisting of carbon and nitrogen;
  • R 4 , R 5 , Re and R 7 are independently selected from the group consisting of hydrogen, allcyl, an aromatic or heteroaromatic ring, an aliphatic or heteroaliphatic ring, an amine, a nitro group, a halogen or frihalomethyl group, a lcetone, a carboxylic acid or ester, an alcohol or an
  • R] and R 2 are selected from the group consisting of hydrogen, methyl, ethyl, propyl and butyl groups optionally substituted with halogen, trihalomethyl, cyano and nitro groups; phenyl optionally substituted with 1-3 substituents independently selected from the group consisting of allcyl, alkoxy, halogen and nifro groups; an amine of formula -(X ⁇ ) nl -NX 2 X 3 where X 2 and X 3 are independently selected from the group consisting of hydrogen and optionally substituted saturated allcyl, and X) is optionally substituted saturated allcyl, and wherein ni is 0 or 1; a nitro group; a halogen or trihalomethyl; a lcetone of formula -CO-X 4 , where X 4 is selected from the group consisting of methyl, ethyl, propyl and butyl; a carboxylic acid of fonnula -
  • R 6 and R 7 are selected from the group consisting of hydrogen, methyl, ethyl, propyl and butyl groups optionally substituted with halogen, frihalomethyl, cyano and nitro groups; an amine of formula -(X,) nl -NX 2 X 3 where X 2 and X 3 are independently selected from the group consisting of hydrogen and optionally substituted saturated alkyl, and Xi is optionally substitufred saturated allcylene, and wherein m .
  • Y may be a 6-7 membered aromatic or heteroaromatic ring or a 6- membered aliphatic or heteroaliphatic ring.
  • G, J, and L are independently nifrogen.
  • X may be oxygen, nifrogen optionally substituted with allcyl or may be S, SO or S0 2 .
  • A, B, D and E are independently selected from the group consisting of carbon and nifrogen where it is understood that when A, B, D or E is nitrogen, R 6 , R 7 , R 8 or R C, respectively, does not exist and there is no bond;
  • G and I are selected from nifrogen and carbon such that when G is nifrogen, J is carbon and when J is nifrogen, G is carbon and when either G or J is nifrogen, then either R 5 or R 5 . does not exist;
  • R 2 and the imidazolyl ring may exchange places on the double bond so that the compound may exist in either the E or the Z configuration about the double bond at the 3-position;
  • R, and R 3 are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, hydroxy, alkoxy, C-carboxy, O-carboxy, C-amido, C-thioamido, sulfonyl and frihalomethylsulfonyl; 114 -
  • R 4 and R 5 are selected from the group consisting of hydrogen, methyl, ethyl, propyl and butyl groups optionally substituted with halogen, frihalomethyl, cyano and nitro groups; an amine of formula -(X 1 ) nl -NX 2 X 3 where X 2 and X 3 are independently selected from the group consisting of hydrogen and optionally substituted saturated allcyl, and Xi is optionally substituted saturated allcyl, and wherein n, is 0 or 1 or where X 2 and X 3 taken together form a 5-6-membered aliphatic or heteroaliphatic ring, optionally substituted at a ring carbon atom or heteroatom with a substituent selected from the group consisting of methyl, ethyl, propyl, phenyl and alkoxyphenyl; a nitro group; a halogen or trihalomethyl; a lcetone of formula -CO-X 4 , where
  • R 2 is selected from the group consisting of hydrogen, alkyl, cycloallcyl, aryl, heteroaryl and halo
  • R t , R 5 and R 5' are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkenyl, allcynyl, aryl, heteroaryl, heteroalicyclic, halo, trihalomethyl, hydroxy, alkoxy, aryloxy, C- carboxy, O-carboxy, carbonyl, nitro, cyano, S-sulfonamido, amino and NR ]0 R ⁇
  • R 10 and R n are independently selected from the group consisting of alkyl, cycloallcyl, aryl, carbonyl, sulfonyl, trihalomethanesulfonyl or may be combined to form a 5-6 membered heteroalicyclic ring
  • R t , R 5 and R 5' are independently selected from the group consisting of
  • R 6 , R 7 , R 8 and R 9 are independently selected from the group consisting of hydrogen, allcyl, trihaloalkyl, cycloallcyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, S-sulfonamido, N-sulfonamido, N-trihalomethanesulfonamido, carbonyl, C-carboxy, O-carboxy, cyano, nitro, halo, cyanato, isocyanato, thiocyanato, isothiocyanato, O-carbamyl, N-carbamyl, O-thiocarbamyl, C-amido, N-amido, amino and NR ⁇ 0
  • R 6 , R 7 , R 8 and R 9 are independently selected from the group consisting of hydrogen, lower allcyl optionally substituted with halo, C-carboxy and -NR ⁇ 0 Rn; lower alkoxy optionally substituted with halo, C-carboxy and -NRioR ⁇ !
  • frihalomethyl alkenyl, allcynyl, aryl optionally substituted with one or more groups selected from lower allcyl, lower allcyl substituted with one or more halo, C- carboxy, alkoxy, amino, S-sulfonamido or -NR ]0 Rn; heteroalicyclic optionally substituted with one or more allcyl optionally substituted with one or more halo groups, aldehyde, lower alkoxy carbonyl, hydroxy, alkoxy optionally substituted with one or more halo, C-carboxy, amino, S- sulfonamido or -NR 10 Rn; aryloxy optionally substituted with one or more of lower allcyl, frihalomethyl, halo, hydroxy, amino, sulfonamido or -NR 10 R ⁇ ; thiohydroxy, thioalkoxy, thioaryloxy optionally substituted one or more halo, hydroxy, amino, S
  • n are independently selected from hydrogen, lower allcyl optionally substituted on the furthest C from the point of attachment with a C-carboxy group, trihalomethyl, halo, hydroxy, alkoxy, O-carboxy, C-carboxy, amino, C-amido, N-amido, S-sulfonamido, nitro, amino and - NRio n.
  • A, B and D are independently selected from the group consisting of carbon and nifrogen where it is understood that when A, B or D is nifrogen, R 3 , R 4 , R 8 or R 5 respectively, does not exist;
  • R is selected from the group consisting of hydrogen, allcyl, cycloallcyl, aryl, heteroaryl, hydroxy, alkoxy, C-carboxy, O-carboxy, C-amido, C-thioamido, sulfonyl and frihalomethylsulfonyl;
  • R 2 is selected from the group consisting of hydrogen, allcyl, cycloalkyl, aryl and heteroaryl;
  • R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and Rj 0 are independently selected from the group consisting of hydrogen, allcyl, frihalomethyl, cycloallcyl, alkenyl, allcynyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, S-sulfonamido, N-sulfonamido,
  • Rn and R i2 are independently selected from the group consisting of hydrogen, allcyl, cycloallcyl, aryl, carbonyl, acetyl, sulfonyl, trihalomethanesulfonyl or may be combined to form a 5-6 membered heteroalicyclic ring;
  • R 3 and R 4 or R 6 and R 7 or R 7 and R 8 or R 8 and R 9 or R 9 and R ⁇ 0 may combine to form a methylenedioxy or ethylenedioxy group; and Q is selected from the group consisting of aryl, heteroaryl and fused heteroaryl cycloalkyl/heteroahcychc groups.
  • Ri and R 2 are hydrogen; A, B and D are carbon; R 3 , R 4 and R 5 are hydrogen,
  • R 6 ⁇ R 7 , Rs, R9 and R 10 are independently selected from hydrogen and lower allcyl;
  • Q is aryl optionally substituted with one or more hydrogen, lower alkyl, lower alkoxy and heteroalicyclic, especially 4-formylp ⁇ perazm-l-yl; heteroaryl, especially pyrrol-2-yl, ⁇ m ⁇ dazo-4-yl and th ⁇ ophen-2-yl; or heteroaryhcycloalkyl/heteroahcychc group in which the heteroaryl moiety is selected from pyrrolo, thiopheno, furano, thizolo, oxazolo, py ⁇ dmo and imadazolo.
  • a panticularly prefe ⁇ ed Q is 4,5,6,7-tefrahydromdol-2-yl. Q may also be optionally substituted with one or more hydrogen, lower allcyl, lower alkoxy, carboxy, carboxy salt, carboxyalkyl and carboxyallcyl salt.
  • Suitable sugars, ohgosaccharides and carbohydrates include:
  • Ca ⁇ ageenans especially lambda, appa and iota ca ⁇ ageenans or de ⁇ vatives thereof, most especially iota ca ⁇ ageenan and derivatives thereof.
  • Derivatives of ca ⁇ ageenan include those prepared by chemical or enzymatic hydrolysis using mild acid or ca ⁇ ageenase. The de ⁇ vatives may also have varying degrees of sulfation.
  • Ca ⁇ ageenans, their denvatives, methods for their preparation and their biological activity are disclosed in WO 94/05267. Ca ⁇ ageenans and derivatives thereof are described as having growth factor antagonist activity, especially FGF-2 antagonist activity.
  • Salts or complexes of sulfated saccha ⁇ des especially salts or complexes with an alkali metal or alkaline earth metal.
  • Particularly prefe ⁇ ed salts and complexes are formed with sodium, potassium, bismuth, calcium, magnesium, barium, aluminium, zmc, copper, titanium, manganese or osmium.
  • the salt or complex may be formed with an organic base, for example, an ammo acid.
  • Prefe ⁇ ed sulfated saccharides are mono or ohgosaccharides and include xylose, fructose, glucose sucrose, lactose, maltose cellobiose, maltot ⁇ ose, maltotetrose, altopentose and maltohexose or fragments of hepann small enough not to bind more than one hepa ⁇ n-bmding growth factor at a time.
  • the sacchandes may be monosulfated, polysulfated or persulfated.
  • Complexes and salts of sulfated sacchandes, methods for their preparation and their biological activity are disclosed in WO 95/34313. The disclosed complexes and salts are described as inhibitors of heparin-binding growth factor activity.
  • Sulfomannans having varying degrees of sulfation and varying chain length and optionally a terminal non-reducing phosphate group.
  • Prefe ⁇ ed sulfomannans have the formula:
  • n is 0 or an integer from 1 to 4, each R is independently selected from S0 3 Na or H;
  • R is H, P0 3 Na 2 or S0 3 Na.
  • the agent interferes with binding of FGF-2 with FGFR by binding with high affinity and specificity to FGF-2.
  • the agent is penfraxin PTX3 or a derivative thereof.
  • Non-limiting examples of this type of agent are disclosed in WO 02/38169.
  • ohgosaccharides that have an antagonistic effect on FGF are described in WO 93/19096.
  • Illustrative ohgosaccharides consist essentially of oligosaccharide chains which are substantially homogeneous with respect to FGF binding affinity and which contain at least four, preferably at least six, disaccharide units including sulphated disaccharide units, preferably a ⁇ anged as a contiguous sequence, that are each composed of an N-sulfated glucosamine residue (+6S) and a 2- O-sulphated iduronic acid residue.
  • the oligosaccharide is characterized in that: (a) it is composed predominantly of a molecular species: where X is ⁇ HexA-GlcNS0 3 (+6S), Y is IdoA(2S)-GlcNS0 3 ( ⁇ 6S), Z is IdoA-GlcR (+6S) or IdoA(2S)-GlcR (+6S) where R is NOS0 3 or NAc, and n is in the range of 4-7 ; (b) the content, if any, of monosaccharide residues having a 6-O-sulfphate group is less than 20%; (c) it is obtainable by a process comprising the steps of digesting a heparan sulphate with heparitinase so as to bring about partial depolymerisation thereof to the fullest extent, followed by size fractionating the oligosaccharide mixture produced using for example gel filtration size exclusion chromatography, collecting a
  • Suitable oligonucleotides, proteins, peptides or polypeptides that impair or interfere with a FGF signalling pathway include: a) Peptides having the following sequence: Phe-Phe-Phe-Glu-Arg-Leu-Glu-Ser-Asn-Asn-Tyr-Asn-Thr-Tyr-Arg-Ser-Arg-Lys-Tyr- Xaa 2 o-Xaa 2 ⁇ -Xaa 22 -Xaa 23 -Val-Ala-Leu-Lys-Arg-Thr-Gly-Gln-Tyr-Lys-Leu-Gly-Xaa 36 - Lys-Thr-Gly-Pro-Gly-Gln-Lys-Ala-Ile-Leu-Phe-Leu-Pro-Met-Ser-Ala-Lys-Ser [SEQ ID NO: 1] wherein Xaa 20 is Ser, Thr or D
  • Prefe ⁇ ed sequences include: Glu-Cys-Phe-Phe-Phe-Glu-Arg-Leu-Glu-Ser-Asn-Asn-Tyr-Asn-Thr-Tyr-Arg-Ser-Arg- Lys-Tyr-Xaa 20 -Ser-T ⁇ -Tyr-Val-Ala-Leu-Lys-Arg wherein Xaa 20 is Thr or Ser [SEQ ID NO: 2]; Tyr-Asn-Thr-Tyr-Arg-Ser-Arg-Lys-Tyr-Xaa 2 o-Ser-T ⁇ -Tyr-Val-Ala-Leu-Lys-Arg wherein Xaa 20 is Thr or Ser [SEQ ID NO: 3]; Tyr-Arg-Ser-Arg-Lys-Tyr-Xaa 20 -Ser-T ⁇ -Tyr wherein Xaa 20 is Thr or Ser [SEQ ID NO: 4]
  • Xaai is Gly or Sar and or Xaa 2 is Ser or Ala, especially those in which Xaai is Gly and Xaa 2 is Ser and Xaai is Sar and Xaa 2 is Ala.
  • Peptides having the sequence: H-Phe-Phe-Phe-Glu-Arg-Leu-Glu-Ser-Asn-Asn-Tyr-Asn-Thr-Tyr-Arg-Ser-Arg-Lys-Tyr- Ser-Ser-T ⁇ -Tyr-Val-Ala-Leu-Lys-Arg-Thr-Gly-Gln-Tyr-Lys-Leu-Gly-Pro-Lys-Thr-Gly- Pro-Gly-Gln-Lys-Y [SEQ ID NO: 8] wherein Y is OH or NH 2 , or a biologically active C-terminally shortened fragment thereof which contains the sequence Tyr-Arg-Ser-Arg-Lys-Tyr-Ser-Ser-T ⁇ -Tyr.
  • Prefe ⁇ ed embodiments have the following sequences: H-Tyr-Arg-Ser-Arg-Lys-Tyr-Ser-Ser-T ⁇ -Tyr-NH 2 [SEQ ID NO: 9]; H-Tyr-Arg-Ser-Arg-Lys-Tyr-Ser-Ser-T ⁇ -Tyr-Val-Ala-Leu-Lys-Arg-Y [SEQ ID NO: 10] wherein Y is OH or NH 2 ; H-Tyr-Arg-Ser-Arg-Lys-Tyr-Ser-Ser-T ⁇ -Tyr-Val-Ala-Leu-Y [SEQ ID NO: 11] wherein Y is OH or NH 2 ; H-Tyr-Arg-Ser-Arg-Lys-Tyr-Ser-Ser-T ⁇ -Tyr-Val-Ala-Leu-Lys-Arg-Thr-Gly-Gln-Tyr-
  • Xaa is any amino acid structure andb is an integer from 1-15 inclusive;
  • Xaaj is a tyrosine residue, a phenylalanine residue, a pyridylalanine residue or a homophenylalanine residue;
  • Xaa 2 is a leucine residue, a norleucine residue, a phenylalanine residue, a pyridylalanine residue, a homophenylalanine residue or a isoleucine residue;
  • Xaa 3 is an arginine residue, an aspartic acid residue, a glutamic acid residue, a serine residue, a tyrosine residue, or a glutamine residue;
  • Xaa 4 is a glutamine residue, a leucine residue, a norleucine residue or a tyrosine residue;
  • Xaa 5 is a tyrosine residue
  • Xaa 6 is a methionine residue, a leucine residue, a norleucine residue, a lysine residue or an arginine residue
  • Xaa 7 is a leucine residue, a norleucine residue, a methionine residue, an aspartic acid residue, a glutamic acid residue, an asparagine residue or a serine residue;
  • Xaa 8 is an arginine residue, a leucine residue, a norleucine residue, a serine residue or a threonine residue;
  • Xaa 9 is a leucine residue, a norleucine residue, a phenylalanine residue, a pyridylalanine residue, a homophenylalanine residue, a methionine residue or a valine residue, or an inverso or refro-inverso isomer thereof.
  • Prefe ⁇ ed sequences include: Asp-Val-Phe-Leu-Asp-Met-Tyr-Gln-Phe-Ser-Val-Ile [SEQ ID NO: 17];

Abstract

L'invention concerne des méthodes et des agents servant à moduler le potentiel de différenciation et/ou la prolifération de préadipocytes. Plus spécifiquement, cette invention a trait à des méthodes et à des agents permettant de moduler une voie de signalisation du facteur de croissance de fibroblaste, notamment, la voie de signalisation FGF-1 ou FGF-2, dans le traitement ou la prévention de troubles liés à l'adiposité, notamment mais non seulement, à l'obésité, au lipome et à la lipomatose.
PCT/AU2005/000008 2004-01-07 2005-01-07 Agents de modulation de la differenciation et utilisations associees WO2005065686A1 (fr)

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US9714421B2 (en) 2013-05-01 2017-07-25 Ionis Pharmaceuticals, Inc. Compositions and methods
WO2018071640A1 (fr) * 2016-10-13 2018-04-19 Lubrizol Advanced Materials, Inc. Composés utiles dans le traitement et/ou le soin de la peau, des cheveux, des ongles et/ou des muqueuses
US9994855B2 (en) 2014-05-01 2018-06-12 Ionis Pharmaceuticals, Inc. Compositions and methods for modulating growth hormone receptor expression
US11471455B2 (en) 2018-10-05 2022-10-18 Annapurna Bio, Inc. Compounds and compositions for treating conditions associated with APJ receptor activity

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WO2003035644A1 (fr) * 2001-10-26 2003-05-01 Aventis Pharma S.A. Derives de benzimidazoles et_leur utilisation comme inhibiteurs de proteine kinase kdr
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WO2003035644A1 (fr) * 2001-10-26 2003-05-01 Aventis Pharma S.A. Derives de benzimidazoles et_leur utilisation comme inhibiteurs de proteine kinase kdr
WO2003035065A1 (fr) * 2001-10-26 2003-05-01 Aventis Pharmaceuticals Inc Benzimidazoles et analogues et leur utilisation comme inhibiteurs de proteines kinases
WO2004041277A1 (fr) * 2002-11-01 2004-05-21 Merck & Co., Inc. Derives de carbonylamino-benzimidazole utilises comme modulateurs du recepteur androgene

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Publication number Priority date Publication date Assignee Title
US9714421B2 (en) 2013-05-01 2017-07-25 Ionis Pharmaceuticals, Inc. Compositions and methods
US9994855B2 (en) 2014-05-01 2018-06-12 Ionis Pharmaceuticals, Inc. Compositions and methods for modulating growth hormone receptor expression
US10793862B2 (en) 2014-05-01 2020-10-06 Ionis Pharmaceuticals, Inc. Compositions and methods for modulating growth hormone receptor expression
US11312964B2 (en) 2014-05-01 2022-04-26 Ionis Pharmaceuticals, Inc. Compositions and methods for modulating growth hormone receptor expression
WO2018071640A1 (fr) * 2016-10-13 2018-04-19 Lubrizol Advanced Materials, Inc. Composés utiles dans le traitement et/ou le soin de la peau, des cheveux, des ongles et/ou des muqueuses
US11471455B2 (en) 2018-10-05 2022-10-18 Annapurna Bio, Inc. Compounds and compositions for treating conditions associated with APJ receptor activity
US11944622B2 (en) 2018-10-05 2024-04-02 Annapurna Bio, Inc. Compounds and compositions for treating conditions associated with APJ receptor activity

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