WO2001008669A2 - Therapeutic agents for lymphoedema - Google Patents

Therapeutic agents for lymphoedema Download PDF

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Publication number
WO2001008669A2
WO2001008669A2 PCT/GB2000/002950 GB0002950W WO0108669A2 WO 2001008669 A2 WO2001008669 A2 WO 2001008669A2 GB 0002950 W GB0002950 W GB 0002950W WO 0108669 A2 WO0108669 A2 WO 0108669A2
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flk
kdr
agent
ligand
lymphoedema
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PCT/GB2000/002950
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French (fr)
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WO2001008669A3 (en
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David Bates
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University Of Bristol
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Publication of WO2001008669A3 publication Critical patent/WO2001008669A3/en

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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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • 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
    • 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
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/235—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids having an aromatic ring attached to a carboxyl group
    • 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
    • A61K31/275—Nitriles; Isonitriles
    • 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
    • A61K31/275—Nitriles; Isonitriles
    • A61K31/277—Nitriles; Isonitriles having a ring, e.g. verapamil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • 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
    • 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
    • A61K31/415—1,2-Diazoles
    • 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
    • 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
    • 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
    • A61K31/502—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with carbocyclic ring systems, e.g. cinnoline, phthalazine
    • 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
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/10—Antioedematous agents; Diuretics
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/71—Assays involving receptors, cell surface antigens or cell surface determinants for growth factors; for growth regulators

Definitions

  • the present invention relates to the treatment of lymphoedema .
  • Lymphoedema arises from an imbalance between transvascular fluid filtration and lymph drainage capacity, but the pathophysiology is poorly understood. It is defined as a swelling associated with lymphatic insufficiency, lymphatic damage or lymphatic pathology and can result from cancer or cancer treatment, congenital disorders or infection by filarial worms, filariasis. Chronic lymphoedema of the arm is a common and distressing late complication of the successful treatment of breast cancer. Surgery and/or radiotherapy to axillary lymph nodes during breast cancer treatment is often followed by post mastectomy oedema (PMO) . There is currently no effective drug treatment for this condition.
  • PMO post mastectomy oedema
  • Receptor tyrosine kinases comprise a large family of transmembrane receptors for growth factors with diverse biological activities. Interaction of the RTK with growth factor results in phosphorylation of the receptor and triggering of an RTK-specific signal transduction pathway.
  • Flk-l/KDR is an RTK found on vascular endothelial cells and has a role in normal vasculogenesis and angiogenesis.
  • Normal vasculogenesis and angiogenesis play important roles in various physiological processes such as embryonic development, wound healing, organ regeneration and reproductive processes such as follicle development in the corpus luteum during ovulation and placental growth after pregnancy.
  • Flk- l/KDR has also been implicated in tumour-associated angiogenesis and increased vascular permeability.
  • a number of ligands for flk-l/KDR have been identified. For example the vascular endothelial growth factors VEGF-A and VEGF-C.
  • VEGF-C is a ligand for flkl/KDR and at least one other receptor known as flt-4 (found on lymphatic endothelial cells) .
  • VEGF-C is known to be produced by cancer cells (Salven P. et al (1998) Am. J. Pathol 153 103-8; Tsurusaki T. et al (1999) Br. J. Cancer 80 309-313; Valtola (1999) Am. J. Pathol. 154 1381-90; Lymboussaki (1998) Am. J. Pathol. 153 395-403; Fielder (1997) Leukemia 11 1234-1237), and is involved in tumour-related angiogenesis.
  • a number of inhibitors of flk-l/KDR have been isolated, such as those disclosed in US-A-5712395, US- A-5710173, US-A-5849742, US-A-5650415 , US-A-5851999, US-A-5792771 and US-A-5763441. These inhibitors are intended for the treatment of cell proliferative and cell differentiation disorders and disorders related to vasculogenesis and angiogenesis (such as cancer, diabetes, haemangioma and Karposi's sarcoma) and disorders associated with increased vascular permeability.
  • vasculogenesis and angiogenesis such as cancer, diabetes, haemangioma and Karposi's sarcoma
  • Activation of the flk-l/KDR receptor is known to activate phospholipase Cy, which results in production of diacylglycerol (DAG) and inositol triphosphate (IP3) .
  • DAG diacylglycerol
  • IP3 inositol triphosphate
  • DAG has been shown to result in calcium entry through DAG- activated calcium channels, and by stimulation of arachidonic acid-dependent calcium channels after degradation by DAG lipase. The precise nature of these calcium channels are unknown, but they are neither voltage operated calcium channels, nor store operated calcium channels.
  • VEGF-C The action of VEGF-C was previously thought to be limited to a few specific roles in an adult (namely wound healing and organ regeneration) and to tumour- associated angiogenesis. Underlying the present invention is the hypothesis that VEGF-C is secreted in response to lymph damage and acts in the lymphatic VEGF-C receptor, fit and that swelling associated with lymphatic insufficiency is brought about because the secreted VEGF-C also stimulates flk-1 on vascular endothelial cells. The inventor has shown that this results in a net increase in the filtration rate of water across the vasculature by a variety of mechanism, but without increasing the size of protein permeable pores in the vessel walls, and hence increased protein permeability.
  • lymphoedema is not a pathology dependent on increased vascular permeability or angiogenesis, but one that is dependent on lymphatic insufficiency, inhibiting the flk-l/KDR receptor is predicted to be an effective therapy for this condition.
  • Lymphoedema is caused by increased water flux without increased protein permeability (Bates).
  • VEGF-A which activates both flk-l/KDR and flt-1, increases water flux without increasing protein permeability (Bates (1998) J. Physiol (Lond) 513: 225-233) .
  • Lymphoedema is associated with lymphangiogenesis (growth of new lymphatics) .
  • Fig. 1 shows a plot of radial lymphatic density as a function of distance from tracer injection site in patients with lymphoedema in the swollen arm and the contralateral normal arm. Radial lymphatic density was measured by intradermal injection of FITC dextran and subsequent imaging on an intravital microscope. The Density was calculated as the number of lymphatics crossing a superimposed grip, per annulus away from the injection site. The area under the curve indicates total lymphatic density and is significantly increased in the ipsilateral arm of post-mastectomy patients, compared to their contralateral, normal arm (Dr Peter Mortimer, St. George's Hospital Medical School - unpublished results) . The fact that lymphatic density is higher in the swollen limb than in the non-swollen limb indicates that lymphangiogenesis has occurred.
  • Lymphangiogenesis is caused by VEGF-C acting on flt-4 ( Cao et al . (1998) Proc Natl Acad Sci USA 95: 14389-14394) .
  • VEGF-C stimulates flk-1 and flt-4 activation (Joukov et al . (1996) EMBO Journal 14:290-298).
  • FIG. 2 is a graph showing the acute and chronic effect of growth factors. Hydraulic conductivity (water flux per unit pressure) was measured in perfused mesenteric capillaries in vivo from the frog, Rana temporaria, using the Landis Michel Technique modified for chronic permeability measurements
  • VEGF-C black bars
  • % BSA control
  • 10 nM PIGF a flt-1 specific agonist
  • FIG. 3 is a graph showing the change in hydraulic conductivity over time in the absence (solid line) or presence (dashed line) of the flkl/KDR inhibitor, ZM323881. Hydraulic conductivity was measured in mesenteric microvessels of the frog, Rana temporaria as previously described. The results show that ZM323881 inhibits the VEGF-mediated increase in water flux per unit measurement.
  • lymphoedema is the result of VEGF-C stimulating flt-4 on lymph vessels causing lymphangiogenesis (to overcome the lymphatic insufficiency) , coupled with the activation of flk-1 on vascular endothelial cells which results in an increase in the ratio of water flux to protein flux, and hence swelling of the arm - this is shown schematically in Figure 4. Therefore, it can be concluded that flk-1 specific inhibitors will block the increase in water flux, and therefore the swelling in the arm after lymphoedema.
  • the basis of this invention therefore is that selective blocking of this receptor, or the signal pathway associated with it, will be effective in treating PMO and other lymphodemas.
  • an agent which selectively inhibits, prevents or interferes with the signal transduced by flk-l/KDR when activated by a ligand, in the manufacture of a medicament for the treatment of lymphoedema.
  • the ligand may be any ligand which is capable of interacting with flk-l/KDR and activating the signal transduction pathway.
  • the ligand may be a growth factor, particularly a vascular endothelial growth factor.
  • vascular endothelial growth factors which interact with flk-l/KDR include VEGF-A and VEGF-C.
  • the agent inhibits, prevents, or interferes with the interaction between a ligand and flk-l/KDR.
  • the agent is an antagonist of the flk-l/KDR receptor which thus prevents access by the normal ligand to that receptor.
  • the agent inhibits, prevents or interferes with the flk-l/KDR-associated signal transduction pathway.
  • the inhibition, prevention or interference activity mediated by the agent is substantially restricted to the flk-l/KDR receptor or its associated signal transduction pathway.
  • the agent does not cross-react and inhibit, prevent or interfere with other receptors or their signal transduction pathways to a significant extent. Significant cross-reaction would reduce the efficiency of the agent and make the use of the agent unpredictable and/or cause undesired side-effects.
  • the agent may be any moiety, for example a molecule or macromolecule, capable of specifically interacting with flk-l/KDR, the ligand, or one or more of the cellular components involved in flk-l/KDR receptor signal transduction.
  • the agent may be a polypeptide (or modified polypeptide, including glycopolypeptides and lipopolypeptides) , such as an antibody or antibody fragment, a soluble receptor molecule, or a peptide.
  • an agent is a protein as described in US-A-5851999 which has a functional flk-l/KDR extracellular and transmembrane domain and a deleted or mutated cytoplasmic domain so that the polypeptide is signalling incompetent.
  • Flk-l/KDR receptors dimerise, and the signalling incompetent polypeptide may dimerise with wild-type receptor and prevent signalling.
  • the agent may be a nucleic acid sequence (including a modified nucleic acid sequence) .
  • the agent may be a polynucleotide which encodes for a polypeptide as described above.
  • the agent may also be an expression vector which comprises such a polynucleotide.
  • an agent is described in US-A-5851999.
  • the agent may be an oligonucleotide, for example an antisense oligonucleotide. Such an oligonucleotide may inhibit prevent or interfere with the transcription or translation of a gene or polypeptide involved in ligand-activated flk-l/KDR signal transduction.
  • the agent may be a mutant ligand or a molecule which is a molecular mimic for a ligand.
  • a molecular mimic is a molecule which is not derived or derivable from the ligand, but mimics the binding capacity of the ligand by having the same or an equivalent binding pocket.
  • a mutant ligand or molecular mimic for a ligand may have the capacity to bind flk-l/KDR, but may lack the capacity to induce flk-l/KDR-associated signal transduction.
  • the mutant ligand or molecular mimic may inhibit ligand-induced signal transduction by competing with the wild-type ligand for the receptor, and/or by directly blocking interaction of the wild-type ligand with the receptor (if the mutant ligand or molecular mimic form a more stable association with the receptor that the wild type ligand) .
  • the agent may be a small organic molecule.
  • Such molecules are already known as agents for use in the treatment of cell proliferative and cell differentiation disorders and disorders related to vasculogenesis and angiogenesis (such as cancer, diabetes, haemangioma and Karposi ' s sarcoma).
  • thienyl compounds such as those disclosed in US-A-5710173 , derivatives of quinoline such as those disclosed in US-A-5650415, and derivatives of quinazoline, quinoxaline, substituted aniline, isoxazoles, acrylonitrile or phenylacrylonitrile compounds such as those disclosed in US-A-5763441, US- A-5792771, US-A-5849742 and US-A-5712395.
  • Agents which inhibit, prevent or interfere with flk-1/KDR-ligand interaction according to the first embodiment of the first aspect of the invention may be identified by a number of methods which are known in the art.
  • antibodies which specifically bind flk-l/KDR can be produced using conventional techniques and screened using a phage display library or by ELISA.
  • Isolated flk-l/KDR or its ligand can be used to screen for potentially useful agents by immobilising the flk-1/KDR/ligand for example on a microtitre plate, a column or on magnetic beads, selectively binding the agent to the immobilised flk- 1/KDR/ligand, and then identifying the bound agent.
  • Agents which inhibit, prevent or interfere with flk- 1/KDR-ligand interaction may be screened by conducting a competitive binding assay for flk-l/KDR with the ligand.
  • inhibitors including: ZD4190
  • the agent inhibits, prevents or interferes with the flk-l/KDR-associated signal transduction pathway.
  • the agent may inhibit, prevent or interfere with flk-l/KDR-associated activation of phospholipase C ⁇ , formation of diacyl glycerol, production of arachidonic acid by DAG-lipase and/or DAG-mediated calcium entry through non-store operated, non-voltage-gated, calcium channels.
  • An example of an agent which inhibits calcium entry through non-store operated, non-voltage-gated, calcium channels is carboxyamido-triazole (CAI, as decribed in USSN 5359078) (Kohn et al (1992) Cancer Res. 52 3208- 3212) .
  • Agents which inhibit, prevent or interfere with the flk-l/KDR-associated signal transduction pathway may be identified by methods known in the art.
  • agents include (1- [6- ( [17 ⁇ ) -3- Methoxyestra-1,3,5 (10) trien-17-yl] amino) hexyl] -1H- pyrrole-2, 5-dione (U73122, an inhibitor of phospholipase C) and 1,6- bis (cyclohexyloximinocarboynlamino)hexane (RHC-80267, an inhibitor of DAG lipase)
  • lymphoedema may be due to surgery for cancer treatment where lymph nodes of, for example, the axilla, neck or inguinal nodes have been partially or wholly removed or destroyed by surgery. Also, lymphoedema may arise due to congenital disorders, or infection by filarial worms, including lymphoedema of the arm, leg, trunk, scrotum, face or other part of the body.
  • the agent defined in the first aspect of the invention may be formulated into a pharmaceutical composition and may be used in conjunction with a pharmaceutically acceptable carrier.
  • the pharmaceutical composition may be in a unit- dose form, such as a tablet, capsule, ampoule or suppository, methods for the preparation of which are known per se .
  • the carrier may act as a filler, binder, disintegrant , stabilizer or lubricant.
  • a method for treating lymphoedema in a mammalian patient which comprises the step of administering an effective amount of a agent as defined in the first aspect of the invention to the mammalian patient .
  • the composition may be administered enterally or parenterally, for example by topical, oral or nasal, administration, or by intracavernous, intravenous, intramucsular or subcutaneous injection.
  • an "effective amount” of the composition to be delivered in one administration will depend on the individual patient characteristics (severity of disease, size, age etc.) as well as the characteristics of the compound. Generally an “effective amount” is that amount necessary to substantially inhibit, prevent or interfere with the stimulation of flk-l/KDR by a ligand, or the flk-1/KDR-associated signal transduction pathway. Unique or continued application of such an effective amount results in reduction or amelioration of the symptoms with lymphoedema.
  • a method for reducing the hydraulic conductivity of blood vessels in a mammalian patient which comprises the step of administering an effective amount of an agent as defined in the first aspect of the invention to the mammalian patient.
  • the hydraulic conductivity may be reduced in particular blood vessels in the patient or in blood vessels in a particular area of the patient.
  • the method of the third aspect of the invention may be used to reduce the hydraulic conductivity of the blood vessels in at least one arm of the mammalian patient.
  • a method for reducing the filtration rate from blood to tissue in at least one arm of a mammalian patient which comprises the step of administering an effective amount of an agent as defined in the first aspect of the invention to the mammalian patient.
  • the filtration rate from blood to tissue may be reduced in particular blood vessels in the patient, in a particular tissue of the patient, or in a particular area of the patient.
  • the method of the fourth aspect of the invention may be used to reduce the filtration rate from blood to tissue in at least one arm of the mammalian patient.
  • a method for identifying compounds useful in the treatment of lymphoedema which comprises the step of screening the compounds for their capacity to selectively inhibit, prevent or interfere with the stimulation of flk-l/KDR by a ligand, or the flk-l/KDR-associated signal transduction pathway and then selecting the compounds which have such a capacity.
  • a compound identified by a method according to the fifth aspect of the invention is provided.

Abstract

The use of an agent which selectively inhibits, prevents or interferes with the signal transduced by flk-1/KDR when activated by a ligand, in the manufacture of a medicament for the treatment of lymphoedema and in a method for reducing the filtration rate from blood to tissue. Also provided is a method for identifying compounds useful in the treatment of lymphoedema, and compounds identified thereby.

Description

THERAPETJTIC AGENTS
The present invention relates to the treatment of lymphoedema .
Lymphoedema arises from an imbalance between transvascular fluid filtration and lymph drainage capacity, but the pathophysiology is poorly understood. It is defined as a swelling associated with lymphatic insufficiency, lymphatic damage or lymphatic pathology and can result from cancer or cancer treatment, congenital disorders or infection by filarial worms, filariasis. Chronic lymphoedema of the arm is a common and distressing late complication of the successful treatment of breast cancer. Surgery and/or radiotherapy to axillary lymph nodes during breast cancer treatment is often followed by post mastectomy oedema (PMO) . There is currently no effective drug treatment for this condition.
Receptor tyrosine kinases (RTKs) comprise a large family of transmembrane receptors for growth factors with diverse biological activities. Interaction of the RTK with growth factor results in phosphorylation of the receptor and triggering of an RTK-specific signal transduction pathway.
Flk-l/KDR is an RTK found on vascular endothelial cells and has a role in normal vasculogenesis and angiogenesis. Normal vasculogenesis and angiogenesis play important roles in various physiological processes such as embryonic development, wound healing, organ regeneration and reproductive processes such as follicle development in the corpus luteum during ovulation and placental growth after pregnancy. Flk- l/KDR has also been implicated in tumour-associated angiogenesis and increased vascular permeability. A number of ligands for flk-l/KDR have been identified. For example the vascular endothelial growth factors VEGF-A and VEGF-C. VEGF-C is a ligand for flkl/KDR and at least one other receptor known as flt-4 (found on lymphatic endothelial cells) . VEGF-C is known to be produced by cancer cells (Salven P. et al (1998) Am. J. Pathol 153 103-8; Tsurusaki T. et al (1999) Br. J. Cancer 80 309-313; Valtola (1999) Am. J. Pathol. 154 1381-90; Lymboussaki (1998) Am. J. Pathol. 153 395-403; Fielder (1997) Leukemia 11 1234-1237), and is involved in tumour-related angiogenesis.
A number of inhibitors of flk-l/KDR have been isolated, such as those disclosed in US-A-5712395, US- A-5710173, US-A-5849742, US-A-5650415 , US-A-5851999, US-A-5792771 and US-A-5763441. These inhibitors are intended for the treatment of cell proliferative and cell differentiation disorders and disorders related to vasculogenesis and angiogenesis (such as cancer, diabetes, haemangioma and Karposi's sarcoma) and disorders associated with increased vascular permeability.
Activation of the flk-l/KDR receptor is known to activate phospholipase Cy, which results in production of diacylglycerol (DAG) and inositol triphosphate (IP3) . However, IP3-mediated release from intracellular stores is not necessary for KDR-mediated increased vascular permeability (Pocock et al . American Journal of Phsyiology 2000, in press) . DAG however, has been shown to result in calcium entry through DAG- activated calcium channels, and by stimulation of arachidonic acid-dependent calcium channels after degradation by DAG lipase. The precise nature of these calcium channels are unknown, but they are neither voltage operated calcium channels, nor store operated calcium channels.
The action of VEGF-C was previously thought to be limited to a few specific roles in an adult (namely wound healing and organ regeneration) and to tumour- associated angiogenesis. Underlying the present invention is the hypothesis that VEGF-C is secreted in response to lymph damage and acts in the lymphatic VEGF-C receptor, fit and that swelling associated with lymphatic insufficiency is brought about because the secreted VEGF-C also stimulates flk-1 on vascular endothelial cells. The inventor has shown that this results in a net increase in the filtration rate of water across the vasculature by a variety of mechanism, but without increasing the size of protein permeable pores in the vessel walls, and hence increased protein permeability. The increase in filtration rate adds to the lymphatic load on the tissue, hence negating the positive effects of VEGF-C on lymphangiogenesis . Therefore, although lymphoedema is not a pathology dependent on increased vascular permeability or angiogenesis, but one that is dependent on lymphatic insufficiency, inhibiting the flk-l/KDR receptor is predicted to be an effective therapy for this condition.
This prediction is based on the following observations :
1. Lymphoedema is caused by increased water flux without increased protein permeability (Bates
(1992) Ph.D. Thesis St. George's Hospital Medical School, University of London) .
2. VEGF-A, which activates both flk-l/KDR and flt-1, increases water flux without increasing protein permeability (Bates (1998) J. Physiol (Lond) 513: 225-233) .
3. Lymphoedema is associated with lymphangiogenesis (growth of new lymphatics) . Fig. 1 shows a plot of radial lymphatic density as a function of distance from tracer injection site in patients with lymphoedema in the swollen arm and the contralateral normal arm. Radial lymphatic density was measured by intradermal injection of FITC dextran and subsequent imaging on an intravital microscope. The Density was calculated as the number of lymphatics crossing a superimposed grip, per annulus away from the injection site. The area under the curve indicates total lymphatic density and is significantly increased in the ipsilateral arm of post-mastectomy patients, compared to their contralateral, normal arm (Dr Peter Mortimer, St. George's Hospital Medical School - unpublished results) . The fact that lymphatic density is higher in the swollen limb than in the non-swollen limb indicates that lymphangiogenesis has occurred.
4. Lymphangiogenesis is caused by VEGF-C acting on flt-4 ( Cao et al . (1998) Proc Natl Acad Sci USA 95: 14389-14394) .
5. VEGF-C stimulates flk-1 and flt-4 activation (Joukov et al . (1996) EMBO Journal 14:290-298).
6. VEGF-C activation of flk-l/KDR causes increased water flux, but PIGF activation of flt-1 does not. This is demonstrated by Figure 2, which is a graph showing the acute and chronic effect of growth factors. Hydraulic conductivity (water flux per unit pressure) was measured in perfused mesenteric capillaries in vivo from the frog, Rana temporaria, using the Landis Michel Technique modified for chronic permeability measurements
(Bates & Curry (1996) American Journal of Physiology Heart and Circulatory Physiology 271: H2520-2528) . The vessels were exposed to 10 nM VEGF-C (black bars), 1 % BSA (control) (white bars) or 10 nM PIGF (a flt-1 specific agonist) (grey bars) and hydraulic conductivity measured acutely, 20 minutes later, and the following day. VEGF-C, but not BSA or PIGF chronically increased hydraulic conductivity.
7. Flk inhibitors block the increase in water flux caused by VEGF-A. This is demonstrated by Figure 3 , which is a graph showing the change in hydraulic conductivity over time in the absence (solid line) or presence (dashed line) of the flkl/KDR inhibitor, ZM323881. Hydraulic conductivity was measured in mesenteric microvessels of the frog, Rana temporaria as previously described. The results show that ZM323881 inhibits the VEGF-mediated increase in water flux per unit measurement.
Therefore, it follows that lymphoedema is the result of VEGF-C stimulating flt-4 on lymph vessels causing lymphangiogenesis (to overcome the lymphatic insufficiency) , coupled with the activation of flk-1 on vascular endothelial cells which results in an increase in the ratio of water flux to protein flux, and hence swelling of the arm - this is shown schematically in Figure 4. Therefore, it can be concluded that flk-1 specific inhibitors will block the increase in water flux, and therefore the swelling in the arm after lymphoedema.
The basis of this invention therefore is that selective blocking of this receptor, or the signal pathway associated with it, will be effective in treating PMO and other lymphodemas. In particular, and according to a first aspect of the present invention, there is provided the use of an agent which selectively inhibits, prevents or interferes with the signal transduced by flk-l/KDR when activated by a ligand, in the manufacture of a medicament for the treatment of lymphoedema.
The ligand may be any ligand which is capable of interacting with flk-l/KDR and activating the signal transduction pathway. For example, the ligand may be a growth factor, particularly a vascular endothelial growth factor. Examples of vascular endothelial growth factors which interact with flk-l/KDR include VEGF-A and VEGF-C.
In a first embodiment of the first aspect of the invention, the agent inhibits, prevents, or interferes with the interaction between a ligand and flk-l/KDR. In one preferred example the agent is an antagonist of the flk-l/KDR receptor which thus prevents access by the normal ligand to that receptor. In a second embodiment of the first aspect of the invention, the agent inhibits, prevents or interferes with the flk-l/KDR-associated signal transduction pathway.
By "selectively inhibits, prevents or interferes with the signal transduced by flk-l/KDR when activated by a ligand", it is meant that the inhibition, prevention or interference activity mediated by the agent is substantially restricted to the flk-l/KDR receptor or its associated signal transduction pathway. In other words, the agent does not cross-react and inhibit, prevent or interfere with other receptors or their signal transduction pathways to a significant extent. Significant cross-reaction would reduce the efficiency of the agent and make the use of the agent unpredictable and/or cause undesired side-effects. The agent may be any moiety, for example a molecule or macromolecule, capable of specifically interacting with flk-l/KDR, the ligand, or one or more of the cellular components involved in flk-l/KDR receptor signal transduction. For example, the agent may be a polypeptide (or modified polypeptide, including glycopolypeptides and lipopolypeptides) , such as an antibody or antibody fragment, a soluble receptor molecule, or a peptide. An example of such an agent is a protein as described in US-A-5851999 which has a functional flk-l/KDR extracellular and transmembrane domain and a deleted or mutated cytoplasmic domain so that the polypeptide is signalling incompetent. Flk-l/KDR receptors dimerise, and the signalling incompetent polypeptide may dimerise with wild-type receptor and prevent signalling.
Alternatively the agent may be a nucleic acid sequence (including a modified nucleic acid sequence) . For example, the agent may be a polynucleotide which encodes for a polypeptide as described above. The agent may also be an expression vector which comprises such a polynucleotide. An example of such an agent is described in US-A-5851999. Alternatively, the agent may be an oligonucleotide, for example an antisense oligonucleotide. Such an oligonucleotide may inhibit prevent or interfere with the transcription or translation of a gene or polypeptide involved in ligand-activated flk-l/KDR signal transduction.
Alternatively the agent may be a mutant ligand or a molecule which is a molecular mimic for a ligand. A molecular mimic is a molecule which is not derived or derivable from the ligand, but mimics the binding capacity of the ligand by having the same or an equivalent binding pocket. For example, a mutant ligand or molecular mimic for a ligand may have the capacity to bind flk-l/KDR, but may lack the capacity to induce flk-l/KDR-associated signal transduction. In this way, the mutant ligand or molecular mimic may inhibit ligand-induced signal transduction by competing with the wild-type ligand for the receptor, and/or by directly blocking interaction of the wild-type ligand with the receptor (if the mutant ligand or molecular mimic form a more stable association with the receptor that the wild type ligand) .
Alternatively the agent may be a small organic molecule. Such molecules are already known as agents for use in the treatment of cell proliferative and cell differentiation disorders and disorders related to vasculogenesis and angiogenesis (such as cancer, diabetes, haemangioma and Karposi ' s sarcoma). For example thienyl compounds, such as those disclosed in US-A-5710173 , derivatives of quinoline such as those disclosed in US-A-5650415, and derivatives of quinazoline, quinoxaline, substituted aniline, isoxazoles, acrylonitrile or phenylacrylonitrile compounds such as those disclosed in US-A-5763441, US- A-5792771, US-A-5849742 and US-A-5712395.
Agents which inhibit, prevent or interfere with flk-1/KDR-ligand interaction according to the first embodiment of the first aspect of the invention may be identified by a number of methods which are known in the art. For example, antibodies which specifically bind flk-l/KDR can be produced using conventional techniques and screened using a phage display library or by ELISA. Isolated flk-l/KDR or its ligand can be used to screen for potentially useful agents by immobilising the flk-1/KDR/ligand for example on a microtitre plate, a column or on magnetic beads, selectively binding the agent to the immobilised flk- 1/KDR/ligand, and then identifying the bound agent. Agents which inhibit, prevent or interfere with flk- 1/KDR-ligand interaction may be screened by conducting a competitive binding assay for flk-l/KDR with the ligand.
A number of such inhibitors are available, including: ZD4190
(Wedge et al (2000) Cancer Res. 60, 970-975;] US- A-5814630 and US-A-5821246) ; (E) - (3 , 5, -Diisopropyl-4- hydroxyphenyl) -2- [ (3-phenyl-n- propyl) aminocarbonyl) acrylonitrile (SU1498) ; and 3- [2 , 4-dimethylpyrrol-5-yl) methylidene] indolin-2-one (SU5416, Fong et al (1999) Cancer Res. 59 99-106) . In the second embodiment of this first aspect of the invention, the agent inhibits, prevents or interferes with the flk-l/KDR-associated signal transduction pathway. For example, the agent may inhibit, prevent or interfere with flk-l/KDR-associated activation of phospholipase Cγ , formation of diacyl glycerol, production of arachidonic acid by DAG-lipase and/or DAG-mediated calcium entry through non-store operated, non-voltage-gated, calcium channels. An example of an agent which inhibits calcium entry through non-store operated, non-voltage-gated, calcium channels is carboxyamido-triazole (CAI, as decribed in USSN 5359078) (Kohn et al (1992) Cancer Res. 52 3208- 3212) .
Agents which inhibit, prevent or interfere with the flk-l/KDR-associated signal transduction pathway may be identified by methods known in the art. Examples of such agents include (1- [6- ( [17β) -3- Methoxyestra-1,3,5 (10) trien-17-yl] amino) hexyl] -1H- pyrrole-2, 5-dione (U73122, an inhibitor of phospholipase C) and 1,6- bis (cyclohexyloximinocarboynlamino)hexane (RHC-80267, an inhibitor of DAG lipase)
The lymphoedema may be due to surgery for cancer treatment where lymph nodes of, for example, the axilla, neck or inguinal nodes have been partially or wholly removed or destroyed by surgery. Also, lymphoedema may arise due to congenital disorders, or infection by filarial worms, including lymphoedema of the arm, leg, trunk, scrotum, face or other part of the body. The agent defined in the first aspect of the invention may be formulated into a pharmaceutical composition and may be used in conjunction with a pharmaceutically acceptable carrier.
The pharmaceutical composition may be in a unit- dose form, such as a tablet, capsule, ampoule or suppository, methods for the preparation of which are known per se . The carrier may act as a filler, binder, disintegrant , stabilizer or lubricant.
According to a second aspect of the present invention, there is provided a method for treating lymphoedema in a mammalian patient, which comprises the step of administering an effective amount of a agent as defined in the first aspect of the invention to the mammalian patient . The composition may be administered enterally or parenterally, for example by topical, oral or nasal, administration, or by intracavernous, intravenous, intramucsular or subcutaneous injection.
The "effective amount" of the composition to be delivered in one administration will depend on the individual patient characteristics (severity of disease, size, age etc.) as well as the characteristics of the compound. Generally an "effective amount" is that amount necessary to substantially inhibit, prevent or interfere with the stimulation of flk-l/KDR by a ligand, or the flk-1/KDR-associated signal transduction pathway. Unique or continued application of such an effective amount results in reduction or amelioration of the symptoms with lymphoedema. According to a third aspect of the invention, there is provided a method for reducing the hydraulic conductivity of blood vessels in a mammalian patient, which comprises the step of administering an effective amount of an agent as defined in the first aspect of the invention to the mammalian patient. The hydraulic conductivity may be reduced in particular blood vessels in the patient or in blood vessels in a particular area of the patient. For example, the method of the third aspect of the invention may be used to reduce the hydraulic conductivity of the blood vessels in at least one arm of the mammalian patient.
According to a fourth aspect of the present invention, there is provided a method for reducing the filtration rate from blood to tissue in at least one arm of a mammalian patient, which comprises the step of administering an effective amount of an agent as defined in the first aspect of the invention to the mammalian patient.
The filtration rate from blood to tissue may be reduced in particular blood vessels in the patient, in a particular tissue of the patient, or in a particular area of the patient. For example, the method of the fourth aspect of the invention may be used to reduce the filtration rate from blood to tissue in at least one arm of the mammalian patient.
According to a fifth aspect of the present invention there is provided a method for identifying compounds useful in the treatment of lymphoedema which comprises the step of screening the compounds for their capacity to selectively inhibit, prevent or interfere with the stimulation of flk-l/KDR by a ligand, or the flk-l/KDR-associated signal transduction pathway and then selecting the compounds which have such a capacity. According to a sixth aspect of the invention, there is provided a compound identified by a method according to the fifth aspect of the invention.
The content of all the references cited herein is incorporated by reference in its entirety.

Claims

1. The use of an agent which selectively inhibits, prevents or interferes with the signal transduced by flk-l/KDR when activated by a ligand, in the manufacture of a medicament for the treatment of lymphoedema .
2. The use according to claim 1, wherein the ligand is VEGF-C.
3. The use according to claim 1 or 2 , wherein the agent inhibits, prevents, or interferes with the interaction between a ligand and flk-l/KDR.
4. The use according to claim 1 or 2 , wherein the agent inhibits, prevents or interferes with the flk-l/KDR-associated signal transduction pathway.
5. The use according to any preceding claim, wherein the agent is a small organic molecule and is selected from the group consisting of: thienyl compounds, as disclosed in US-A-5710173; derivatives of quinoline as disclosed in US-A-5650415; and derivatives of quinazoline, quinoxaline, substituted aniline, isoxazoles, acrylonitrile or phenylacrylonitrile compounds as disclosed in US-A-5763441, US-A-5792771, US-A-5849742 and US-A-5712395.
6. A method for reducing the hydraulic conductivity of blood vessels in a mammalian patient, which comprises the step of administering an effective amount of an agent as defined in any preceding claim to the mammalian patient.
7. A method for reducing the filtration rate from blood to tissue in at least one arm of a mammalian patient, which comprises the step of administering an effective amount of an agent as defined in any of claims 1 to 5 to the mammalian patient.
8. A method for identifying compounds useful in the treatment of lymphoedema which comprises the step of screening the compounds for their capacity to selectively inhibit, prevent or interfere with the stimulation of flk-l/KDR by a ligand, or the flk-l/KDR- associated signal transduction pathway and then selecting the compounds which have such a capacity.
9. A compound identified by a method according to claim 8.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7723349B2 (en) 2003-04-24 2010-05-25 Incyte Corporation Aza spiro alkane derivatives as inhibitors of metalloproteases

Citations (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992014748A1 (en) * 1991-02-22 1992-09-03 American Cyanamid Company Identification of a novel human receptor tyrosine kinase gene
WO1994011499A1 (en) * 1992-11-13 1994-05-26 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Flk-1 is a receptor for vascular endothelial growth factor
WO1995021613A1 (en) * 1994-02-09 1995-08-17 Sugen, Inc. Compounds for the treatment of disorders related to vasculogenesis and/or angiogenesis
WO1996040675A1 (en) * 1995-06-07 1996-12-19 Sugen, Inc. Thienyl compounds for inhibition of cell proliferative disorders
WO1996040629A1 (en) * 1995-06-07 1996-12-19 Sugen, Inc. Tyrphostin-like compounds for the treatment of cell proliferative disorders or cell differentiation disorders
WO1996040648A1 (en) * 1995-06-07 1996-12-19 Sugen, Inc. Quinazolines and pharmaceutical compositions
WO1997009427A1 (en) * 1995-09-08 1997-03-13 Genentech, Inc. Vegf-related protein
US5650415A (en) * 1995-06-07 1997-07-22 Sugen, Inc. Quinoline compounds
WO1997034920A1 (en) * 1996-03-21 1997-09-25 Sugen, Inc. Assays for kdr/flk-1 receptor tyrosine kinase inhibitors
US5712395A (en) * 1992-11-13 1998-01-27 Yissum Research Development Corp. Compounds for the treatment of disorders related to vasculogenesis and/or angiogenesis
US5763441A (en) * 1992-11-13 1998-06-09 Sugen, Inc. Compounds for the treatment of disorders related to vasculogenesis and/or angiogenesis
US5766863A (en) * 1993-11-23 1998-06-16 Genentech, Inc. Kinase receptor activation assay
US5766860A (en) * 1992-11-23 1998-06-16 American Cyanamid Company Screening method using a recombinant kinase insert domain containing receptor and gene encoding same
US5773459A (en) * 1995-06-07 1998-06-30 Sugen, Inc. Urea- and thiourea-type compounds
US5792771A (en) * 1992-11-13 1998-08-11 Sugen, Inc. Quinazoline compounds and compositions thereof for the treatment of disease
WO1998039035A1 (en) * 1997-03-07 1998-09-11 The Wistar Institute Of Anatomy & Biology Method and compositions for healing tissue defects and inducing hypervascularity in mammalian tissue
WO1998055638A1 (en) * 1997-06-03 1998-12-10 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Regulatory sequences capable of conferring expression of a heterologous dna sequence in endothelial cells in vivo and uses thereof
WO1998058053A1 (en) * 1997-06-18 1998-12-23 Merck & Co., Inc. Human receptor tyrosine kinase, kdr
WO1999010537A1 (en) * 1997-08-26 1999-03-04 Board Of Regents, The University Of Texas System A tumor suppressor designated ts10q23.3
US5891650A (en) * 1993-11-23 1999-04-06 Genentech Incorporated Kinase receptor activation assay
WO1999017769A1 (en) * 1997-10-06 1999-04-15 Basf Aktiengesellschaft INDENO[1,2-c]-, NAPHTHO[1,2-c]- AND BENZO[6,7]CYCLOHEPTA[1,2-c]PYRAZOLE DERIVATIVES
WO1999021859A1 (en) * 1997-10-10 1999-05-06 Glaxo Group Limited Azaoxindole derivatives
WO1999024440A1 (en) * 1997-11-11 1999-05-20 Pfizer Products Inc. Thienopyrimidine and thienopyridine derivatives useful as anticancer agents
US5916763A (en) * 1995-11-09 1999-06-29 The Regents Of The University Of California Promoter for VEGF receptor
WO2000058511A1 (en) * 1999-03-26 2000-10-05 Ludwig Institute For Cancer Research Screening and therapy for lymphatic disorders involving the flt4 receptor tyrosine kinase (vegfr-3)

Patent Citations (30)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992014748A1 (en) * 1991-02-22 1992-09-03 American Cyanamid Company Identification of a novel human receptor tyrosine kinase gene
US5861301A (en) * 1992-02-20 1999-01-19 American Cayanamid Company Recombinant kinase insert domain containing receptor and gene encoding same
US5763441A (en) * 1992-11-13 1998-06-09 Sugen, Inc. Compounds for the treatment of disorders related to vasculogenesis and/or angiogenesis
WO1994011499A1 (en) * 1992-11-13 1994-05-26 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Flk-1 is a receptor for vascular endothelial growth factor
US5849742A (en) * 1992-11-13 1998-12-15 Sugen Inc. Compounds for the treament of disorders related to vasculogenesis and/or angiogenesis
US5792771A (en) * 1992-11-13 1998-08-11 Sugen, Inc. Quinazoline compounds and compositions thereof for the treatment of disease
US5712395A (en) * 1992-11-13 1998-01-27 Yissum Research Development Corp. Compounds for the treatment of disorders related to vasculogenesis and/or angiogenesis
US5766860A (en) * 1992-11-23 1998-06-16 American Cyanamid Company Screening method using a recombinant kinase insert domain containing receptor and gene encoding same
US5891650A (en) * 1993-11-23 1999-04-06 Genentech Incorporated Kinase receptor activation assay
US5766863A (en) * 1993-11-23 1998-06-16 Genentech, Inc. Kinase receptor activation assay
WO1995021613A1 (en) * 1994-02-09 1995-08-17 Sugen, Inc. Compounds for the treatment of disorders related to vasculogenesis and/or angiogenesis
US5773459A (en) * 1995-06-07 1998-06-30 Sugen, Inc. Urea- and thiourea-type compounds
WO1996040675A1 (en) * 1995-06-07 1996-12-19 Sugen, Inc. Thienyl compounds for inhibition of cell proliferative disorders
US5710173A (en) * 1995-06-07 1998-01-20 Sugen, Inc. Thienyl compounds for inhibition of cell proliferative disorders
US5650415A (en) * 1995-06-07 1997-07-22 Sugen, Inc. Quinoline compounds
US5891917A (en) * 1995-06-07 1999-04-06 Sugen, Inc. Certain acrylonitrile-sulfonamide derivatives
US5886195A (en) * 1995-06-07 1999-03-23 Sugen, Inc. Thienyl compounds for inhibition of cell proliferative disorders
WO1996040629A1 (en) * 1995-06-07 1996-12-19 Sugen, Inc. Tyrphostin-like compounds for the treatment of cell proliferative disorders or cell differentiation disorders
WO1996040648A1 (en) * 1995-06-07 1996-12-19 Sugen, Inc. Quinazolines and pharmaceutical compositions
WO1997009427A1 (en) * 1995-09-08 1997-03-13 Genentech, Inc. Vegf-related protein
US5916763A (en) * 1995-11-09 1999-06-29 The Regents Of The University Of California Promoter for VEGF receptor
WO1997034920A1 (en) * 1996-03-21 1997-09-25 Sugen, Inc. Assays for kdr/flk-1 receptor tyrosine kinase inhibitors
WO1998039035A1 (en) * 1997-03-07 1998-09-11 The Wistar Institute Of Anatomy & Biology Method and compositions for healing tissue defects and inducing hypervascularity in mammalian tissue
WO1998055638A1 (en) * 1997-06-03 1998-12-10 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Regulatory sequences capable of conferring expression of a heterologous dna sequence in endothelial cells in vivo and uses thereof
WO1998058053A1 (en) * 1997-06-18 1998-12-23 Merck & Co., Inc. Human receptor tyrosine kinase, kdr
WO1999010537A1 (en) * 1997-08-26 1999-03-04 Board Of Regents, The University Of Texas System A tumor suppressor designated ts10q23.3
WO1999017769A1 (en) * 1997-10-06 1999-04-15 Basf Aktiengesellschaft INDENO[1,2-c]-, NAPHTHO[1,2-c]- AND BENZO[6,7]CYCLOHEPTA[1,2-c]PYRAZOLE DERIVATIVES
WO1999021859A1 (en) * 1997-10-10 1999-05-06 Glaxo Group Limited Azaoxindole derivatives
WO1999024440A1 (en) * 1997-11-11 1999-05-20 Pfizer Products Inc. Thienopyrimidine and thienopyridine derivatives useful as anticancer agents
WO2000058511A1 (en) * 1999-03-26 2000-10-05 Ludwig Institute For Cancer Research Screening and therapy for lymphatic disorders involving the flt4 receptor tyrosine kinase (vegfr-3)

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
M.H.WITTE ET AL.: "PHENOTYPIC AND GENOTYPIC HETEROGENEITY IN FAMILIAL MILROY LYMPHEDEMA" LYMPHOLOGY, vol. 31, no. 4, December 1998 (1998-12), pages 145-155, XP000990600 *
M.J.KARKKAINEN ET AL.: "Vascular endothelial growth factor receptors in the regulation of angiogenesis and lymphangiogenesis" ONCOGENE, vol. 19, no. 49, 20 November 2000 (2000-11-20), pages 5598-5605, XP000990610 *
R.E.FERRELL ET AL.: "Hereditary lymphedema: evidence for linkage and genetic heterogeneity" HUMAN MOLECULAR GENETICS, vol. 7, no. 13, December 1998 (1998-12), pages 2073-2078, XP000990647 *
T. M[KINEN ET AL.: "Inhibition of lymphangiogenesis with resulting lymphedema in transgenic mice expressing soluble VEGF receptor-3" NAT.MED., vol. 7, no. 2, February 2001 (2001-02), pages 199-205, XP000990590 *
V. JOUKOV ET AL.: "A novel vascular endothelial growth factor, VEGF-C, is a ligand for the Flt4 (VEGFR-3) and KDR (VEGFR-2) receptor tyrosine kinases" THE EMBO JOURNAL, vol. 15, no. 2, 1996, pages 290-298, XP000100201 cited in the application *
Y. CAO ET AL.: "Vascular endothelial growth factor C induces angiogenesis in vivo" PROC.NATL.ACAD.SCI. USA, vol. 95, no. 24, 24 November 1998 (1998-11-24), pages 14389-14394, XP000994696 cited in the application *

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