WO2005008250A1 - Composes et methodes pour favoriser l'angiogenese, au moyen d'un inhibiteur de la gamma secretase et en inhibant la voie de la gamma secretase - Google Patents

Composes et methodes pour favoriser l'angiogenese, au moyen d'un inhibiteur de la gamma secretase et en inhibant la voie de la gamma secretase Download PDF

Info

Publication number
WO2005008250A1
WO2005008250A1 PCT/SE2004/001146 SE2004001146W WO2005008250A1 WO 2005008250 A1 WO2005008250 A1 WO 2005008250A1 SE 2004001146 W SE2004001146 W SE 2004001146W WO 2005008250 A1 WO2005008250 A1 WO 2005008250A1
Authority
WO
WIPO (PCT)
Prior art keywords
gamma
secretase
angiogenesis
cells
group
Prior art date
Application number
PCT/SE2004/001146
Other languages
English (en)
Inventor
Mats HELLSTRÖM
Linda Karlsson
Elisabet Wallgard
Original Assignee
Angiogenetics Sweden Ab
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from SE0302111A external-priority patent/SE0302111D0/xx
Application filed by Angiogenetics Sweden Ab filed Critical Angiogenetics Sweden Ab
Priority to EP04749182A priority Critical patent/EP1671129A1/fr
Publication of WO2005008250A1 publication Critical patent/WO2005008250A1/fr
Priority to US11/306,973 priority patent/US20060264380A1/en
Priority to US12/378,731 priority patent/US20090209473A1/en

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase

Definitions

  • the invention relates to the field of angiogenesis and treatment of diseases or conditions related to angiogenic abnormalities.
  • Angiogenesis is a fundamental process required for the normal growth and development of tissues, and involves the proliferation of new capillaries from preexisting blood vessels. Under normal physiological conditions, humans or animals only undergo angiogenesis in very specific situations and angiogenesis is tightly controlled through a highly regulated system of angiogenic stimulators and inhibitors. Deviation from such a tight control often leads to or is associated with disease.
  • Angiogenesis is a prerequisite for the development and differentiation of the vascular tree, as well as for a wide variety of fundamental physiological processes including embryogenesis, somatic growth, tissue and organ repair and regeneration, cyclical growth of the corpus luteum and endometrium, and development and differentiation of the nervous system.
  • angiogenesis occurs in the follicle during its development, in the corpus luteum following ovulation and in the placenta to establish and maintain pregnancy.
  • Angiogenesis additionally occurs as part of the body's repair processes, e.g. in the healing of wounds and fractures.
  • Angiogenesis is also a factor in tumor growth, since a tumor must continuously stimulate growth of new capillary blood vessels in order to grow.
  • Endothelial cells and pericytes surrounded by a basement membrane, form capillary blood vessels.
  • Angiogenesis begins with the erosion of the basement membrane by enzymes released by endothelial cells and leukocytes.
  • the endothelial cells which line the lumen of blood vessels, then protrude through the basement membrane.
  • Angiogenic stimulants induce the endothelial cells to migrate through the eroded basement membrane.
  • the migrating cells form a "sprout" off the parent blood vessel, where the endothelial cells undergo mitosis and proliferate.
  • the endothelial sprouts merge with each other to form capillary loops, creating the new blood vessel. See e.g. Folkman et al, Adv. Cancer Res., Vol. 43, pp. 175-203 (1985), and Ingber et al, Cell, Vol. 58, pp. 803-805 (1985).
  • angiogenesis While persistent, unregulated angiogenesis occurs in numerous disease states, insufficient or nonexistent angiogenesis can also be a serious medical problem. Promoting angiogenesis is desirable in situations where vascularization is to be established or extended, for example after tissue or organ transplantation, or to stimulate establishment of collateral circulation in tissue infarction or arterial stenosis, such as in coronary heart disease and thromboangitis obliterans. Enhancing angiogenic activity may also be useful in treating ischemic conditions, including cardiovascular and limb ischemia. Finally, materials or methods that initiate or increase angiogenesis could potentially also be used to create research models with greater-fhan-normal angiogenesis.
  • the protease gamma-secretase is a complex of at least four proteins: presenilin 1 (PS 1), nicastrin, APH-1, and PEN-2.
  • PS 1 presenilin 1
  • Gamma-secretase has more than one enzymatic activity cleaving multiple substrates. It is also involved in processing the Notch receptor, part of a signalling pathway critical for embryonic development.
  • DAPT a cell-permeable dipeptide protease inhibitor, one of the known gamma- secretase inhibitors which blocks Notch signaling (Micchelli, CA. et al, 2003, gamma-secretase/presenilin inhibitors for Alzheimer's disease phenocopy Notch mutations in Drosophila, FASEB J. 17, 79-81).
  • Alzheimer's is characterized by the formation of plaques.
  • the plaques are mainly comprised of beta-amyloid peptides (A beta).
  • a beta beta-amyloid peptides
  • a beta are produced during normal cellular metabolism.
  • the peptides are 40-42 amino acids in length and are derived from larger amyloid precursor proteins. To date, this family of proteins is understood to contain three members: amyloid precursor protein (APP), amyloid precursor-like protein 1 (APLP 1), and amyloid precursor-like protein 2 (APLP2).
  • APP amyloid precursor protein
  • APLP 1 amyloid precursor-like protein 1
  • APLP2 amyloid precursor-like protein 2
  • At least two pathways are involved in the processing of these proteins, which can result in, inter alia, the formation of A beta.
  • APP may be cleaved extracellularly by alpha-secretase to create a membrane-bound intermediate. This intermediate is subsequently cleaved by gamma-secretase to release a non- amyloidogenic fragment.
  • BACE1 beta-secretase
  • This intermediate is subsequently cleaved by gamma-secretase to release the amyloidogenic A beta fragment (Scheinfeld et al, 2002, J. Biol. Chem. 277:44195- 201).
  • gamma-secretase inhibitors have surprisingly been shown to increase angiogenesis. This novel elucidation of activity is exploited in treatments for angiogenesis and related conditions.
  • an angiogenesis initiator or increaser which comprises a pharmaceutically-effective amount of a gamma- secretase inhibitor.
  • the gamma-secretase inhibitor may be DAPT.
  • the angiogenesis initiator or increaser may further include a pharmaceutically acceptable carrier or adjuvant.
  • a method of influencing a disease state in a cell, a group of cells, or an organism comprises administering at least one of a gamma-secretase inhibitor or a gamma-secretase pathway inhibitor to the cell, group of cells, or organism.
  • the disease thus influenced can be selected from the group consisting of atherosclerosis, hemangioma, hemangioendothelioma, vascular malformations, warts, pyogenic granulomas, hair growth, Kaposi's sarcoma, scar keloids, allergic edema, neoplasms, psoriasis, decubitus or stasis ulcers, gastrointestinal ulcers, dysfunctional uterine bleeding, follicular cysts, ovarian hyperstimulation, endometriosis, neoplasms, preeclampsia, placental insufficiency, respiratory distress, ascites, peritoneal sclerosis, adhesion formation, metastatic spreading, coronary artery disease, ischemic heart disease, ischemic limb disease, obesity, rheumatoid arthritis, synovitis, bone destruction, cartilage destruction, osteomyelitis, pannus growth, osterphyte formation, cancer, as
  • the gamma-secretase inhibitor could be DAPT or an analogue of DAPT.
  • a method of increasing the angiogenic process in a cell, a group of cells, or an organism comprises administering a pharmaceutical composition comprising a pharmaceutically effective amount of at least one gamma-secretase inhibitor or gamma-secretase pathway inhibitor to the cell, group of cells, or organism.
  • the gamma-secretase inhibitor could be DAPT or an analogue of DAPT.
  • the pharmaceutical composition may be administered to prevent, treat, or cure a condition selected from any of the above-noted disease states.
  • a method for initiating or increasing angiogenesis in a cell, a group of cells, a tissue, or an organism comprises inhibiting the gamma-secretase pathway in the cell, group of cells, tissue or organism.
  • a further method is provided which initiates or increases angiogenesis in a cell, a group of cells, a tissue, or an organism by inhibiting gamma- secretase in the cell, group of cells, tissue or organism.
  • the methods may comprise reducing the level of expression of gamma-secretase, administering DAPT, administering an antibody against gamma-secretase, or delivering a vector to the target, wherein the vector comprises a polynucleotide encoding at least one gamma- secretase inhibitor, operatively linked to a suitable promoter.
  • the promoter may be a tissue or organ specific promoter specific for a tissue or organ in which angiogenesis is to be initiated or increased.
  • a method for screening for a substance which initiates or increases angiogenesis comprises measuring an activity of the gamma-secretase pathway in the presence of a candidate compound, measuring an activity of the gamma-secretase pathway in the absence of a candidate compound, and then comparing the activity measured in the presence of a candidate compound with the activity measured in the absence of the candidate compound. A change in activity indicates that that candidate initiates or increases angiogenesis.
  • gamma-secretase inhibitor means any material or compound that, e.g., binds to, partially or totally blocks activity, decreases, prevents, delays activation, inactivates, desensitizes, or down regulates the activity or expression of gamma- secretase or the gamma-secretase pathway.
  • Inhibitors include genetically modified versions of gamma-secretase proteins, e.g., versions with altered activity, as well as naturally occurring and synthetic ligands, antagonists, agonists, antibodies, small chemical molecules and the like. Inhibitor, as the term is used herein, includes but is not limited to an antagonist.
  • the present invention encompasses compounds and compositions which have are pharmaceuticals or have a pharmaceutical effect.
  • the compounds of the invention may be admixed, encapsulated, conjugated or otherwise associated with other molecules, molecule structures or mixtures of compounds for assisting in uptake, distribution and/or absorption. They encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other compound which, upon administration to an animal including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to prodrugs and pharmaceutically acceptable salts of the compounds of the invention, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents.
  • compositions of the present invention may additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art- established usage levels.
  • the compositions may contain additional, compatible, pharmaceutically-active materials such as, for example, local anesthetics, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers.
  • additional materials useful in physically formulating various dosage forms of the compositions of the present invention such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers.
  • such materials when added, should not unduly interfere with the biological activities of the components of the compositions of the present invention.
  • the formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings and/or aromatic substances and the like which do not deleteriously interact with the active compound.
  • auxiliary agents e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings and/or aromatic substances and the like which do not deleteriously interact with the active compound.
  • phrases “pharmaceutically acceptable” refers to molecular entities and compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, dizziness and the like, when administered to a patient.
  • phrases "pharmaceutically effective amount” is used herein to mean an amount sufficient to initiate or increase to some beneficial degree, preferably to increase by at least about 30 percent, more preferably by at least 40 percent, more preferably by at least 50 percent, more preferably by at least 60 percent, more preferably by at least 70 percent, more preferably by at least 80 percent, most preferably by at least 90 percent, angiogenesis as compared to untreated controls.
  • the compounds and compositions disclosed herein may be administered by any route, including intradermally, subcutaneously, orally, intraarterially or intravenously.
  • the concentration of a disclosed compound in a pharmaceutically acceptable mixture will vary depending on several factors, including the dosage of the compound to be administered, the pharmacokinetic characteristics of the compound(s) employed, and the route of administration. Skilled workers can extrapolate the mouse data presented herein, which is based on lOOmg/kg, 0.1-l ⁇ M plasma concentration, to reach the desired effect in the organism of interest.
  • the agent may be administered in a single dose or in repeat doses.
  • organ refers to animals, preferably mammals, more preferably mammals such as experimental mammals or humans.
  • subject to be treated by the inventive methods can mean either a human or non-human animal.
  • vector refers to a nucleic acid construct, generated recombinantly or synthetically, with a series of specified nucleic acid elements that permit transcription of a particular nucleic acid in a host cell as known in the art.
  • the expression vector can be part of a plasmid, virus, or nucleic acid fragment.
  • the expression vector includes a nucleic acid to be transcribed operably linked to a promoter.
  • Figure 1 shows the central region of a control mouse retina
  • Figure 2 shows the central region of a treated mouse retina
  • Figure 3 shows the capillary enclosed areas of a control mouse retina
  • Figure 4 shows the capillary enclosed areas of a treated mouse retina
  • Figure 5 depicts quantitative data on retinal vessel density
  • Figure 6 shows labeled astrocytes in a control mouse retina
  • Figure 7 shows labeled astrocytes in a treated mouse retina
  • Figure 8 shows a retinal cross section with asterisks marking vascular tufts
  • Figure 9 shows vascular tufts in a control mouse retina
  • Figure 10 shows the absence of vascular tufts in a treated mouse retina
  • Figure 11 depicts quantitative data on retinal vascular tuft formation
  • Figure 12 depicts quantitative measurements of VEGF-A levels in mouse retinas
  • Figure 13 shows vascularization of a control mouse retina
  • Figure 14 shows vascularization of a control mouse retina
  • Figure 15 shows vascularization of a treated mouse retina
  • Figure 16 shows vascularization of a treated mouse retina.
  • the findings of the present invention are particularly useful in modulating conditions characterized by systemic or local abno ⁇ nalities in angiogenic activity.
  • examples include, but are not limited to, eye diseases (e.g., AMD, retinopathy of prematurity, diabetic retinopathy), responses to organ transplantation, coronary artery disease, ischemic heart disease, wound healing, peripheral vascular disease, tumorogenesis/cancer, and inflammatory conditions (e.g., rheumatoid arthritis).
  • mice employed murine cells or tissues because they provided a convenient way to analyze factors upregulated in mammalian embryos.
  • inventive methods and compositions are also applicable to other mammals including, but not limited to, mice, rats, rabbits, dogs, pigs, and humans.
  • murine models have been extrapolated to Alzheimer's in humans.
  • C. elegans and Drosophila species have been used to elucidate Alzheimer's related pathways with good reproducibility in mice.
  • the close homology between mammalian genes when compared to the non-mammal models is further evidence that the data from one species of mammal is applicable to other mammals.
  • the present disclosure that the gamma-secretase pathway and factors active therein influence angiogenesis forms the basis for treatment methods of many human and animal diseases.
  • the invention also encompasses kits and reagents adapted to the subject methods.
  • DAPT is used as an inhibitor of the gamma- secretase pathway. Additional inhibitors of gamma-secretase pathway can be identified. For example, medicinal and combinatorial chemistry methods well-known to those skilled in the art can be used to modify known PS-1 antagonists to form new gamma-secretase inhibitors with improved efficacy for the purposes of the present invention. Further, as DAPT has already been identified as one useful compound according to the invention, analogs of DAPT may be used.
  • assays are conducted to evaluate their ability to initiate or increase angiogenesis.
  • assays are well-known to those of ordinary skill in the art, see e.g. Murray, Angiogenesis protocols, in Murray, Methods in Molecular Medicine, 2001, ISBN 0-89603-687-7.
  • Known gamma-secretase inhibitors rely on the previously-elucidated understanding of the role of the gamma-secretase pathway, making some inhibitors more and some inhibitors less effective at influencing angiogenesis. Accordingly, known factors may also be evaluated for their ability to create the results desired for the novel application disclosed herein.
  • Assays for a gamma-secretase inhibitor that creates the desired effect on angiogenesis may also rely on its role on tumor growth.
  • Cell lines or animal models with a known propensity for tumorogenesis can be subjected to treatment with a candidate inhibitor. Tumor growth can be monitored and evaluations can be made of vascular parameters in the tumor and vascular density and morphology in biopsies from the tumor. These results can be compared with known or control values to indicate the efficacy of the gamma-secretase inhibitor/angiogenesis increaser tested.
  • Administration may preferentially be oral. Parenteral administration could also be utilized, particularly where the properties of the gamma-secretase inhibitor and any vehicles or diluents employed are not compatible with oral uptake and distribution. Dosing of gamma-secretase inhibitors would be based on the pharmacology of the inhibitor or inhibitor mixture, including consideration of IC50 values, metabolism, excretion and toxicity values. Administration may be for the purpose of managing disease, treating disease, preventing disease, research or other purposes.
  • diseases which may be particularly affected by compounds and methods of the present invention include atherosclerosis, hemangioma, hemangioendothelioma, vascular malformations, warts, pyogenic granulomas, hair growth, Kaposi's sarcoma, scar keloids, allergic edema, neoplasms, psoriasis, decubitus or stasis ulcers, gastrointestinal ulcers, dysfunctional uterine bleeding, follicular cysts, ovarian hyperstimulation, endometriosis, neoplasms, preeclampsia, placental insufficiency, respiratory distress, ascites, peritoneal sclerosis, adhesion formation, metastatic spreading, coronary artery disease, ischemic heart disease, ischemic limb disease, obesity, rheumatoid arthritis, synovitis, bone destruction, cartilage destruction, osteomyelitis, pannus growth, osterphyte formation, cancer
  • monitoring activity of the gamma-secretase pathway could be accomplished by creating a substrate for the gamma-secretase that can be detected in various assays.
  • Kinoshita et al. 2002, J Neurochem.
  • APP-CT gamma secretase- generated carboxyl-terminal domain of APP
  • FRET fluorescence resonance energy transfer
  • BRET2 Bioluminescence Resonance Energy Transfer
  • a fluorescent dye and a quencher are attached to either side of the gamma-secretase substrate cleavage site of a gamma-secretase substrate.
  • the dye and quencher are in close proximity and no signal is produced from the assay.
  • the quencher is removed from the dye, a signal results which can be monitored and quantified.
  • This assay could be performed as an isolated biochemical assay in vitro, in cells and in animal models in vivo.
  • An alternate assay that could be used to monitor the efficacy of the drug in cells, animal models or human patients involves taking biopsies from diseased tissue and monitoring the cleavage of gamma-secretase substrates using conventional techniques such as detecting the presence and quantities of the substrates with antibodies.
  • mice with oxygen induced retinopathy are a commonly-used model and are described in the literature. They are at times referred to as retinopathy of prematurity models or ROP mice.
  • the model takes advantage of the fact that full term mice pups are born with an immature retinal vascularization which matures during the first three weeks of postnatal growth.
  • mice from the same litter were placed, with their nursing mother, in a hyperoxic environment (75% oxygen) at age day seven. After exposure to the hyperoxic environment for 5 days, the 12-day old pups were removed to normal air. In the treated mice, DAPT was administered once daily during age days 12-16. Control pups were treated the same way but injected with the vehicle. At post natal day 17 the mice were euthanized and the retinas were prepared for whole mount hnmunohistochemistry .
  • mice pups at 17 post natal days after oxygen induced retinopathy exhibit formation of avascular zones in the central, i.e., close to the optic nerve, areas of the retina. At the same time there is an increased vascular density in the peripheral parts of the retina. This can be quantified in the superficial capillary plexus by counting the number of capillary enclosed areas in the peripheral part of the retina.
  • Example 2 Evaluation of retinal vascularization and vessel density in gamma- secretase inhibitor treated OIR mice
  • FIG. 3 shows a control mouse retina, with asterisks pointing out capillary enclosed areas.
  • the peripheral retina shown in Figure 4 is from a DAPT-treated mouse, showing a significant increase in capillary enclosed areas and therefore an angiogenesis- increasing effect. To further evaluate these results, the capillary enclosed areas were counted.
  • the data presented in Figure 5 (p ⁇ 0.001) reflects the near doubling of such areas in DAPT-treated mice.
  • GFAP Glial Acidic Fibrillary Protein
  • Vascular tufts form in the retinas of OIR mice models.
  • the tufts consist of endothelial cells growing in a small localized cluster above the inner limiting membrane and pouching into the vitreous.
  • Figure 8 shows a retinal cross section with asterisks marking vascular tufts.
  • Figure 9 shows a no ⁇ nal OIR mouse retina and numerous vascular tufts. Despite the increased angiogenic response seen in DAPT-treated OIR mice, a significant reduction in capillary tufts was observed as shown in Figure 10.
  • the numbers of capillary tufts can be quantified as a measurement of the pathological angiogenic response in the OIR model. This can be done, for example, with whole retinas stained with isolectin using a Nikon Microphot-FXA microscope and 4x magnification lens. Quantification of the results in this case are reflected in Figure 11 (p-value ⁇ 0.01).
  • Example 4 Evaluation of possible VEGF-A role in gamma-secretase inhibitor treated OIR mice
  • VEGF-A Vascular Endothelial Growth Factor A
  • VEGF-A Vascular Endothelial Growth Factor A
  • Example 5 Evaluation of retinal vessel density and architecture in gamma-secretase inhibitor treated non-OIR mice
  • Example 6 Evaluation of gamma-secretase inhibitor in animal models of myocardial and ischemia, myocardial infarction, and peripheral ischemia
  • a gamma-secretase inhibitor e.g. DAPT
  • DAPT gamma-secretase inhibitor
  • infarcted hearts are processed for morphometric analysis after immunostaining for endothelial thrombomodulin, which stains all vessels, or for smooth muscle x-actin, which stains mature SMC-covered vessels.
  • endothelial thrombomodulin which stains all vessels
  • smooth muscle x-actin which stains mature SMC-covered vessels.
  • Perfusion averaged from 3 images per mouse in the upper hind limb (adductor region where collaterals enlarge) or in total hind limb, is expressed as a ratio of right (ischemic) to left (normal) limb.
  • Spontaneous mobility is scored by monitoring the gait abnormalities, the position of right foot in rest and after manipulation, and the "tail-abduction-reflex.” Mice are scored 0 when one observation is abnormal and 1 when normal. Based on the results of the present invention, it is expected that such models of myocardial and limb ischemia will reveal DAPT-treated mice exhibit increased angiogenesis resulting in increased perfusion and formation of collateral vessels leading to increased healing/decreased tissue damage and increased function of the tissue.
  • mice are conditioned for 9 days to swim in a 31°C controlled swimming pool under non-stressed conditions.
  • baseline exercise time for each mouse is determined using a counter-current swimming pool kept at 31°C; flow at 0.2 m/s. See, for example, Matsumoto, K., et al, 1996 An adjustable-current swimming pool for the evaluation of endurance capacity of mice. J. Appl. Physiol. 81, 1843-1849.
  • maximal endurance exercise i.e., the total swimming period until fatigue, the failure to rise to the surface of the water to breathe within 7 seconds is assessed.
  • the femoral artery is occluded as described above.
  • minipumps are removed under isoflurane anesthesia before endurance exercise.
  • Fluorescent microspheres (yellow-green, 15 ⁇ m, 1 x 10 beads per ml, Molecular Probes, Eugene, Oregon) are administered after maximal vasodilation (sodium nitroprusside, 50 ng/ml, Sigma), processed, and flow was calculated (Carmeliet, P. et al. 1999, Impaired myocardial angiogenesis and ischemic cardiomyopathy in mice lacking the vascular endothelial growth factor isoforms VEGF164 and VEGF188. Nature Med. 5, 495-502). Bismuth gelatino-angiography is performed (see, for example, Carmeliet, P.
  • Collateral side branches are categorized as follows: second-generation collateral arterioles directly branch off from the main collateral, whereas third-generation collateral arterioles are orientated perpendicularly to the second-generation branches. The number of collateral branches per cm length of the primary collateral arteriole is counted. Fluoroangiography is performed with a modified version of a described protocol (Carmeliet, P.
  • Total perfusion area is calculated using the total sum of the side branch luminal areas.
  • Capillary density is determined by immunostaining for thrombomodulin.
  • Wall thickness of fully SMC-covered vessels is morphometrically measured on histological sections, after smooth muscle -x-actin staining. For all treatment groups, six cross- sections (150 ⁇ m apart) are analyzed per main collateral. Only second-generation collateral arterioles larger than 300 ⁇ m 2 are included in the analysis. At least 10 measurements of wall thickness of the second-generation collateral arterioles are obtained. See, generally, Luttun A, et al, Aug 2002, Revascularization of ischemic tissues by PIGF treatment, and inhibition of tumor angiogenesis, arthritis and atherosclerosis by anti-Fltl., Nat Med.

Abstract

L'angiogenèse peut être déclenchée ou augmentée, par l'utilisation d'inhibiteurs de la gamma sécrétase. L'inhibiteur de la gamma sécrétase DAPT peut déclencher et augmenter l'angiogenèse. L'invention concerne des méthodes permettant de déclencher et d'augmenter l'angiogenèse. Ces méthodes sont utilisées pour prévenir et pour traiter une maladie, ainsi que pour générer des modèles de recherche.
PCT/SE2004/001146 2003-07-21 2004-07-21 Composes et methodes pour favoriser l'angiogenese, au moyen d'un inhibiteur de la gamma secretase et en inhibant la voie de la gamma secretase WO2005008250A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP04749182A EP1671129A1 (fr) 2003-07-21 2004-07-21 Composes et methodes pour favoriser l'angiogenese, au moyen d'un inhibiteur de la gamma secretase et en inhibant la voie de la gamma secretase
US11/306,973 US20060264380A1 (en) 2003-07-21 2006-01-18 Compounds and Methods for Promoting Angiogenesis
US12/378,731 US20090209473A1 (en) 2003-07-21 2009-02-19 Componds and methods for pormoting angiogenesis

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US48834503P 2003-07-21 2003-07-21
US60/488,345 2003-07-21
SE0302111A SE0302111D0 (sv) 2003-07-21 2003-07-21 Compounds and methods for inhibiting or promoting angiogenesis field of the invention
SE0302111-0 2003-07-21

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US11/306,973 Continuation-In-Part US20060264380A1 (en) 2003-07-21 2006-01-18 Compounds and Methods for Promoting Angiogenesis

Publications (1)

Publication Number Publication Date
WO2005008250A1 true WO2005008250A1 (fr) 2005-01-27

Family

ID=34082463

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE2004/001146 WO2005008250A1 (fr) 2003-07-21 2004-07-21 Composes et methodes pour favoriser l'angiogenese, au moyen d'un inhibiteur de la gamma secretase et en inhibant la voie de la gamma secretase

Country Status (2)

Country Link
EP (1) EP1671129A1 (fr)
WO (1) WO2005008250A1 (fr)

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1666068A1 (fr) * 2003-09-24 2006-06-07 Santen Pharmaceutical Co., Ltd. Remede pour maladies des yeux accompagnees de blessures du nerf optique
WO2007103114A2 (fr) * 2006-03-07 2007-09-13 The Brigham & Women's Hospital, Inc. Inhibition de notch dans le traitement ou la prévention d'athérosclérose
EP1865001A1 (fr) * 2006-06-08 2007-12-12 Gsf-Forschungszentrum Für Umwelt Und Gesundheit, Gmbh Inhibiteurs spécifiques de protéase et leur utilisation dans la thérapie de cancer
WO2007141029A1 (fr) * 2006-06-08 2007-12-13 Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt (Gmbh) Inhibiteurs spécifiques de protéase et leur utilisation pour le traitement du cancer
WO2008055945A1 (fr) 2006-11-09 2008-05-15 Probiodrug Ag Dérivés 3-hydr0xy-1,5-dihydr0-pyrr0l-2-one utiles en tant qu' inhibiteurs de la glutaminyl-cyclase dans le traitement des ulcères, du cancer et d'autres maladies
WO2008065141A1 (fr) 2006-11-30 2008-06-05 Probiodrug Ag Nouveaux inhibiteurs de glutaminylcyclase
WO2008104580A1 (fr) 2007-03-01 2008-09-04 Probiodrug Ag Nouvelle utilisation d'inhibiteurs de la glutaminyl cyclase
WO2011029920A1 (fr) 2009-09-11 2011-03-17 Probiodrug Ag Dérivés hétérocycliques en tant qu'inhibiteurs de glutaminyle cyclase
WO2011107530A2 (fr) 2010-03-03 2011-09-09 Probiodrug Ag Nouveaux inhibiteurs
WO2011110613A1 (fr) 2010-03-10 2011-09-15 Probiodrug Ag Inhibiteurs hétérocycliques de la glutaminyl cyclase (qc, ec 2.3.2.5)
WO2011131748A2 (fr) 2010-04-21 2011-10-27 Probiodrug Ag Nouveaux inhibiteurs
WO2012123563A1 (fr) 2011-03-16 2012-09-20 Probiodrug Ag Dérivés de benzimidazole en tant qu'inhibiteurs de la glutaminyl cyclase
US8343923B2 (en) * 2007-11-09 2013-01-01 Washington University Use of notch signaling regulators for modulating osteogenesis
KR101330184B1 (ko) 2010-10-15 2013-11-15 성균관대학교산학협력단 감마-세크레타제 저해제를 유효성분으로 함유하는 류마티스성 관절염의 예방 또는 치료용 조성물
US8859628B2 (en) 2003-02-27 2014-10-14 JoAnne McLaurin Method for preventing, treating and diagnosing disorders of protein aggregation
US8889131B2 (en) 2009-11-01 2014-11-18 The Brigham And Women's Hospital, Inc. NOTCH inhibition in the treatment and prevention of a metabolic disease or disorder and cardiovascular complications thereof
EP2865670A1 (fr) 2007-04-18 2015-04-29 Probiodrug AG Dérivés de thio-urée utilisés comme inhibiteurs de la glutaminyl cyclase
US9567396B2 (en) 2006-03-07 2017-02-14 Evonik Degussa Gmbh Notch inhibition in the prevention of vein graft failure
US9803013B2 (en) 2005-12-12 2017-10-31 Bioinvent International Ab Biological materials and uses thereof
EP3461819A1 (fr) 2017-09-29 2019-04-03 Probiodrug AG Inhibiteurs de la glutaminyl-cyclase

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002018544A2 (fr) * 2000-08-31 2002-03-07 Loyola University Chicago Procedes et reactifs permettant la formation d'une barriere epitheliale et traitement d'affections cutanees malignes et benignes par modulation de la voie notch

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040229816A1 (en) * 2003-02-18 2004-11-18 Daniel Paris Anti-angiogenic and anti-tumoral properties of beta and gamma secretase inhibitors

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002018544A2 (fr) * 2000-08-31 2002-03-07 Loyola University Chicago Procedes et reactifs permettant la formation d'une barriere epitheliale et traitement d'affections cutanees malignes et benignes par modulation de la voie notch

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
BLOOD, vol. 100, no. 11, 16 November 2002 (2002-11-16), PG-ABSTRACTS NO. 594 *
DATABASE BIOSIS [online] FRANZISKA JUNDT ET AL.: "Novel gamma-Secretase inhibitor DAPT blocks activated Notch signaling and controls tumor cell growth in Hodgkin and anaplastic large cell lymphoma", XP002903868, Database accession no. PREV200300356701 *
JO D.-G. ET AL.: "Contribution of presenilin/gamma-secretase to calsenilin-mediated apoptosis", BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, vol. 305, 2003, pages 62 - 66, XP001155084 *
LUTGENS E ET AL.: "Chronic myocardial infarction in the mouse: cardiac structural and functional changes", CARDIOVASC RES., vol. 41, no. 3, March 1999 (1999-03-01), pages 586 - 93
LUTTUN A ET AL.: "Revascularization of ischemic tissues by PIGF treatment, and inhibition of tumor angiogenesis, arthritis and atherosclerosis by anti-Fltl.", NAT MED., vol. 8, no. 8, August 2002 (2002-08-01), pages 831 - 40
MANN D.M.A. ET AL.: "Amyloid angiopathy and variability in amyloid Beta deposition is determined by mutation position in presenilin-1-linked Alzheimer's disease", AMERICAN JOURNAL OF PATHOLOGY, vol. 158, no. 6, June 2001 (2001-06-01), pages 2165 - 2175, XP002903866 *
MICCHELLI C.A. ET AL.: "Gamma-Secretase/presenilin inhibitors for Alzheimer's disease phenocopy Notch mutations in Drosophila", THE FASEB JOURNAL, vol. 17, January 2003 (2003-01-01), pages 79 - 81, XP002903867 *
NAKAJIMA M. ET AL.: "Abnormal blood vessel development in mice lacking presenilin-1", MECHANISMS OF DEVELOPMENT, vol. 120, 2003, pages 657 - 667, XP002903865 *
VAGNUCCI A.H. ET AL.: "Alzheimer's disease and angiogenesis", THE LANCET, vol. 361, 15 February 2003 (2003-02-15), pages 605 - 608, XP004410316 *

Cited By (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8859628B2 (en) 2003-02-27 2014-10-14 JoAnne McLaurin Method for preventing, treating and diagnosing disorders of protein aggregation
US9833420B2 (en) 2003-02-27 2017-12-05 JoAnne McLaurin Methods of preventing, treating, and diagnosing disorders of protein aggregation
EP1666068A1 (fr) * 2003-09-24 2006-06-07 Santen Pharmaceutical Co., Ltd. Remede pour maladies des yeux accompagnees de blessures du nerf optique
EP1666068A4 (fr) * 2003-09-24 2010-08-04 Santen Pharmaceutical Co Ltd Remede pour maladies des yeux accompagnees de blessures du nerf optique
US9803013B2 (en) 2005-12-12 2017-10-31 Bioinvent International Ab Biological materials and uses thereof
WO2007103114A3 (fr) * 2006-03-07 2007-11-29 Brigham & Womens Hospital Inhibition de notch dans le traitement ou la prévention d'athérosclérose
US8133857B2 (en) 2006-03-07 2012-03-13 The Brigham and Women's FHospital, Inc. NOTCH inhibition in the treatment of atherosclerosis
US9567396B2 (en) 2006-03-07 2017-02-14 Evonik Degussa Gmbh Notch inhibition in the prevention of vein graft failure
US9289489B2 (en) 2006-03-07 2016-03-22 The Brigham And Women's Hospital, Inc. NOTCH inhibition in the treatment of cardiovascular disease
WO2007103114A2 (fr) * 2006-03-07 2007-09-13 The Brigham & Women's Hospital, Inc. Inhibition de notch dans le traitement ou la prévention d'athérosclérose
WO2007141029A1 (fr) * 2006-06-08 2007-12-13 Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt (Gmbh) Inhibiteurs spécifiques de protéase et leur utilisation pour le traitement du cancer
EP1865001A1 (fr) * 2006-06-08 2007-12-12 Gsf-Forschungszentrum Für Umwelt Und Gesundheit, Gmbh Inhibiteurs spécifiques de protéase et leur utilisation dans la thérapie de cancer
WO2008055945A1 (fr) 2006-11-09 2008-05-15 Probiodrug Ag Dérivés 3-hydr0xy-1,5-dihydr0-pyrr0l-2-one utiles en tant qu' inhibiteurs de la glutaminyl-cyclase dans le traitement des ulcères, du cancer et d'autres maladies
WO2008065141A1 (fr) 2006-11-30 2008-06-05 Probiodrug Ag Nouveaux inhibiteurs de glutaminylcyclase
WO2008104580A1 (fr) 2007-03-01 2008-09-04 Probiodrug Ag Nouvelle utilisation d'inhibiteurs de la glutaminyl cyclase
EP2481408A2 (fr) 2007-03-01 2012-08-01 Probiodrug AG Nouvelle utilisation d'inhibiteurs glutaminyle cyclase
EP2865670A1 (fr) 2007-04-18 2015-04-29 Probiodrug AG Dérivés de thio-urée utilisés comme inhibiteurs de la glutaminyl cyclase
US8343923B2 (en) * 2007-11-09 2013-01-01 Washington University Use of notch signaling regulators for modulating osteogenesis
WO2011029920A1 (fr) 2009-09-11 2011-03-17 Probiodrug Ag Dérivés hétérocycliques en tant qu'inhibiteurs de glutaminyle cyclase
US8889131B2 (en) 2009-11-01 2014-11-18 The Brigham And Women's Hospital, Inc. NOTCH inhibition in the treatment and prevention of a metabolic disease or disorder and cardiovascular complications thereof
US10988530B2 (en) 2009-11-01 2021-04-27 The Brigham And Women's Hospital, Inc. Notch inhibition in the treatment and prevention of nonalcoholic fatty liver disease
WO2011107530A2 (fr) 2010-03-03 2011-09-09 Probiodrug Ag Nouveaux inhibiteurs
WO2011110613A1 (fr) 2010-03-10 2011-09-15 Probiodrug Ag Inhibiteurs hétérocycliques de la glutaminyl cyclase (qc, ec 2.3.2.5)
WO2011131748A2 (fr) 2010-04-21 2011-10-27 Probiodrug Ag Nouveaux inhibiteurs
KR101330184B1 (ko) 2010-10-15 2013-11-15 성균관대학교산학협력단 감마-세크레타제 저해제를 유효성분으로 함유하는 류마티스성 관절염의 예방 또는 치료용 조성물
WO2012123563A1 (fr) 2011-03-16 2012-09-20 Probiodrug Ag Dérivés de benzimidazole en tant qu'inhibiteurs de la glutaminyl cyclase
EP3461819A1 (fr) 2017-09-29 2019-04-03 Probiodrug AG Inhibiteurs de la glutaminyl-cyclase

Also Published As

Publication number Publication date
EP1671129A1 (fr) 2006-06-21

Similar Documents

Publication Publication Date Title
US20060264380A1 (en) Compounds and Methods for Promoting Angiogenesis
WO2005008250A1 (fr) Composes et methodes pour favoriser l'angiogenese, au moyen d'un inhibiteur de la gamma secretase et en inhibant la voie de la gamma secretase
Cai et al. FUNDC1-dependent mitophagy induced by tPA protects neurons against cerebral ischemia-reperfusion injury
Monnier et al. Involvement of caspase-6 and caspase-8 in neuronal apoptosis and the regenerative failure of injured retinal ganglion cells
Haddadin et al. Thrombospondin-1 (TSP1)–null and TSP2-null mice exhibit lower intraocular pressures
CA2458798A1 (fr) Methode permettant de reduire l'hypertension et l'insuffisance cardiaque
JP2013529181A (ja) 神経障害を治療するための組成物および方法
Qu et al. Stimulation of sigma-1 receptor protects against cardiac fibrosis by alleviating IRE1 pathway and autophagy impairment
Hu et al. Deficiency of ROS-activated TRPM2 channel protects neurons from cerebral ischemia-reperfusion injury through upregulating autophagy
Hao et al. Soluble epoxide hydrolase inhibition alleviated cognitive impairments via NRG1/ErbB4 signaling after chronic cerebral hypoperfusion induced by bilateral carotid artery stenosis in mice
KR101133289B1 (ko) Ａｋａｐ１２를 함유하는 조성물 및 동물 모델로서ａｋａｐ１２ 돌연변이 제브라피쉬의 용도
TWI814760B (zh) 脂質運載蛋白型前列腺素ｄ２合成酵素產生促進劑
KR101174333B1 (ko) 심근 경색증의 치료 방법
US10751324B2 (en) Treatment of TNF- alpha cytotoxicity
EP3139944B1 (fr) Peptide dérivé de dsg2 pour utilisation dans le traitement des maladies angiogéniques oculaires
WO2019129179A1 (fr) Utilisation d'un composé dans la préparation d'un médicament pour le traitement de la maladie des petits vaisseaux cérébraux
Fan et al. Celastrol relieves myocardial infarction-induced cardiac fibrosis by inhibiting NLRP3 inflammasomes in rats
WO2007094193A1 (fr) Agent anti-angiogenique, agent prophylactique ou therapeutique pour une maladie accompagnee d'une angiogenese et aliment
Pereira Focusing on metabolomic dysregulation and modulation of retinal metabolism to develop novel therapeutic strategies for diabetic retinopathy
TWI759739B (zh) 新穎性醫藥組合物於修復視網膜損傷與治療視網膜病變之用途
Sun et al. Juglanin alleviates myocardial injury in rats with acute myocardial infarction through modulating MAPK signaling pathway
CN116115619B (zh) 利培酮在治疗器官纤维化中的应用
Snodgrass Investigating the roles of endoglin in cardiovascular development, disease and repair
JP2006502142A (ja) 虚血性損傷を減少させる方法
Geist Canonical and noncanonical action of thyroid hormone receptor α in the cardiovascular system

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DPEN Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed from 20040101)
WWE Wipo information: entry into national phase

Ref document number: 11306973

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: 2004749182

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 2004749182

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 11306973

Country of ref document: US