EP2004210A2 - Antiangiogenic peptide and therapeutic uses thereof - Google Patents

Antiangiogenic peptide and therapeutic uses thereof

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Publication number
EP2004210A2
EP2004210A2 EP07734153A EP07734153A EP2004210A2 EP 2004210 A2 EP2004210 A2 EP 2004210A2 EP 07734153 A EP07734153 A EP 07734153A EP 07734153 A EP07734153 A EP 07734153A EP 2004210 A2 EP2004210 A2 EP 2004210A2
Authority
EP
European Patent Office
Prior art keywords
peptide
hnr
peptide according
antiangiogenic
angiostatin
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP07734153A
Other languages
German (de)
French (fr)
Inventor
Claudio Brigati
Raffaella Dell'eva
Erika Nieddu
Roberto Benelli
Douglas M. Noonan
Adriana Albini
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Istituto Nazionale per La Ricerce Sul Cancro
Istituto Nazionale per la Ricerca Sul Cancro
Original Assignee
Istituto Nazionale per La Ricerce Sul Cancro
Istituto Nazionale per la Ricerca Sul Cancro
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Application filed by Istituto Nazionale per La Ricerce Sul Cancro, Istituto Nazionale per la Ricerca Sul Cancro filed Critical Istituto Nazionale per La Ricerce Sul Cancro
Publication of EP2004210A2 publication Critical patent/EP2004210A2/en
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/06—Antiglaucoma agents or miotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0806—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6435—Plasmin (3.4.21.7), i.e. fibrinolysin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/21—Serine endopeptidases (3.4.21)
    • C12Y304/21007—Plasmin (3.4.21.7), i.e. fibrinolysin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to an angiostatin peptide fragment having antichemotactic, antiangiogenic and antitumor activities, useful for the preparation of medicaments for the treatment of inflammatory, connective degenerative, vascular and neoplastic conditions.
  • Angiostatin is an antiangiogenic molecule produced in the body under specific conditions, such as solid tumors, and it represents a homeostatic response to the underlying pathologic condition. It is produced as a fragment of plasminogen, and induces apoptosis in endothelial cells and on other cellular targets (1), blocking vessel neoformation and growth in both primitive and metastatic tumors. In angiostatin-treated animals, neither toxic effects nor development of resistance have been observed.
  • a fragment has been isolated from angiostatin molecule's Kringle 3, which is able to reproduce inhibition of neutrophils chemotaxis, induction of apoptosis in the endothelium and angiogenesis inhibition, i.e. the effects typical of angiostatin itself.
  • Fig. 1-a illustrates the antichemotactic activity of HNR in human polymorphonucleate (PMN) -enriched fractions.
  • PMN human polymorphonucleate
  • X the peptide activity is very high compared to both angiostatin and IL-8 (p ⁇ 0.004; Mann Whitney).
  • EC endothelial HUVEC
  • mononuclear cells the peptide did not show significant activity.
  • the antichemotactic action of HNR is therefore quite selective for PMN.
  • AST is known to induce endothelial apoptosis through inactivation of ATP synthase.
  • the pro-apoptotic action of HNR on HUVEC was therefore tested.
  • Fig 1 - shows a significant increase in free nucleosomes 8 hrs after treatment with 2.5 ⁇ g/ml of peptide HNR.
  • PMN or PBMC no significant results were obtained when using either PMN or PBMC. Therefore, the induction of apoptosis by HNR is selective for endothelial cells.
  • Fig. 2-a Angiogenesis induced in mice is highly inhibited by HNR (p ⁇ 0.02), although to a slightly lower extent than angiostatin.
  • HNR histological control of the sponges was also carried out after completion of the experiment, to verify the cellularity, the extension of vessel formation and their morphology (Fig. 2-b).
  • KS Kaposi sarcoma
  • the peptide HNR is preferably combined with pharmaceutically acceptable vehicles and/or carriers.
  • the invention concerns pharmaceutical compositions containing the peptide HNR, suitable for the oral or parenteral administration, particularly through the subcutaneous, intravenous, intramuscular, rectal, ophthalmic, nasal, topical, intrauterine or vaginal routes.
  • the compositions can be solid, e.g. capsules, tablets or granules; liquid, e.g. solutions, suspensions, syrups, drops, tinctures, sprays or aerosols; semi-solid, e.g. creams, ointments or gels.
  • the dosage of the peptide will depend on the severity of the disease or disorder to be treated, as well as on factors such as age and weight of the subject or administration route. In principle, for the treatment of humans or animals, an amount of peptide ranging from 0.01 to 250 mg/Kg, preferably from 0.1 to 100 mg/Kg, will be used.
  • the peptide of the invention and its pharmaceutical compositions can be used for the preventive or therapeutic treatment of chronic inflammation, diabetes mellitus, connective tissue degeneration, including rheumatoid arthritis and atherosclerosis, primitive vascular conditions, particularly eye disorders such as macular degeneration, premature retinopathy, corneal rejection, vascular glaucoma, retrolental fibroplasia, rubeosis, cardiovascular and cerebrovascular conditions, tumors, particularly solid tumors and leukemias.
  • the peptide of the invention is more stable, is highly bioavailable, reaching all target tissues, it is easy to produce in high purity degrees (absence of bacterial contaminants) and has low-cost.
  • the HNR peptide can be synthesized according to established procedures (Stuart and Young, 1984, Solid Phase Peptide Synthesis, 2 nd ed., Pierce Chemical Co.; Tarn et al., Am Soc, 1983 105: 6442; Merrifield, 1979, The Peptides, Gross and Meihofer eds NY Academic Press, 1-284), in solution, solid phase or using an automated synthesizer.
  • the peptide may be produced by recombinant DNA techniques (Sambrook et al., Molecular Cloning, a laboratory manual, CSH Press, CSH, NY, 1982 or Ausbel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Inc. NY, 1987).
  • amino acid residues within the HNR sequence may be in D or L configuration, or they can be chemically modified, for instance by amidation of the carboxy-terminus, linkage to lipophilic groups (i.e. fatty acid residues) glycosylation or conjugation to other molecules, so as to improve the peptide activity profile or bioavailability.
  • Fig. 1-a Chemotaxis on polymorphonucleates
  • Migrated PMN cells were counted (y-axis) and the count was confirmed by densitometry (not shown).
  • SFM serum free medium
  • IL-8 Interleukin 8 used as the chemotactic.
  • Fig. 1-b HUVEC-Induced apoptosis
  • HNR 2.5 and 5 represent HNR concentrations at 2.5 and 5 ⁇ g/ml.
  • Fig. 2-a In vivo angiogenesis
  • FIG. 3 shows sections stained with hematoxylin-eosin; lack of vessels and poor cell infiltrate in the HNR sample compared with chemoattractant VTH alone (VTH CTRL) can be observed.
  • Fig. 3 Tumor growth inhibition
  • CTRL-peptide is a control peptide with sequence AlaSerAla.
  • PBS PBS+0,1% BSA, used as the vehicle.
  • CDl nude mice were split in three groups: one was injected peritoneally with 50 ⁇ g HNR peptide/mouse (Calbiochem, La JoIIa 5 California) in 100 ⁇ l PBS-BSA, the control group was injected with vehicle, a third group with 50 ⁇ g of random sequence peptide.
  • AST was injected on alternate days, HNR every four days. On day zero, 5 millions KS IMM cells were injected. Mice were sacrificed at day 31.
  • the caliper-measured volume was calculated by means of the formula ⁇ /6 x length x width.
  • Matrigel sponge assay The matrigel sponge angiogenesis assay in C57bl mice was performed as described (3) using VTH or KS cells supernatant as the chemotactic.
  • Control peptide ("random” in Fig. 2-a): AlaSerAla. Histology of matrigel sponges
  • pellets were fixed in 4% PAF and paraffin-embedded; 4 ⁇ M sections were stained with hematoxylin-eosin standard procedures.
  • Chemotaxis - apoptosis This assay is conducted in 48 wells chambers as described in (4).
  • the apoptosis assay was conducted with a Cell Death Detection Kit (Roche) according to the producer's instructions.

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Abstract

An angiostatin peptide fragment having antichemotactic, antiangiogenic and antitumor activities, pharmaceutical compositions containing it and their uses for the treatment of inflammatory, connective degenerative, vascular and neoplastic conditions.

Description

ANTIANGIOGENIC PEPTIDE AND THERAPEUTIC USES THEREOF
The present invention relates to an angiostatin peptide fragment having antichemotactic, antiangiogenic and antitumor activities, useful for the preparation of medicaments for the treatment of inflammatory, connective degenerative, vascular and neoplastic conditions. BACKGROUND OF THE INVENTION
Angiostatin (AST) is an antiangiogenic molecule produced in the body under specific conditions, such as solid tumors, and it represents a homeostatic response to the underlying pathologic condition. It is produced as a fragment of plasminogen, and induces apoptosis in endothelial cells and on other cellular targets (1), blocking vessel neoformation and growth in both primitive and metastatic tumors. In angiostatin-treated animals, neither toxic effects nor development of resistance have been observed.
A feature of the molecule is the presence of functional domains named
Kringles. The properties of angiostatin and its Kringles have been described in a number of publications and patent applications, inter alia You W-K et al.,
Protein Expression and Purification, 36 (2004); US2004/002459; US5837682;
Bouma-Ter Stegee J C A et al., Critical Reviews in Eukaryotic Gene
Expression, 11 (2001). Other patents disclose angiostatin sequence variants and methods to isolate, purify and synthetically produce it, see e.g. US patents No. 5861372; 5639725; 5792845; 5885795; 5854205; 5854221; 6024688.
DESCRIPTION OF THE INVENTION
A fragment has been isolated from angiostatin molecule's Kringle 3, which is able to reproduce inhibition of neutrophils chemotaxis, induction of apoptosis in the endothelium and angiogenesis inhibition, i.e. the effects typical of angiostatin itself.
The peptide subject of this invention, having HNR sequence (HisAsnArg), has been investigated in an in vitro chemotaxis assay, in an angiogenesis assay in vivo utilizing matrigel sponges and in an animal tumor model. The procedures used are described in the section "Materials and Methods". Fig. 1-a illustrates the antichemotactic activity of HNR in human polymorphonucleate (PMN) -enriched fractions. As it can be observed in the graph (column "X"), the peptide activity is very high compared to both angiostatin and IL-8 (p<0.004; Mann Whitney). Conversely, when the peptide was assayed on endothelial HUVEC (EC) and mononuclear cells, the peptide did not show significant activity. The antichemotactic action of HNR is therefore quite selective for PMN.
AST is known to induce endothelial apoptosis through inactivation of ATP synthase. The pro-apoptotic action of HNR on HUVEC was therefore tested. Fig 1 -shows a significant increase in free nucleosomes 8 hrs after treatment with 2.5 μg/ml of peptide HNR. On the other hand, no significant results were obtained when using either PMN or PBMC. Therefore, the induction of apoptosis by HNR is selective for endothelial cells.
The antiangiogenic activity is shown in Fig. 2-a. Angiogenesis induced in mice is highly inhibited by HNR (p<0.02), although to a slightly lower extent than angiostatin. A histological control of the sponges was also carried out after completion of the experiment, to verify the cellularity, the extension of vessel formation and their morphology (Fig. 2-b). Cellularity, sample vascularization and the results obtained by measurement of hemoglobin in matrigel sponges were perfectly matching (Fig. 2-b). Finally, the ability of the HNR peptide to counteract tumor growth in a
Kaposi sarcoma (KS) cell-graft model in nude mice was investigated. It was observed that the intraperitoneal administration of the peptide every four day after cells inoculation slowed down tumor growth compared with the medium alone (ρ<0.02, Mann Whitney, Fig. 3, curve HNR). The tumor weight at the end of the experiments confirmed the result.
For use in therapy, the peptide HNR is preferably combined with pharmaceutically acceptable vehicles and/or carriers. In a further aspect, the invention concerns pharmaceutical compositions containing the peptide HNR, suitable for the oral or parenteral administration, particularly through the subcutaneous, intravenous, intramuscular, rectal, ophthalmic, nasal, topical, intrauterine or vaginal routes. The compositions can be solid, e.g. capsules, tablets or granules; liquid, e.g. solutions, suspensions, syrups, drops, tinctures, sprays or aerosols; semi-solid, e.g. creams, ointments or gels. The dosage of the peptide will depend on the severity of the disease or disorder to be treated, as well as on factors such as age and weight of the subject or administration route. In principle, for the treatment of humans or animals, an amount of peptide ranging from 0.01 to 250 mg/Kg, preferably from 0.1 to 100 mg/Kg, will be used. The peptide of the invention and its pharmaceutical compositions can be used for the preventive or therapeutic treatment of chronic inflammation, diabetes mellitus, connective tissue degeneration, including rheumatoid arthritis and atherosclerosis, primitive vascular conditions, particularly eye disorders such as macular degeneration, premature retinopathy, corneal rejection, vascular glaucoma, retrolental fibroplasia, rubeosis, cardiovascular and cerebrovascular conditions, tumors, particularly solid tumors and leukemias.
Compared with angiostatin, the peptide of the invention is more stable, is highly bioavailable, reaching all target tissues, it is easy to produce in high purity degrees (absence of bacterial contaminants) and has low-cost.
The HNR peptide can be synthesized according to established procedures (Stuart and Young, 1984, Solid Phase Peptide Synthesis, 2nd ed., Pierce Chemical Co.; Tarn et al., Am Soc, 1983 105: 6442; Merrifield, 1979, The Peptides, Gross and Meihofer eds NY Academic Press, 1-284), in solution, solid phase or using an automated synthesizer. Alternatively, the peptide may be produced by recombinant DNA techniques (Sambrook et al., Molecular Cloning, a laboratory manual, CSH Press, CSH, NY, 1982 or Ausbel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Inc. NY, 1987). The amino acid residues within the HNR sequence may be in D or L configuration, or they can be chemically modified, for instance by amidation of the carboxy-terminus, linkage to lipophilic groups (i.e. fatty acid residues) glycosylation or conjugation to other molecules, so as to improve the peptide activity profile or bioavailability.
Description of the Figures
Fig. 1-a. Chemotaxis on polymorphonucleates
Migrated PMN cells were counted (y-axis) and the count was confirmed by densitometry (not shown). SFM = serum free medium, IL-8 = Interleukin 8 used as the chemotactic.
Fig. 1-b. HUVEC-Induced apoptosis
After 8 hrs treatment, free nucleosomes were examined. AST= Angiostatin, Vinc= Vincristin 25 mM (positive control). CTL= untreated cells (blank). HNR 2.5 and 5 represent HNR concentrations at 2.5 and 5 μg/ml.
Fig. 2-a. In vivo angiogenesis
The hemoglobin contents in the matrigel pellets is reported in abscissa, as the vascularization index. VTH= Vegf, TNF, Heparin used as proangiogenetic agents. AST= Angiostatin, X = HNR Fig. 2-b. Matrigel sponge histological analysis
The Figure shows sections stained with hematoxylin-eosin; lack of vessels and poor cell infiltrate in the HNR sample compared with chemoattractant VTH alone (VTH CTRL) can be observed. Fig. 3. Tumor growth inhibition
Experiments were stopped at day 25; the CTRL-peptide is a control peptide with sequence AlaSerAla. PBS= PBS+0,1% BSA, used as the vehicle.
Materials methods Tumor Inhibition
CDl nude mice were split in three groups: one was injected peritoneally with 50 μg HNR peptide/mouse (Calbiochem, La JoIIa5 California) in 100 μl PBS-BSA, the control group was injected with vehicle, a third group with 50 μg of random sequence peptide. AST was injected on alternate days, HNR every four days. On day zero, 5 millions KS IMM cells were injected. Mice were sacrificed at day 31.
The caliper-measured volume was calculated by means of the formula π/6 x length x width.
Matrigel sponge assay The matrigel sponge angiogenesis assay in C57bl mice was performed as described (3) using VTH or KS cells supernatant as the chemotactic.
Peptide sequences
Peptide: HNR.
Control peptide ("random" in Fig. 2-a): AlaSerAla. Histology of matrigel sponges
After removal from the animals, pellets were fixed in 4% PAF and paraffin-embedded; 4 μM sections were stained with hematoxylin-eosin standard procedures.
Chemotaxis - apoptosis This assay is conducted in 48 wells chambers as described in (4).
The apoptosis assay was conducted with a Cell Death Detection Kit (Roche) according to the producer's instructions. Bibliography
1. Benelli, R., Morini, M., Carrozzino, F., Ferrari, N., Minghelli, S., Santi, L., Cassatella, M., Noonan, D., and Albini, A. Neutrophils as a key cellular target for angiostatin: implications for regulation of angiogenesis and inflammation. FASEB J5 16: 267-269, 2002.
2. Schnurr, M., Then, F., Galambos, P., Scholz, C, Siegmund, B., Endres, S., and Eigler, A. EHNRtracellular ATP and TNF-alpha synergize in the activation and maturation of human dendritic cells. J Immunol, 165: 4704-4709, 2000.
3. Albini, A., Fontanini, G., Masiello, L., Tacchetti C, B., D, Luzzi, P., Noonan, D. M., and Stetler-Stevenson, W.G. Angiogenic potential in vivo by Kaposi's sarcoma cell-free supernatants and HIV-I tat product: inhibition of KS-like lesions by tissue inhibitor of metalloproteinase-2., 8: 1237-1244, 1994.
4. FaIk, W., Goodwin, R.H., Jr., and Leonard, E.J.A 48-well micro chemotaHNRis assembly for rapid and accurate measurement of leukocyte migration. J Immunol Methods, 33: 239-247, 1980.
5. Albini A, Benelli R, Noonan DM, Brigati C. The "chemoinvasion assay": a tool to study tumor and endothelial cell invasion of basement membranes. Int J Dev Biol. 2004;48(5-6):563-71.

Claims

1. Isolated antiangiogenic peptide having sequence HisAsnArg (HNR).
2. The peptide according to claim 1, in which the amino acid residues have D- or L- configuration, or they are chemically modified by amidation of the carboxy-terminus, linkage to lipophylic groups, particularly fatty acid residues, or glycosylation.
3. The antiangiogenic peptide according to claim 1 or 2, for use as a medicament.
4. Pharmaceutical composition containing a peptide according to claims 1-2.
5. The use of the peptide according to claims 1-2 or of a pharmaceutical composition according to claim 4, for the preparation of a medicament for the preventive or therapeutic treatment of: chronic inflammation, diabetes mellitus, connective tissue degenerations, including rheumatoid arthritis and atherosclerosis, primitive vascular conditions, particularly eye disorders such as macular degeneration, premature retinopathy, corneal rejection, vascular glaucoma, retrolental fibroplasia, rubeosis, cardiovascular and cerebrovascular conditions, tumors, particularly solid tumors and leukemias.
EP07734153A 2006-03-31 2007-03-30 Antiangiogenic peptide and therapeutic uses thereof Withdrawn EP2004210A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IT000625A ITMI20060625A1 (en) 2006-03-31 2006-03-31 ANTIANGIOGENIC PEPTIDE AND ITS THERAPEUTIC USES
PCT/IB2007/000831 WO2007116278A2 (en) 2006-03-31 2007-03-30 Antiangiogenic peptide and therapeutic uses thereof

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EP2004210A2 true EP2004210A2 (en) 2008-12-24

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Publication number Priority date Publication date Assignee Title
EP3643318A1 (en) * 2013-06-07 2020-04-29 The Johns Hopkins University A biomimetic peptide and biodegradable delivery platform for the treatment of angiogenesis- and lymphangiogenesis-dependent diseases

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Publication number Priority date Publication date Assignee Title
SG87828A1 (en) * 1999-09-03 2002-04-16 Univ Singapore Small peptides having potent anti-angiogenic activity
ITMI20041962A1 (en) * 2004-10-15 2005-01-15 Istituto Naz Per La Ricerca S "PEPTIDE OF ANGIOSTATIN AND ITS THERAPEUTIC EMPLOYEES"

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