EP1680136A1 - Peptides having, for example, an antiangiogenic activity and applications thereof in therapeutics - Google Patents

Peptides having, for example, an antiangiogenic activity and applications thereof in therapeutics

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Publication number
EP1680136A1
EP1680136A1 EP04787446A EP04787446A EP1680136A1 EP 1680136 A1 EP1680136 A1 EP 1680136A1 EP 04787446 A EP04787446 A EP 04787446A EP 04787446 A EP04787446 A EP 04787446A EP 1680136 A1 EP1680136 A1 EP 1680136A1
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EP
European Patent Office
Prior art keywords
seq
motif
ggxrgdmfgx
peptides
crgdmfg
Prior art date
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Application number
EP04787446A
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German (de)
French (fr)
Inventor
Etienne Jacotot
Annie Borgne-Sanchez
Sylvie Dupont
Dominique Rebouillat
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Theraptosis SA
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Theraptosis SA
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Publication of EP1680136A1 publication Critical patent/EP1680136A1/en
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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/78Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00Drugs for skeletal disorders
    • A61P19/02Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00Drugs for disorders of the cardiovascular system
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00Drugs for disorders of the cardiovascular system
    • A61P9/10Drugs 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
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/001Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides

Definitions

  • the invention relates to peptides having in particular an anti-angiogenic activity and to their applications in therapy.
  • the inventors' research into therapeutically active peptides led them to develop constructs which have proved to be of great interest with regard to their anti-angiogenic properties.
  • the invention therefore relates to such peptides and the use of their therapeutic properties to develop drugs. It thus relates to pharmaceutical compositions containing these peptides as active principle. It also relates to the use of these peptides to manufacture drugs with anti-angiogenic effect, for the treatment of pathologies linked to hypervascularization.
  • the peptides according to the invention are characterized in that they are cyclized peptides, corresponding to the sequence SEQ ID No.
  • Xi is either a G or a GG whose amino-terminal end is free, alkylated, acylated, in particular acetylated, or comprises a labeling group such as the biotinyl group
  • X 3 is either a motif M or a nor-Leucine motif
  • X is either a motif or a succession of two di-, tri- or tetra-peptide motifs composed of G or of a combination of G and S, such as GG, GGG, GGGG, GGS, GGGS, GGSGGS, or even
  • X 5 is a C motif whose side chain (thiol function) serves as a covalent bonding point with a 3-nitro-2-pyridinesulfenyl group (Npys; Drijfhout et al ., 1988 Int J Peptide protein Res, 32: 161-166) located on the N-terminal end of the following amino acid (L),
  • X 6 is either an R motif or a K motif - X 7 is either an R motif, either a KX 8 motif is either an R motif, or a K Xg motif is an aliphatic amino acid (such as G, or A) whose C-terminal end is amid
  • peptides contain from 25 to 35 amino acids.
  • a peptide of this type responds to the sequence SEQ ID No. 2: GG * CRGDMFG * CGGLLFIHFRIGSRHSRIG ( ⁇ indicates a disulfide bridge connecting the two motifs C).
  • Other peptides are as defined above and have an alkyl group at their N-terminal end.
  • one or more amino acids are replaced by their dextrorotatory form ( D aa).
  • Other peptides according to the invention correspond to SEQ ID No. 1 above, but have one or more peptide bonds to form bioisosteres.
  • peptides of the invention include a sequence SEQ ID No. 11: XRGDMF-GX 'exposing the RGD motif by a lactam bridge between the amino acids X (X) -CO-NH- (X'), X and X 'being amino acids such that one carries an acid group, and the other carries an amino.
  • Preferred peptides of this group correspond to the sequences SEQ ID No. 12 to SEQ ID No. 23:
  • the peptides of the invention are further characterized in that they induce apoptosis of human endothelial cells expressing the ⁇ V ⁇ 3 receptors.
  • the specificity of the peptides of the invention results from the addition of the mitochondrio-toxic part to ligands of the integrins to exert a toxicity by the way of the mitochondrio-toxicity, the ligands of the integrins being present for purposes of targeting and not themselves having angiostatic activity.
  • the treatment of primary human endothelial cells with doses of peptides of the order of a micromolar leads to a dissipation of the mitochondrial transmembrane potential ( ⁇ m), to the release of mitochondrial cytochrome c, to l 'exposure of phosphatidyl-serine and condensation of nuclear chromatin.
  • compositions according to the invention are characterized in that they contain a therapeutically effective amount of at least one peptide, as defined above, in combination with a phar aceutically acceptable vehicle.
  • These compositions are advantageously in the dosage forms suitable for their administration by injectable route. Mention will in particular be made of injectable solutions intended for administration by the intravenous route.
  • the invention further relates to the use of peptide constructs as defined above for manufacturing anti-angiogenic drugs for the treatment of pathologies due to hypervascularization. Mention will in particular be made of the treatment of solid tumors such as pulmonary tumors, adenomas, melanomas, cancers of the prostate, breast, colon, pancreas, osteosarcomas.
  • the invention also applies to the treatment of diabetic retinopathies and polyarthritis.
  • the dosages of the administration forms and the treatments will be determined by a person skilled in the art depending on the pathology to be treated and the condition of the patient. Other characteristics and advantages of the invention will be given in the examples which follow relating to the construction SEQ ID No. 2 (hereafter TEAM-VP) SEQ ID No.
  • FIGS. 1 to 3 represent, respectively, FIG. 1, the analysis of cytotoxicity on endothelial cells, FIG. 2, the recognition of the CycRGD motif by the integrins ⁇ V ⁇ 3, FIG. 3, the effects of TEAM-VP on isolated itochondria and HUVEC cells.
  • the HUVEC cells were pre-incubated or not for 30 min at room temperature with 25 ⁇ M of CycRGD, CycRAD, GRGDS and GRGES peptide before addition of FITC-CycRGD peptide (0.5 ⁇ M), then analyzed by flow cytometry (Figure 2c).
  • d Correlation between expression of integrins, binding and toxicity of peptides: The cells HUVEC, HMVECd, MCF-7, MDA, HeLa, HT-29, Jurkat, CEM and PBMC were labeled with the antibodies directed against the integrins ⁇ V ⁇ 3 and ⁇ V ⁇ 5 and analyzed by flow cytometry. The binding of the CycRGD peptide and the induction of apoptosis by TEAM-VP on the different cell types were measured (Figure 2d).
  • the FITC-CycRGD and TEAM-VP (FITC) peptides enter the HUVECs and co-locate with the dextran beads (5 hours of co-treatment).
  • the entry of TEAM-VP and dextran beads is inhibited by the treatment of sodium azide + deoxyglucose, indicating entry of the peptide by endocytosis.
  • No entry of the FITC-CycRAD peptide was observed in the HUVECs, nor was there any entry of the FITC-CycRGD peptide in the HeLa. + TEAM-VP intracellular routing:
  • HUVECs treated with TEAM-VP are observed for 8, 24 and 32 hours.
  • TEAM-VP revealed with Streptavidin-Texas Red co-distributes with lysosomes (anti-Lamp2-FITC) at 8 h of treatment and seems to leave these organelles at 24 h.
  • the isolated mitochondria were incubated with the peptide CycRGD or TEAM-VP in the presence or not of bongkrekic acid (BA, 50 ⁇ M), cyclosporin A (CsA, 10 ⁇ M), and DIDS (8 ⁇ M). Induction of the mitochondrial membrane potential drop: The isolated mitochondria were incubated with 1 ⁇ M of TEAM-VP or its controls (C1, C2, C3), labeled with JC-1 and analyzed by flow cytometry (FIG. 3a),
  • HUVEC cells were incubated for 24 hours with 15 ⁇ M of TEAM-VP peptide, CycRGD and C4 then labeled with JC-1 or with an anti-PARP or Annexin-V-FITC and analyzed by flow cytometry (Figure 3d).

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Animal Behavior & Ethology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Public Health (AREA)
  • Engineering & Computer Science (AREA)
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  • Gastroenterology & Hepatology (AREA)
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  • Heart & Thoracic Surgery (AREA)
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  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention relates to cyclised peptides corresponding to sequence SEQ ID N°1: X1X2RGDX3FGX4X5LLFIHFX6IGSX7HSX8IX9, wherein: the letters without any numerical index correspond to amino acids defined by the single-letter international code; X1 is G or GG having an amino-terminal end which may or may not be free; X2 is either a C, in which case X2 = X4, the two Cs being connected by a disulphide bridge, or X2 is capable of forming a lactam bridge with X4, either X2 or X4 being an amino acid bearing an acid group, such as A or D, and the other bearing an amino function such as Q, N; X3 is either an M pattern or a norleucine pattern; X5 is one or several di-, tri-, or tetra-peptide patterns comprising G or a combination of G and S, or X5 is a C pattern having a side-chain which serves as a covalent linkage point with a 3-nitro-2-pyridinesulphenyl group which is located at the N-terminal end of the next amino acid (L); X6 is either an R pattern or a K pattern; X7 is either an R pattern or a K pattern; X8 is either an R pattern or a K pattern; and X9 is an aliphatic amino acid (such as G or A) having an amide C-terminal end. The inventive peptides can be used as active ingredients in medicaments, for example, for the treatment of pathologies linked to hypervascularisation.

Description

" PEPTIDES POSSÉDANT NOTAMMENT UNE ACTIVITÉ ANTI-ANGIOGÉNIQUE ET LEURS APPLICATIONS EN THÉRAPEUTIQUE " "PEPTIDES WITH ESPECIALLY ANTI-ANGIOGENIC ACTIVITY AND THEIR THERAPEUTIC APPLICATIONS"
L'invention se rapporte à des peptides possédant notamment une activité anti-angiogénique et à leurs applications en thérapeutique . Les recherches des inventeurs concernant des peptides thérapeutiquement actifs les ont amené à élaborer des constructions qui se sont révélées de grand intérêt au regard de leurs propriétés anti-angiogéniques . L'invention vise donc de tels peptides et la mise à profit de leurs propriétés thérapeutiques pour élaborer des médicaments. Elle vise ainsi les compositions pharmaceutiques renfermant ces peptides à titre de principe actif. Elle vise également l'utilisation de ces peptides pour fabriquer des médicaments à effet anti-angiogénique, pour le traitement de pathologies liées à une hypervascularisation. Les peptides selon l'invention sont caractérisés en ce qu'il s'agit de peptides cyclisés, répondant à la séquence SEQ ID N°l : XιX2RGDX3FGX4X5LLFIHFX5IGSX7HSX8IX9 dans laquelle : les lettres sans indice numérique correspondent aux acides aminés définis par le code international à une lettre . Xi est soit un G soit un GG dont l'extrémité amino- terminale est libre, alkylée, acylée, notamment acétylée, ou comporte un groupe de marquage comme le groupe biotinyle, X2 est soit un C, auquel cas X2 = X4, les 2 C étant alors réunis par un pont disulfure, ou X2 est capable de former un pont lactame avec X4, l'un de X2 ou X4 étant un acide aminé porteur d'un groupement acide, comme A ou D, l'autre portant une fonction aminée, comme Q, N. X3 est soit un motif M, soit un motif nor-Leucine X est soit un motif, soit une succession de deux motifs di-, tri- ou tetra-peptidiques composée de G ou d'une combinaison de G et de S, comme GG, GGG, GGGG, GGS, GGGS, GGSGGS, ou encore X5 est un motif C dont la chaîne latérale (fonction thiol) sert de point de liaison covalente avec un groupe 3-nitro-2-pyridinesulfényle (Npys; Drijfhout et al., 1988 Int J Peptide protein Res, 32:161-166) situé sur l'extrémité N-terminale de l'acide aminé suivant (L) , X6 est soit un motif R, soit un motif K - X7 est soit un motif R, soit un motif K X8 est soit un motif R, soit un motif K Xg est un acide aminé aliphatique (comme G, ou A) dont l'extrémité C-terminale est amidée. Ces peptides comportent de 25 à 35 acides aminés. Un peptide de ce type répond à la séquence SEQ ID N°2 : GG*CRGDMFG*CGGLLFIHFRIGSRHSRIG (^indique un pont disulfure reliant les deux motifs C) . D'autres peptides sont tels que définis ci-dessus et présentent un groupe alkylé à leur extrémité N-terminale. Dans d'autres peptides encore, un ou plusieurs acides aminés sont remplacés par leur forme dextrogyre (Daa) . D'autres peptides selon l'invention répondent à la SEQ ID N°l ci-dessus, mais comportent une ou plusieurs liaisons peptidiques pour former des bioisostères . On citera par exemple la réduction d'un pont amide en -CH2NH-,ou une réaction de rétro-inverso, tels que définis par Goodman et Ro (1995, dans Burger's Médicinal Chemistry. Fifth ed vol.l pages 803- 861, édité par ME olff) . Comme variants du peptide de séquence SEQ ID N°2 exposant le motif RGD par un pont disulfure entre deux cystéines, on citera les peptides de séquences SEQ ID N°3 à 10: SEQ ID N°3 : GG*CRGDMFG*CGGLLRIHFRIGSRHSRIGThe invention relates to peptides having in particular an anti-angiogenic activity and to their applications in therapy. The inventors' research into therapeutically active peptides led them to develop constructs which have proved to be of great interest with regard to their anti-angiogenic properties. The invention therefore relates to such peptides and the use of their therapeutic properties to develop drugs. It thus relates to pharmaceutical compositions containing these peptides as active principle. It also relates to the use of these peptides to manufacture drugs with anti-angiogenic effect, for the treatment of pathologies linked to hypervascularization. The peptides according to the invention are characterized in that they are cyclized peptides, corresponding to the sequence SEQ ID No. 1: XιX 2 RGDX 3 FGX4X5LLFIHFX 5 IGSX 7 HSX 8 IX9 in which: the letters without numerical index correspond amino acids defined by the international one-letter code. Xi is either a G or a GG whose amino-terminal end is free, alkylated, acylated, in particular acetylated, or comprises a labeling group such as the biotinyl group, X 2 is either a C, in which case X 2 = X 4 , the 2 Cs being then united by a disulfide bridge, or X 2 is capable of forming a lactam bridge with X 4 , one of X 2 or X 4 being an amino acid carrying an acid group, such as A or D , the other carrying an amino function, like Q, N. X 3 is either a motif M or a nor-Leucine motif X is either a motif or a succession of two di-, tri- or tetra-peptide motifs composed of G or of a combination of G and S, such as GG, GGG, GGGG, GGS, GGGS, GGSGGS, or even X 5 is a C motif whose side chain (thiol function) serves as a covalent bonding point with a 3-nitro-2-pyridinesulfenyl group (Npys; Drijfhout et al ., 1988 Int J Peptide protein Res, 32: 161-166) located on the N-terminal end of the following amino acid (L), X 6 is either an R motif or a K motif - X 7 is either an R motif, either a KX 8 motif is either an R motif, or a K Xg motif is an aliphatic amino acid (such as G, or A) whose C-terminal end is amidated. These peptides contain from 25 to 35 amino acids. A peptide of this type responds to the sequence SEQ ID No. 2: GG * CRGDMFG * CGGLLFIHFRIGSRHSRIG (^ indicates a disulfide bridge connecting the two motifs C). Other peptides are as defined above and have an alkyl group at their N-terminal end. In still other peptides, one or more amino acids are replaced by their dextrorotatory form ( D aa). Other peptides according to the invention correspond to SEQ ID No. 1 above, but have one or more peptide bonds to form bioisosteres. Examples include the reduction of an amide bridge to -CH 2 NH-, or a retro-inverso reaction, as defined by Goodman and Ro (1995, in Burger's Medicinal Chemistry. Fifth ed vol.l pages 803- 861 , edited by ME olff). As variants of the peptide of sequence SEQ ID No. 2 exhibiting the RGD motif by a disulfide bridge between two cysteines, mention will be made of the peptides of sequence SEQ ID No. 3 to 10: SEQ ID N ° 3: GG * CRGDMFG * CGGLLRIHFRIGSRHSRIG
SEQ ID N°4 : GG*CRGDMFG*CGG-LFIHFRIGSRHSRIGSEQ ID N ° 4: GG * CRGDMFG * CGG-LFIHFRIGSRHSRIG
SEQ ID N°5 : GG*CRGDMFG*CGGSLFIHFRIGSRHSRIGSEQ ID N ° 5: GG * CRGDMFG * CGGSLFIHFRIGSRHSRIG
SEQ ID N°6 : GG*CRGDMFG*CGGLLFIHFKIGSRHSRIGSEQ ID N ° 6: GG * CRGDMFG * CGGLLFIHFKIGSRHSRIG
SEQ ID N°7 : GG*CRGDMFG*CGGLLFIHFNRIGSRHSRIG (NR représentant un motif N-alkylarginine)SEQ ID N ° 7: GG * CRGDMFG * CGGLLFIHF N RIGSRHSRIG ( N R representing an N-alkylarginine motif)
SEQ ID N°8 : GG*CRGDMFG*CGGLLSRHFRIGSRHSRIGSEQ ID N ° 8: GG * CRGDMFG * CGGLLSRHFRIGSRHSRIG
SEQ ID N°9 : GG*CRGDMFG*CGGLLSIHFRIGSRHSRIGSEQ ID N ° 9: GG * CRGDMFG * CGGLLSIHFRIGSRHSRIG
SEQ ID N°10 : GG*CRGDMFG*CGGLLFRHFRIGSRHSRIGSEQ ID N ° 10: GG * CRGDMFG * CGGLLFRHFRIGSRHSRIG
D'autres peptides de l'invention comportent une séquence SEQ ID N°ll : X-R-G-D-M-F-GX' exposant le motif RGD par un pont lactame entre les acides aminés X (X) -C-O-NH- (X' ) , X et X' étant des acides aminés tels que l'un porte un groupement acide, et l'autre porte une aminé . Des peptides préférés de ce groupe répondent aux séquences SEQ ID N°12 à SEQ ID N°23 :Other peptides of the invention include a sequence SEQ ID No. 11: XRGDMF-GX 'exposing the RGD motif by a lactam bridge between the amino acids X (X) -CO-NH- (X'), X and X 'being amino acids such that one carries an acid group, and the other carries an amino. Preferred peptides of this group correspond to the sequences SEQ ID No. 12 to SEQ ID No. 23:
SEQ ID N°12 : GGXRGDMFGX' GGLLFIHFRIGCRHSRIGSEQ ID N ° 12: GGXRGDMFGX 'GGLLFIHFRIGCRHSRIG
SEQ ID N°13 : GGXRGDMFGX' GGLLFIFFRIGCRFSRIGSEQ ID N ° 13: GGXRGDMFGX 'GGLLFIFFRIGCRFSRIG
SEQ ID N°14 : GGXRGDMFGX' GGLLFIHFRIGSRHSRIGSEQ ID N ° 14: GGXRGDMFGX 'GGLLFIHFRIGSRHSRIG
SEQ ID N°15 : GGXRGDMFGX' GGLLRIHFRIGSRHSRIGSEQ ID N ° 15: GGXRGDMFGX 'GGLLRIHFRIGSRHSRIG
SEQ ID N°16 : GGXRGDMFGX' GG-LFIHFRIGSRHSRIGSEQ ID N ° 16: GGXRGDMFGX 'GG-LFIHFRIGSRHSRIG
SEQ ID N°17 : GGXRGDMFGX' GGSLFIHFRIGSRHSRIGSEQ ID N ° 17: GGXRGDMFGX 'GGSLFIHFRIGSRHSRIG
SEQ ID N°18 : GGXRGDMFGX' GGLLFIHFKIGSRHSRIGSEQ ID N ° 18: GGXRGDMFGX 'GGLLFIHFKIGSRHSRIG
SEQ ID N°19 : GGXRGDMFGX' GGLLFIHFRIGSRHSRIG (NR représentant un motif N-alkylarginine)SEQ ID N ° 19: GGXRGDMFGX 'GGLLFIHFRIGSRHSRIG ( N R representing an N-alkylarginine motif)
SEQ ID N°20 : GGXRGDMFGX' GGLLSRHFRIGSRHSRIGSEQ ID N ° 20: GGXRGDMFGX 'GGLLSRHFRIGSRHSRIG
SEQ ID N°21 : GGXRGDMFGX' GGLLSIHFRIGSRHSRIGSEQ ID N ° 21: GGXRGDMFGX 'GGLLSIHFRIGSRHSRIG
SEQ ID N°22 : GGXRGDMFGX' GGLLFRHFRIGSRHSRIGSEQ ID N ° 22: GGXRGDMFGX 'GGLLFRHFRIGSRHSRIG
SEQ ID N°23 : GGXRGDMFGX' GGLLFIHFRIGSRHSRIG Lesdites séquences peuvent être modifiées, c'est-à-dire correspondre au peptide natif mais comporter un ou plusieurs acides différents, modifiés chimiquement, des lors que ces modifications n'affecte pas la fonction recherchée. On citera notamment le remplacement de Met par nor-Leu, Arg par N-alkyl Arg, qui permet en particulier de stabiliser la construction. Ces modifications comprennent également un groupe acyle, notamment acétyle en N-terminal. Les peptides de l'invention sont encore caractérisés en ce qu'ils induisent l'apoptose de cellules endotheliales humaines exprimant les récepteurs αVβ3. Ils sont en outre avantageusement caractérisés en ce qu'ils subissent une endocytose par des cellules endotheliales humaines exprimant les récepteurs αVβ3, se localisent au niveau du compartiment itochondrial et exercent un effet mitochondriotoxique . Lorsqu'on met en contact un peptide tel qu'élaboré ci- dessus, ou tel que défini ci-dessus, avec des cellules endotheliales, on observe une reconnaissance spécifique des intégrines Vβ3 à la surface des cellules endotheliales, ce qui permet l' endocytose du peptide chimère. Une fois internalisé, le peptide se localise transitoirement dans les lysosomes, comme montré en microscopie confocale et se distribue progressivement dans le compartiment mitochondrial. On notera que la spécificité des peptides de l'invention résulte de l'ajout de la partie mitochondrio-toxique à des ligands des intégrines pour exercer une toxicité par la voie de la mitochondrio-toxicité, les ligands des intégrines étant présents à des fins de ciblage et n'ayant pas eux-mêmes d'activité angiostatique. Comme illustré par les exemples donnés ci-après, le traitement de cellules endotheliales primaires humaines avec des doses de peptides de l'ordre du micromolaire conduit à une dissipation du potentiel transmembranaire mitochondrial (Δψm) , à la libération de cytochrome c mitochondrial, à l'exposition de phosphatidyl-sérine et à la condensation de chromatine nucléaire. Ces constructions peptidiques présentent l'avantage d'une absence de toxicité sur les cellules Vβ3 négatives. L'invention vise donc également la mise à profit de ces propriétés pour induire sélectivement PMM et l'apoptose de cellules endotheliales angiogéniques dans le cadre de stratégies thérapeutiques, notamment anti-cancer, traitement de la polyarthrite, de la rétinopathie diabétique. Les compositions pharmaceutiques selon l'invention sont caractérisées en ce qu'elles renferment une quantité thérapeutique ent efficace d'au moins un peptide, tel que défini ci-dessus, en association avec un véhicule phar aceutiquement acceptable. Ces compositions se présentent avantageusement sous les formes galéniques appropriées pour leur administration par voie injectable. On citera en particulier les solutions injectables destinées à une administration par voie intra veineuse. L'invention vise en outre l'utilisation de constructions peptidiques telles que définies ci-dessus pour fabriquer des médicaments anti-angiogéniques pour le traitement de pathologies dues à une hypervascularisation. On citera en particulier le traitement de tumeurs solides telles que les tumeurs pulmonaires, les adénomes, les mélanomes, les cancers de la prostate, du sein, du colon, du pancréas, les ostéosarcomes . L'invention s'applique également au traitement de rétinopathies diabétiques et de polyarthrites . Les posologies des formes d'administration et les traitements seront déterminés par l'homme du métier en fonction de la pathologie à traiter et de l'état du patient. D'autres caractéristiques et avantages de l'invention seront donnés dans les exemples qui suivent se rapportant à la construction SEQ ID N°2 ( ci-après TEAM-VP) SEQ ID N°2 : GG*CRGDMFG*C-GG-LLFIHFRIGSRHSRIG-amide avec ou sans biotine, "*" indiquant une cyclisation par formation d'un pont disulfure. Il sera fait référence aux figures 1 à 3, qui représentent, respectivement, la figure 1, l'analyse de la cytotoxicité sur des cellules endotheliales, la figure 2, la reconnaissance du motif CycRGD par les intégrines αVβ3, la figure 3, les effets de TEAM-VP sur des itochondries isolées et des cellules HUVEC.SEQ ID N ° 23: GGXRGDMFGX 'GGLLFIHFRIGSRHSRIG Said sequences can be modified, that is to say correspond to the native peptide but contain one or more different acids, chemically modified, as long as these modifications do not affect the desired function. These include the replacement of Met by nor-Leu, Arg by N-alkyl Arg, which in particular allows the construction to be stabilized. These modifications also include an acyl group, in particular N-terminal acetyl. The peptides of the invention are further characterized in that they induce apoptosis of human endothelial cells expressing the αVβ3 receptors. They are also advantageously characterized in that they undergo endocytosis by human endothelial cells expressing the αVβ3 receptors, are localized in the itochondrial compartment and exert a mitochondriotoxic effect. When a peptide as developed above, or as defined above, is brought into contact with endothelial cells, a specific recognition of the Vβ3 integrins is observed on the surface of the endothelial cells, which allows endocytosis chimeric peptide. Once internalized, the peptide is temporarily located in the lysosomes, as shown in confocal microscopy and is gradually distributed in the mitochondrial compartment. It will be noted that the specificity of the peptides of the invention results from the addition of the mitochondrio-toxic part to ligands of the integrins to exert a toxicity by the way of the mitochondrio-toxicity, the ligands of the integrins being present for purposes of targeting and not themselves having angiostatic activity. As illustrated by the examples given below, the treatment of primary human endothelial cells with doses of peptides of the order of a micromolar leads to a dissipation of the mitochondrial transmembrane potential (Δψm), to the release of mitochondrial cytochrome c, to l 'exposure of phosphatidyl-serine and condensation of nuclear chromatin. These peptide constructions have the advantage of an absence of toxicity on negative Vβ3 cells. The invention therefore also aims to take advantage of these properties to selectively induce PMM and apoptosis of angiogenic endothelial cells in the context of therapeutic strategies, in particular anti-cancer, treatment of polyarthritis, diabetic retinopathy. The pharmaceutical compositions according to the invention are characterized in that they contain a therapeutically effective amount of at least one peptide, as defined above, in combination with a phar aceutically acceptable vehicle. These compositions are advantageously in the dosage forms suitable for their administration by injectable route. Mention will in particular be made of injectable solutions intended for administration by the intravenous route. The invention further relates to the use of peptide constructs as defined above for manufacturing anti-angiogenic drugs for the treatment of pathologies due to hypervascularization. Mention will in particular be made of the treatment of solid tumors such as pulmonary tumors, adenomas, melanomas, cancers of the prostate, breast, colon, pancreas, osteosarcomas. The invention also applies to the treatment of diabetic retinopathies and polyarthritis. The dosages of the administration forms and the treatments will be determined by a person skilled in the art depending on the pathology to be treated and the condition of the patient. Other characteristics and advantages of the invention will be given in the examples which follow relating to the construction SEQ ID No. 2 (hereafter TEAM-VP) SEQ ID No. 2: GG * CRGDMFG * C-GG-LLFIHFRIGSRHSRIG -amide with or without biotin, "*" indicating cyclization by formation of a disulfide bridge. Reference will be made to FIGS. 1 to 3, which represent, respectively, FIG. 1, the analysis of cytotoxicity on endothelial cells, FIG. 2, the recognition of the CycRGD motif by the integrins αVβ3, FIG. 3, the effects of TEAM-VP on isolated itochondria and HUVEC cells.
Exemples : Analyse de la cytotoxicité de TEAM-VP sur cellules endotheliales a. Les cellules HUVEC ont été incubées pendant 24h avec 5-30 μM de peptide CycRGD, LLFIHFRIGSRHSRIG-amide (C4) ou TEAM-VP, puis marquées avec la 7-AAD et analysées par cytométrie en flux (Figure la) . b. Les cellules HUVEC ont été incubées pendant 24, 48, 72 et 96h avec 15 μM de TEAM-VP puis marquées avec la 7-AAD et analysées par cytométrie en flux (Figure lb) .Examples: Analysis of the cytotoxicity of TEAM-VP on endothelial cells a. The HUVEC cells were incubated for 24 h with 5-30 μM of peptide CycRGD, LLFIHFRIGSRHSRIG-amide (C4) or TEAM-VP, then labeled with 7-AAD and analyzed by flow cytometry (Figure la). b. The HUVEC cells were incubated for 24, 48, 72 and 96 h with 15 μM of TEAM-VP and then labeled with 7-AAD and analyzed by flow cytometry (Figure lb).
Reconnaissance du motif CycRGD par les intégrines αVβ3 a. Analyse de la liaison cellulaire : Les cellules HUVEC ont été incubées pendant 45 min à température ambiante avec les peptides suivants : GGCRGDMFGCGG-amide (RGD linéaire) , GG*CRADMFG*CGG-amide (CycRAD) et GG*CRGDMFG*CGG-amide (CycRGD) (0,5 à 2 μM) marqués FITC et analysées par cytométrie en flux (Figure 2a) . b. Chasse du peptide CycRGD par lui-même: Les cellules HUVEC ont été pré-incubées ou non pendant 30 min à température ambiante avec 200 μM de peptide CycRGD non marqué avant ajout de peptide FITC-CycRGD (10 μM) pendant 45 min et analysées par cytométrie en flux (Figure 2b) . c. Compétition pour les sites intégrines.:Recognition of the CycRGD motif by the αVβ3 integrins a. Cell binding analysis: HUVEC cells were incubated for 45 min at room temperature with the following peptides: GGCRGDMFGCGG-amide (linear RGD), GG * CRADMFG * CGG-amide (CycRAD) and GG * CRGDMFG * CGG-amide ( CycRGD) (0.5 to 2 μM) marked FITC and analyzed by flow cytometry (Figure 2a). b. Hunt for the CycRGD peptide by itself: The HUVEC cells were pre-incubated or not for 30 min at room temperature with 200 μM of unlabeled CycRGD peptide before addition of FITC-CycRGD peptide (10 μM) for 45 min and analyzed by flow cytometry (Figure 2b). vs. Competition for integral sites:
Les cellules HUVEC ont été pré-incubées ou non pendant 30 min à température ambiante avec 25 μM de peptide CycRGD, CycRAD, GRGDS et GRGES avant ajout de peptide FITC-CycRGD (0,5 μM) , puis analysées par cytométrie en flux (Figure 2c) . d. Corrélation entre expression des intégrines, liaison et toxicité des peptides : Les cellules HUVEC, HMVECd, MCF-7, MDA, HeLa, HT-29, Jurkat, CEM et PBMC ont été marquées avec les anticorps dirigés contre les intégrines αVβ3 et αVβ5 et analysées par cytométrie en flux. La liaison du peptide CycRGD et l'induction d' apoptose par TEAM-VP sur les différents types cellulaire ont été mesurées (Figure 2d) .The HUVEC cells were pre-incubated or not for 30 min at room temperature with 25 μM of CycRGD, CycRAD, GRGDS and GRGES peptide before addition of FITC-CycRGD peptide (0.5 μM), then analyzed by flow cytometry (Figure 2c). d. Correlation between expression of integrins, binding and toxicity of peptides: The cells HUVEC, HMVECd, MCF-7, MDA, HeLa, HT-29, Jurkat, CEM and PBMC were labeled with the antibodies directed against the integrins αVβ3 and αVβ5 and analyzed by flow cytometry. The binding of the CycRGD peptide and the induction of apoptosis by TEAM-VP on the different cell types were measured (Figure 2d).
+ Etude du processus d'entrée des peptides :+ Study of the peptide entry process:
Les peptides FITC-CycRGD et TEAM-VP (FITC) rentrent dans les HUVEC et co-localisent avec les billes de dextran (5h de co- traitement) . L'entrée de TEAM-VP et des billes de dextran est inhibée par le traitement Azide de sodium + Déoxyglucose, indiquant une entrée du peptide par endocytose. On n'observe pas d'entrée du peptide FITC-CycRAD dans les HUVEC, ni d'entrée du peptide FITC-CycRGD dans les HeLa. + Routage intracellulaire de TEAM-VP :The FITC-CycRGD and TEAM-VP (FITC) peptides enter the HUVECs and co-locate with the dextran beads (5 hours of co-treatment). The entry of TEAM-VP and dextran beads is inhibited by the treatment of sodium azide + deoxyglucose, indicating entry of the peptide by endocytosis. No entry of the FITC-CycRAD peptide was observed in the HUVECs, nor was there any entry of the FITC-CycRGD peptide in the HeLa. + TEAM-VP intracellular routing:
On observe au confocal des HUVEC traitées avec TEAM-VP pendant 8, 24 et 32h.In the confocal, HUVECs treated with TEAM-VP are observed for 8, 24 and 32 hours.
TEAM-VP révélé avec la Streptavidine-Texas Red co-distribue avec les lysosomes (anti-Lamp2-FITC) à 8h de traitement et semblerait quitter ces organelles à 24h. TEAM-VP révélé avec la Streptavidine-FITC co-distribue partiellement avec les mitochondries (anti-VDAC) à 24h et totalement à 32h. On n'observe pas de co-distribution avec l'appareil de Golgi (anti-Golgin) tout au long du traitement.TEAM-VP revealed with Streptavidin-Texas Red co-distributes with lysosomes (anti-Lamp2-FITC) at 8 h of treatment and seems to leave these organelles at 24 h. TEAM-VP revealed with Streptavidin-FITC partially co-distributes with the mitochondria (anti-VDAC) at 24h and totally at 32h. We does not observe co-distribution with the Golgi apparatus (anti-Golgin) throughout the treatment.
Effets de TEAM-VP sur des mitochondries isolées et des cellules HUVEC a. Effet sur les mitochondries isolées : Induction du gonflement mitochondrial :Effects of TEAM-VP on isolated mitochondria and HUVEC cells a. Effect on isolated mitochondria: Induction of mitochondrial swelling:
Les mitochondries isolées ont été incubées avec le peptide CycRGD ou TEAM-VP en présence ou non d'acide bongkrékique (BA, 50 μM) , de cyclosporine A (CsA, 10 μM) , et de DIDS (8 μM) . Induction de la chute de potentiel membranaire mitochondrial : Les mitochondries isolées ont été incubées avec 1 μM de TEAM- VP ou ses contrôles (C1,C2,C3), marquées au JC-1 et analysées en cytométrie de flux (Figure 3a) ,The isolated mitochondria were incubated with the peptide CycRGD or TEAM-VP in the presence or not of bongkrekic acid (BA, 50 μM), cyclosporin A (CsA, 10 μM), and DIDS (8 μM). Induction of the mitochondrial membrane potential drop: The isolated mitochondria were incubated with 1 μM of TEAM-VP or its controls (C1, C2, C3), labeled with JC-1 and analyzed by flow cytometry (FIG. 3a),
SEQ ID N°24 Cl= GG*CRADMFG*CGGLLFlHFRIGSRHSRIGamide SEQ ID N°25 Cl= GG*CRGDMFG*CGGLLFIHFAIGSRHSAIGamide SEQ ID N°26 C3= RKKRRQRRRGGLLFIHFRIGSRHSRIGamideSEQ ID N ° 24 Cl = GG * CRADMFG * CGGLLFlHFRIGSRHSRIGamide SEQ ID N ° 25 Cl = GG * CRGDMFG * CGGLLFIHFAIGSRHSAIGamide SEQ ID N ° 26 C3 = RKKRRQRRRGGLLFIHFRIGSRHSRIGamide
b. Relargage de cytochrome c :b. Release of cytochrome c:
Les mitochondries isolées ont été incubées avec l'Alaméthicine (5 ug/ml) ou TEAM-VP (10 μM) et le surnageant analysé en Western Blot avec un anti-cytochrome c (Figure 3b) . c. Analyse de l'apoptose nucléaire :The isolated mitochondria were incubated with Alamethicin (5 ug / ml) or TEAM-VP (10 μM) and the supernatant analyzed in Western Blot with an anti-cytochrome c (Figure 3b). vs. Analysis of nuclear apoptosis:
Les cellules HUVEC traitées avec TEAM-VP (15-40 μM) en présence ou non d'inhibiteur de caspases pendant 8, 16, 24 et 48h ont été marquées au Hoechst et observées au microscope inversé. Les pourcentages de cellules présentant des noyaux intacts, de stade I ou de stade II sont reportés (Figure 3c) . d. Induction du la chute de potentiel membranaire mitochondrial et exposition des phosphatidyl serines in cellula :HUVEC cells treated with TEAM-VP (15-40 μM) in the presence or absence of a caspase inhibitor for 8, 16, 24 and 48 h were labeled with Hoechst and observed with an inverted microscope. The percentages of cells with intact nuclei, stage I or stage II are reported (Figure 3c). d. Induction of the mitochondrial membrane potential drop and exposure of phosphatidyl serines in cellula:
Les cellules HUVEC ont été incubées pendant 24h avec 15 μM de peptide TEAM-VP, CycRGD et C4 puis marquées au JC-1 ou avec un anti-PARP ou à l' Annexin-V-FITC et analysées par cytométrie en flux (Figure 3d) .The HUVEC cells were incubated for 24 hours with 15 μM of TEAM-VP peptide, CycRGD and C4 then labeled with JC-1 or with an anti-PARP or Annexin-V-FITC and analyzed by flow cytometry (Figure 3d).
+ Relargage cytochrome C in cellula :+ Cytochrome C in cellula release:
Relargage de cytochrome c observé en microscopie après 24h de traitement avec TEAM-VP (10 μM) et double marquage des cellules fixées avec un anti-cytochrome c et un anti-VDAC.Release of cytochrome c observed under microscopy after 24 hours of treatment with TEAM-VP (10 μM) and double labeling of fixed cells with an anti-cytochrome c and an anti-VDAC.
+ Chute de potentiel membranaire mitochondrial in cellula :+ Loss of mitochondrial membrane potential in cellula:
Chute de potentiel observée au microscope après marquage auFall in potential observed under the microscope after labeling with
JC-1 des cellules HUVEC traitées pendant 16-24h avec 15 μM de peptide TEAM-VP.JC-1 of the HUVEC cells treated for 16-24 hours with 15 μM of TEAM-VP peptide.
Modèles animaux :Animal models:
Les modèles animaux mis en œuvre pour démontrer l'efficacité des produits correspondent à ceux classiquement utilisés (voir notamment Kisher et al., 2001 Cancer Research 61:7669-7674,The animal models used to demonstrate the efficacy of the products correspond to those conventionally used (see in particular Kisher et al., 2001 Cancer Research 61: 7669-7674,
Galaup et al., 2003 Mol Therapy 7:731-740). Galaup et al., 2003 Mol Therapy 7: 731-740).

Claims

REVENDICATIONS
1 . Peptides possédant notamment une activité anti- angiogénique, caractérisés en ce qu' il s ' agit de peptides cyclisés , répondant à la séquence SEQ ID N° l : XιX2RGDX3FGX4X5LLFIHFX6IGSX7HSX8IXg dans laquelle : les lettres sans indice numérique correspondent aux acides aminés définis par le code international à une lettre. Xi est soit un G soit un GG dont l'extrémité amino- terminale est libre, alkylée, acylée, notamment acétylée, ou comporte un groupe de marquage comme le groupe biotinyle, X2 est soit un C, auquel cas X2 = X4, les 2 C étant alors réunis par un pont disulfure, ou X2 est capable de former un pont lactame avec X4, l'un de X2 ou X4 étant un acide aminé porteur d'un groupement acide, comme A ou D, l'autre portant une fonction aminée, comme Q, N. X3 est soit un motif M, soit un motif nor-Leucine X5 est soit un motif, soit une succession de deux motifs di-, tri- ou tetra-peptidiques composée de G ou d'une combinaison de G et de S, comme GG, GGG, GGGG, GGS, GGGS, GGSGGS, ou encore X5 est un motif C dont la chaîne latérale (fonction thiol) sert de point de liaison covalente avec un groupe 3-nitro-2-pyridinesulfényle situé sur l'extrémité N-terminale de l'acide aminé suivant (L) . X6 est soit un motif R, soit un motif K X7 est soit un motif R, soit un motif K X8 est soit un motif R, soit un motif K Xg est un acide aminé aliphatique (comme G, ou A) dont l'extrémité C-terminale est amidée. 1. Peptides having in particular an anti-angiogenic activity, characterized in that they are cyclized peptides, corresponding to the sequence SEQ ID No 1: XιX 2 RGDX 3 FGX 4 X 5 LLFIHFX 6 IGSX 7 HSX 8 IXg in which: letters without a numerical index correspond to the amino acids defined by the international one-letter code. Xi is either a G or a GG whose amino-terminal end is free, alkylated, acylated, in particular acetylated, or comprises a labeling group such as the biotinyl group, X 2 is either a C, in which case X 2 = X 4 , the 2 Cs being then united by a disulfide bridge, or X 2 is capable of forming a lactam bridge with X 4 , one of X 2 or X 4 being an amino acid carrying an acid group, such as A or D , the other carrying an amino function, such as Q, N. X 3 is either a motif M or a nor-Leucine motif X 5 is either a motif or a succession of two di-, tri- or tetra-peptide motifs composed of G or a combination of G and S, such as GG, GGG, GGGG, GGS, GGGS, GGSGGS, or even X 5 is a C motif whose side chain (thiol function) serves as a covalent bonding point with a 3-nitro-2-pyridinesulfenyl group located on the N-terminal end of the next amino acid (L). X 6 is either an R motif, or a KX 7 motif is either an R motif, or a KX 8 motif is either an R motif, or a K motif Xg is an aliphatic amino acid (such as G, or A) whose C-terminal end is amidated.
2. Peptide selon la revendication 1, caractérisé en ce qu'il répond à la séquence SEQ ID N°2 : GG*CRGDMFG*CGGLLFIHFRIGSRHSRIG (*indique un pont disulfure reliant les deux motifs C) . 2. Peptide according to claim 1, characterized in that it responds to the sequence SEQ ID No. 2: GG * CRGDMFG * CGGLLFIHFRIGSRHSRIG (* indicates a disulfide bridge connecting the two C motifs).
3. Peptides selon la revendication 1 ou 2, caractérisés en ce qu'ils sont modifiés par rapport au peptide natif et présentent notamment un groupe alkylé à leur extrémité N- terminale et/ou en ce qu'un ou plusieurs acides aminés sont remplacés par leur forme dextrogyre (Daa) et/ou en ce qu'ils comportent une ou plusieurs liaisons peptidiques pour former des bioisosteres, par exemple la réduction d'un pont amide en -CH2NH-, ou une réaction de rétro-inverso. 3. Peptides according to claim 1 or 2, characterized in that they are modified with respect to the native peptide and in particular have an alkyl group at their N-terminal end and / or in that one or more amino acids are replaced by their dextrorotatory form ( D aa) and / or in that they comprise one or more peptide bonds to form bioisosteres, for example the reduction of an amide bridge into -CH 2 NH-, or a retro-inverso reaction.
4. Peptides selon la revendication 2, dans lesquels le motif RGD est exposé par un pont disulfure entre deux cystéines, en particulier les peptides de séquences SEQ ID N°3 à 10:4. Peptides according to claim 2, in which the RGD motif is exposed by a disulfide bridge between two cysteines, in particular the peptides of sequences SEQ ID N ° 3 to 10:
SEQ ID N°3 : GG*CRGDMFG*CGGLLRIHFRIGSRHSRIGSEQ ID N ° 3: GG * CRGDMFG * CGGLLRIHFRIGSRHSRIG
SEQ ID N°4 GG*CRGDMFG*CGG-LFIHFRIGSRHSRIG SEQ ID N°5 GG*CRGDMFG*CGGSLFIHFRIGSRHSRIG SEQ ID N°6 GG*CRGDMFG*CGGLLFIHFKIGSRHSRIG SEQ ID N°7 TR représentant un motif N-alkylarginine)SEQ ID N ° 4 GG * CRGDMFG * CGG-LFIHFRIGSRHSRIG SEQ ID N ° 5 GG * CRGDMFG * CGGSLFIHFRIGSRHSRIG SEQ ID N ° 6 GG * CRGDMFG * CGGLLFIHFKIGSRHSRIG SEQ ID N ° 7 TR representing an N-alkylarginine motif)
SEQ ID N°8 : GG*CRGDMFG*CGGLLSRHFRIGSRHSRIG SEQ ID N°9 : GG*CRGDMFG*CGGLLSIHFRIGSRHSRIG SEQ ID N°10 : GG*CRGDMFG*CGGLLFRHFRIGSRHSRIG 5. Peptides selon la revendication 1, caractérisés en ce qu'ils comportent une séquenceSEQ ID N ° 8: GG * CRGDMFG * CGGLLSRHFRIGSRHSRIG SEQ ID N ° 9: GG * CRGDMFG * CGGLLSIHFRIGSRHSRIG SEQ ID N ° 10: GG * CRGDMFG * CGGLLFRHFRIGSRHSRIG 5. Peptides according to claim 1, characterized in that they contain a sequence
SEQ ID N°ll : X-R-G-D-M-F-G-X' exposant le motif RGD par un pont lactame entre les acides aminés X (X) -C-O-NH- (X' ) , X et X' étant des acides aminés tels que l'un porte un groupement acide, et l'autre porte une aminé. 6. Peptides selon la revendication 5, caractérisés en ce qu'ils répondent aux séquences SEQ ID N°12 à SEQ ID N°23 : SEQ ID N°12 GGXRGDMFGX ' GGLLFIHFRIGCRHSRIG SEQ ID N°13 GGXRGDMFGX ' GGLLFIFFRIGCRFSRIG SEQ ID N°14 GGXRGDMFGX ' GGLLFIHFRIGSRHSRIG SEQ ID N°15 GGXRGDMFGX ' GGLLRIHFRIGSRHSRIG SEQ ID N°16 GGXRGDMFGX ' GG-LFIHFRIGSRHSRIG SEQ ID N°17 GGXRGDMFGX ' GGSLFIHFRIGSRHSRIG SEQ ID N°18 GGXRGDMFGX ' GGLLFIHFKIGSRHSRIG SEQ ID N°19 GGXRGDMFGX ' GGLLFIHFNRIGSRHSRIG (NR représentant un motif N-alkylarginine)SEQ ID No. 11: XRGDMFGX 'exposing the RGD motif by a lactam bridge between the amino acids X (X) -CO-NH- (X'), X and X 'being amino acids such that one carries a group acid, and the other carries an amino. 6. Peptides according to claim 5, characterized in that they correspond to the sequences SEQ ID No. 12 to SEQ ID No. 23: SEQ ID NO: 12 GGXRGDMFGX 'GGLLFIHFRIGCRHSRIG SEQ ID NO: 13 GGXRGDMFGX' GGLLFIFFRIGCRFSRIG SEQ ID NO: 14 GGXRGDMFGX 'GGLLFIHFRIGSRHSRIG SEQ ID NO: 15 GGXRGDMFGX' GGLLRIHFRIGSRHSRIG SEQ ID NO: 16 GGXRGDMFGX GG-LFIHFRIGSRHSRIG SEQ ID NO: 17 GGXRGDMFGX 'GGSLFIHFRIGSRHSRIG SEQ ID N ° 18 GGXRGDMFGX 'GGLLFIHFKIGSRHSRIG SEQ ID N ° 19 GGXRGDMFGX' GGLLFIHF N RIGSRHSRIG ( N R representing an N-alkylarginine motif)
SEQ ID N°20 GGXRGDMFGX ' GGLLSRHFRIGSRHSRIG SEQ ID N°21 GGXRGDMFGX ' GGLLSIHFRIGSRHSRIG SEQ ID N°22 GGXRGDMFGX ' GGLLFRHFRIGSRHSRIG SEQ ID N°23 GGXRGDMFGX ' GGLLFIHFRIGSRHSRIG 7. Peptides selon l'une quelconque des revendications 1 àSEQ ID N ° 20 GGXRGDMFGX 'GGLLSRHFRIGSRHSRIG SEQ ID N ° 21 GGXRGDMFGX' GGLLSIHFRIGSRHSRIG SEQ ID N ° 22 GGXRGDMFGX 'GGLLFRHFRIGSRHSRIG SEQ ID N ° 23 GGXIGGGFRGIFRGFRGFGRGFRGFGRGFRGFRGFRGFRGFRGGFRGFGRGFGRGFRGFGRGFGRGFGRGFGGFRGFRGFGGFRGFGFRGFGFRGGFGFRGGFGFRGGFGFRGFGFRGGFGFRGFGFRGFGFRGGFGFRGFGFGGFGGFGGFGGFGGFGGFGGFGGFGGFGGFGGFGGFGGFGGFGGFGG
6, caractérisés qu'ils induisent l'apoptose de cellules endotheliales humaines exprimant les récepteurs αVβ3. 8. Peptides selon l'une quelconque des revendications 1 à6, characterized that they induce apoptosis of human endothelial cells expressing the αVβ3 receptors. 8. Peptides according to any one of claims 1 to
7, caractérisés en ce qu'ils subissent une endocytose par des cellules endotheliales humaines exprimant les récepteurs αVβ3, se localisent au niveau du compartiment mitochondrial et exercent un effet mitochondriotoxique. 9. Compositions pharmaceutiques, caractérisées en ce qu'elles renferment une quantité therapeutiquement efficace d'au moins un peptide tel que défini dans l'une quelconque des revendications 1 à 8, en association avec un véhicule pharmaceutiquement acceptable. 10. Compositions pharmaceutiques selon la revendication 9, caractérisées en ce qu'elles se présentent sous les formes galeniques appropriées pour leur administration par voie injectable, en particulier sous forme de solutions injectables destinées à une administration par voie intra veineuse. 11. Utilisation de peptides selon l'une quelconque des revendications 1 à 8, pour fabriquer des médicaments anti- angiogéniques pour le traitement de pathologies dues à une hypervascularisation. 12. Utilisation selon la revendication 11, pour la fabrication de médicaments pour le traitement de tumeurs solides telles que les tumeurs pulmonaires, les adénomes, les mélanomes, les cancers de la prostate, du sein, du colon, du pancréas, les ostéosarcomes, le traitement de rétinopathies diabétiques et de polyarthrites. 7, characterized in that they undergo endocytosis by human endothelial cells expressing the αVβ3 receptors, are located in the mitochondrial compartment and exert a mitochondriotoxic effect. 9. Pharmaceutical compositions, characterized in that they contain a therapeutically effective amount of at least one peptide as defined in any one of claims 1 to 8, in association with a pharmaceutically acceptable vehicle. 10. Pharmaceutical compositions according to claim 9, characterized in that they are in dosage forms suitable for their administration by injectable route, in particular in the form of injectable solutions intended for administration by intravenous route. 11. Use of peptides according to any one of claims 1 to 8, for manufacturing anti-drugs angiogenic drugs for the treatment of pathologies due to hypervascularisation. 12. Use according to claim 11, for the manufacture of medicaments for the treatment of solid tumors such as lung tumors, adenomas, melanomas, cancers of the prostate, breast, colon, pancreas, osteosarcomas, treatment of diabetic retinopathies and polyarthritis.
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