Construct comprising or consisting of a peptide sequence capable of inhibiting the interaction of filamin a with the ut receptor - pharmaceutical composition and related products

Abstract

The present invention relates to a construct comprising a cell-penetrating peptide sequence coupled by a peptide bond to at least one first binding molecule which is coupled by a peptide bond to the N-terminal end of an interfering amino acid sequence selected from the following sequences: SEQ ID NO: 1 which forms the 2104-2206 domain of the human FlnA protein and SEQ ID NO: 2 which defines the 330-350 domain of the UT receptor, the sequences having at least 90% identity, and preferably at least 95% identity, with the aforementioned sequences.

Description

[0001] CONSTRUCTION COMPRISING OR CONSTITUTING A PEPTIDE SEQUENCE CAPABLE OF INHIBITING THE INTERACTION OF FILAMIN A WITH THE UT RECEPTOR - PHARMACEUTICAL COMPOSITION AND ASSOCIATED PRODUCTS

[0002] Technical field

[0003] The present invention relates to synthetic constructs containing peptide sequences which are capable of penetrating into cells (passing the plasma membrane) and associating with the amino acid sequence of the C-terminal domain of the urotensin II (Ull) receptor, namely the UT receptor, or capable of penetrating into cells and associating with the amino acid sequence of the Filamin A (FlnA) domain involved in the interaction of Filamin A with the UT receptor.

[0004] These constructs can be used as a drug or as a diagnostic agent, particularly for the treatment / diagnosis of cancer and more specifically the treatment of glioblastoma.

[0005] The present invention also relates to a pharmaceutical composition containing such constructs as well as other related products.

[0006] Technological background

[0007] The expression and involvement of FlnA in processes associated with tumorigenesis and tumor aggressiveness have already been widely described in the literature. Although its expression is not systematically increased, variations in FlnA expression are, however, associated with a poor prognosis and are correlated with tumor aggressiveness, and the metastatic process, in particular. Indeed, FlnA is overexpressed in breast cancers (see the publication “Inhibition of Filamin-A reduces cancer metastatic potential” by Jiang et al., 2012 published in International Journal of Biological Science and “Differential expression of filamin A and its clinical significance in breast cancer” by Tian et al., 2012 published in Oncology Letters), in lung adenocarcinomas (see the publication “A positive relationship between filamin and VEGF in patients with lung cancer” by Uramoto et al., 2010 published in Anticancer Research), in pancreatic cancers (see the publication “Targeting mechanoresponsive proteins in pancreatic cancer: 4-hydroxyacetophenone blocks dissemination and invasion by activating MYH14” by Surcel et al., 2018 published in Cancer Research) and in melanomas (see the publication “Filamin A expression correlates with proliferation and invasive properties of human metastatic melanoma tumors: implications for survival in patients” by Zhang et al., 2014 published in the Journal of Cancer Research and Clinical Oncology), showing the highest levels systematically associated with a poor prognosis in terms of survival, characterized by a higher number of metastases and earlier recurrence.

[0008] The decrease in the expression level of FlnA has been observed in prostate cancer (see the publication “Filamin A regulates MMP-9 expression and suppresses prostate cancer cell migration and invasion” by Sun et al., 2014 published in Tumor Biology), in colorectal adenocarcinomas (see the publication “New cancer suppressor gene for colorectal adenocarcinoma: Filamin A” by Tian et al., 2015 published in World Journal of Gastroenterology) as well as in gastric, renal and nasopharyngeal carcinomas (see the publications “An antiproliferative gene FLNA regulates migration and invasion of gastric carcinoma cell in vitro and its clinical significance” by Sun et al., 2014 published in Tumor Biology; “Interactions between Filamin A and MMP-9 Regulate Proliferation and Invasion in Renal Cell Carcinoma” by Sun et al., 2014 published in the Asian Pacific Journal of Cancer Prevention and “Absent expression of FLNA is correlated with poor prognosis of nasopharyngeal cancer” published by Sun et al., 2014 in the journal Tumor Biology). In these cases, decreased expression of FlnA is very often associated with a poor prognosis.

[0009] Taken together, the effects of FlnA expression variations in tumors suggest that this protein plays a central role in regulating mechanisms associated with tumorigenesis. It appears to be a good diagnostic and prognostic marker, potentially useful in tumor management. Thus, the use of FlnA as a rapid diagnostic marker of treatment response and / or recurrence may also be a strategy for non-invasive monitoring of therapeutic response during patient management.

[0010] Diffuse gliomas in adults are the most common and aggressive primary brain tumors, particularly anaplastic gliomas (grade III) and glioblastomas (GB). The standard first-line treatment used to date is the Stupp protocol (see the publication "Radiotherapy plus Concomitant and Adjuvant Temozolomide for Glioblastoma" by Stupp et al., 2005 published in The New England Journal of Medicine), which is based on surgical resection of the tumor as completely as possible, followed by concomitant radio-chemotherapy, and adjuvant chemotherapy with temolozomide (TMZ). However, recurrence is almost inevitable and occurs within 7 to 10 months after diagnosis, which explains the median patient survival of 15 to 17 months. This is mainly due to the highly invasive diffuse nature of GB which does not allow complete resection of the tumor, pathological neo-angiogenesis and resistance to treatments.

[0011] At relapse, no treatment can significantly improve patient survival and the use of chemotherapy is associated with response rates of approximately 5%. Although many different therapeutic strategies have been developed in recent years, unfortunately the many targeted therapies developed have not been effective in improving overall patient survival. It should be noted, however, that anti-angiogenic therapies (Bevacizumab, Afilbercept, Sunitib and Cediranib), including Bevacizumab, an antibody directed against VEGF-A, allow a response rate of 30% at relapse, as well as an increase in progression-free survival and quality of life for patients with GB.

[0012] Thus, although some progress has been made in the management of diffuse high-grade gliomas or GB, recurrence remains the rule, and must therefore be treated.

[0013] Invasion of healthy brain parenchyma by GB cells, at the edge or at a distance from the resection area, constitutes a major therapeutic challenge. These invasion mechanisms are driven by cellular transformations linked to environmental constraints such as hypoxia and angiogenesis, responsible for a mesenchymal transition (MT) mainly controlled by the transcription factors STAT3 and CEBPa which stimulate 70% of secondary mesenchymal genes. Hypoxia associated with TM notably leads to the overexpression of 7-transmembrane domain G protein-coupled receptors (GPCRs) of chemokines. The most widely studied chemotactic GPCR in GB is CXCR4 relaying its promigratory effects via couplings and the Gai / PI3K / Akt and Ga12 / 13 / Rho / ROCK / Myosin pathways as well as its clathrin-dependent endocytosis and recycling relayed by the activity of p-arrestins.Previously published work has also demonstrated that the UT receptor of the neuropeptide Ull behaves as a chemokine receptor that can stimulate the directional migration of GB cells via the sequential activation of Gai / PI3K then Ga13 / Rho / ROCK allowing cell polarization, the emission of lamellipodia, the polymerization of actin stress fibers, and cell contraction (See the publication “Signaling switch of the urotensin II vasosactive peptide GPCR: prototypic chemotaxic mechanism in glioma” by Lecointre et al., published in 2015 in Oncogene and the thesis summary of Mr. Alexandre Mutel accessible via the following link: https: / / www.theses.fr / 2019NORMR081).

[0014] Technical problem to be solved

[0015] An object of the present invention is to provide constructs comprising or consisting of (essentially) peptide sequence(s) capable of penetrating into cells and interacting with the UT receptor of the human neuropeptide Ull or the FlnA protein so as to inhibit the interaction between FlnA and the UT receptor.

[0016] Another aim of the invention is to provide constructions as mentioned above which can be used as a medicament or prognostic agent or diagnostic agent, in particular for cancer and more particularly for diffuse glioma, in particular in adults, anaplastic glioma (grade III) and glioblastoma.

[0017] Another aim of the invention is to propose constructions as mentioned above which make it possible to reduce or even prevent the invasion of healthy parenchyma relayed by the activation of the UT receptor, and other chemokine receptors following resection of a tumor of the diffuse glioma or glioblastoma type.

[0018] Another aim of the invention is to propose constructions as mentioned above which make it possible to counteract or limit resistance to current treatments for glioblastoma, namely in particular Temozolomide.

[0019] Another aim is to propose constructs that allow synergy of action with anticancer drugs in other peripheral cancers, notably tyrosine kinase inhibitors such as Gefitinib or DNA alkylating agents such as Temozolomide, particularly for the treatment of glioblastomas and lung, breast and prostate cancers.

[0020] Another aim of the present invention is to provide peptide sequences which make it possible to obtain the aforementioned constructions.

[0021] Brief summary of the invention

[0022] According to a first aspect, the present invention relates to a construct comprising a cell penetrating peptide sequence coupled by a peptide bond to at least one first linker molecule which is coupled by a peptide bond to the N-terminal end of an interfering amino acid sequence chosen from: a) sequences comprising or consisting of the following sequence: VTYCPTEPGN YIINIKFADQHVPGSPFSVKVTGEGRVKESITRRRRAPSVANVGSHCDLSLKIPEISIQD MTAQVTSPSGKTHEAEIVEGENHTYCIRFVPAE (SEQ ID NO: 1 or FlnA 2104-2206) which forms domain 2104-2206 of the human FlnA protein;b) sequences comprising or consisting of a fragment of the sequence SEQ ID NO: 1, said sequences or said fragments being capable of associating with the sequence VRGPGSGGGRGPVPSLQPRAR (SEQ ID NO: 2 or UT-330-350) which defines domain 330-350 of the UT receptor or with a fragment of this sequence SEQ ID NO: 2, in particular with the fragment of sequence GGGRGPVPSLQPRAR (SEQ ID NO: 3 or UT-336-350) forming domain 336-350 of the UT receptor or with the fragment RGPVPSLQPRAR (SEQ ID NO: 4 or UT-339-350) defining domain 339-350 of the UT receptor; c) sequences comprising or consisting of a fragment of the sequence SEQ ID NO: 2, in particular the fragment forming domain 330-350 of the UT receptor (SEQ ID NO: 2) or the fragment forming domain 336-350 of the UT receptor (SEQ ID NO: 3) or the fragment forming domain 339-350 of the UT receptor (SEQ ID NO: 4) and capable of associating with a fragment of the sequence SEQ ID NO: 1;(d) sequences having at least 90% identity and preferably at least 95% identity with the sequences defined in subparagraphs (a) to (c); and (e) sequences substantially homologous to the sequences defined in subparagraphs (a) to (d) and preferably deriving from the latter by one or more conservative substitutions.;

[0023] It is the inventors' merit to have shown that the above-mentioned constructs, some of which may be constructs, interfere with the Filamin A protein and the UT receptor of Ull and can thus be used as a drug. The above-mentioned constructs have in vitro activity on glioblastoma cells and prevent the spread of glioblastoma cancer cells after tumor resection.

[0024] Detailed description

[0025] According to a particular embodiment, the first linking molecule is an amino acid, optionally modified and in particular a beta-alanine molecule. The bond with the permeant sequence and the first linking molecule is a peptide bond and the bond between the first linking molecule and the interfering sequence is a peptide bond.

[0026] According to a particular embodiment, the interfering amino acid sequence is chosen from the amino acid sequences defined in paragraph b) and in particular from the following sequences:

[0027] MTAQVTSPSGKTHEAEIVE (SEQ ID NO: 5 or FlnA-D20-CD),

[0028] GNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 6 or FlnA-D19-FG),

[0029] THEAEIVEGENHTYCIRF (SEQ ID NO: 7 or FlnA-D20-DE), the peptide sequences having at least 90% and preferably at least 95% identity with said sequences and the substantially homologous peptide sequences, preferably deriving from the sequences SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7 by one or more conservative substitutions.

[0030] - RGPVPSLQPRAR (SEQ ID NO: 4 or UT-339-350) and GGGRGPVPSLQPRAR (SEQ ID NO: 3 or UT-336-350) and the peptide sequences having at least 90% and preferably at least 95% identity with said sequences SEQ ID NO: 3 and SEQ ID NO: 4 and the substantially homologous peptides, preferably deriving from the sequences SEQ ID NO: 3 or SEQ ID NO: 4 by one or more conservative substitutions.

[0031] The cell penetration sequence is not limited according to the invention. Advantageously, the cell penetration sequence is chosen from the following sequences:

[0032] -TCTWLKYH (SEQ ID NO: 8 or Glio) and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 8 and the substantially homologous sequences, preferably deriving from the sequence SEQ ID NO: 8 by one or more conservative substitutions;

[0033] - the peptide sequence GRRRQRRKKRKCCKRKKRRQRRRG (SEQ ID NO: 9 or CycloGlio) in which residues 12 and 13 (cysteine) are linked by a disulfide bridge and in which residues 2 to 10 and 15 to 23 are dextrorotatory (D) amino acids and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 9 and the substantially homologous sequences, preferably deriving from the sequence SEQ ID NO: 9 by one or more conservative substitutions, the two ends of said penetration sequence each being linked by a peptide bond to a first linking molecule which is an amino acid, optionally modified, said linking molecule itself being linked by a peptide bond to an interfering sequence, the two interfering sequences being identical or different.

[0034] These two penetration sequences exhibit rapid cell penetration kinetics; constructs containing these sequences enter cells within 5 minutes of the start of in vitro incubation.

[0035] The cell penetration sequence SEQ ID NO: 8 will reduce potential penetration into other cell types, and therefore undesirable side effects, when administered to the patient.

[0036] The cell penetration sequence SEQ ID NO: 9 or CycloGlio is a U-shaped molecule due to the disulfide bridge that links the two cysteine ​​molecules. It allows the synthesis of hairpin-shaped molecules. The two interfering sequences each linked to one end of the penetration sequence may or may not be identical. They may be, for example, two identical or different fragments of FlnA or two identical or different fragments of the UT receptor. It follows that when one arm is linked, either to FlnA or to the UT receptor, the other arm being free, it repels any molecule approaching FlnA or the UT receptor, respectively. There is therefore a double action of inhibition of the relationship between FlnA and the UT receptor: inhibition due to the binding with the construct of the invention and inhibition due to a steric effect due to the U-shape of the construct of the invention.

[0037] Advantageously, said cell penetration sequence is chosen from the peptide sequence GRRRQRRKKRKCCKRKKRRQRRRG (SEQ ID NO: 9 or CycloGlio) in which residues 12 and 13 (cysteine) are linked by a disulfide bridge and in which residues 2 to 10 and 15 to 23 are dextrorotatory (D) amino acids and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 9 and the substantially homologous sequences, preferably deriving from the sequence SEQ ID NO: 9 by one or more conservative substitutions, in that the first two linker molecules are identical and in particular beta alanine molecules; and one of the interfering sequences is chosen from the sequences defined in paragraphs a) and b) of claim 1, in particular from the following interfering sequences: MTAQVTSPSGKTHEAEIVE (SEQ ID NO: 5 or FlnA-D20-CD),

[0038] GNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 6 or FlnA-D19-FG), THEAEIVEGENHTYCIRF (SEQ ID NO: 7 or FlnA-D20-DE) and the peptide sequences having at least 90% and preferably at least 95% identity with said sequences and the substantially homologous sequences, preferably deriving from the sequences SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7 by one or more conservative substitutions, while the other interfering sequence is chosen from the sequences defined in paragraph c) of claim 1 and in particular from the following interfering sequences: RGPVPSLQPRAR (SEQ ID NO: 4 or UT-339-350) and GGGRGPVPSLQPRAR (SEQ ID NO: 3 or UT-336-350) and the peptide sequences having at least 90% and preferably at least 95% identity with said sequences SEQ ID NO: 3 and SEQ ID NO: 4 and the substantially homologous sequences, preferably deriving from the sequences SEQ ID NO: 3 or SEQ ID NO: 4 by one or more conservative substitutions.Furthermore, this penetration sequence SEQ ID NO: 9 allows endosomal release of the construction of the invention, allowing cytosolic incorporation greater than 90%, thereby potentiating the interfering effects of the interfering sequences.

[0039] Advantageously, when said cell penetration sequence is chosen from the peptide sequence GRRRQRRKKRKCCKRKKRRQRRRG (SEQ ID NO: 9 or CycloGlio) in which residues 12 and 13 (cysteine) are linked by a disulfide bridge and in which residues 2 to 10 and 15 to 23 are dextrorotatory (D) amino acids and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 9 and the substantially homologous sequences, preferably deriving from the sequence SEQ ID NO: 9 by one or more conservative substitutions, the lysines in positions 11 and 14 of said cell penetration sequence are acetylated. This acetylation avoids any risk of peptide bonding with another sequence, since lysines still have a free amine function, they can still bond even when they are part of a peptide sequence.

[0040] The present invention also relates to an interfering construct selected from the following sequence constructs: TVKVSFPSGPVHQDAFKINHYNGXGrrrqrrkkrKCCKrkkrrqrrrGXGNYIINIKFADQHVPGS PFSVKVT (SEQ ID: 10, also referred to as FlnA-CPPD19-FG-CycloGlio) in which residues 37 and 38 are linked by a disulfide bridge, X is in both positions an optionally modified amino acid and preferably beta-alanine in both positions and residues 27 to 35 and 40 to 48 are dextrorotatory (D) amino acids and in which the lysines in positions 36 and 39 are acetylated;

[0041] EVIEAEHTKGSPSTVQATMXGrrrqrrkkrKCCKrkkrrqrrrGXMTAQVTSPSGKTHEAElVE (SEQ ID NO: 11; also referred to as FlnA-CPPD20-CD-CycloGlio) wherein residues 32 and 33 are linked by a disulfide bridge, X is an optionally modified amino acid and preferably beta-alanine in both positions and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and wherein the lysines at positions 31 and 34 are acetylated; FRISYTHNEGEVIEAEHTXGrrrqrrkkrKCCKrkkrrqrrrGXTHEAEIVEGENHTYSIRF (SEQ ID NO: 12; also referred to as FlnA-CPPD20-DE-CycloGlio) wherein residues 31 and 32 are linked by a disulfide bridge, X is an optionally modified amino acid and preferably beta-alanine and residues 21 to 29 and 34 to 42 are dextrorotatory (D) amino acids; wherein the lysines at positions 30 and 33 are acetylated; TCTWLKYHXGNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 13;also referred to as FlnA-CPPD19-FG-Glio) in which X is an optionally modified amino acid and preferably beta-alanine;

[0042] TCTWLKYHXMTAQVTSPSGKTHEAEIVE (SEQ ID NO: 14; also referred to as FlnA-CPPD20-CD-Glio) wherein X is an optionally modified amino acid and preferably beta-alanine;

[0043] TCTWLKYHXTHEAEIVEGENHTYCIRF (SEQ ID NO: 15; also referred to as FlnA-CPPD20-DE-Glio) wherein X is an optionally modified amino acid and preferably beta-alanine;

[0044] RARPQLSPVPGRGGGXGrrrqrrkkrKCCKrkkrrqrrrGXGGGRGPVPSLQPRAR (SEQ ID NO: 16, also referred to as UT-CPP336-350-CycloGlio) wherein residues 28 and 29 are linked by a disulfide bridge, X is an optionally modified amino acid and preferably beta-alanine in both positions and residues 18 to 26 and 31 to 39 are dextrorotatory (D) amino acids wherein the lysines at positions 27 and 30 are acetylated;

[0045] RARPQLSPVPGRXGrrrqrrkkrKCCKrkkrrqrrrGXRGPVPSLQPRAR SEQ ID NO: 17; also referred to as UT-CPP339-350-CycloGlio) wherein residues 25 and 26 are linked by a disulfide bridge, X is an optionally modified amino acid and preferably beta-alanine in both positions and residues 15 to 23 and 28 to 36 are dextrorotatory (D) amino acids and wherein the lysines at positions 24 and 27 are acetylated,

[0046] TCTWLKYHXGGGRGPVPSLQPRAR (SEQ ID NO: 18; also referred to as UT-CPP336-350-Glio) wherein X is an optionally modified amino acid and preferably beta-alanine; - TCTWLKYHXRGPVPSLQPRAR (SEQ ID NO: 19; also referred to as UT-CPP339-350-Glio) wherein X is an optionally modified amino acid and preferably beta-alanine;

[0047] - constructs exhibiting at least 90% identity and preferably at least 95% identity with the aforementioned constructs; and constructs which are substantially homologous and preferably derived from the sequences SEQ ID NO: 10 to SEQ ID NO: 19 by one or more conservative substitutions.

[0048] The constructs of sequence SEQ ID NO: 19, SEQ ID NO: 11, SEQ ID NO: 18, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 13, SEQ ID NO: 17 prove to be particularly effective on several types of solid cancer (glioblastoma, prostate, lung and breast) with regard to cell penetration and inhibition of cancer cell migration. The same is true for the marked constructs containing these sequences.

[0049] In particular, the present invention relates to the following sequence constructs: The present invention also relates to the following sequence constructs: TVKVSFPSGPVHQDAFKINHYNGAGrrrqrrkkrKCCKrkkrrqrrrGAGNYIINIKFADQHVPGS PFSVKVT (SEQ ID: 28, also referred to as FlnA-CPPD19-FG-CycloGlio-syn) wherein residues 37 and 38 are disulfide bonded, alanines at positions 25 and 50 are beta-alanine and residues 27 to 35 and 40 to 48 are dextrorotatory (D) amino acids and wherein lysines at positions 36 and 39 are acetylated;

[0050] EVIEAEHTKGSPSTVQATMAGrrrqrrkkrKCCKrkkrrqrrrGAMTAQVTSPSGKTHEAElVE (SEQ ID NO: 29; also referred to as FlnA-CPPD20-CD-CycloGlio-syn) wherein residues 32 and 33 are disulfide bonded, alanines at positions 20 and 45 are beta-alanine, and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids; and wherein lysines at positions 31 and 34 are acetylated;

[0051] FRISYTHNEGEVIEAEHTAGrrrqrrkkrKCCKrkkrrqrrrGATHEAElVEGENHTYSIRF (SEQ ID NO: 30; also referred to as FlnA-CPPD20-DE-CycloGlio-syn) wherein residues 31 and 32 are disulfide bonded, alanines at positions 19 and 44 are beta-alanine, and residues 21 to 29 and 34 to 42 are dextrorotatory (D) amino acids; TCTWLKYHAGNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 31; also referred to as FlnA-CPPD19-FG-Glio-syn) wherein alanine at position 9 is beta-alanine) and wherein lysines at positions 30 and 33 are acetylated;

[0052] TCTWLKYHAMTAQVTSPSGKTHEAEIVE (SEQ ID NO: 32; also referred to as FlnA-CPPD20-CD-Glio-syn) wherein the alanine at position 9 is a beta-alanine;

[0053] TCTWLKYHATHEAEIVEGENHTYCIRF (SEQ ID NO: 33; also referred to as FlnA-CPPD20-DE-Glio-syn) wherein the alanine at position 9 is a beta-alanine;

[0054] RARPQLSPVPGRGGGAGrrrqrrkkrKCCKrkkrrqrrrGAGGGRGPVPSLQPRAR (SEQ ID NO: 34; also referred to as UT-CPP336-350-CycloGlio-syn) wherein residues 28 and 29 are disulfide bonded, alanines at positions 16 and 41 are beta-alanine, and residues 18 to 26 and 31 to 39 are dextrorotatory (D) amino acids, and wherein lysines at positions 27 and 30 are acetylated;

[0055] RARPQLSPVPGRAGrrrqrrkkrKCCKrkkrrqrrrGARGPVPSLQPRAR (SEQ ID NO: 35; also referred to as UT-CPP339-350-CycloGlio-syn) wherein residues 25 and 26 are disulfide bonded, alanines at positions 13 and 38 are beta-alanines and residues 15 to 23 and 28 to 36 are dextrorotatory (D) amino acids and wherein lysines at positions 24 and 27 are acetylated;

[0056] TCTWLKYHAGGGRGPVPSLQPRAR (SEQ ID NO: 36; also referred to as UT-CPP336-350-Glio-syn) wherein the alanine at position 9 is a beta-alanine;

[0057] - TCTWLKYHARGPVPSLQPRAR (SEQ ID NO: 37; also referred to as UT-CPP339-350-Glio-syn) wherein the alanine at position 9 is a beta-alanine; the sequences having at least 90% identity and, preferably, at least 95% identity with the aforementioned sequences and the sequences substantially homologous to the aforementioned sequences and preferably deriving from the latter by one or more conservative substitutions.

[0058] When the constructs comprise the Cycloglio sequence (SEQ ID NO: 9) or a sequence identical to 90% or 95% of this sequence or a substantially homologous sequence and preferably deriving from this sequence by one or more conservative substitutions, the lysines located on either side of the disulfide bridge are preferably acetylated, that is to say linked to a CH3CO radical, when they are not linked to a molecule of a labeling agent. The construct of sequence SEQ ID NO: 35 or UT-339-350-CycloGlio provides effects which are systematically found in various glioblastoma cell lines.This interfering and penetrating construct (peptide) inhibits the motility of glioblastoma cells, chemotactic migration in response to UH of glioblastoma cells, reduces the viability of glioblastoma cells when applied alone or in combination with TMZ (Temozolomide) chemotherapy, reduces the viability and promotes the effects of chemotherapy with Gefitinib on a lung cancer cell line.

[0059] The constructs of sequences SEQ ID NO: 37, SEQ ID NO: 47, SEQ ID NO: 29, SEQ ID NO: 39, SEQ ID NO: 36, SEQ ID NO: 46, SEQ ID NO: 32, SEQ ID NO: 42, SEQ ID NO: 33, SEQ ID NO: 43, SEQ ID NO: 31, SEQ ID NO: 41, SEQ ID NO: 35 and SEQ ID: 45 are particularly effective on several types of solid cancer (glioblastoma, prostate, lung and breast) with regard to cell penetration and inhibition of cancer cell migration.

[0060] The present invention also relates to constructs as mentioned above which further comprise a labeling agent capable of penetrating into the cell. In particular, the penetration sequence is linked to at least one labeling agent. The labeling agent may be a fluorophore.

[0061] The labeling agent is preferably a particular permeant fluorophore, 5-carboxytetramethylrhodamine or 7-diethylaminocoumarin-3-carboxylic acid.

[0062] Advantageously, when the penetration sequence is chosen from the peptide sequence GRRRQRRKKRKCCKRKKRRQRRRG (SEQ ID NO: 9 or CycloGlio) in which residues 12 and 13 (cysteine) are linked by a disulfide bridge and in which residues 2 to 10 and 15 to 23 are dextrorotatory (D) amino acids and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 9 and the substantially homologous sequences, preferably deriving from the sequence SEQ ID NO: 9 by one or more conservative substitutions, a molecule of said labeling agent is linked to the lysines in position 11 and 14 and said labeling agent is in particular and preferably 5-carboxytetramethylrhodamine.Thus, the present invention also relates to a construct which comprises or consists of a peptide sequence chosen from the sequences SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 18, SEQ ID NO: 19, the constructs having at least 90% identity and preferably at least 95% identity with the abovementioned constructs and the constructs which are substantially homologous and preferably derived from the abovementioned sequences by one or more conservative substitutions in which the first residue of the permeant sequence is linked by a peptide bond to the carboxylic acid function of beta alanine which is linked by a peptide bond with the carboxyl group of 7-diethylaminocoumarin-3-carboxylic acid.

[0063] The present invention also relates to a construct which comprises or consists of a peptide sequence selected from the sequences SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 16, SEQ ID NO: 17, the constructs having at least 90% identity and preferably at least 95% identity with the abovementioned constructs and the constructs which are substantially homologous and preferably derived from the abovementioned sequences by one or more conservative substitutions in which residues 11 and 14 (K: lysine) of the permeant sequence are each linked by a peptide bond with the carboxylic acid group of 5-carboxytetramethylrhodamine.

[0064] Thus, the present invention also relates to the following sequences, which can be used for the treatment and diagnosis / prognosis of the pathologies mentioned below: TVKVSFPSGPVHQDAFKINHYNGAGrrrqrrkkrKCCKrkkrrqrrrGAGNYIINIKFADQHVPGS PFSVKVT (SEQ ID: 38, also called Mq-FlnA-CPPD19-FG-CycloGlio-syn) in which residues 37 and 38 are linked by a disulfide bridge, the alanines at positions 25 and 50 are beta-alanine and residues 27 to 35 and 40 to 48 are dextrorotatory (D) amino acids and residues 36 and 39 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine;

[0065] EVIEAEHTKGSPSTVQATMAGrrrqrrkkrKCCKrkkrrqrrrGAMTAQVTSPSGKTHEAElVE (SEQ ID NO: 39; also referred to as Mq-FlnA-CPPD20-CD-CycloGlio-syn) wherein residues 32 and 33 are linked by a disulfide bridge, the alanines at positions 20 and 45 are beta-alanine and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and residues 31 and 34 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine;

[0066] FRISYTHNEGEVIEAEHTAGrrrqrrkkrKCCKrkkrrqrrrGATHEAEIVEGENHTYSIRF (SEQ ID NO: 40; also referred to as Mq-FlnA-CPPD20-DE-CycloGlio-syn) wherein residues 31 and 32 are linked by a disulfide bridge, the alanines at positions 19 and 44 are beta-alanine and residues 21 to 29 and 34 to 42 are dextrorotatory (D) amino acids and residues 30 and 33 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine;

[0067] ATCTWLKYHAGNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 41; also referred to as Mq-FlnA-CPPD19-FG-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid;

[0068] ATCTWLKYHAMTAQVTSPSGKTHEAEIVE (SEQ ID NO: 42; also referred to as Mq-FlnA-CPPD20-CD-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid;

[0069] ATCTWLKYHATHEAEIVEGENHTYCIRF (SEQ ID NO: 43; also referred to as Mq-FlnA-CPPD20-DE-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid;

[0070] RARPQLSPVPGRGGGAGrrrqrrkkrKCCKrkkrrqrrrGAGGGRGPVPSLQPRAR (SEQ ID NO: 44; also referred to as Mq-UT-CPP336-350-CycloGlio-syn) wherein residues 28 and 29 are disulfide bonded, alanines at positions 16 and 41 are beta-alanine, and residues 18 to 26 and 31 to 39 are dextrorotatory (D) amino acids, and residues 27 and 30 are each peptide bonded to the carboxylic acid group of tetramethylrhodamine; RARPQLSPVPGRAGrrrqrrkkrKCCKrkkrrqrrrGARGPVPSLQPRAR SEQ ID NO: 45; also referred to as Mq-UT-CPP339-350-CycloGlio-syn) in which residues 25 and 26 are linked by a disulfide bridge, the alanines at positions 13 and 38 are beta-alanine and residues 15 to 23 and 28 to 36 are dextrorotatory (D) amino acids residues 24 and 27 are each linked by a peptide bond to the carboxylic acid group of tetramethylrhodamine,

[0071] ATCTWLKYHAGGGRGPVPSLQPRAR (SEQ ID NO: 46; also referred to as Mq-UT-CPP336-350-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid;

[0072] ATCTWLKYHARGPVPSLQPRAR (SEQ ID NO: 47; also referred to as Mq-UT-CPP339-350-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid;

[0073] - sequences having at least 90% identity and, preferably, at least 95% identity with the aforementioned sequences; and sequences substantially homologous to the aforementioned sequences and preferably deriving from the latter by one or more conservative substitutions.

[0074] The present invention also relates to a construct according to the invention for its use as a medicament or diagnostic / prognostic agent, in particular for cancer, in particular solid cancerous tumors, breast cancer, lung cancer more particularly in the treatment of pulmonary adenocarcinomas, melanomas, prostate cancer, colon cancer in particular colorectal adenocarcinomas, stomach cancer, in particular gastric carcinomas, kidney cancer, in particular renal carcinomas, nasopharyngeal cancers in particular nasopharyngeal carcinomas, pancreatic cancer, diffuse glioma and glioblastoma in a patient / subject having such a need, in particular a human. Y1

[0075] The present invention also relates to a pharmaceutical composition which comprises, as active ingredient (active principle), at least one construction according to the invention and a pharmaceutically acceptable solvent, excipient and / or vehicle.

[0076] According to a particular embodiment, the pharmaceutical composition of the invention may also comprise as a second active ingredient, an anti-cancer agent chosen in particular from tyrosine kinase inhibitors, in particular Gefitinib and alkylating agents causing double-strand breaks in DNA and cell death, in particular TMZ for their simultaneous, delayed or sequenced use.

[0077] The composition of the invention may be in the form of a solid, in particular a powder, a gel or an injectable solution, possibly colloidal.

[0078] The composition can be taken orally, injected intravenously or administered intranasally after tumor resection, partly in the case of glioblastomas, concomitantly or at a later time than the injection of a chemotherapy product and / or radiotherapy.

[0079] When the pharmaceutical composition of the invention is in liquid, gel, or solid form, it can be introduced into the space created by the resection of the tumor, in particular in the case of blastomas and glioblastomas, before the patient is treated with radiotherapy and / or chemotherapy. The constructs and peptides of the invention will then penetrate into the surrounding cells and block the invasion of the glioblastoma.

[0080] The composition of the invention can also be injected into a tumor.

[0081] The present invention also relates to a nucleic acid coding for one of the sequences corresponding to the sequences according to the invention, in particular for a construction chosen from the following sequences: SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7.

[0082] The present invention also relates to a vector comprising such a nucleic acid, preferably an adenoviral or lentiviral vector.

[0083] The present invention also relates to a combination product (kit) containing at least one construct according to the invention and in particular a construct chosen from the constructs of sequence SEQ: ID NO: 16, SEQ ID NO: 34, SEQ ID NO: 17, SEQ ID NO: 35, SEQ ID NO: 10, SEQ ID NO: 28, SEQ ID NO: 11, SEQ ID NO: 29, SEQ ID NO: 12, SEQ ID NO: 30, SEQ ID NO: 18, SEQ ID NO: 36, SEQ ID NO: 19, SEQ ID NO: 37, SEQ ID NO: 13, SEQ ID NO: 31, SEQ ID NO: 14, SEQ ID NO: 32, SEQ ID NO: 15, SEQ ID NO: 33 and an anticancer agent chosen in particular from tyrosine kinase inhibitors, in particular Gefitinib and alkylating agents causing double-strand breaks in DNA and cell death, particularly TMZ, the use of the construct and the anti-cancer agent may be simultaneous, delayed or sequenced.

[0084] The present invention also discloses the following sequences which are used for comparison: ALQQSRLPPQVSPQDGALAAGrrrqrrkkrKCCKrkkrrqrrrGAALAGDQPSVQPPLRSQQLA (SEQ ID NO: 20 or FlnA-H1a-Cycloglio) in which residues 32 and 33 are linked by a disulfide bridge, the alanine at position 20 and position 45 is a beta alanine (in both positions) and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids; WTQQGGQAYTYQPALQQSRAGrrrqrrkkrKCCKrkkrrqrrrGARSQQLAPQYTYAQGGQQT W (SEQ ID NO: 21 or FlnA-H1 b-Cycloglio) in which residues 32 and 33 are linked by a disulfide bridge, the alanine at positions 20 and 45 is a beta alanine (in both positions) and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids;AKTLSDVFVSSTEHLSHNSAGrrrqrrkkrKCCKrkkrrqrrrGASNHSLHETSSVFVDSLTKA (SEQ ID NO: 22 or FlnA-H2a-CycloGlio) in which residues 32 and 33 are linked by a disulfide bridge, the alanine at position 20 and that at position 45 is a beta alanine (in both positions) and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids;

[0085] PGPGPAGHQPASTAKTLSDAGrrrqrrkkrKCCKrkkrrqrrrGADSLTKATSAPQHGAPGPGP (SEQ ID NO: 23 or FlnA-H2b-CycloGlio) wherein residues 32 and 33 are linked by a disulfide bridge, the alanine at position 20 and that at position 45 is a beta alanine and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids;

[0086] TCTWLKYHAALAGDQPSVQPPLRSQQLA (SEQ ID NO: 24 or FlnA-H1a-Glio) wherein the alanine at position 9 is a beta-alanine.

[0087] TCTWLKYHARSQQLAPQYTYAQGGQQTW (SEQ ID NO: 25 or FlnA-H1 b-Glio) wherein the alanine at position 9 is a beta-alanine.

[0088] TCTWLKYHASNHSLHETSSVFVDSLTKA (SEQ ID NO: 26 or FlnA-H2a-Glio) wherein the alanine at position 9 is a beta-alanine. TCTWLKYHADSLTKATCAPQHGAPGPGP (SEQ ID NO: 27 or FlnA-H2b-Glio) wherein the alanine at position 9 is a beta-alanine.

[0089] The present invention discloses, for comparison, the following constructs: ALQQSRLPPQVSPQDGALAAGrrrqrrkkrKCCKrkkrrqrrrGAALAGDQPSVQPPLRSQQLA (SEQ ID NO: 48 or Mq-FlnA-H1a-Cycloglio-syn) wherein residues 32 and 33 are linked by a disulfide bridge, the alanines at positions 20 and 45 are beta-alanine and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and residues 31 and 34 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine,

[0090] WTQQGGQAYTYQPALQQSRAGrrrqrrkkrKCCKrkkrrqrrrGARSQQLAPQYTYAQGGQQT W (SEQ ID NO: 49 or Mq-FlnA-H1 b-Cycloglio-syn) wherein residues 32 and 33 are linked by a disulfide bridge, the alanines at positions 20 and 45 are beta-alanine and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and residues 31 and 34 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine;

[0091] AKTLSDVFVSSTEHLSHNSAGrrrqrrkkrKCCKrkkrrqrrrGASNHSLHETSSVFVDSLTKA (SEQ ID NO: 50 or Mq-FlnA-H2a-CycloGlio-syn) wherein residues 32 and 33 are linked by a disulfide bridge, the alanines at positions 20 and 45 are beta-alanine and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and residues 31 and 34 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine;

[0092] PGPGPAGHQPASTAKTLSDAGrrrqrrkkrKCCKrkkrrqrrrGADSLTKATSAPQHGAPGPGP (SEQ ID NO: 51 or Mq-FlnA-H2b-CycloGlio-syn) wherein residues 32 and 33 are linked by a disulfide bridge, the alanines at positions 20 and 45 are beta-alanine and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and residues 31 and 34 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine;

[0093] ATCTWLKYHAALAGDQPSVQPPLRSQQLA (SEQ ID NO: 52 or Mq-FlnA-H1a-Glio- syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid; ATCTWLKYHARSQQLAPQYTYAQGGQQTW (SEQ ID NO: 53 or Mq-FlnA-H1 b-Glio- syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid;

[0094] ATCTWLKYHASNHSLHETSSVFVDSLTKA (SEQ ID NO: 54 or Mq-FlnA-H2a-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid;

[0095] ATCTWLKYHADSLTKATCAPQHGAPGPGP (SEQ ID NO: 55 or Mq-FlnA-H2b-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid.

[0096] Definitions

[0097] The term “patient / subject” refers throughout this application to a mammal, male or female, adult or not (child or adolescent) and particularly to a human.

[0098] The term "treatment" includes curative and preventive treatment. Curative treatment consists of at least one of the following: alleviating, improving and / or eliminating, reducing or stabilizing at least one symptom as well as delaying the progression or onset of a symptom of an altered health condition. Preventive treatment consists of at least one of the following: avoiding the deterioration of the health condition, reducing the risk of onset of the altered health condition, reducing the incidence, development and delaying the onset of at least one symptom of an altered health condition.

[0099] The term "penetrating sequence" refers to a peptide or peptide sequence capable of transferring into a cell without causing substantial damage to the cell membrane and which can be used as a vector for other molecules to which it is bound.

[0100] The term "interfering sequence" refers to a peptide sequence capable of binding to a given peptide sequence possibly forming part of a protein and thus capable of modifying the mechanisms in which the given peptide sequence or the protein of which the latter is a part is involved. The term "sequence derived from" refers to a peptide sequence longer than the sequence from which it is derived or a sequence homologous to the sequence from which it is derived.

[0101] The term "percent identity" refers to the number of identical residues in two sequences to be compared, obtained after the best alignment, this percentage being purely statistical and the differences between the two sequences being distributed randomly and over their entire length. By "best alignment" or "optimal alignment" is meant the alignment for which the percentage identity determined as below is the highest. Sequence comparisons between two are traditionally carried out by comparing these sequences after having aligned them optimally, said comparison being carried out by segment or by "comparison window" to identify and compare local regions of sequence similarity. For example, the BLOSUM matrices and the local homology algorithm of Smith and Waterman (1981, J. Mol Evol., 18:38-46) can be used. Any other known method can also be used.

[0102] The term "conservative substitution" refers to the exchange of one amino acid for another without altering the overall conformation of the peptide and without altering the peptide's function. Amino acids with similar properties are well known. For example, arginine, histidine, and lysine are hydrophilic and basic amino acids that are interchangeable. Isoleucine can be replaced by leucine, methionine, or valine. Asparagine, glutamine, serine, and threonine can be interchangeable.

[0103] The term "essentially peptide" means that more than 50% of the bonds connecting amino acids are peptide bonds but that some residues may be linked to molecules other than amino acids or to amino acids but for example by a disulfide bridge.

[0104] The acronym GB stands for glioblastoma.

[0105] The term "pharmaceutically acceptable" refers to compounds, materials, excipients, compositions or dosage forms which are, within the scope of good medical judgment, suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic response or other problematic complications corresponding to an acceptable benefit / risk ratio.

[0106] The terms "treat", "treated" or "treatment", as used in the context of the invention, refer to a therapeutic treatment whose aim is to eliminate or alleviate symptoms.

[0107] Beneficial or desired clinical outcomes include, but are not limited to, elimination of symptoms, alleviation of symptoms, decrease in the extent of the condition, stabilized (i.e., not worsened) health status, delay or slowing of the progression of the condition.

[0108] A therapeutically effective amount can be readily determined by the treating diagnostician, as a person skilled in the art, using conventional techniques and observing results obtained under analogous circumstances.

[0109] In determining the therapeutically effective amount, a number of factors are considered by the attending diagnostician, including, but not limited to: the species of the subject; the subject's size, age, and general health; the specific disease involved; the degree of involvement or severity of the disease; the response of the individual subject; the particular compound administered; the mode of administration; the bioavailability characteristic of the preparation administered; the dosage regimen selected; the use of concomitant medications; and other relevant circumstances.

[0110] The term “active ingredient” indicates that the molecule is present in a therapeutically effective quantity.

[0111] The association between the peptide and the protein can be measured by any method known to the person skilled in the art, for example, dual polarization interferometry, static light scattering (SLS), light scattering (DLS), surface plasmon resonance (SPR), fluorescence polarization / anisotropy, fluorescence correlation spectrometry, fluorescent intermolecular energy transfer (FRET), isothermal titration calorimetry, microscale thermophoresis, Figures

[0112] Fig. 1 schematically represents the mechanism of the interaction between FlnA and the UT receptor. Figs. 2 and 3 schematically represent the mechanism of the inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the UT-CPP336-350-Cycloglio family (namely the constructs corresponding to the sequences SEQ ID NO: 16, SEQ ID NO: 34 and SEQ ID NO: 44) and the results obtained; Figs. 4 and 5 schematically represent the mechanism of the inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the UT-CPP339-350-Cycloglio family (namely the constructs corresponding to the sequences SEQ ID NO: 17, SEQ ID NO: 35 and SEQ ID NO: 45) and the results obtained; Figs.6 and 7 schematically represent the mechanism of inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the FlnA-CPPD19-FG-Cycloglio- family (namely the constructs corresponding to the sequences SEQ ID NO: 10, SEQ ID NO: 28 and SEQ ID NO: 38) as well as the results obtained;.

[0113] Figs. 8 and 9 schematically represent the mechanism of inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the FlnA-CPPD20-CD-Cycloglio- family (namely the constructs corresponding to the sequences SEQ ID NO: 11, SEQ ID NO: 29 and SEQ ID NO: 39) as well as the results obtained;

[0114] Figs. 10 and 11 schematically represent the mechanism of inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the FlnA-CPPD20-DE-Cycloglio- family (namely the constructs corresponding to the sequences SEQ ID NO: 12, SEQ ID NO: 30 and SEQ ID NO: 40) and the results obtained; Figs. 12 and 13 schematically represent the mechanism of inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the UTCPP339-350-Glio- family (namely the constructs corresponding to the sequences SEQ ID NO: 19, SEQ ID NO: 37 and SEQ ID NO: 47) and the results obtained; Figs.14 and 15 schematically represent the mechanism of inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the UTCPP336-350-Glio- family (namely the constructs corresponding to the sequences SEQ ID NO: 18, SEQ ID NO: 36 and SEQ ID NO: 46) and the results obtained; Figs. 16 and 17 schematically represent the mechanism of inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the FlnACPP-D19-FG-Glio- family (namely the constructs corresponding to the sequences SEQ ID NO: 13, SEQ ID NO: 31 and SEQ ID NO: 41) and the results obtained; Figs.18 and 19 schematically represent the mechanism of inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the FlnACPP-D20-CD-Glio- family (namely the constructs corresponding to the sequences SEQ ID NO: 14, SEQ ID NO: 32 and SEQ ID NO: 42) and the results obtained; Figs. 20 and 21 schematically represent the mechanism of inhibition of the interaction between FlnA and the UT receptor provided by the constructs of the invention of the FlnACPP-D20-DE-Glio- family (namely the constructs corresponding to the sequences SEQ ID NO: 15, SEQ ID NO: 33 and SEQ ID NO: 43) and the results obtained) and the results obtained;.

[0115] EXAMPLES

[0116] Description of examples of implementation

[0117] Summaries of the constructions according to the invention as well as constructions used for comparison.

[0118] The syntheses of the compounds corresponding to the following sequences: SEQ ID NO: 9, SEQ ID NO: 28, SEQ ID NO: 38, SEQ ID NO: 29, SEQ ID NO: 39, SEQ ID NO: 30,

[0119] SEQ ID NO: 40, SEQ ID NO: 34, SEQ ID NO: 44, SEQ ID NO: 35, SEQ ID NO: 45,

[0120] SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 48,

[0121] SEQ ID NO: 49, SEQ ID NO: 50 and SEQ ID NO: 51 are performed on 2-chlorotrityl chloride resin using a Symphony-X automated synthesizer (Gyros Protein Technologies) according to the Fmoc strategy. Briefly, the C-terminal amino acid of each sequence is anchored on 2-chlorotrityl resin to obtain a substitution rate of approximately 0.25 mmol / g in the presence of diisopropylethylamine in dichloromethane. Subsequently, the excess reactive sites are quenched by treatment with methanol in the presence of diisopropylethylamine. These resins are used for automated synthesis according to the protocols recommended by the Symphony-X manufacturer. For each synthesis cycle, deprotections are performed using 20% ​​piperidine in dimethylformamide (DMF) for 2 treatments of 5 minutes.After washing the resin with DMF (4x), the couplings were performed by treating with a mixture of the amino acid Fmoc (5 eq), HAUT (5 eq), pure oxyma (5 eq) and diisopropylethylamine (15 eq) for 1 hour. Finally, the resin was washed with DMF (4x). The last two amino acids of the sequences that were coupled were Fmoc-Lys(Dde)-OH and Boc-Cys(Trt)-OH. Thus, the Dde protecting group was cleaved while preserving the peptides on resin by treatment 3 x 2 minutes with 2% hydrazine in DMF. The desired carboxy tetramethylrhodamine was optionally coupled on the side chains of the N-terminal lysines. Monomeric peptides were cleaved by treatment with TFA / water / TIPS / EDT (92.5%, 2.5%, 2.5%, 2.5%) for 5 hours and precipitated in ether. Once centrifuged and decanted, the peptides were dissolved in water and purified by reverse-phase preparative HPLC.Pure monomeric peptides are dissolved in 50 mmol ammonium bicarbonate buffered to pH 10 with dilute ammonium hydroxide. They are stirred overnight in the presence of atmospheric oxygen to provide the desired symmetrical dimers. The solutions are finally lyophilized.

[0122] The syntheses of the compounds corresponding to the following sequences: SEQ ID NO: 8, SEQ ID NO: 31, SEQ ID NO: 41, SEQ ID NO: 32, SEQ ID NO: 42, SEQ ID NO: 33,

[0123] SEQ ID NO: 43, SEQ ID NO: 36, SEQ ID NO: 46 and SEQ ID NO: 37, SEQ ID NO: 47,

[0124] SEQ ID NO: 24, SEQ ID NO: 52, SEQ ID NO: 25 and SEQ ID NO: 53, SEQ ID NO: 26,

[0125] SEQ ID NO: 54, SEQ ID NO: 27 and SEQ ID NO: 55, are performed on 2-chlorotrityl chloride resin using a Symphony-X automated synthesizer (Gyros Protein Technologies) according to the Fmoc strategy. Briefly, the C-terminal amino acid of each sequence was anchored on 2-chlorotrityl resin to obtain a substitution rate of approximately 0.25 mmol / g in the presence of diisopropylethylamine in dichloromethane. Subsequently, the excess reactive sites are deactivated by treatment with methanol in the presence of diisopropylethylamine. These resins are used for automated synthesis according to the protocols recommended by the Symphony-X manufacturer. For each synthesis cycle, deprotections were performed using 20% ​​piperidine in dimethylformamide (DMF) for 2 treatments of 5 minutes.After washing the resin with DMF (4x), the couplings are performed by treating with a mixture of the amino acid Fmoc (5 eq), HAUT (5 eq), pure oxyma (5 eq) and diisopropylethylamine (15 eq) for 1 hour. Finally, the resin was washed with DMF (4x). The last coupled acid is DEAC if necessary. The peptides are cleaved by treatment with TFA / water / TIPS / EDT (92.5%, 2.5%, 2.5%, 2.5%) for 3 hours and precipitated in ether. Once centrifuged and decanted, the peptides are dissolved in water and purified by reverse-phase preparative HPLC.

[0126] Similarly, it is possible to synthesize a construct from the cycloglio family that includes two different interfering sequences, one capable of binding to the UT receptor and the other to FlnA.

[0127] In vitro biological effects of the constructs of the invention

[0128] Example 1: Study of the incorporation of the construct SEQ ID NO: 35 mimicking a UT sequence binding FlnA in the 8MG glioblastoma (GB), A549 lung cancer, MDA-MB-231 breast cancer and LnCAP prostate cancer lines. At t0 100,000 cells of each line are seeded in the wells of a 12-well plate previously coated with fibronectin (2 μg / ml; 1 h; 37°C). After 24 hours of adhesion, the cells are treated with 100 nM of the construct SEQ ID NO: 35 which carries 5-carboxytetramethylrhodamine groups grafted onto the K2 residues (construct of sequence SEQ ID NO: 35) which will allow the incorporation of the peptide to be followed by video microscopy during 12 hours of acquisition.

[0129] Table 1 groups together the results relating to the penetration kinetics of the construct SEQ ID NO 35 in the 8MG glioblastoma, A549 lung cancer, MDA-MB-231 breast cancer and LnCAP prostate cancer lines.

[0130] Table 1

[0131] The construct of sequence SEQ ID NO: 35 mimicking a sequence of the FlnA-binding UT rapidly penetrates into the glioblastoma 8MG, lung cancer A549, breast cancer MDA-MB-231 and prostate cancer LnCAP lines with a first penetration time of less than 3 min. In addition, this construct of sequence SEQ ID NO: 35 diffuses completely into the cytosol of the solid cancer cells tested with very similar kinetics between 303 min and 416.3 min, thus the construct of sequence SEQ ID NO: 35 rapidly penetrates into the subcellular compartment in which FlnA and UT interact and can in fact relay its interfering effects rapidly after administration. In addition, the construct of sequence SEQ ID NO: 35 persists in the cytosol of all solid cancer lines at least 774 min after treatment, allowing the interfering properties of this construct to be maintained over time.

[0132] Example 2: Effect of the construct of the invention mimicking a segment of UT involved in binding to FlnA (SEQ ID NO: 35) on cell migration of GB lines 8MG, 42MG, U87 and GL15, A549 lung cancer, MDA-MB-231 breast cancer and LnCAP prostate cancer in response to 10' 9 M of peptide Ull.

[0133] At tO 30,000 GB 8MG, 42MG, U87 and GL15 cells and 75,000 A549 lung cancer cells, 100,000 MDA-MB-231 breast cancer cells and 75,000 LnCAP prostate cancer cells were seeded onto the porous membrane (8 μm) of the upper Boyden chamber in the presence of 100 nM of the sequence construct SEQ ID NO: 35. A homogeneous concentration of Ull 10' 9M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed and those in the lower chamber are fixed with DAPI (4',6-diamino-2-phenylindole), then the membranes are cut and mounted between slide and coverslip and 8 microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results (Statistical test: Two-way ANOVA; Sidak post-test, 2 membranes per condition and n=8 fields per membrane) are grouped in the following Table 2.

[0134] Table 2 The sequence construct SEQ ID NO: 35 completely and significantly prevents directional migration evoked by 10' 9M of Ull in 8MG, 42MG, U87 and GL15 GB and LnCAP prostate cancer lines, with responses to UH in control condition ranging from -57.47%, p = 0.0028; -69.30%, p < 0.0001; -62.11%, p < 0.0001; -93.13%, p < 0.0001 to -153.46%, p < 0.0001, of migration stimulation, respectively. The construct of sequence SEQ ID NO: 35 is indeed an effective compound that can inhibit the directional migration of solid cancer lines responding to chemoattractant gradients of Ull peptide.

[0135] Example 3: Effect of the construct of the invention mimicking a segment of UT involved in binding to FlnA (SEQ ID NO: 35) on cell migration of GB lines 8MG, 42MG, U87 and GL15, A549 lung cancer, MDA-MB-231 breast cancer and LnCAP prostate cancer in response to 50 ng / ml EGF (Epidermal Growth Factor).

[0136] At T0, 30,000 8MG, 42MG, U87 and GL15 cells and 75,000 A549 lung cancer cells, 100,000 MDA-MB-231 breast cancer cells and 75,000 LnCAP prostate cancer cells were seeded on the porous membrane (8 μm) of the upper Boyden chamber in the presence of 100 nM of the construct of sequence SEQ ID NO: 35. A homogeneous concentration of EGF 50 ng / ml is placed in the lower chambers. After 24 h, the cells of the upper chamber are removed, and those of the lower chamber are fixed and the nuclei labeled with DAPI (4',6-diamino-2-phenylindole), then the membranes are cut and mounted between slide and coverslip and eight microphotographs are made per membrane. The number of cells / field was quantified semi-automatically using ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are summarized in Table 3 below. Table 3

[0137] The sequence construct SEQ ID NO: 35 strongly and significantly prevents directional migration evoked by 50 ng / ml EGF in the 8MG, 42MG and

[0138] U87 of GB and LnCAP of prostate cancer that migrate in response to EGF in control condition when applied alone (-40.55%, p = 0.0332; -49.48%, p < 0.0001; -103.18%, p < 0.0001 and -256.54%, p < 0.0001 respectively). In GL15 of GB, A549 of lung cancer and MDA-MB-231 of breast cancer the construct of sequence SEQ ID NO: 35 partially prevents the directional migration evoked by 50 ng / ml of EGF (-27.61%, p = 0.0002; -22.14%, p < 0.0001 and -56.54%, p < 0.0001 respectively). The construct of sequence SEQ ID NO: 35 is indeed an effective compound capable of inhibiting the directional migration of solid cancer lines responding to EGF chemoattractant gradients.

[0139] Example 4: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 29) on cell migration of GB lines 8MG, 42MG, U87 and GL15, A549 lung cancer, MDA-MB-231 breast cancer and LnCAP prostate cancer in response to 10' 9 M of peptide Ull.

[0140] At T0, 30,000 GB cells 8MG, 42MG, U87 and GL15 and 75,000 lung cancer cells A549, 100,000 breast cancer cells MDA-MB-231 and 75,000 prostate cancer cells LnCAP were seeded on the porous membrane (8 μm) of the upper Boyden chamber in the presence of 100 nM of the construct of sequence SEQ ID NO: 29. A homogeneous concentration of Ull 10' 9M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed and the nuclei are labeled with DAPI (4',6-diamino-2-phenylindole) then the membranes are cut and mounted between slide and coverslip and eight microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the image analysis software ImageJ (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 4. As for all other experiments, Statistical test: Two-way ANOVA; Sidak post-test, 2 membranes per condition and n = 8 fields per membrane.

[0141] Table 4

[0142] The sequence construct SEQ ID NO: 29 completely and significantly prevents directional migration evoked by 10' 9M of Ull in 8MG, U87 and GL15 lines of GB and LnCAP of prostate cancer which respond positively to Ull in control condition when applied alone (-68.41%, p = 0.0007; -63.17%, p < 0.0001; -91.80%, p < 0.0001 and -138.15%, p = 0.0022 respectively). In addition, it partially prevents the migration evoked by 10' 9 M of Ull in the 42MG GB line (-26.72%, p = 0.0015). The sequence construct SEQ ID NO: 29 is indeed an effective compound that can inhibit the directional migration of solid cancer lines responding to chemoattractant gradients of Ull peptide.

[0143] Example 5: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 29) on cell migration of GB lines 8MG, 42MG, U87 and GL15, A549 lung cancer, MDA-MB-231 breast cancer and LnCAP prostate cancer in response to 50 ng / ml EGF (Epidermal Growth Factor). At T0, 30,000 GB 8MG, 42MG, U87 and GL15 cells and 75,000 A549 lung cancer cells, 100,000 MDA-MB-231 breast cancer cells and 75,000 LnCAP prostate cancer cells were seeded on the porous membrane (8 μm) of the upper Boyden chamber in the presence of 100 nM of the construct of sequence SEQ ID NO: 29. A homogeneous concentration of EGF 50 ng / ml is placed in the lower chambers.After 24 hours, the cells in the upper chamber were removed, and those in the lower chamber were fixed and the nuclei were labeled with DAPI (4',6-diamino-2-phenylindole), then the membranes were cut and mounted between slide and coverslip and eight microphotographs were taken per membrane. The number of cells / field was quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 5. Table 5.

[0144] The construct of sequence SEQ ID NO: 29 partially prevents directional migration evoked by 50 ng / ml EGF in U87 and GL15 GB lines that respond positively to EGF in control conditions when applied alone (-63.59%, p = 0.0022 and -37.75%, p < 0.0001 respectively). The construct of sequence SEQ ID NO: 29 is indeed a partially effective compound that can inhibit directional migration of solid cancer lines responding to chemoattractant gradients of EGF.

[0145] Example 6: Effect of the construct of the invention mimicking a segment of UT involved in binding to FlnA (SEQ ID NO: 37) on cell migration of GB lines 8MG, 42MG, U87 and U251 in response to 10' 9M of Ull peptide. At T0, 30,000 GB cells 8MG, 42MG, U87 and U251 were seeded on the porous membrane (8 μm) of the upper Boyden chamber in the presence of 100 nM of the construct of sequence SEQ ID NO: 37. A homogeneous concentration of Ull 10' 9 M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed and the nuclei are labeled with DAPI (4',6-diamino-2-phenylindole) then the membranes are cut and mounted between slide and coverslip and eight microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 6. Table 6

[0146] The sequence construct SEQ ID NO: 37 completely and significantly prevents directional migration evoked by 10' 9 M of Ull in GB lines 8MG, 42MG, U87 and U251 that respond positively to Ull in control condition when applied alone (-123.98%, p < 0.0001; -115.36%, p < 0.0001; -110.96%, p < 0.0001 and -60.50%, p < 0.0001 respectively). The construct of sequence SEQ ID NO: 37 is indeed an effective compound that can inhibit the directional migration of GB lines responding to chemoattractant gradients of Ull peptide.

[0147] Example 7: Effect of the construct of the invention mimicking a segment of UT involved in binding to FlnA (SEQ ID NO: 36) on cell migration of GB lines 8MG, 42MG, U87 and U251 in response to 10' 9M of Ull peptide. At T0, 30,000 8MG, 42MG, U87 and U251 glioblastoma cells were seeded onto the porous membrane (8 μm) of the upper Boyden chamber in the presence of 100 nM of the construct with sequence SEQ ID NO 36. A homogeneous concentration of Ull 10' 9 M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed and the nuclei labeled with DAPI (4',6-diamino-2-phenylindole) then the membranes are cut and mounted between slide and coverslip and eight microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 7.

[0148] Table 7

[0149] The sequence construct SEQ ID NO: 36 completely and significantly prevents directional migration evoked by 10' 9 M of Ull in the 8MG and 42MG lines that migrate in response to Ull in control condition when applied alone (-155.78%, p < 0.0001 and -100.85%, p < 0.0001 respectively). In addition, the sequence construct SEQ ID NO: 36 partially prevents the migration evoked by Ull 10' 9 M in the U87 line (-66.45%, p = 0.0211). The construct of sequence SEQ ID NO: 36 is in fact a partially effective compound capable of inhibiting the directional migration of GB lines responding to chemoattractant gradients of Ull peptide. Example 8: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to FUT (SEQ ID NO: 32) on the cell migration of GB lines 8MG, 42MG, U87 and U251 in response to 10' 9 M of peptide Ull.

[0150] At TO, 30,000 GB 8MG, 42MG, U87 and U251 cells were seeded onto the porous membrane (8 pm) of the upper Boyden chamber in the presence of 100 nM of the construct with sequence SEQ ID NO: 32. A homogeneous concentration of Ull 10' 9 M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed and the nuclei are labeled with DAPI (4',6-diamino-2-phenylindole) then the membranes are cut and mounted between slide and coverslip and eight microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 8. Table 8

[0151] The sequence construct SEQ ID NO: 32 completely and significantly prevents directional migration evoked by 10' 9 M of Ull in GB lines 8MG, 42MG, U87 and U251 that respond positively to Ull in control condition when applied alone (-116.32%, p < 0.0001; -97.40%, p = 0.0009; -119.71%, p < 0.0001 and -46.12%, p < 0.0001 respectively). The construct of sequence SEQ ID NO: 32 is indeed an effective compound that can inhibit the directional migration of GB lines responding to chemoattractant gradients of Ull peptide.

[0152] Example 9: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 33) on cell migration of GB lines 8MG, 42MG, U87 and U251 in response to 10' 9M of Ull peptide. At T0, 30,000 cells of 8MG, 42MG, U87 and U251 were seeded on the porous membrane (8 pm) of the upper Boyden chamber in the presence of 100 nM of the construct of sequence SEQ ID NO: 33. A homogeneous concentration of Ull 10' 9 M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed and the nuclei are labeled with DAPI (4',6-diamino-2-phenylindole) then the membranes are cut and mounted between slide and coverslip and eight microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 9:

[0153] Table 9 The sequence construct SEQ ID NO: 33 completely and significantly prevents directional migration evoked by 10' 9 M of Ull in the 8MG, 42MG and U251 lines of GB that respond positively to Ull in control condition when applied alone (-105.48%, p < 0.0001; -104.29%, p = 0.0037 and -58.53%, p < 0.0001 respectively). In addition, the sequence construct SEQ ID NO: 33 partially prevents the migration evoked by Ull 10' 9 M in the U87 line (-24.83%, p = 0.0037). The construct with sequence SEQ ID NO: 33 is indeed an effective compound that can inhibit the directional migration of GB lines responding to chemoattractant gradients of Ull peptide.

[0154] Example 10: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 31) on cell migration of GB lines 8MG, 42MG, U87 and U251 in response to 10' 9 M of peptide Ull.

[0155] At T0, 30,000 GB 8MG, 42MG, U87 and U251 cells were seeded onto the porous membrane (8 pm) of the upper Boyden chamber in the presence of 100 nM of the construct with sequence SEQ ID NO: 31. A homogeneous concentration of Ull 10' 9 M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed with DAPI (4',6-diamino-2-phenylindole), then the membranes are cut and mounted between slide and coverslip and 8 microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 10.

[0156] Table 10

[0157] The sequence construct SEQ ID NO: 31 completely and significantly prevents directional migration evoked by UH 10' 9 M in the GB U251 line that responds positively to UH in control condition when applied alone (-93.60%, p < 0.0001). In addition, the sequence construct SEQ ID NO: 31 partially prevents the migration evoked by UH 10' 9 M in the U87 line (-37.17%, p = 0.0164). The construct SEQ ID NO 31 is indeed a partially effective compound that can inhibit the directional migration of GB lines responding to chemo-attractant gradients of UII peptide.

[0158] Example 11: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 53) on cell migration of GB lines 8MG and U87 in response to UH 10' 9 Mr.

[0159] At T0, 30,000 GB 8MG and U87 cells were seeded onto the porous membrane (8 pm) of the upper Boyden chamber in the presence of 10 pM of the construct with sequence SEQ ID NO: 53. A homogeneous concentration of Ull 10' 9 M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed with DAPI (4',6-diamino-2-phenylindole), then the membranes are cut and mounted between slide and coverslip and 8 microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 11: Table 11

[0160] The sequence construct SEQ ID NO: 53 significantly prevents directional migration evoked by UH 10' 9M in GB lines 8MG and U87 that respond positively to UH in control condition when applied alone (-127.06%, p < 0.0001 and -93.65%, p < 0.0001 respectively). The construct SEQ ID NO: 53 is indeed an effective compound that can inhibit the directional migration of GB lines responding to chemoattractant gradients of Ull peptide independently of binding to the UT receptor.

[0161] Example 12: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 54) on cell migration of GB lines 8MG and U87 in response to UH 10' 9 M. At T0, 30,000 GB 8MG and U87 cells were seeded onto the porous membrane (8 pm) of the upper Boyden chamber in the presence of 10 pM of the construct with sequence SEQ ID NO: 54. A homogeneous concentration of Ull 10' 9M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed with DAPI (4',6-diamino-2-phenylindole), then the membranes are cut and mounted between slide and coverslip and 8 microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 12. Table 12

[0162] The sequence construct SEQ ID NO: 54 significantly prevents directional migration evoked by UH 10' 9M in GB lines 8MG and U87 that respond positively to UH in control condition when applied alone (-192.90%, p < 0.0001 and -106.12%, p = 0.0002 respectively). The construct SEQ ID NO: 54 is indeed an effective compound that can inhibit the directional migration of GB lines responding to chemoattractant gradients of Ull peptide independently of binding to the UT receptor.

[0163] Example 13: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 55) on cell migration of GB lines 8MG and U87 in response to 10' 9 M of UII peptide.

[0164] At T0, 30,000 GB 8MG and U87 cells were seeded onto the porous membrane (8 pm) of the upper Boyden chamber in the presence of 10 pM of the construct with sequence SEQ ID NO: 55. A homogeneous concentration of Ull 10' 9M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed with DAPI (4',6-diamino-2-phenylindole), then the membranes are cut and mounted between slide and coverslip and 8 microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 13.

[0165] Table 13

[0166] The sequence construct SEQ ID NO: 55 significantly prevents directional migration evoked by UH 10' 9M in GB lines 8MG and U87 that respond positively to UH in control condition when applied alone (-173.77%, p < 0.0001 and -96.41%, p < 0.0001 respectively). The construct SEQ ID NO: 55 is indeed an effective compound that can inhibit the directional migration of GB lines responding to chemoattractant gradients of Ull peptide independently of binding to the UT receptor.

[0167] Example 14: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 48) on cell migration of the GB U87 line in response to UH 10' 9 M. At T0, 30,000 GB U87 cells were seeded onto the porous membrane (8 pm) of the upper Boyden chamber in the presence of 10 pM of the construct with sequence SEQ ID NO: 48. A homogeneous concentration of Ull 10' 9M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed with DAPI (4',6-diamino-2-phenylindole), then the membranes are cut and mounted between slide and coverslip and 8 microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the Imaged image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 14.

[0168] Table 14

[0169] Sequence construct SEQ ID NO: 48 partially prevents directional migration evoked by UH 10' 9M in the GB line U87 which responds positively to UH in control condition when applied alone (-19.08%, p = 0.3674). The construct SEQ ID NO: 48 is in fact a non-effective compound for the inhibition of directional migration of GB lines responding to chemo-attractant gradients of Ull peptide independently of binding to the UT receptor.

[0170] Example 15: Effect of the construct of the invention mimicking a segment of FlnA involved in binding to UT (SEQ ID NO: 51) on cell migration of the GB U87 line in response to UH 10' 9 M. At T0, 30,000 GB U87 cells were seeded onto the porous membrane (8 pm) of the upper Boyden chamber in the presence of 10 pM of the construct with sequence SEQ ID NO: 51. A homogeneous concentration of Ull 10' 9M is placed in the lower chambers. After 24 hours, the cells in the upper chamber are removed, and those in the lower chamber are fixed with DAPI (4',6-diamino-2-phenylindole), then the membranes are cut and mounted between slide and coverslip and 8 microphotographs are taken per membrane. The number of cells / field is quantified semi-automatically using the ImageJ image analysis software (8 fields analyzed per membrane, 2 membranes per condition). The results are grouped in the following Table 15. Table 15

[0171] Sequence construct SEQ ID NO: 51 partially prevents Ull 10-evoked directional migration -9M in the GB U87 line that responds positively to Ull in control conditions when applied alone (-40.16%, p = 0.3674). The construct SEQ ID NO: 51 is in fact a non-effective compound for the inhibition of directional migration of GB lines responding to chemoattractant gradients of Ull peptide independently of binding to the UT receptor.

[0172] Example 16: Influence of the construct SEQ ID NO: 35 mimicking a sequence of the UT binding FlnA on the viability of U251, 8MG, 42MG, U87 and GL15 cells of GB, A549, HCC827 and PC9 of lung cancer, MDA-MB-231 of breast cancer and LnCAP of prostate cancer. At T0, 2x10 3 cells are seeded in the wells of a 96-well plate, in the presence of increasing concentrations of sequence construct SEQ ID NO: 35 tested from 10' 5 M to 10' 12M. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are summarized in Table 16.

[0173] Table 16 Breast

[0174] The sequence construct SEQ ID NO: 35 exhibits inhibitory effects that increase over time on all of the solid cancer lines cited above, with particularly significant inhibitory effects at 72 hours. These inhibitory effects are maximal at the concentration of 10 pM with percentages of inhibition of viability of 61.14% (p < 0.0001), 74.99% (p < 0.0001), 58.07% (p = 0.0021), 72.57% (p < 0.0001), 79.60% (p = 0.0006), 58.16% (p = 0.0008), 77.72% (p < 0.0001), 66.92% (p < 0.0001), 60.26% (p < 0.0001), 74.63% (0.0093) for the lines U251, 8MG, 42MG, U87, GL15, A549, HCC827, PC9, MDA-MB-231 and LnCAP respectively.The minimal inhibition effects are observed at 1 pM in U251, 8MG, 42MG, U87, GL15, A549, HCC827 and MDA-MB-231 lines, they are more specifically 33.18% (p = 0.0073), 48.87% (p < 0.0001), 35.04% (p = 0.0128), 41.36% (p = 0.0011), 56.63% (p = 0.0059), 39.22% (p = 0.0034), 21.48% (p = 0.0006), 35.92% (p = 0.0004) respectively. The inhibition is 34.46% (p = 0.0104) at 0.01 pM in the PC9 line. At 72h, the construct of sequence SEQ ID NO: 35 presents IC50s in the order of pM between 1.06 and 6.23 pM. The construct of sequence SEQ ID NO: 35 shows inhibitory effects on the viability of GB, lung and breast cancer lines.

[0175] Example 17: Influence of the sequence construction SEQ ID NO: 29 mimicking a UT-binding FlnA sequence on the viability of GB U251, 8MG, 42MG, U87 and GL15 cells, A549, HCC827 and PC9 lung cancer cells, MDA-MB-231 breast cancer cells and LnCAP prostate cancer cells.

[0176] At T0, 2x10 3 cells are seeded in the wells of a 96-well plate, in the presence of increasing concentrations of construct SEQ ID NO: 29 tested from 10' 5 M to 10' 12M. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are summarized in Table 17. The construct with sequence SEQ ID NO: 29 exhibits inhibitory effects that increase over time on all the solid cancer lines cited above, with particularly significant inhibitory effects at 72h. These inhibitory effects are maximal at the concentration of 10 pM with inhibition percentages of 49.48% (p = 0.0036), 46.28% (p < 0.0001), 44.39% (p = 0.0035), 46.41% (p = 0.0209), 43.73% (p = 0.0014), 53.19% (p < 0.0001), 49.25% (p = 0.002), 61.68% (p = 0.0024) for the U251, 8MG, U87, A549, HCC827, PC9, MDA-MB-231 and LnCAP lines respectively. The construct shows no effect at 72h in the 42MG and GL15 lines.A significant inhibition of 21.94% (p = 0.0084), 18.76% (p = 0.0007) and 19.77% (p = 0.0251) was observed in the 8MG, PC9 and LnCAP lines, respectively, from a concentration of 1 pM. At 72 hours, the construct of sequence SEQ ID NO: 29 presented IC50s of the order of ten pM, between 5.17 and 25.18 pM for the responding lines. Table 17 groups together the experimental results relating to the effects of the construct SEQ ID NO 29 on the viability of different GB lines (U251, 8MG, 42MG, U87, GL15), lung cancers (A549, HCC827 PC9), breast cancer (MDA-MB-231) and prostate cancer (LnCAP). Statistical test: Kruskal-Wallis; Dunn post-test, n = 2 or 3 independent experiments per line in triplicate. Table 17.

[0177] Example 18: Influence of the sequence construct SEQ ID NO: 37 mimicking a UT sequence binding FlnA on the viability of GB U251, 8MG, 42MG, U87 and GL15 cells and A549 lung cancer cells. At TO, 2x10 3 cells are seeded in the wells of a 96-well plate, in the presence of increasing concentrations of construct of sequence SEQ ID NO: 37 tested from 10' 5 M to 10- 12M. Cell density was assessed at 24h, 48h and 72h using the CelITiter-Glo 3D kit (Promega). The results are summarized in Table 18. The construct with sequence SEQ ID NO: 37 exhibits inhibitory effects increasing with time only in the GB U251 cell line with a maximum effect at 72h, 10 pM of 89.40% (p = 0.0017) of inhibition of viability. The inhibitory properties of the construct of sequence SEQ ID NO: 37 are not very specific and suggest that the fusion of the constructs of sequence SEQ ID NO: 4 and SEQ ID NO: 8 is less effective than for its counterpart of sequence SEQ ID NO: 35 resulting from the fusion of the constructs SEQ ID NO: 4 and SEQ ID NO: 9. The permeant sequence SEQ ID NO: 9 seems, in fact, decisive for relaying the inhibitory effects on the cell viability of this construct.Table 18 summarizes the experimental results of the effects of construct SEQ ID NO 37 on the viability of different GB lines (U251, 8MG, 42MG, U87, GL15) and lung cancer (A549). Statistical test: Kruskal-Wallis; Dunn's post-test, n = 2 or 3 independent experiments per line in triplicate.

[0178] Table 18

[0179] Example 19: Influence of the construct SEQ ID NO: 32 mimicking a sequence of FlnA binding UT on the viability of U251, 8MG, 42MG, U87 and GL15 cells of GB and A549 lung cancer. At TO, 2x10 3 cells are seeded in the wells of a 96-well plate, in the presence of increasing concentrations of construct SEQ ID NO: 32 tested from 10' 5 M to 10' 12M. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are summarized in Table 19. Construct SEQ ID NO: 32 showed no significant inhibitory effect on the tested lines. The absence of inhibitory properties of construct SEQ ID NO: 32 suggests that the association of constructs SEQ ID NO: 5 and SEQ ID NO: 8 of which it is the fusion is less effective than its counterpart SEQ ID NO: 29 resulting from the fusion of constructs SEQ ID NO: 5 and SEQ ID NO: 9. The permeant sequence SEQ ID NO: 9 also seems to be decisive in relaying the inhibitory effects on cell viability of this construct. Statistical test: Kruskal-Wallis; Dunn's post-test, n = 2 or 3 independent experiments per line in triplicate.

[0180] Table 19

[0181] Example 20: Influence of the sequence construct SEQ ID NO: 30 mimicking a UT-binding FlnA sequence on the viability of GB U251, 8MG and 42MG cells. At T0, 2x10 3 cells are seeded in the wells of a 96-well plate, in the presence of increasing concentrations of construct SEQ ID NO: 30 tested from 10' 5 M to 10' 12 M. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are summarized in Table 20. The construct with sequence SEQ ID NO: 30 exhibits inhibitory effects that increase over time only in the GB U251 cell line with a maximum effect at 72h, 10 pM of 73.67% (p = 0.0017) inhibition of viability. The lack of efficacy of the construct SEQ ID NO: 30 suggests that the interfering sequence with sequence SEQ ID NO: 5 that is part of its structure is not sufficient to relay inhibitory effects on the viability of GB cell lines.

[0182] Table 20 groups the effects of the construct SEQ ID NO 30 on the viability of different GB lines (U251, 8MG, 42MG). Statistical test: Kruskal-Wallis; Dunn post-test, n = 2 or 3 independent experiments per line in triplicate.

[0183] Table 20

[0184] Example 21: Influence of the sequence construct SEQ ID NO: 34 mimicking a UT sequence binding FlnA on the viability of GB U251 cells.

[0185] At TO, 2x10 3 cells are seeded in the wells of a 96-well plate, in the presence of increasing concentrations of construct SEQ ID NO: 34 tested from 10' 5 M to 10' 12M. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are grouped in Table 21. The construct with sequence SEQ ID NO: 34 exhibits inhibitory effects on early viability on the U251 line from 24h at 10 pM (50.15%, p = 0.0095), however, unlike the other effective compounds of the invention, its effects diminish at 72h and are no longer significant (p = 0.0513). Table 21 summarizes the effect of construct SEQ ID NO: 34 on the viability of the U251 GB line. Statistical test: Kruskal-Wallis; Dunn's post-test, n = 3 independent experiments in triplicate. Table 21 Example 22: Influence of the construction of sequence SEQ ID NO: 36 mimicking a sequence of the UT binding FlnA on the viability of GB U251 cells.

[0186] At TO, 2x10 3cells are seeded in the wells of a 96-well plate, in the presence of increasing concentrations of the tested sequence construct SEQ ID NO: 36 from 10' 5 M to 10- 12 M. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are summarized in Table 22. The construct with sequence SEQ ID NO: 36 exhibits inhibitory effects that increase over time on the viability of the GB U251 line with a maximum effect at 72h, 10 pM of 82.41% (p = 0.0025) inhibition of viability.

[0187] Table 22 summarizes the experimental results relating to the effect of construct SEQ ID NO 36 on the viability of the GB U251 line. Statistical test: Kruskal-Wallis; Dunn's post-test, n = 3 independent experiments in triplicate.

[0188] Table 22

[0189] Example 23: Influence of the sequence construct SEQ ID NO: 35 mimicking a UT sequence binding FlnA on the viability of U251, 8MG, 42MG, U87 and GL15 GB, A549, HCC827 and PC9 lung cancer cells, MDA-MB-231 breast cancer and LnCAP prostate cancer cells in combination with standard therapies used in the clinic. At TO, 2x10 3 cells are seeded into the wells of a 96-well plate, in the presence of increasing concentrations of the tested sequence construct SEQ ID NO: 35 from 10- 5 M to 10- 12M. In addition, GB lines U251, 8MG, 42MG, U87 and GL15 received treatment with 950 pM of TMZ and lung cancer lines A549, HCC827, PC9 and breast cancer lines MDA-MB-231 were treated with 20 pM, 0.008 pM, 0.020 pM and 20 pM of gefitinib, respectively. Finally, the prostate cancer line LnCAP received treatment with 1 nM of docetaxel. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are grouped in Table 23. The construct of sequence SEQ ID NO: 35 administered at 10 pM shows significant additive inhibitory effects compared to the effects of TMZ in the GB lines U251, 8MG and U87, particularly significant at 72 hours, bringing the effect of TMZ alone from 38.3% to 86.51% (p < 0.0001); 80.11% to 92.80% (p = 0.0199) and 79.95% to 91.86% (p = 0.0398).

[0190] Table 23 summarizes the experimental results relating to the effect of construct SEQ ID NO 35 on the viability of different GB (U251, 8MG, 42MG, U87, GL15), lung (A549, HCC827 PC9), breast (MDA-MB-231) and prostate (LnCAP) cancer lines in combination with the corresponding conventional therapies. GB lines U251, 8MG, 42MG, U87, GL15 were treated with 950 pM TMZ; lung cancer lines A549, HCC827, PC9 and breast MDA-MB-231 were treated with 20 pM, 0.008 pM, 0.020 pM and 20 pM gefitinib respectively; Finally, the LnCAP prostate cancer line was treated with 1 nM Docetaxel. Statistical test: Kruskal-Wallis; Dunn's post-test, n = 2 or 3 independent experiments per line in triplicate.

[0191] Table 23

[0192]

[0193] In HCC827 and PC9 lung cancer lines, the construct also exhibits an additive effect to the effect of gefitinib at 72h, increasing the inhibition of viability induced by gefitinib from 5.58% to 76.47% (p = 0.0028) and from 76.00% to 91.44% (p = 0.0009), respectively. The additive effect of the compound of sequence SEQ ID NO: 35 to TMZ and gefitinib is particularly interesting for a combined application with chemotherapy or with targeted therapy in the clinic in a patient.

[0194] Example 24: Influence of the sequence construct SEQ ID NO: 29 mimicking a UT-binding FlnA sequence on the viability of U251, 8MG, 42MG, U87 and GL15 GB, A549, HCC827 and PC9 lung cancer cells, MDA-MB-231 breast cancer and LnCAP prostate cancer cells in combination with standard therapies used in the clinic.

[0195] At T0, 2x10 3cells are seeded into the wells of a 96-well plate, in the presence of increasing concentrations of the tested sequence construct SEQ ID NO: 29 from 10- 5 M to 10- 12M. In addition, GB lines U251, 8MG, 42MG, U87 and GL15 received treatment with 950 pM of TMZ and lung cancer lines A549, HCC827, PC9 and breast cancer lines MDA-MB-231 were treated with 20 pM, 0.008 pM, 0.020 pM and 20 pM of gefitinib, respectively. Finally, the prostate cancer line LnCAP received treatment with 1 nM of docetaxel. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are grouped in Table 24. The sequence construct SEQ ID NO: 29 administered at 10 pM presents on the GB lines U251 and 8MG significant additive inhibitory effects to the effect of TMZ, particularly significant at 72 hours, bringing the effect of TMZ alone from 41.68% to 80.07% (p = 0.0137) and 78.26% to 88.86% (p = 0.0108) in the lines U251 and 8MG respectively.In the PC9 lung cancer line, the construct also exhibits an additive effect to gefitinib at 72h, increasing the inhibition of gefitinib-induced viability from 76.00% to 89.80% (p = 0.0168). The additive effect of compound SEQ ID NO: 29 to TMZ and Gefitinib is not systematic across GB and lung cancer lines.

[0196] Table 24 summarizes the experimental results relating to the effect of construct SEQ ID NO 29 on the viability of different GB (U251, 8MG, 42MG, U87, GL15), lung (A549, HCC827 PC9), breast (MDA-MB-231) and prostate (LnCAP) cancer lines in combination with the corresponding conventional therapies. GB lines U251, 8MG, 42MG, U87, GL15 were treated with 950 pM TMZ; lung cancer lines A549, HCC827, PC9 and breast MDA-MB-231 were treated with 20 pM, 0.008 pM, 0.020 pM and 20 pM gefitinib respectively; Finally, the LnCAP prostate cancer line was treated with 1 nM Docetaxel. Statistical test: Kruskal-Wallis; Dunn's post-test, n = 2 or 3 independent experiments per line in triplicate.

[0197] Table 24 Pulmonary

[0198] Example 25: Influence of the sequence construct SEQ ID NO: 37 mimicking a sequence of the UT binding FlnA on the viability of GB cells U251, 8MG, 42MG, U87 and GL15 in combination with standard therapies used in the clinic which is TMZ. At tO 2x10 3 cells are seeded into the wells of a 96-well plate, in the presence of increasing concentrations of the tested sequence construct SEQ ID NO: 37 from 10- 5 M to 10- 12M. In addition, GB lines U251, 8MG, 42MG, U87 and GL15 received treatment with 950 pM TMZ. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are summarized in Table 25. The sequence construction SEQ ID NO: 37 only shows significant additive effects to TMZ at 72h, 10 pM in the U251 line, bringing the effects of TMZ from 61.65% to 94.36% (p = 0.0066). Such effects are not reproduced in the other GB lines 8MG, 42MG, U87 and GL15 suggesting that the sequence construct SEQ ID NO: 37 is not the most optimal for application in combination with TMZ in the clinic in a patient.

[0199] Table 25 summarizes the experimental results relating to the effect of construct SEQ ID NO 37 on the viability of different GB lines (U251, 8MG, 42MG, U87, GL15) in combination with the corresponding conventional therapy, TMZ. GB lines U251, 8MG, 42MG, U87, GL15 were treated with 950 pM TMZ. Statistical test: Kruskal-Wallis; Dunn's post-test, n = 2 or 3 independent experiments per line in triplicate. Table 25

[0200] Example 26: Influences the construction of sequence SEQ ID NO: 32 mimicking a sequence of FlnA binding UT on the viability of GB U251, 8MG, 42MG, U87 and GL15 cells in combination with the standard therapy used in the clinic which is TMZ. At TO, 2x10 3 cells are seeded into the wells of a 96-well plate, in the presence of increasing concentrations of the tested sequence construct SEQ ID NO: 32 from 10- 5 M to 10- 12M. In addition, the GB lines U251, 8MG, 42MG, U87 and GL15 received a treatment of 950 pM of TMZ. The cell density was evaluated at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are grouped in Table 26. The construct with sequence SEQ ID NO: 32 does not show additive effects to TMZ in all the lines tested, suggesting that it is not suitable for application in combination with TMZ in a patient's clinic. Table 26 groups the experimental results relating to the effect of the construct SEQ ID NO: 32 on the viability of different GB lines (U251, 8MG, 42MG, U87, GL15) in combination with the corresponding conventional therapy, TMZ. GB lines U251, 8MG, 42MG, U87, GL15 were treated with 950 pM TMZ. Statistical test: Kruskal-Wallis; Dunn's posttest, n = 2 or 3 independent experiments per line in triplicate.

[0201] Table 26

[0202]

[0203] Example 27: Influence of the sequence construct SEQ ID NO: 30 mimicking a UT-binding FlnA sequence on the viability of GB U251, 8MG and 42MG cells in combination with the standard therapy used clinically which is TMZ. At T0, 2x10 3 cells are seeded into the wells of a 96-well plate, in the presence of increasing concentrations of the tested sequence construct SEQ ID NO: 30 from 10' 5 M to 10- 12M. In addition, GB lines U251, 8MG and 42MG received treatment with 950 pM of TMZ. Cell density was assessed at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are grouped in Table 27. The construct SEQ ID NO: 30 exhibited a significant additive effect to transient TMZ at 48h, at 10pM, in the U251 line, increasing its effect from 22.12% to 59.01% (p = 0.0193). This weak transient effect, not reproduced in the other two GB lines 8MG and 42MG, does not allow us to consider the construct of sequence SEQ ID NO: 30 as usable in combination with TMZ for clinical application in a patient. Table 27 summarizes the experimental results relating to the effect of construct SEQ ID NO 30 on the viability of different GB lines (U251, 8MG and 42MG) in combination with the corresponding conventional therapy, TMZ. GB lines U251, 8MG and 42MG were treated with 950 pM TMZ.Statistical test: Kruskal-Wallis; Dunn post-test, n = 2 or 3 independent experiments per line in triplicate.

[0204] Table 27

[0205] Example 28: Influence of the sequence construct SEQ ID NO: 34 mimicking a sequence of the UT binding FlnA on the viability of U251 glioblastoma cells in combination with the standard therapy used in the clinic which is TMZ. At TO, 2x10 3 cells are seeded in the wells of a 96-well plate, in the presence of increasing concentrations of construct SEQ ID NO: 34 tested from 10' 5 M to 10' 12M. In addition, the U251 GB line receives treatment with 950 pM of TMZ. Cell density is evaluated at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are grouped in Table 28. The construct with sequence SEQ ID NO: 34, administered at 10 pM, shows significant additive inhibitory effects to TMZ in the U251 GB line, particularly significant at 72h, bringing the effect of TMZ alone from 55.82% to 83.33% (p = 0.0045). It is therefore possible to use the construct with sequence SEQ ID NO: 34 in combination with TMZ in clinical practice on patients with GB from the same subgroup as the U251 line. Table 28 summarizes the experimental results relating to the effect of construct SEQ ID NO 34 on the viability of the GB U251 line (in combination with the corresponding conventional therapy, TMZ). The glioblastoma U251 line was treated with 950 pM of TMZ.Statistical test: Kruskal-Wallis; Dunn post-test, n = 2 or 3 independent experiments per line in triplicate.

[0206] Table 28

[0207] Example 29: Influences the construction of sequence SEQ ID NO: 36 mimicking a sequence of the UT binding FlnA on the viability of GB U251 cells in combination with the standard therapy used in the clinic which is TMZ.

[0208] At T0, 2x10 3 cells are seeded into the wells of a 96-well plate, in the presence of increasing concentrations of the tested sequence construct SEQ ID NO: 36 from 10' 5 M to 10- 12M. In addition, the U251 GB line receives treatment with 950 pM of TMZ. Cell density is evaluated at 24h, 48h and 72h using the CelITiter-GIo 3D kit (Promega). The results are grouped in Table 29. The construct with sequence SEQ ID NO: 36, administered at 10 pM, shows significant additive inhibitory effects to TMZ in the U251 GB line, particularly significant at 72h, bringing the effect of TMZ alone from 69.77% to 95.83 (p = 0.0031). It is therefore possible to use the construct with sequence SEQ ID NO: 36 in combination with TMZ in clinical practice on patients with the same GB subgroup as the U251 line. Table 29 summarizes the experimental results relating to the effect of construct SEQ ID NO 36 on the viability of the GB U251 line (in combination with the corresponding conventional therapy, TMZ). The GB U251 line was treated with 950 pM of TMZ.Statistical test: Kruskal-Wallis; Dunn post-test, n = 2 or 3 independent experiments per line in triplicate.

[0209] Table 29

Claims

Claims 1. A construct comprising a cell-penetrating peptide sequence coupled by a peptide bond to at least one first linker molecule, which is coupled by a peptide bond to the N-terminal end of an interfering amino acid sequence selected from: a) sequences comprising or consisting of the following sequence VTYCPTEPGNYIINIKFADQHVPGSPFSVKVTGEGRVKESITRRRRAPSVANVGSHCDL SLKIPEISIQDMTAQVTSPSGKTHEAEIVEGENHTYCIRFVPAE (SEQ ID NO: 1 or FlnA 2104-2206) which forms domain 2104-2206 of the human FlnA protein; b) sequences comprising or consisting of a fragment of the sequence SEQ ID NO: 1, said sequences or said fragments being capable of associating with the sequence VRGPGSGGGRGPVPSLQPRAR (SEQ ID NO: 2 or UT-330-350) which defines the domain 330-350 of the UT receptor or with a fragment of this sequence SEQ ID NO: 2, in particular with the fragment of sequence GGGRGPVPSLQPRAR (SEQ ID NO: 3 or UT-336-350) forming the domain 336-350 of the UT receptor or with the fragment RGPVPSLQPRAR (SEQ ID NO: 4 or UT-339-350) defining the domain 339-350 of the UT receptor; c) the sequences comprising or consisting of a fragment of the sequence SEQ ID NO: 2, in particular the fragment forming domain 330-350 of the UT receptor (SEQ ID NO: 2) or the fragment forming domain 336-350 of the UT receptor (SEQ ID NO: 3) or the fragment forming domain 339-350 of the UT receptor (SEQ ID NO: 4) and capable of associating with a fragment of the sequence SEQ ID NO: 1; d) the sequences having at least 90% identity and, preferably, at least 95% identity with the sequences defined in paragraphs a) to c); 2. Construction according to claim 1, characterized in that said at least one first binding molecule is an amino acid, optionally modified and in particular a beta-alanine molecule.

3. Construction according to claim 1 or 2, characterized in that said interfering amino acid sequence is chosen from the amino acid sequences defined in paragraph b) and in particular from the following interfering sequences: MTAQVTSPSGKTHEAEIVE (SEQ ID NO: 5 or FlnA-D20-CD), GNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 6 or FlnA-D19-FG), THEAEIVEGENHTYCIRF (SEQ ID NO: 7 or FlnA-D20-DE); - peptide sequences having at least 90% and preferably at least 95% identity with said sequences and substantially homologous sequences, preferably deriving from the sequences SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7 by one or more conservative substitutions. - RGPVPSLQPRAR (SEQ ID NO: 4 or UT-339-350) and GGGRGPVPSLQPRAR (SEQ ID NO: 3 or UT-336-350) and the peptide sequences having at least 90% and preferably at least 95% identity with said sequences SEQ ID NO: 3 and SEQ ID NO:

4.

4. Construction according to any one of the preceding claims, characterized in that said cell penetration sequence is chosen from the following sequences: -TCTWLKYH (SEQ ID NO: 8 or Glio) and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 8 and the substantially homologous sequences, preferably deriving from the sequence SEQ ID NO: 8 by one or more conservative substitutions; - the peptide sequence GRRRQRRKKRKCCKRKKRRQRRRG (SEQ ID NO: 9 or CycloGlio) in which residues 12 and 13 (cysteine) are linked by a disulfide bridge and in which residues 2 to 10 and 15 to 23 are dextrorotatory (D) amino acids and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 9, the two ends of said penetration sequence each being linked by a peptide bond to a first linking molecule which is an amino acid, optionally modified, said linking molecule itself being linked by a peptide bond to an interfering sequence, the two interfering sequences being identical or different.

5. Construction according to claim 4, characterized in that said cell penetration sequence is chosen from the peptide sequence GRRRQRRKKRKCCKRKKRRQRRRG (SEQ ID NO: 9 or CycloGlio) in which residues 12 and 13 (cysteine) are linked by a disulfide bridge and in which residues 2 to 10 and 15 to 23 are dextrorotatory (D) amino acids and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 9, in that the first two linker molecules are identical and in particular beta alanine molecules; and in that -one of the interfering sequences is chosen from the sequences defined in paragraphs a) and b) of claim 1, in particular from the following interfering sequences: MTAQVTSPSGKTHEAEIVE (SEQ ID NO: 5 or FlnA-D20-CD), GNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 6 or FlnA-D19-FG), THEAEIVEGENHTYCIRF (SEQ ID NO: 7 or FlnA-D20-DE) and peptide sequences having at least 90% and preferably at least 95% identity with said sequences; and - the other interfering sequence is chosen from the sequences defined in paragraph c) of claim 1 and in particular from the following interfering sequences: RGPVPSLQPRAR (SEQ ID NO: 4 or UT-339-350) and GGGRGPVPSLQPRAR (SEQ ID NO: 3 or UT-336-350) and the peptide sequences having at least 90% and preferably at least 95% identity with said sequences SEQ ID NO: 3 and SEQ ID NO:

4.

6. Interfering construct according to any one of claims 4 and 5, characterized in that said cell penetration sequence being chosen from the peptide sequence GRRRQRRKKRKCCKRKKRRQRRRG (SEQ ID NO: 9 or CycloGlio) in which residues 12 and 13 (cysteine) are linked by a disulfide bridge and in which residues 2 to 10 and 15 to 23 are dextrorotatory amino acids (D) and the peptide sequences having at least 90% and preferably at least 95% identity with said sequence SEQ ID NO: 9, the lysines in positions 11 and 14 of said cell penetration sequence are acetylated.

7. Interfering construct according to any one of the preceding claims, characterized in that it is chosen from the following sequence constructs: TVKVSFPSGPVHQDAFKINHYNGXGrrrqrrkkrKCCKrkkrrqrrrGXGNYIINIKFADQHVPGS PFSVKVT (SEQ ID NO: 10, also called FlnA-CPPD19-FG-CycloGlio) in which residues 37 and 38 are linked by a disulfide bridge, X is in both positions an optionally modified amino acid and preferably beta-alanine in both positions and residues 27 to 35 and 40 to 48 are dextrorotatory (D) amino acids and in which the lysines in positions 36 and 39 are acetylated; EVIEAEHTKGSPSTVQATMXGrrrqrrkkrKCCKrkkrrqrrrGXMTAQVTSPSGKTHEAElVE (SEQ ID NO: 11; also referred to as FlnA-CPPD20-CD-CycloGlio) wherein residues 32 and 33 are linked by a disulfide bridge, X is an optionally modified amino acid and preferably beta-alanine in both positions and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and wherein the lysines at positions 31 and 34 are acetylated; FRISYTHNEGEVIEAEHTXGrrrqrrkkrKCCKrkkrrqrrrGXTHEAElVEGENHTYSIRF (SEQ ID NO: 12; also referred to as FlnA-CPPD20-DE-CycloGlio) wherein residues 31 and 32 are linked by a disulfide bridge, X is an optionally modified amino acid and preferably beta-alanine and residues 21 to 29 and 34 to 42 are dextrorotatory (D) amino acids and wherein the lysines at positions 30 and 33 are acetylated; TCTWLKYHXGNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 13; also referred to as FlnA-CPPD19-FG-Glio) wherein X is an optionally modified amino acid and preferably beta-alanine; TCTWLKYHXMTAQVTSPSGKTHEAEIVE (SEQ ID NO: 14; also referred to as FlnA-CPPD20-CD-Glio) wherein X is an optionally modified amino acid and preferably beta-alanine; TCTWLKYHXTHEAEIVEGENHTYCIRF (SEQ ID NO: 15; also referred to as FlnA-CPPD20-DE-Glio) wherein X is an optionally modified amino acid and preferably beta-alanine; RARPQLSPVPGRGGGXGrrrqrrkkrKCCKrkkrrqrrrGXGGGRGPVPSLQPRAR (SEQ ID NO: 16, also referred to as UT-CPP336-350-CycloGlio) wherein residues 28 and 29 are linked by a disulfide bridge, X is an optionally modified amino acid and preferably beta-alanine in both positions and residues 18 to 26 and 31 to 39 are dextrorotatory (D) amino acids and wherein the lysines at positions 27 and 30 are acetylated; RARPQLSPVPGRXGrrrqrrkkrKCCKrkkrrqrrrGXRGPVPSLQPRAR SEQ ID NO: 17; also referred to as UT-CPP339-350-CycloGlio) in which residues 25 and 26 are linked by a disulfide bridge, X is an optionally modified amino acid and preferably beta-alanine in both positions and residues 15 to 23 and 28 to 36 are dextrorotatory (D) amino acids and in which the lysines in positions 24 and 27 are acetylated, TCTWLKYHXGGGRGPVPSLQPRAR (SEQ ID NO: 18; also referred to as UT-CPP336-350-Glio) wherein X is an optionally modified amino acid and preferably beta-alanine; - TCTWLKYHXRGPVPSLQPRAR (SEQ ID NO: 19; also referred to as UT-CPP339-350-Glio) in which X is an optionally modified amino acid and preferably beta-alanine; - constructions presenting at least 90% identity and preferably at least 95% identity with the aforementioned constructions.

8. Construction according to any one of the preceding claims, characterized in that it is chosen from the following sequence constructions: TVKVSFPSGPVHQDAFKINHYNGAGrrrqrrkkrKCCKrkkrrqrrrGAGNYIINIKFADQHVPGS PFSVKVT (SEQ ID NO: 28, also called FlnA-CPPD19-FG-CycloGlio-syn) in which residues 37 and 38 are linked by a disulfide bridge, the alanines in positions 25 and 50 are beta-alanine and residues 27 to 35 and 40 to 48 are dextrorotatory (D) amino acids and in which the lysines in positions 36 and 39 are acetylated; EVIEAEHTKGSPSTVQATMAGrrrqrrkkrKCCKrkkrrqrrrGAMTAQVTSPSGKTHEAElVE (SEQ ID NO: 29; also referred to as FlnA-CPPD20-CD-CycloGlio-syn) wherein residues 32 and 33 are disulfide bonded, alanines at positions 20 and 45 are beta-alanine and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and wherein lysines at positions 31 and 34 are acetylated; FRISYTHNEGEVIEAEHTAGrrrqrrkkrKCCKrkkrrqrrrGATHEAElVEGENHTYSIRF (SEQ ID NO: 30; also referred to as FlnA-CPPD20-DE-CycloGlio-syn) wherein residues 31 and 32 are disulfide bonded, alanines at positions 19 and 44 are beta-alanine and residues 21 to 29 and 34 to 42 are dextrorotatory (D) amino acids and wherein lysines at positions 30 and 33 are acetylated; TCTWLKYHAGNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 31; also referred to as FlnA-CPPD19-FG-Glio-syn) wherein the alanine at position 9 is a beta-alanine; TCTWLKYHAMTAQVTSPSGKTHEAEIVE (SEQ ID NO: 32; also referred to as FlnA-CPPD20-CD-Glio-syn) wherein the alanine at position 9 is beta-alanine; TCTWLKYHATHEAEIVEGENHTYCIRF (SEQ ID NO: 33; also referred to as FlnA-CPPD20-DE-Glio-syn) wherein the alanine at position 9 is beta-alanine; RARPQLSPVPGRGGGAGrrrqrrkkrKCCKrkkrrqrrrGAGGGRGPVPSLQPRAR (SEQ ID NO: 34; also referred to as UT-CPP336-350-CycloGlio-syn) wherein residues 28 and 29 are disulfide bonded, alanines at positions 16 and 41 are beta-alanine, residues 18 to 26 and 31 to 39 are dextrorotatory (D) amino acids and wherein lysines at positions 27 and 30 are acetylated; RARPQLSPVPGRAGrrrqrrkkrKCCKrkkrrqrrrGARGPVPSLQPRAR SEQ ID NO: 35; also referred to as UT-CPP339-350-CycloGlio-syn) wherein residues 25 and 26 are disulfide bonded, alanines at positions 13 and 38 are beta-alanine and residues 15 to 23 and 28 to 36 are dextrorotatory (D) amino acids and wherein lysines at positions 24 and 27 are acetylated; TCTWLKYHAGGGRGPVPSLQPRAR (SEQ ID NO: 36; also referred to as UT-CPP336-350-Glio-syn) wherein the alanine at position 9 is a beta-alanine; TCTWLKYHARGPVPSLQPRAR (SEQ ID NO: 37; also referred to as UT-CPP339-350-Glio-syn) wherein the alanine at position 9 is a beta-alanine; the sequences having at least 90% identity and, preferably, at least 95% identity with the aforementioned sequences.

9. Construction according to any one of claims 1 to 5, characterized in that it further comprises a marking agent capable of penetrating into the cell.

10. Construction according to claim 9, characterized in that it is chosen from the following sequence constructions: TVKVSFPSGPVHQDAFKINHYNGAGrrrqrrkkrKCCKrkkrrqrrrGAGNYIINIKFADQHVPGS PFSVKVT (SEQ ID: 38, also referred to as Mq-FlnA-CPPD19-FG-CycloGlio-syn) wherein residues 37 and 38 are disulfide bonded, alanines at positions 25 and 50 are beta-alanine, and residues 27 to 35 and 40 to 48 are amino acids dextrorotatory (D) and residues 36 and 39 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine; EVIEAEHTKGSPSTVQATMAGrrrqrrkkrKCCKrkkrrqrrrGAMTAQVTSPSGKTHEAElVE (SEQ ID NO: 39; also referred to as Mq-FlnA-CPPD20-CD-CycloGlio-syn) wherein residues 32 and 33 are linked by a disulfide bridge, the alanines at positions 20 and 45 are beta-alanine and residues 22 to 30 and 35 to 43 are dextrorotatory (D) amino acids and residues 31 and 34 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine; FRISYTHNEGEVIEAEHTAGrrrqrrkkrKCCKrkkrrqrrrGATHEAElVEGENHTYSIRF (SEQ ID NO: 40; also referred to as Mq-FlnA-CPPD20-DE-CycloGlio-syn) wherein residues 31 and 32 are linked by a disulfide bridge, the alanines at positions 19 and 44 are beta-alanine and residues 21 to 29 and 34 to 42 are dextrorotatory (D) amino acids and residues 30 and 33 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine; ATCTWLKYHAGNYIINIKFADQHVPGSPFSVKVT (SEQ ID NO: 41; also referred to as Mq-FlnA-CPPD19-FG-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid; ATCTWLKYHAMTAQVTSPSGKTHEAEIVE (SEQ ID NO: 42; also referred to as Mq-FlnA-CPPD20-CD-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid; ATCTWLKYHATHEAEIVEGENHTYCIRF (SEQ ID NO: 43; also referred to as Mq-FlnA-CPPD20-DE-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid; RARPQLSPVPGRGGGAGrrrqrrkkrKCCKrkkrrqrrrGAGGGRGPVPSLQPRAR (SEQ ID NO: 44; also referred to as Mq-UT-CPP336-350-CycloGlio-syn) in which the residues 28 and 29 are linked by a disulfide bridge, the alanines at positions 16 and 41 are beta-alanine and residues 18 to 26 and 31 to 39 are dextrorotatory (D) amino acids and residues 27 and 30 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine; RARPQLSPVPGRAGrrrqrrkkrKCCKrkkrrqrrrGARGPVPSLQPRAR SEQ ID NO: 45; also referred to as Mq-UT-CPP339-350-CycloGlio-syn) wherein residues 25 and 26 are linked by a disulfide bridge, the alanines at positions 13 and 38 are beta-alanine and residues 15 to 23 and 28 to 36 are dextrorotatory (D) amino acids residues 24 and 27 are each linked by a peptide bond to the carboxylic acid group of 5-carboxytetramethylrhodamine, ATCTWLKYHAGGGRGPVPSLQPRAR (SEQ ID NO: 46; also referred to as Mq-UT-CPP336-350-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid; ATCTWLKYHARGPVPSLQPRAR (SEQ ID NO: 47; also referred to as Mq-UT-CPP339-350-Glio-syn) wherein the alanine at position 1 and the alanine at position 10 are beta-alanine and wherein the alanine at position 1 is linked by a peptide bond to the carboxylic acid group of 7-diethylaminocoumarin-3-carboxylic acid; - sequences exhibiting at least 90% identity and, preferably, at least 95% identity with the aforementioned sequences 11. Construction according to any one of claims 1 to 10, for its use as a medicament or diagnostic / prognostic agent, in particular for cancer, in particular solid cancerous tumors including breast cancer, lung cancer more particularly in the treatment of pulmonary adenocarcinomas, melanomas, prostate cancer, colon cancer in particular colorectal adenocarcinomas, stomach cancer, in particular gastric carcinomas, kidney cancer, in particular renal carcinomas, nasopharyngeal cancers in particular nasopharyngeal carcinomas, pancreatic cancer, diffuse glioma and glioblastoma in a subject having such a need, in particular a human.

12. Pharmaceutical composition which comprises, as active ingredient, at least one construction according to any one of claims 1 to 10, and a pharmaceutically acceptable solvent, excipient and / or vehicle.

13. Composition according to claim 12, characterized in that it is in the form of a solid, in particular a powder, a gel or an injectable solution, possibly colloidal.

14. Nucleic acid coding for one of the sequences according to any one of claims 1 to 10, in particular for a construction chosen from the following sequences: SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO:

7.

15. Viral vector comprising a nucleic acid according to claim 14, preferably an adenoviral or lentiviral vector.

16. Combination product containing at least one construct according to the invention and in particular a construct chosen from the constructs of sequence SEQ ID NO: 16, SEQ ID NO: 34, SEQ ID NO: 17, SEQ ID NO: 35, SEQ ID NO: 10, SEQ ID NO: 28, SEQ ID NO: 11, SEQ ID NO: 29, SEQ ID NO: 12, SEQ ID NO: 30, SEQ ID NO: 18, SEQ ID NO: 36, SEQ ID NO: 19, SEQ ID NO: 37, SEQ ID NO: 13, SEQ ID NO: 31, SEQ ID NO: 14, SEQ ID NO: 32, SEQ ID NO: 15, SEQ ID NO: 33 and an anticancer agent chosen in particular from tyrosine kinase inhibitors, in particular Gefitinib and alkylating agents causing double strand breaks in DNA and cell death, in particular TMZ.

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