Patent Description:
Aneurysms are outward bulging of blood vessels and are caused by a localized, abnormal, weakness of the blood vessels walls. This is generally due to the scarcity of the cells on the blood vessels walls, leading to the decay of the extracellular matrix which can't withstand the pressure. Weakened walls can suddenly leak, which can lead to dramatic consequences.

Current treatments aim at improving the healing of the aneurysm opening by occluding the aneurysm pocket. For this, commercial intrasaccular devices are used such as platinum coils and Endosaccular Flow Disrupter such as WEB (Woven EndoBridge, self-expanding intra expandable device), made of Nitinol.

Vascular disorders are generally treated by stents coated with active ingredients. However, the treatment of aneurysms is highly specific: it differs mainly in that a pro-aggregation/pro-thrombotic effect is desired to enhance proliferation of the cells on the walls of the endoprothesis, whereas an anti-coagulation effect is generally desired in the treatment of vascular conditions, such as atherosclerosis.

Additionally, intravascular devices used for aneurysms differ from vascular stents in that they have a distinctive structure/geometry and are made of different materials.

Further, coils and webs used in cerebral aneurysms aim at healing by cicatrization, whereas stents used in vascular conditions aim at endothelialization.

Also, the vascular stents are generally coated with active ingredients that are released in the body, which is neither desired nor required in the case of aneurysms.

In any case, the current treatments of aneurysms with endovascular devices are generally ineffective in <NUM>% of cases due to a lack of cells to promote healing of the aneurysm as the material does not foster the cicatrization.

There is therefore a need to provide an improved device to enhance cell proliferation and healing for treating aneurysms.

<CIT> discloses medical devices implanted in vessels or hollow organs, said devices being coated in order to promote progenitor endothelial cells to adhere, grow and differentiate on the surface of the implanted device to form a functional endothelium.

Fucoidans are sulphated polysaccharides that can be found inter alia in the extracellular matrix of brown marine algae.

Originally named fucoidin in <NUM> by Kylin, it was renamed fucoidan by IUPAC. The fucoidans, mostly composed of sulphated fucose, belong to the fucan family which also includes ascophyllans (branched polyuronics) and sargassans (compounds of galactose, fucose and uronic acid). Fucoidans can be found in more of <NUM> species of marine algae.

Fucoidans have been reported to exhibit various biological activities, such as pro/antithrombotic, angiogenic, anti-cancer, anti-inflammatory and anti-oxidant activities.

<NPL>») disclosed the extraction and purification of fucoidans, their affinity for P-selectin which is used for thrombose imaging. Specifically, it was investigated whether atherosclerosis could be diagnosed with fucoidans-grafted gold chips.

It has now been discovered that an improved aneurysms endoprothesis grafted with fucoidans exhibit improved healing properties.

The present invention thus provides an aneurysms endoprothesis grafted with fucoidans.

According to an object, the present invention provides an aneurysms endoprothesis comprising.

Where * represents the covalent attachment with a polyaryl chain ; and.

As used therein,the term « biologically active » refers to the activity exerted by the fucoidan molecules that are grafted to the endoprothesis.

Said activity includes in particular the healing effect by increasing the cell proliferation.

According to the invention, the Fucoidan groups are covalently attached to the metallic support through the polyaryl chains, and as such are not released upon their implantation.

According to an embodiment, the film comprises at least one one chain of formula (II) :.

Said Linker has a terminal (proximal) end that is covalently grafted on the metallic support and one or more terminal (distal) ends that are covalenly bound with the Fucoidan group. Typically, said Linker comprises at its terminal distal end the terminal functional group comprising the group of formula (I) as defined above for covalently attachment with the Fucoidan.

According to an embodiment, said Linker may be represented by Formula (III) :
<CHM>.

As used herein, "Aryl" means an aromatic monocyclic or multicyclic ring system of about <NUM> to about <NUM> carbon atoms, preferably of about <NUM> to about <NUM> carbon atoms. Exemplary aryl groups include phenyl or naphthyl, or phenyl substituted or naphthyl substituted.

"Heteroaryl" means an aromatic monocyclic or multicyclic ring system of about <NUM> to about <NUM> carbon atoms, preferably about <NUM> to about <NUM> carbon atoms, in which one or more of the carbon atoms in the ring system is/are hetero element(s) other than carbon, for example nitrogen, oxygen or sulfur. Preferred ring sizes of rings of the ring system include about <NUM> to about <NUM> ring atoms. Exemplary heteroaryl and substituted heteroaryl groups include pyrazinyl, thienyl, isothiazolyl, oxazolyl, pyrazolyl, furazanyl, pyrrolyl, <NUM>,<NUM>,<NUM>-thiadiazolyl, pyridazinyl, quinoxalinyl, phthalazinyl, imidazo[<NUM>,<NUM>-a]pyridine, imidazo[<NUM>,<NUM>-b]thiazolyl, benzofurazanyl, azaindolyl, benzimidazolyl, benzothienyl, thienopyridyl, thienopyrimidyl, pyrrolopyridyl, imidazopyridyl, benzoazaindole, <NUM>,<NUM>,<NUM>-triazinyl, benzthiazolyl, furanyl, imidazolyl, indolyl, indolizinyl, isoxazolyl, isoquinolinyl, isothiazolyl, oxadiazolyl, pyrazinyl, pyridazinyl, pyrazolyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, <NUM>,<NUM>,<NUM>-thiadiazolyl, thiazolyl, thienyl and triazolyl. Preferred heteroaryl groups include pyrazinyl, thienyl, pyridyl, pyrimidinyl, isoxazolyl and isothiazolyl.

According to an embodiment, Ar represents a phenyl group optionally substituted by at least one substituent chosen from alkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, aldehyde, hydroxyl, ketone, carbonyl, carboxyl, ester, ether, amine, amide, nitrile, sulfonic and/or halogen atoms; more particularly nitro.

Particularly preferably, the aryl group is a phenyl group, optionally bearing one or more substituents, whereby the polyarylene film which is grafted is a film of a polyphenylene. Even more specifically, the aryl group is a phenyl group substituted by a nitro group, in which case the film formed is a film of a poly (nitrophenylene).

More particularly, Ar (also referred to as the aryl unit in the polyaryl chain) is an optionally substituted nitrophenyl unit, such as an optionally substituted <NUM>-nitrophenyl, of formula (IV) :
<CHM>
Where.

Accordingly, where the aryl unit is a group of formula (IV), in the proximal terminal unit,typically represents the covalent bond with the metallic support and in the one or more distal terminal units, one or more of R<NUM>, R<NUM> typically represents the covalent bond with the functional terminal group as defined above.

Said Fucoidan may be a synthetic or natural polysaccharide.

In particular it may be extracted from brown algae, such as Chorda filum, Ascophyllum nodosum, Fucus vesiculosus, Fucus evanescens, Laminaria saccharina, Fucus serratus L. and Cladosiphon okamuranus.

It can also be extracted from marine invertibrate animals such as Ludwigothuria grisea; Strongylocentrotus droebachiensis and Strongylocentrotus franciscanus.

The structure of fucoidans from different algae has been extensively studied during the last century. It depends on both the species and the parameters (ecosystem, season, location), as well as the extraction method.

The first structure was highlighted in <NUM> by Conchy and Percival from a fucoidan from Fucus vesiculosus. The main chain consists of a linear skeleton of α (<NUM> → <NUM>) -L-fucose with a substitution of the sulfate function in C4 but also occasional bonds α (<NUM> → <NUM>). In <NUM> the model was corrected by Patankar et al. : a linear chain of fucoses linked in α (<NUM> → <NUM>) with a C4 sulfate and a few branches in C4 or C2, in the end about one modification for two or three fucose residues within the chain. Subsequently, studies have shown the existence of recurrent units for purified fractions from different species of algae. The structures are linear α (<NUM>-><NUM>) -L-fucose skeletons with alternately α (<NUM>-><NUM>) bonds, the sulphate groups being mainly found on positions O-<NUM> and O-<NUM>. Fucoidans extracted from marine animals exhibit more regular structures with little or no branching. Representative structures of fucoidans are depicted below:
<CHM>.

Or the pharmaceutically acceptable salts thereof.

According to an embodiment, the fucoidan is an extract from Ascophyllum nodosum, in particular a depolymerized fraction thereof.

Typically, the fucoidan is a mixture having an average (weight) molar weight comprised between <NUM> and <NUM>/mol (kDa), typically between <NUM> and <NUM> kDa.

Suitable fucoidans for the present invention may be commercially available. A suitable fucoidan composition may be purchased from Algues et Mer under the trade name Ascophyscient®. Ascophyscient® is a low molecular weight depolymerized fraction extracted from Ascophyllum nodosum.

According to an embodiment, said fucoidan is a compound comprising the repetition of one or more units of formula (V) :
<CHM>.

Where R<NUM> is chosen from H ; SO<NUM>X, where X represents H or a cation, typically an alkali metal, such as Na+ ; or a lateral sugar chosen from xylose, glucose, galactose, mannose, or a glucoronic acid ;.

The compounds of formula (V) can be provided in the form of a free base or in the form of addition salts with acids, which also form part of the invention.

These salts are advantageously prepared with pharmaceutically acceptable acids, but salts with other acids, useful for example for the purification or for the isolation of the compounds of formula (V), also form part of the invention.

According to an embodiment, the endoprothesis may be any of the devices that are implanted for healing aneurysms, such as cerebral aneurysms. Typical endoprothesis are coils or WEBs (Woven EndoBridge, self-expanding intra expandable device). They may be typically made of platinum, nickel, titane, and alloys thereof, such as Nitinol (nickel/titane).

According to another object, the present invention concerns the process of manufacture of the endoprothesis according to the invention.

Said process comprises the following steps :.

According to an embodiment, the functionalization step comprises :.

Reductive amination consists in grafting a primary amine at the end of the polysaccharide chain onto the aldehyde function of the fucoidan.

According to an embodiment, diaminopropane may be chosen as a suitable primary amine. Typically, the reductive amination step may be conducted by reacting the fucoidan with said primary amine, followed by reacting the reactional mixture with dimethylborane (C<NUM>H<NUM>B). The reaction with the primary amine can be conducted in protic conditions, typically in an aqueous acidic solution, such as acetic acid in water. The reaction may be conducted under heating at a temperature comprised between the room temperature and the reflux temperature of the reactional mixture.

Typically, dimethylborane may be used as a diluted aqueous solution.

Typically, the metacrylic acid group introduced at the fucoidan is formula:
<CHM>.

The methacrylation step comprises reacting the aminated fucoidan with methacrylic anhydride. This reaction may be typically conducted in aprotic conditions, such as in DMF.

The functionalized fucoidan that is so obtained in then grafted onto the metallic support. The grafting may be achieved by using as an anchor an aryldiazonium salt of formula (VI):.

Where Y- represents a counter ion and Ar is defined as in formula (III) above.

Said aryldiazonium salt may be a nitrobenzodiazonium salt of formula (VII):
<CHM>.

Where R<NUM> and R<NUM> are defined as in formula (IV), and Y- represents a counter ion.

The counter ion may be an inorganic anion such as a halide ion such as a bromide (Br), iodide (I-) or chloride (Cl-) ion, a tetrahaloborate ion such as a tetrafluoroborate ion (BF<NUM>-), a hydrogen sulphate ion (HSO<NUM>-), a dihydrogen phosphate ion (H<NUM>PO<NUM>-), a nitrate ion (NO<NUM>-) or a chlorate ion (ClO<NUM>-). However, it can also be an organic anion such as an acetate ion (CH<NUM>CO<NUM>-) or a formate ion (HCO<NUM>).

Suitable aryldiazonium salts may include phenyldiazonium tetrafluoroborate, <NUM>-methylphenyldiazonium tetrafluoroborate, <NUM>-nitrophenyldiazonium tetrafluorobrate, <NUM>-carboxyphenyldiazonium tetrafluoroborate, tetrafluoroborate of <NUM>-carboxyphenyldiazonium tetrafluoroborate. <NUM>-carboxymethylphenyl-diazonium, <NUM>-chloromethylphenyldiazonium tetrafluoroborate, <NUM>-bromophenyldiazonium tetrafluoroborate, tetrafluoroborate <NUM>-diethylaminophenyldiazonium, <NUM>-cyanophenyldiazonium tetrafluoroborate, <NUM>-aminophenyldiazonium chloride, <NUM>-aminomethylphenyldiazonium chloride, <NUM>-methoxyphenyldiazonium tetrafluoroborate, tetrafluoroborate <NUM>-heptadecylfluorooctylphenyldiazonium and <NUM>-acetamido-phenyldiazonium tetrafluoroborate.

Specifically, the counter ion may be a tetrafluoroborate ion (BF<NUM>-), so that the aryldiazonium salt is NBDT (nitrobenzodiazonium tetrafluoroborate):
<CHM>.

The grafting onto the metallic support may be typically achieved by reduction of the aryldiazonium salt leading to aryldiazonium radicals. This reduction may be typically conducted by either of the two following alternatives procedures:
According to a first alternative, grafting is conducted by electrochemical reduction of the aryldiazonium salt, including electrografting.

Electrografting is an electrochemical method for surface modification, that can be implemented by application or adaptation of the method disclosed by <NPL>. It may be commercially available under the tradename eG® :
Cyclic voltammetry is used to reduce the diazonium salt, causing the formation of radicals. Some radicals graft on the metallic support and some radicals react with the methacrylated Fucoidan. (Figure 2B).

According a second alternative, grafting is conducted in a chemically induced reduction of the aryldiazonium salt, in the presence of ascorbic acid as a reducing agent. According to this alternative, the metallic support is contacted with a solution of ascorbic acid, the diazonium salt and the functionalized fucoidan. The mechanism of grafting relies on a key step, which is the formation of aryl radicals from the ascorbic acid-induced reduction of the diazonium salt in solution. Indeed, those radicals have a double role. They can graft onto the substrate and form a primer-layer (rich in aromatic rings) essential for the subsequent grafting of functionalized fucoidan; meanwhile they initiate the radical polymerization of the vinyl cgain of the functionalized fucoidan in solution. Finally, the growing radical polymer chains eventually graft by a "grafting to" pathway on the aromatic rings present on the surface to form a grafted polymer films on the substrate.

This may be conducted by application or adaptation of the Graftfast® procedure.

In the process above, starting compounds and reactants, unless otherwise indicated, are commercially available or described in literature, or can be prepared according to methods described in literature or known to one of skill in the art.

Accordig to a further object, the present invention concerns an endoprothesis as defined above for its use for the treatment and/or prevention of aneurysms, such as cerebral aneurysms.

According to an embodiment, the endoprothesis of the invention is suitable for preventing the recidivism of aneurysms.

The following examples are illustrative of the present invention.

In order to graft polysaccharides on metals, two processes are considered : Graftfast® and electrografting such as eG®, requiring in both cases the introduction of a methacrylate chemical group at the reducing end of the polysaccharide chain In the presence of a diazonium salt the radical polymerization of the modified polysaccharides occurs forming a polymer layer at the surface of metals.

The modification of the polysaccharide was performed in two steps :.

It is to be understood that although the starting fucoidan is generally represented with a terminal cyclic form, the opened form form which is in equilibrium with said cyclic form has been depicted above as it is locked in this form by the reaction with the diamine.

In a <NUM> flask, about <NUM> of polysaccharide previously dried overnight under vacuum at <NUM> ° C is dissolved in a solution prepared with <NUM> of diaminopropane <NUM>, <NUM> of water and <NUM> of acetic acid. The flask is heated at <NUM> ° C for <NUM> hours. After cooling the flask in an ice bath, add <NUM> of <NUM> dimethylborane solution freshly prepared. The solution is heated for <NUM> at <NUM>. After cooling, the solution is dialyzed (MWCO 1kDa) successively against <NUM> carbonate buffer at pH9. <NUM> with <NUM> NaCl (5x1L), a water/ethanol mixture (<NUM>:<NUM>, v/v) with <NUM> NaCl (5x1L), and osmotic water (5x1L). The final solution is frozen and lyophilized. The product is obtained as a colorless foam in <NUM>-<NUM>% yield. The amount of primary amines introduced per polymer chain evaluated by colorimetric assay with o-phthalaldehyde using bromopropylamine as standard is ranging from <NUM> to <NUM>.

<NUM>) The second step is the methacrylation of the primary amine introduced in the polymer chain. An exemple is provided below with representative fucoidan.

In a <NUM> flask, about <NUM> of aminated polysaccharide previously dried overnight under vacuum at <NUM> ° C is dissolved under nitrogen in <NUM> of anhydrous DMF. Then <NUM> of <NUM> methacrylic anhydride in anydrous DMF are added and kept under stirring and nitrogen bubbling for <NUM> at room temperature. Then, the solution is dialyzed (MWCO 1kDa) successively against <NUM> carbonate buffer at pH9. <NUM> with <NUM> NaCl (5x1L), a water/ethanol mixture (<NUM>:<NUM>, v/v) with <NUM> NaCl (5x1L), and osmotic water (5x1L). The final solution is frozen and lyophilized. The product is obtained as a colorless foam in <NUM>-<NUM>% yield.

<NUM>-NMR analysis allows to validate the presence of the methacrylic group (see <FIG>).

<FIG> presents schematically the grafting of methacrylated fucoidan, as an exemple. With the electrochemical method, the grafting is obtained by electro-initiated radical polymerization using cyclic voltammetry.

The Graftfast® process is a so-called "one-pot" process as the reducing agent is added in the reactional mixture. All the compounds are put into solution from the start. Graftfast® uses ascorbic acid as a reducing agent instead of electrochemistry.

A potential is applied (from + <NUM>. 5V to -<NUM>. 5V) of constant sweep (about <NUM> mV / s) starting from a potential where no reaction is initiated. The potential starts at <NUM>. 5V and decreases to -<NUM>. 5V passing through <NUM>. 2V where the reduction of diazonium salts (NBDT) in solution begins. The potential returns to its initial value. <NUM> cycles are performed. The experiments are carried out in a glass cell with <NUM> electrodes: the metal sample (coil) on which the polysaccharide is to be fixed serves as the working electrode, an Ag/AgNO3 electrode used as a reference electrode, and the counter electrode made of platinum.

The reagents are introduced in the following molar ratios NBDT/ascorbic acid/methacrylated polymer: <NUM>/<NUM>/<NUM>-<NUM>. The sample (coil) to be grafted is placed in contact with the solution and the mixture is left to act for <NUM> minutes at room temperature under nitrogen bubbling. After <NUM> minutes the sample is taken out and washed with osmotic water, ethanol before drying in an oven at <NUM>.

Analyzes were performed on samples grafted with fucoidan using electrografting or graftfast®. In samples, carbon (C), oxygen (O), nitrogen(N), platinum (Pt) and sulfur (S) atoms were evidenced (table <NUM>). Platinum (Pt) was used to calibrate the spectra. The <FIG> presents the charactristic signals from 2p electrons of sulfur.

Since only fucoidan provided sulfur, fucoidan is thus grafted to the surface of the sample with both methods.

An EDS analysis allowed an elementary mapping of the surface of a sample. The <FIG> exhibits images of commercial platinum coil grafted with fucoidan using the graftfast method. As expected, a homogeneous distribution of the platinum and sulfur is observed.

The aneurysms are created on New Zealand white rabbits (<NUM>-<NUM>) under general anesthesia by intramuscular injection of ketamine (<NUM>/kg) + Xylazine (<NUM>/kg) + Acepromazine <NUM>% (<NUM>/kg). After shaving the cervical area, local anesthesia by subcutaneous injection of <NUM>% lidocaine (Xylocaine®, ©AstraZeneca) is performed followed by a median incision to expose the right Common Carotid Artery (CCA). Following dissection of the artery, an arteriotomy is conducted, and a <NUM> French introducer (Mini-Plastic Guide <NUM> Fr. Radiofocus®, Introducer II, Terumo Europe) is advanced retrogradely to <NUM> from the ostium of the right CCA. heparin (Heparin Choay® sodium, <NUM> I. / <NUM>) is administered by intravenous injection to prevent the formation of per procedural arterial thrombi. Under X-ray scopic control (Mobil C-arm - Siemens Arcadis Varic Model No.<NUM> Serial No. <NUM> IVK; Munchen, Germany), a <NUM> French arterial embolectomy catheter (CEM804, Vygon, France)is advanced retrogradely to the origin of the right CCA. At this location, the embolectomy balloon is inflated with a solution containing contrast medium (Hexabrix® <NUM>/mL, Guerbet, Roissy CdG, France) in order to generate a filling chamber. A solution containing <NUM> I. of porcine pancreatic elastase (Classification of the enzyme by IUB System:<NUM>. <NUM>; Reference: Code: ES Cat. No. LS002280 Size 1gm; Worthington Biochemical Corporation) is injected into this filling chamber through the introducer, and is left to incubate for <NUM> minutes. The embolectomy catheter and the introducer are then removed and the carotid is ligated distally.

After suturing the deep muscle planes and the skin plane with a Vicryl <NUM>-<NUM>, the rabbit is reinserted into its cage. The development of an aneurysm requires <NUM> weeks after the surgical creation of the aneurysm. This experimental model provides aneurysms that are histologically similar to human intracranial aneurysms with sizes roughly <NUM>-<NUM> larger in diameter and a collar of approximately <NUM>.

Aneurysms were treated <NUM> weeks after their creation. Under general anesthesia, a <NUM> F Envoy guide catheter (Codman Neurovascular, Raynham, MA, USA) was navigated from the right common femoral artery into the brachiocephalic trunk. Using the coaxial technique, an Excelsior SL <NUM> microcatheter (Stryker, Fremont, CA, USA) was advanced into the aneurysm. Either Axium (Medtronic, Irvine, CA) or Optima (BALT, Montmorency, France) Detachable bare platinum coils were used. Aneurysms were embolized with bare platinum coils (control group, n=<NUM>) or bare platinum coils covered with dextran (control group, n=<NUM>) or bare platinum coils covered with fucoidan (test group, n=<NUM>). No anti-platelet agents were given post embolization. The dextran was a witness to show whether fucoidan actually increases the cell proliferation/healing capacity. After embolization control DSA was performed.

<NUM> weeks after embolization a final control DSA was performed followed by euthanasia of the animals by using a lethal injection of embutramide ensued by tissue harvest. Median sternotomy and pericardiotomy were executed. Access to the left ventricle was secured by direct puncture with a <NUM>-gauge catheter. A small cut was done at the right atrial appendage. With pressure pump, heparinized saline (100U/ml) was constantly perfused into the catheter, until the effluent from the right appendage was light pink. The coil aneurysm was then collected and submerged into phosphate buffer saline (PBS). The parent artery was cut longitudinally to visualize the neck orifice for gross inspection to assess the tissue growth at the neck. After photography, the sample was fixed in <NUM>% glutaraldehyde (G5882, Merck, Germany) for <NUM> hours for further whole tissue mount staining.

A blinded reader with experience estimated aneurysm occlusion and packing density.

Treated tissues were dehydrated in a succession of ethanol baths (<NUM> baths of <NUM> - <NUM> at <NUM>%, <NUM> at <NUM>%, <NUM> at <NUM>% and <NUM> at <NUM>%) and embedded in a poly-(methyl-methacrylate) PMMA resin (<NUM> methyl methacrylate, <NUM>,<NUM> butyl methacrylate, <NUM>µL methyl benzoate, <NUM>µL polyethylene glycol <NUM> (PEG <NUM>), <NUM> Luperox® A75 (benzoyl peroxide) and <NUM>µL N,N-dimethyl-p-toluidine), <NUM> hours at <NUM>, under neutral atmosphere (nitrogen). Before embedding, dehydrated aneurysms were infused <NUM> in a MMA I solution containing the same quantities as above of methyl methacrylate, butyl methacrylate, methyl benzoate and PEG <NUM>, then infused <NUM> in a MMA II solution containing MMA I reagents and <NUM> of benzoyl peroxide.

PMMA blocks were then sectioned longitudinally (<NUM> section) in a microtome (RM2245, Leica) equipped with a tungsten carbide blade. The sections were deposed on ethanol on Superfrost Microscope Slides (Superfrost Plus® Gold, Thermo Scientific, USA) and let dry <NUM>.

Slides were stained in hematoxylin-eosin-safran and Masson trichrome. Immunohistochemistry was also performed using EnVision+ Kit (EnVision+ System-HRP anti-mouse, K4001, Dako), and antibodies targeting CD31 (JC70A clone, M0823, Dako), RAM-<NUM> (RAM-<NUM> clone, M0633, Dako) and alpha-SMA (1A4 clone, M0851, Dako).

The slides were scanned on a NanoZoomer RS2. <NUM> (Hamamatsu Photonics, France) and studied with the NDPView software. Images were reviewed by blinded observers.

In bare coil aneurysm section (<FIG>), as in all other groups, it was observed infiltration of cells and formation of healing tissue inside the aneurysm, that indicates a filling of the aneurysm. However, in the control group, the healing tissue presents a low cellular proliferation (the cellular nuclei appear in darker grey, as examples indicated with clear arrowheads), with a low-level organization, and a dominance of cellular matrix, including around the coil scars. In dextran-covered coil treated aneurysm (<FIG>), almost the same results can be seen as in the untreated control, with an unorganized tissue formed, containing a low cells concentration and mostly filled with extracellular matrix.

In the fucoidan-covered coil treated aneurysm (<FIG>), the staining reveals a more extensive cell proliferation in scar tissue than in the controls. It was also observed an orientation of cellular nuclei (flattened nuclei indicated by full arrowheads) around the coil scars, that would indicate an orientation of cells around the coils.

It follows from these experiments that the healing of the aneurysms is better in the fucoidantreated group than in the control (without fucoidan).

The histological index for aneurysms treated with bare coils was <NUM>/<NUM> versus <NUM>/<NUM> for coils with fucoidan with a significant difference (p=<NUM>) (<FIG>.

Claim 1:
Endoprothesis for aneurysms comprising
a metallic support and
a biologically active film covalently grafted on said metallic support,
characterized in that said film comprises one or more polyaryl chains having a proximal end that is covalently grafted to said metallic support, and
at least one distal end that is covalently bound with a Fucoidan terminal group through a terminal functional group comprising the following group of formula (I):
<CHM>
Where * represents the covalent attachment with a polyaryl chain ; and
** represents the covalent attachment with the Fucoidan terminal group.