EP2152782A1

EP2152782A1 - Polyglutamic acids functionalized by cationic groups and hydrophobic groups and applications thereof, in particular therapeutic applications thereof

Info

Publication number: EP2152782A1
Application number: EP08750064A
Authority: EP
Inventors: You-Ping Chan; Olivier Breyne; Cécile BONNET GONNET
Original assignee: Flamel Technologies SA
Current assignee: Flamel Technologies SA
Priority date: 2007-05-03
Filing date: 2008-05-05
Publication date: 2010-02-17
Also published as: FR2915748B1; BRPI0809208A2; ZA200906941B; WO2008135563A1; AU2008248604B2; IL201017A0; KR20100016280A; CN101663346A; JP2010529214A; NZ580906A; FR2915748A1; MX2009011813A; IL201017A; CA2683741A1; EA200901481A1; AU2008248604A1; US20090012028A1; EA016911B1; US8716217B2; CN101663346B

Abstract

The present invention relates to novel biodegradable materials based on modified polyamino acids that can be used, in particular, for the vectorization of active principle(s) (PA). The invention also relates to novel pharmaceutical, cosmetic, dietetic or phytosanitary compositions based on these polyamino acids. The objective of the invention is to provide a novel polymer starting material that is capable of being used for the AP vectorization and that makes it possible to optimally satisfy all the specifications required in the case in point: biocompatibility, biodegradability, ability to associate easily with numerous active principles or to dissolve them, and to release these active principles in vivo. This objective is achieved by the present invention, which relates to novel polyglutamates modified by cationic groups that, if they can be deprotonated, have a pKa greater than or equal to 7, and by hydrophobic groups comprising from 8 to 30 carbon atoms. These polyglutamates modified by cationic groups are capable of being easily and economically converted into particles for the vectorization of active principles, these particles being themselves capable of forming stable aqueous colloidal suspensions. These modified polyglutamates have the advantage of being less viscous than other similar polymers, while retaining an ability to associate proteins such as insulin. Some are soluble in water at acid pH and become insoluble at physiological pH (7.4) and should therefore, during a subcutaneous injection, precipitate at the injection site.

Description

POLYGLUTAMIC ACIDS FUNCTIONALIZED BY

CATIONIC GROUPS AND HYDROPHOBIC GROUPS

AND THEIR APPLICATIONS, IN PARTICULAR THERAPEUTIC

The present invention relates to novel biodegradable materials based on copolyamino acids, useful in particular for the vectorization of active principle (s) (PA). The invention also relates to novel pharmaceutical, cosmetic, dietetic or phytosanitary compositions based on these modified polyamino acids. These compositions may be of the type of those allowing the vectorization of AP and are preferably in the form of emulsions, micelles, nanoparticles, microparticles, gels, implants or films.

The PAs considered are, advantageously, biologically active compounds that can be administered to an animal or human organism orally, parenterally, nasally, vaginally, ocularly, subcutaneously, intravenously, intramuscularly, intradermally, intraperitoneally, intracerebrally, orally, etc.

PAs more particularly, but not exclusively, concerned by the invention are proteins, glycoproteins, peptides, polysaccharides, lipopolysaccharides, oligo or polynucleotides, and organic molecules. But it can also be cosmetic products or phytosanitary products, such as herbicides, insecticides, fungicides, etc.

In the field of vectorization of the active ingredients, in particular medicinal products, there is a need, in many cases:

• to protect them against degradation (hydrolysis, enzymatic digestion, etc.),

• and / or control their release rate in order to maintain a therapeutic level over a defined period of time,

• and / or to convey them (by protecting them) at the site of action.

For these purposes, several types of polymers have been studied and some are even commercially available. There may be mentioned, for example, polymers of the polylactic, polylactic-glycolic, polyoxyethylene-oxypropylene, polyamino acid or polysaccharide type. These polymers are raw materials for manufacturing, for example, mass implants, microparticles, nanoparticles, vesicles, micelles or gels. In addition to the fact that these polymers must be suitable for the manufacture of such systems, they must also be biocompatible, non-toxic, non-immunogenic, economical and they must be easily removed from the body. and / or be biodegradable. On this last aspect, it is moreover essential that the biodegradation in the organism generates non-toxic products.

As an illustration of the prior art concerning polymers used as raw materials for the production of AP vectorization systems, various patents or patent applications or scientific articles are cited below.

US-B-4,652,441 discloses polylactide microcapsules encapsulating the hormone LH-RH. These microcapsules are produced by preparing a water-in-oil-in-water emulsion and comprise an aqueous inner layer containing the hormone, a substance (gelatin) fixing the same, an oily layer of polylactide, and an aqueous outer layer. (polyvinyl alcohol). The release of the AP can be done over a period of more than two weeks after subcutaneous injection.

US-B-6,153,193 discloses compositions based on amphiphilic poly (oxyethylene) -poly (oxypropylene) micelles, for the vectorization of anticancer agents such as doxorubicin.

Akiyoshi et al. (J. Controlled Release, 1998, 54, 313-320) describe pullulans which are rendered hydrophobic by grafting cholesterol and which form nanoparticles in water. These nanoparticles, capable of reversibly complexing with insulin, form stable colloidal suspensions.

US Pat. No. 4,351,337 describes amphiphilic copolyamino acids based on leucine and glutamate that can be used in the form of implants or microparticles for the controlled release of active principles. The release of these can be done over a very long time depending on the rate of degradation of the polymer.

US-B-4,888,398 discloses polymers based on polyglutamate or polyaspartate, and optionally polyleucine, with pendant groups of alkyloxycarbonyl-methyl type, randomly placed on the polyamino acid chain. These polyamino acids, grafted with side groups e.g. methoxycarbonylmethyl, can be used in the form of biodegradable implants containing a sustained release PA.

US Pat. No. 5,904,936 describes nanoparticles obtained from a polyleucine-polyglutamate block polymer capable of forming stable colloidal suspensions capable of associating spontaneously with biologically active proteins without denaturing them. These can then be released in vivo in a controlled manner over a long period.

US Pat. No. 5,449,513 describes amphiphilic block copolymers comprising a polyoxyethylene block and a polyamino acid block, for example poly (beta-benzyl-L-aspartate). These polyoxyethylene-polybenzylaspartate polymers form micelles which are capable of encapsulating hydrophobic active molecules such as doxorubicin or indomethacin.

The patent application WO-A-99/61512 describes polylysines and polyornithines functionalized with a hydrophobic group (palmitic acid connected to polylysine or ornithine) and a hydrophilic group (polyoxyethylene). These polymers, for example polylysine grafted with polyoxyethylene and palmitoyl chains, form, in the presence of cholesterol, vesicles capable of encapsulating doxorubicin or DNA. These polymers based on polylysines are cationic in a physiological medium.

The patent application EP-A-963 758 describes polyamino acids functionalized with a cationic group. These polymers are capable of forming complexes with a nucleic acid and can be used in gene therapy. The cationic groups are amino derivatives which are not derived from amino acids and the polyamino acids do not contain hydrophobic groups.

Yang et al. {Biotechnology Letters, 2005, 27, 977-982) describe polyaspartates functionalized with linear alkyl groups and oligoarginines. These polymers form nanoparticles from 8 to 40 nm in water and are able to be internalized by cells. It is suggested that these particles can be used for the vectorization of hydrophobic molecules.

In the same field, the Applicant has described, in several patent applications, polymers based on polyglutamate (anionic) with related concepts.

WO-A-03/104303 discloses anionic polyamino acids functionalized with alpha-tocopherol.

The application WO-A-2004/013206 describes anionic polyamino acids having hydrophobic groups and characterized in that these groups are connected to the polymer via a ball joint containing two amide functions, and more specifically via a lysine-type spacer. or ornithine. The application WO-A-2004/060968 describes polyamino acids functionalized with at least one oligoamino acid group based on leucine and / or isoleucine and / or valine and / or phenylalanine.

The article by WC Shen, "Acid-sensitive dissociation between poly (lysine) and histamine-modified poly (glutamate) as a model for drug-releasing from endosomes" Biochim Biophys Acta 1034 (1): 122-124, 1990 discloses a polyglutamate functionalized with 40% histamine. However, no hydrophobized polyglutamate backbone is described. In addition, the developed polymer precipitates between pH 4 and 5 and is soluble at physiological pH. Only the application aimed at forming complexes with a polylysine sensitive to pH is developed. These complexes are based on electrostatic interactions. Indeed, at physiological pH, the polyglutamatehistamine-polylysine complex is formed, while it decomposes at pH 4 - 5, a pH value in the endosome.

Thus, even if a large number of technical solutions are developed and proposed in the prior art for the vectorization of active drug ingredients, the response to all the requirements is difficult to obtain and remains perfectible. More specifically, the invention relates to biodegradable polyamino acids that can be converted into nano- or micro-colloidal vector particles capable of reversibly associating with active principles.

In this context, one of the essential objectives of the present invention is to provide novel amphiphilic copolyglutamates comprising both positive charges at neutral pH or close to neutrality and pendant hydrophobic groups. These polymers represent an improvement over those described in the patents or patent applications cited above in terms of vectorization of an active ingredient such as a peptide or a therapeutic protein, a DNA, an RNA or a small molecule.

Another essential objective of the present invention is that these polymers are capable of being used for the AP active ingredient vectorization and make it possible to optimally satisfy all the specifications of the specification, namely in particular: o capacity:

To form easily and economically stable aqueous colloidal suspensions,

• to associate easily with many active ingredients,

• and release these active ingredients in vivo, o biocompatibility, o biodegradability, o stability to hydrolysis.

This object, among others, is achieved by the present invention, which relates to polyamino acids, or their pharmaceutically acceptable salts, comprising glutamic residues, characterized in that some of the glutamic residues carry a pendant cationic group which, if it is deprotonable, has a pKa greater than or equal to 7, said cationic groups being identical or different from each other, and in that other glutamic residues carry a hydrophobic group (GH) during, said hydrophobic groups ( GH) being identical or different from each other.

In the description which follows, unless otherwise stated, the term "cationic group" will generally refer to cationic groups which are not deprotonable, and cationic groups which are deprotonable and which have a pKa greater than or equal to 7.

The term "pharmaceutically acceptable salts" of a polyamino acid according to the invention means all of the polyamino acids with the counterions associated with the ionized functions of the polymer.

In the following description, the term "small molecule" refers to a molecule of molecular weight less than 1 kDa.

Unless otherwise stated, the alkyl radicals have 1 to 10 carbon atoms.

Each polyglutamate according to the invention is therefore functionalized by a multiplicity of cationic groups and hydrophobic groups (GH), pendant and respectively identical or different from each other.

For the purposes of the invention, the term "multiplicity" means that the polyglutamate is functionalized by: at least 1% of cationic groups (mol% relative to the glutamic acid residues) and up to 99%,

on average, at least two pendant hydrophobic groups (GH) per molecule. It is possible according to the invention that the polyglide glutamic has, in addition to pendant hydrophobic groups (GH), hydrophobic groups (GH) attached to at least one end of the copolymer chains.

For the purposes of the invention, the term "carrier" means that the group carried is pendant, that is to say that said group is a lateral group with respect to glutamic residues and is a substituent of the γ carbonyl function of the glutamic residue that carries it.

Polyglutamate also carries cationic groups. These groups are preferably grafted to glutamic residues through an amide or ester bond.

According to a variant of the invention, other glutamic residues may carry a non-ionizable group during, other than hydrophobic groups (GH), said nonionizable groups being identical or different from each other. Such a pendant nonionizable group may be, for example, the hydroxyethylamino group.

According to another variant of the invention, other glutamic residues may carry, also on their carbonyl in γ, a non-ionized group at neutral pH, different from hydrophobic groups (GH), said nonionized groups at pH neutral being the same or different from each other. Such an un-ionized group at neutral pH may for example satisfy the following formula:

where -R ¹⁰ is -H, -CO ₂ H, alkyl ester (preferably -COOMe or -COOEt), -CH ₂ OH, -C (= O) -NH ₂ , -C (= O) -NH -CH ₃ or -C (= O) -N (CH ₃ ) ₂ .

The polyglutamate, carrying both cationic groups and hydrophobic groups, and optionally also nonionic groups (nonionizable or nonionized at neutral pH) may also comprise negative charges (always at neutral pH) resulting from the ionization of pendant groups of polyglutamic acid that have not been functionalized.

It is the merit of the applicant to have developed a new family of polymers based on polyglutamate, carriers of cationic groups, and functionalized by a multiplicity of hydrophobic groups, which polymers are capable of forming stable colloidal systems. The ability to modulate the polymer load according to the degree of functionalization can be very effective for:

lowering the viscosity of the polymer for easier injection or easier implementation during the formulation step. - ensure a good association with neutral, anionic or cationic active ingredients. Thus, the polymers of the invention can be:

Anionic, cationic, neutral and capable of associating charged active ingredients or unloaded APs;

• cationic at moderately acidic pH (pH = 4-5) and neutral or weakly charged at neutral pH. In this case, this pH dependence allows them, after association with the active ingredient in solution at pH = 4-5, to form a deposit in a physiological medium.

Moreover, they are easily degraded, in the presence of enzymes, into non-toxic catabolites / metabolites (amino acids).

For the purposes of the invention and throughout this presentation, the terms "association" or "associate" used to describe the relationship between one or more active ingredients and the modified polyglutamates, mean, in particular, that the active ingredient (s) are related to the (x) polyglutamate (s) in particular by a hydrophobic interaction, and / or are encapsulated by the polyglutamate (s).

Advantageously, the polyamino acids according to the invention are functionalized homopolymers of L-glutamate or L-glutamic acid; preferably these residues are bonded in the polyamino acid by their carboxyl in the alpha position.

The cationic groups that can be used to functionalize the glutamate residues are identical or different from each other and correspond to the following general formula:

X ^ W ₃ ⁺ Z

in which :

X = O, NH, Y = independently H or CH ₃ ,

Z ^" = chloride, sulphate, phosphate or acetate,

L = linear (C ₂ -C ₆ ) alkylene and optionally substituted by a carboxyl or derivative functional group. According to a preferred characteristic, the cationic groups are obtained from precursors chosen from the group comprising: lysine, pornithine, arginine and their derivatives, choline, ethanolamine (bound by oxygen), putrescine and lysine. agmatine.

The derivatives of lysine, ornithine, and arginine may be, for example, ethyl and methyl esters, amides, and methylated amides.

Thus, cationic groups that can be used in the present invention can be chosen from the following group of radicals:

H H - N - C-CORa

NH ₃ ⁺ , Z-

where Ra represents a hydroxyl, alkoxy or alkylamino group, preferably a group -OMe, -OEt, -NH ₂ , -NHCH ₃ or

-N (CH ₃ ) ₂ , and Z ^" represents a chloride, a sulfate, a phosphate or an acetate, preferably a chloride, or

-NH- (CH ₂ ) ₄ -NH ₃ ⁺ , Z ^" , -NH- (CH ₂ ) ₄ -NH-C (= NH) -NH ₃ ⁺ , Z ^" ,

-O- (CH ₂ ) ₂ -N ⁺ (CH ₃ ) ₃ , Z-, -O- (CH ₂ ) ₂ -NH ₃ ⁺ , Z ^" where Z ^" represents a chloride, a sulfate, a phosphate or an acetate preferably a chloride. For example, the pendant cationic groups may have the following formulas (where the name of the precursor is indicated under each radical): And ester or αmldβ Agmαtine Arghlne ester or αmlde

in which Ra represents a hydroxyl, alkoxy or alkylamino group, preferably a group -OMe, -OEt, -NH ₂ , -NH-CH ₃ or N (CH ₃ ) ₂ .

According to a preferred characteristic, the polyamino acids of the invention comprise on average at least 3 hydrophobic groups (GH) per polymer chain.

Advantageously, at least one of the hydrophobic groups GH is included in a hydrophobic graft comprising at least one spacer (or "spacer") balloon (or pattern) for connecting the hydrophobic group GH to a polyglutamate chain (for example a main chain - skeleton-polyglutamate). This patella may comprise, e.g., at least one direct covalent bond and / or at least one amide bond and / or at least one ester bond. For example, the patella may be of the type belonging to the group comprising, in particular: the amino acid residues different from the constituent monomeric unit of the polyglutamate, the aminoalcohol derivatives, the polyamine derivatives (for example the diamines), the derivatives of the polyols (for example diols) and derivatives of hydroxy acids.

The grafting of GH on the polyglutamate chain may involve the use of precursors of GH, able to bind to the polyglutamate chain.

The precursors of GH are, in practice and without being limited to, selected from the group comprising alcohols and amines, these compounds being easily functionalized by those skilled in the art. The grafting of GH is explained in more detail below in the description of the process for obtaining modified polyamino acids according to the invention.

According to a preferred characteristic, the hydrophobic group GH of the hydrophobic graft comprises from 8 to 30 carbon atoms. These hydrophobic groups GH are advantageously and judiciously selected from the group comprising:

Linear or branched C ₈ to C ₃₀ alkyls which may optionally comprise at least one unsaturation and / or at least one heteroatom,

C ₈ -C ₃₀ alkylaryls or arylalkyls which may optionally comprise at least one unsaturation and / or at least one heteroatom,

And the (poly) cyclic C ₈ to C ₃₀ may optionally comprise at least one unsaturation and / or at least one heteroatom.

The GH-forming patellae of the hydrophobic grafts may be di-, tri- or tetravalent (or even pentavalent and more). In the case of a divalent patella, the hydrophobic graft comprises a single GH group, while a trivalent patella gives the hydrophobic graft a bifid character, that is to say that the graft has two "legs" GH. As an example of a trivalent patella, mention may be made, inter alia, of "amino acid" residues, for example "glutamic acid" or polyol residues, for example glycerol. Thus, two advantageous but non-limiting examples of hydrophobic grafts comprising bifid GHs are dialkylglycerols and dialkylglutamates.

The hydrophobic groups GH can be, for example, derived from groups selected from the following group: octanol, dodecanol, tetradecanol, hexadecanol, octadecanol, oleyl alcohol, tocopherol or cholesterol.

According to another variant, the polyglutamates according to the invention may also carry at least one graft of polyalkylene (preferably ethylene) glycol type bonded to a glutamate residue.

Preferably, the backbone of the polyglutamate according to the present invention comprises alpha-L-glutamate and / or alpha-L-glutamic acid residues.

Even more preferably, the polyglutamates according to the invention have the following formula (I):

(I)

in which :

A represents independently:

RNH- wherein R is H, a linear C ₂ -C _1O, a branched alkyl, C ₃ -C _1O or benzyl, a terminal amino acid residue of the formula:

H -NH - C- -COR ⁷

in which

-R ⁷ is -OH, -OR ⁹ or -NHR ¹⁰ , and

R ⁸ , R ⁹ and R ¹⁰ independently represent H, linear C ₂ -C ₁₀ alkyl, branched C ₃ -C ₁₀ alkyl or benzyl; B is a direct bond, a divalent linking group, trivalent or tetravalent, preferably chosen from the following radicals: -O-, -NH-, -N (C μ ₅ alkyl) -, an amino acid residue (of natural preference), diol, triol, diamine, triamine, aminoalcohol or hydroxyacid having 1 to 6 carbon atoms; D is H, linear C ₂ -C ₁₀ acyl, branched C ₃ -C ₁₀ acyl, or pyroglutamate; the hydrophobic groups GH each independently represent a radical chosen from: • linear or branched C ₈ to C ₃ o alkyls which may optionally comprise at least one unsaturation and / or at least one heteroatom (preferably O and / or N and / or S), or C ₈ -C ₃₀ alkylaryls or arylalkyls which may optionally comprise at least one unsaturation and / or at least one heteroatom (preferably O and / or N and / or S), or

The C ₈ to C ₃₀ (poly) cyclic compounds possibly comprising at least one unsaturation and / or at least one heteroatom (preferably

O and / or N and / or S);

preferably, this radical is selected from the following group: octyloxy-, dodecyloxy-, tetradecyloxy-, hexadecyloxy-, octadecyloxy-, 9-octadecenyloxy-, tocopheryloxy- or cholesteryloxy-, B being then a direct bond;

R ¹ represents a radical chosen from the group having the following formulas:

- -NH- (CH ₂ ) _W -NH ₃ ⁺ , Z ^" with w between 2 and 6, and preferably w is equal to 4,

-NH- (CH ₂ ) ₄ -NH-C (= NH) -NH ₃ ⁺ , Z ^" , -O- (CH ₂ ) ₂ -NH ₃ ⁺ , Z-,

-O- (CH ₂ ) ₂ -N ⁺ (CH ₃ ) ₃ , Z-,

an amino acid residue or an amino acid derivative of formula:

in which :

X is an oxygen atom or a -NH-,

R is H, linear C ₂ to Ci ₀ alkyl, branched C ₃ to Ci ₀ or benzyl,

-R ¹³ is - (CH ₂ ) ₄ -NH ₃ ⁺ , Z ^" , - (CH ₂ ) ₃ -NH-C (= NH) -NH ₃ ⁺ , Z ^" , - (CHa) ₃ -NH ₃ ⁺ , Z; in which the counteranion Z ^" is a chloride, a sulfate, a phosphate or an acetate, preferably a chloride;

R ³ represents a hydroxyethylamino-, an alkylene glycol, a polyalkylene glycol or a radical of formula:

where -R ¹⁰ represents -H, -CO ₂ H, an alkyl ester (preferably -COOMe or -COOEt), -CH ₂ OH, -C (= O) -NH ₂ , -C (= O) - NH-CH ₃ or -C (= O) -N (CH ₃ ) ₂ ;

• p, q, r and s are positive integers; "(p) / (p + q + r + s) is defined as the molar grafting ratio of the hydrophobic groups GH and varies from 2 to 99 mol%, and preferably between 5 and 50%, provided that each copolymer chain has on average at least 3 hydrophobic grafts;

• (q) / (p + q + r + s) is defined as the molar grafting rate of the cationic groups and varies from 1 to 99 mol%;

"(p + q + r + s) varies from 10 to 1000, preferably from 30 to 500;

• (r) / (p + q + r + s) varies from 0 to 98 mol%;

• (s) / (p + q + r + s) varies from 0 to 98 mol%; As well as their pharmaceutically acceptable salts Preferably, the hydrophobic groups GH and the cationic groups are randomly arranged in pendant groups.

In general, the general formula (I) described above should not be interpreted as representing only block copolymers (or blocks), but also random copolymers or multiblock copolymers.

It is also preferable that the molar grafting rate of the polyglutamates according to the invention in the hydrophobic groups is between 2 and 99%, and preferably between 5 and 50%, provided that each polymer chain has an average of at least 3 hydrophobic grafts.

The ratio (q) / (p + q + r + s) of the polyglutamates according to the invention means that they can contain from 1 to about 97 mol% of groups containing a cationic charge.

The ratio (s) / (p + q + r + s) of the polyglutamates according to the invention means that they can be anionic, neutral or cationic at neutral pH.

According to another characteristic of the invention, the polymers according to the invention have a molar mass which is between 2000 and 200000 g / mol, and preferably between 5000 and 100000 g / mol.

Naturally, the invention also covers mixtures of modified polyamino acids as defined above. Remarkably, the polyglutamates of the invention are likely to be used in several ways depending on the nature of the hydrophobic groups and cationic groups, the charge and the degree of polymerization of the polyglutamate. Methods for shaping a polymer for the encapsulation of an active ingredient in the various forms contemplated by the invention are known to those skilled in the art. For more details, we can refer, for example to these few particularly relevant references:

"Microspheres, Microcapsules and Liposomes, vol 1. Preparing and Chemical Applications" Ed R. Arshady, Citas Books 1999. ISBN: 0-9532187-1-6.

"Sustained-Release Injectable Products" Ed. J. Senior and M. Radomsky, Interpharm Press 2000. ISBN: 1-57491-101 -5.

"Colloidal Drug Delivery Systems" Ed. J. Kreuter, Marcel Dekker, Inc. 1994. ISBN:

0-8247-9214-9.

"Handbook of Pharmaceutical Controlled Release Technology" Ed. D. L. Wise, Marcel

Dekker, Inc. 2000. ISBN: 0-8247-0369-3.

These polyglutamates (in the form of particles or not) can encapsulate or easily combine active ingredients such as proteins, peptides, DNA, RNA or small molecules. The preferred shaping is that described in US Pat. No. 6,630,171, which consists of dispersing the copolymer in water and incubating the solution in the presence of an active ingredient (PA). This colloidal solution of vectorization particles, consisting of polyglutamates according to the invention, can then be filtered under 0.2 microns and then directly injected into a patient.

In the case where the polymer is cationic and soluble at acid pH due to an excess of cationic charge and that this charge is partially or completely neutralized at neutral pH, such a polymer is said to be pH dependent. This type of polymer can therefore be used to form a deposit after administration, for example in the subcutaneous tissue.

It should be understood that the carboxylic residual functions of the modified polyglutamate are either neutral (COOH form) or ionized (COO anion), depending on pH and composition. We will therefore speak indifferently of i) glutamate residue or glutamic acid residue, ii) polyglutamic acid or polyglutamate. In aqueous solution, the counter-cation may be a metal cation such as sodium, calcium or magnesium, or an organic cation such as triethanolamine, tris (hydroxymethyl) aminomethane or a polyamine such as polyethyleneimine. If it is divalent, a counter cation can salt two nearby monovalent cationic groups. The counter anion of the cationic groups is preferably chosen from the group comprising a chloride, a sulfate, a phosphate or an acetate. If he is divalent, a counteranion can salify two nearby monovalent cationic groups. As a result, in the present description, the term "pharmaceutically acceptable salts" of the polymer according to the invention all of the polymers with the counter-ions associated with the ionized functions of the polymer. It is also conceivable, for certain structures where there is co-existence of positive and negative charges that there is a total or partial neutralization of charges. A polymer having an equivalent number of positive charges and negative charges (isoelectric point) can exist without the presence of counter-anion or counter-cation.

The copolymers of the invention are for example obtained by methods known to those skilled in the art. First of all, it should be remembered that for the production of alpha-type polyamino acids, the most common technique is based on the polymerization of N-carboxy-amino acid anhydrides (NCA), described, for example, in Biopolymers, 1976, 15, 1869 and in HR Kricheldorf "Alpha-Aminoacid-N-carboxy Anhydride and related Heterocycles" Springer Verlag (1987). The NCA derivative is preferably NCA-GIu-OR ₃ (R ₃ = methyl, ethyl or benzyl). The polymers are then hydrolyzed under appropriate conditions to obtain the polymer in its acid form. These methods are inspired by the description given in patent FR 2801 226. A number of polymers that can be used according to the invention, for example, of the poly (alpha-L-glutamic), poly (alpha-D-glutamic), poly type. (alpha-D, L-glutamate) and poly (gamma-L-glutamic) of variable masses are commercially available. Preferably, the copolymers of the invention are synthesized according to 2 routes. In the first, the cationic group (for example pargininamide) and the group B-GH (for example dodecylamine) are first grafted simultaneously or in sequence on a poly (L-glutamic acid). This reaction can be carried out in a solvent such as DMF, DMSO or NMP according to the following scheme.

In the above mechanism, when q is not zero, the precursor of the R 1 group, such as nitrogen-bound ethanolamine, is introduced during the synthesis together with the cationic group.

In the case where the cationic group contains two chemically undifferentiated amino functions (e.g. linear diamine), it can be introduced in a form in which one of the two functions is protected. A last step of cleavage of the protecting group is then added to the above scheme.

The poly (L-glutamic acid) can be synthesized according to the route described in the patent application FR 2801 226. In the case where the HB-GH group is linked via an ester function, it is easier to graft first. the B-GH group by a conventional coupling reaction using a carbodiimide before grafting the cationic group.

In the above mechanism, when q is not zero, the precursor of the group R 1, such as ethanolamine bonded with nitrogen, is introduced during the synthesis together with the cationic group.

The polymerization chemistry and coupling reactions of the groups are conventional and well known to those skilled in the art (see for example the patents or patent applications of the applicant mentioned above).

These methods will be better understood through the description of the examples.

It should be observed that the degree of polymerization is defined by the molar ratio of the initiator to that of the monomer.

The coupling of the hydrophobic graft GH with an acidic function of the polymer is easily achieved by reacting the polyamino acid in the presence of a carbodiimide as a coupling agent and optionally a catalyst such as 4-dimethylaminopyridine and in a suitable solvent such as dimethylformamide ( DMF), N-methyl pyrrolidone (NMP) or dimethylsulfoxide (DMSO). The carbodiimide is, for example, dicyclohexylcarbodiimide or diisopropylcarbodiimide. Coupling reagents such as chloro Formates can also be used for the formation of amide bonds (see, for example, Bodanszky: "Principles of Peptide Synthesis" Springer Verlag, 1984). The degree of grafting is controlled by the stoichiometry of the constituents and reactants or the reaction time. Hydrophobic grafts functionalized with an amino acid other than that of the polymer are obtained by conventional peptide coupling or by direct condensation by acid catalysis. These techniques are well known to those skilled in the art.

According to another aspect, the invention relates to a pharmaceutical, cosmetic, dietetic or phytosanitary composition comprising at least one polyglutamate as defined above and optionally at least one active ingredient, which may be therapeutic, cosmetic, dietetic or phytosanitary.

According to an advantageous arrangement of the invention, the active principle is associated with (x) polyamino acid (s) modified with a cationic group by one or more bonds other than (or that) chemical bond (s) (s) ( s) covalent (s).

The techniques for associating one or more PAs with modified polyamino acids according to the invention are similar to those described in particular in US Pat. No. 6,630,171. They consist in incorporating at least one active ingredient in the liquid medium containing Vectorization Particles (PV), so as to obtain a colloidal suspension of PV loaded or associated with one or more active principle (s) PA. This incorporation, which leads to PA trapping by the PVs, can be carried out as follows:

• aqueous dissolution of PA, then adding the PV, either as a colloidal suspension or as isolated PV (lyophilisate or precipitate);

Or adding PA, either in solution, or in the pure state or preformulated, to a colloidal suspension of PV particles, optionally prepared extemporaneously by the dispersion of dry PV in a suitable solvent, such as water. Preferably, the active principle is chosen from the group comprising: proteins, glycoproteins, proteins linked to one or more polyalkylene glycol chains [preferably polyethylene glycol (PEG): "PEGylated proteins"], peptides, polysaccharides, liposaccharides, oligonucleotides, polynucleotides and mixtures thereof, and more preferably still in the erythropoietin subgroup, such as epoetin alpha, epoetin beta, darbepoetin, hemoglobin, their analogues or their derivatives; oxytocin, vasopressin, adrenocorticotropic hormone, epidermal growth factor, platelet derived growth factor (PDGF), stimulating factors of hematopoiesis and their mixtures, blood factors, such as alteplase, tenecteplase, factor VΙI (a), factor VII; hemoglobin, cytochromes, albumin, prolactin, luliberin, luteinizing hormone releasing hormone (LHRH) and the like, such as leuprolide, goserelin, triptorelin, buserelin, nafarelin; LHRH antagonists, LHRH agonists, human growth hormones (GH), porcine or bovine hormones, growth hormone releasing factor, insulin, somatostatin, glucagon, interleukins or mixtures thereof (IL-2, IL-1, IL-12), interferons, such as interferon alpha, alpha-2b, beta, beta, or DD gastrin, tetragastrin, pentagastrin, urogastrone, secretin, calcitonin, enkephalins, endomorphins, angiotensins, thyrotropin releasing hormone (TRH), tumor necrosis factor (TNF), nerve growth factor (NGF), growth factors such as beclapermine, trafermine, ancestime, keratinocyte growth factor, granulocyte hematopoietic growth factor (G-CSF), macrophage granulocyte haematopoietic growth factor (GM-CSF), macrophage haematopoietic growth factor ( M-CSF), heparinase, prot bone morphogenetic factors (BMP), human atrial natriuretic factor (hANP), glucagon-like peptide (GLP-I), vascular endothelial growth factor (VEGF), recombinant hepatitis B antigen ( rHBsAg), renin, cytokines, bradykinin, bacitracins, polymyxins, colistins, tyrocidine, gramicidines, etanercept, imiglucerase, alpha drotrecogin, synthetic cyclosporins and analogues, modifications and active fragments pharmaceutically enzymes, cytokines, antibodies, antigens and vaccines, antibodies such as rituximab, infliximab, trastuzumab, adalimumab, omalizumab, tositumomab, efalizumab, and cetuximab. Other active ingredients are polysaccharides (eg, heparin) and oligo- or polynucleotides, DNA, RNA, iRNA, antibiotics, and living cells. Another class of active ingredients includes pharmaceutical substances acting on the central nervous system, for example, risperidone, zuclopenthixol, fluphenazine, perphenazine, flupentixol, haloperidol, fluspirilene, quetiapine, clozapine, amisulprid, sulpiride, ziprasidone, etc.

According to one variant, the active ingredient is a small hydrophobic, hydrophilic or amphiphilic organic molecule. As used herein, a "small" molecule is especially a small nonprotein molecule.

As examples of PA that may be associated with the polyamino acids according to the invention, whether or not in the form of (nano or micro) particles, mention may be made of: o proteins such as insulin, interferons, hormones growth, interleukins, erythropoietin or cytokines; peptides such as leuprolide or cyclosporine; o small molecules such as those belonging to the family of anthracyclines, taxoids or camptothecins; o and their mixtures.

Advantageously, the active principle is chosen from at least one of the following families of active substances: alcohol abuse treatment agents, agents for treating Alzheimer's disease, anesthetics, acromegaly treatment agents, analgesics, anti-asthmatics, allergy-treating agents, anti-cancer agents, anti-inflammatories, anticoagulants and antithrombotics, anticonvulsants, antiepileptics, antidiabetics, antiemetics, anti-glaucoma, antihistamines, antiparasitics, antibiotics, antifungals, antivirals, antiparkinsonians, anticholinergics, antitussives, carbonic anhydrase inhibitors, cardiovascular agents, lipid-lowering agents, anti-arrhythmics, vasodilators, anti-hypertensives, vasoprotective agents, cholinesterase inhibitors, anti-inflammatory agents, disorders of central nervous system, central nervous system stimulants, contraceptives, fertility promoters, uterine labor inducers and inhibitors, cystic fibrosis agents, dopamine receptor agonists, endometriosis, erectile dysfunction agents, fertility agents, gastrointestinal treatment agents, immunomodulators and immunosuppressants, memory disorders agents, anti-migraine medications, muscle relaxants, nucleoside analogues, osteoporosis agents, parasympathomimetics, prostaglandins, psychotherapeutic agents, sedatives, hypnotics and tranquillizers, neuroleptics, anxiolytics, psychostimulants, antidepressants, treatment agents dermatological, steroids and hormones, amphetamines tamines, anorexics, non-analgesic pain relievers, anti-epileptics, barbiturates, benzodiazepines, hypnotics, laxatives, psychotropics and all combinations of these products.

According to one embodiment, the composition of the invention is in the form of a gel, a solution, a suspension, an emulsion, micelles, nanoparticles, microparticles, an implant, a d a powder, a suspension or a film. According to one of its particularly preferred forms, the composition, whether loaded or not with active ingredient (s), is a stable colloidal suspension of nanoparticles and / or microparticles and / or polyamino acid micelles, in an aqueous or oily phase.

Microparticles can be obtained by various methods such as coacervation in the presence of an aggregating agent (divalent or trivalent ions or polyelectrolytes), precipitation by change of pH or ionic strength, extraction / evaporation or by atomization.

In particular, the composition according to the invention may be a colloidal solution of nanoparticles in an aqueous phase at acidic pH and which precipitates at physiological pH.

Advantageously, a polyamino acid of the invention having an excess of cationic charge can condense an anionic active principle such as DNA, a DNA fragment, an RNA or an oligo RNA in the form of nano- or microparticles and these particles can be internalized in a cell.

According to another embodiment, the composition of the invention is in the form of a solution in a biocompatible solvent and can be injected subcutaneously, intramuscularly or into a tumor.

The composition according to the invention, since it is pharmaceutical, can be administered orally, pulmonary, parenterally, nasally, vaginally, ocularly, subcutaneously, intravenously, intramuscularly, intradermally, intraperitoneally, intracerebrally or buccally.

It is also conceivable that the composition is in the form of a solution in a solvent or a mixture of biocompatible solvents that can be injected subcutaneously, intramuscularly or into a tumor.

In another embodiment, the composition may contain an excipient for pH adjustment and / or osmolarity and / or to improve stability (antioxidants) and / or as an antimicrobial agent. These excipients are well known to those skilled in the art (see Injectable Drug Development, P.G. Gupta et al., Interpharm Press, Denver, 1999).

The invention also relates to a preparation process

• drugs, especially for oral, nasal, vaginal, ocular, subcutaneous, intravenous, intramuscular, intradermal, intraperitoneal or intracerebral, the active principles of these drugs may be, in particular, proteins, glycoproteins, proteins linked to one or more polyalkylene glycol chains (for example PolyEthyleneGlycol (PEG), it is called "PEGylated" proteins), peptides hydrophobic, hydrophilic or amphiphilic polysaccharides, liposaccharides, oligonucleotides, polynucleotides and small organic molecules;

• and / or nutrients;

• and / or cosmetic or phytosanitary products; this method being characterized in that it consists essentially in implementing at least one of the polyamino acids as defined above and / or the composition described above.

The invention also relates to a method of therapeutic treatment consisting essentially of administering the composition as described herein, orally, pulmonally, parenterally, nasally, vaginally, ocularly, subcutaneously, intravenously, intramuscularly, intradermally, intraperitoneally, intracerebral or oral.

According to a particular variant of the invention, this method of therapeutic treatment essentially consists in using the composition as described above in the form of a solution in a biocompatible solvent and then injecting it subcutaneously, intramuscularly or into a tumor, preferably so that it forms a deposit on the injection site.

The invention will be better understood and its advantages and implementation variants will emerge from the examples which follow and which describe the synthesis of the polymers of the invention, their transformation into PA vectorization system (stable aqueous colloidal suspension) and the demonstration. the ability of such a system to associate with a protein or nucleic acid to form pharmaceutical compositions.

EXAMPLES Example 1 Synthesis of the Polymer (1)

Indices and groups: T = D, L-α-tocopherol, p = s = 1, q = 198

Ten grams of a poly (glutamic acid) of DP 220 grafted at 5% randomly with racemic α-tocopherol are solubilized in 125 mL of NMP at 80 ^° C. This solution is cooled to 0 ^° C., and 8.7 ml of chlor-butyl chloroformate and 7.35 ml of TV-methyl-morpholine are added. This reaction mixture is stirred for 15 min at 0 ^° C. In parallel, 24.67 g of argininamide dihydrochloride are suspended in 308 mL of NMP and 14.7 mL of triethylamine are added. The suspension obtained is stirred for a few minutes at 20 ^° C. and then cooled to 0 ^° C. The milky suspension of activated polymer is then added to this suspension, and the reaction mixture is stirred for 2 h at 0 ^° C. and then overnight at 20 ^° C. ⁰ C. After addition of 2.1 mL of a solution of 35% HCl then 100 ml of water, the reaction mixture is poured dropwise into 1.6 L of water. The solution obtained is diafiltered against 8 volumes of salt water (0.9%) then 4 volumes of water, and concentrated to a volume of about 250 mL. The percentage of grafted argininamide, determined by proton NMR in D ₂ O, is 90%. Example 2 Synthesis of the Polymer (2)

Indices and groups: T = D, L-α-tocopherol, p = s = 6, q = 108

Three and a half grams of a poly (glutamic acid) of DP 120 grafted at 5% randomly with racemic α-tocopherol are solubilized in 70 ml of NMP at 80 ^° C. This solution is cooled to 0 ^° C. and 3.2 g of n-butyl chloroformate and then 2.37 g of N-methyl-morpholine are added. This reaction mixture is stirred for 10 min at 0 ^° C. In parallel, 4.62 g of 7-tert-butyloxycarbonyl-1,4-butanediamine (BOC-putrescine) are solubilized in 9 ml of NMP and then cooled to ^0.degree. C. The milky suspension of activated polymer is then added to this solution, and the reaction mixture is stirred for 2 h at 0 ^° C. and then overnight at 20 ^° C. After addition of 0.7 ml of a solution of 35% HCl, the reaction mixture is poured dropwise into 317 ml of water. The solution obtained is adjusted to pH = 7.4 with 1N sodium hydroxide and then dialyzed against salt water (0.9%) and then with water. The suspension obtained is lyophilized to give 4.1 g of a white powder. This powder is redissolved in TFA, stirred for 2 h at 20 ⁰ C, then poured dropwise into a foot of water by adjusting the pH to 7 with a 1N sodium hydroxide solution. The solution obtained is diafiltered against 8 salt water volumes (0.9%) then 4 volumes of water, and concentrated to a volume of about 50 mL. The percentage of grafted BOC-putrescine, determined by proton NMR in D ₂ O, is 90%. Example 3 Synthesis of the Polymer (3)

Indices and groups: T = D, L-α-tocopherol, p = 11, q = 88, r = 99, s = 22

Ten grams of a poly (glutamic acid) of DP 220 grafted at 5% randomly with racemic α-tocopherol are solubilized in 125 mL of NMP at 80 ^° C. This solution is cooled to 0 ^° C., and 9.1 ml of chlor-butyl chloroformate and then 7.71 ml of N-methyl morpholine are added. This reaction mixture is stirred for 15 minutes at 0 ^° C. In parallel, 8.2 g of argininamide dihydrochloride are suspended in 103 mL of NMP and 9.31 mL of triethylamine are added. Another 1.6 ml of ethanolamine is added, and the suspension obtained is stirred for a few minutes at 20 ^° C. and then cooled to 0 ^° C. The milky suspension of activated polymer is then added to this suspension, and the reaction mixture is stirred for 2 h to 0 ⁰ C. 1.2 ml of ethanolamine is added, then stirred overnight at 20 ^° C. After adding 2.1 ml of a solution of 35% HCl and then 200 ml of water, the The reaction mixture is poured dropwise into 700 mL of water while adjusting the pH to 7.4. The solution obtained is diafiltered against 8 volumes of salt water (0.9%) then 4 volumes of water, and concentrated to a volume of about 250 mL. The percentage of argininamide and ethanolamine grafted, determined by proton NMR in D ₂ O, are 40 and 45% respectively. Example 4 Synthesis of the Polymer (4)

Indices and groups: T = D, L-α-tocopherol, p = 6, q = 59, r = 52, s = 3

Five grams of a poly (glutamic acid) of DP 120 grafted at 5% randomly with racemic α-tocopherol are solubilized in 63 ml of NMP at 80 ^° C. This solution is cooled to 0 ^° C., and 4.3 g of chlor-butyl chloroformate and then 3.7 g of N-methyl-morpholine are added. This reaction mixture is stirred for 10 min at 0 ^° C. In parallel, 3.15 g of N-tert-butyloxycarbonyl-1,4-butanediamine (BOC-putrescine) are solubilized in 39 ml of ΝMP and then cooled to 0 ^° C. This solution is added to the milky suspension of activated polymer, and the reaction mixture is stirred for 2 hours at

0 ⁰ C. was added 2 ml of ethanolamine and stirred overnight at room temperature. After addition of 1.04 mL of 35% HCl solution, the reaction mixture is dripped into 407 mL of water. The suspension obtained is adjusted to pH = 7.4 with sodium hydroxide

1 Ν, then dialyzed (cutoff threshold of 1 kD) against salt water (0.9%) and then water. The suspension obtained is lyophilized to give a white powder. This powder is redissolved in 100 mL of TFA, stirred 1.25 h at 20 ⁰ C, and then dropped into a foot of water (500 mL) by adjusting the pH to 7 with a solution of sodium hydroxide 1 Ν . After addition of 600 mL of ethanol, the resulting solution is diafiltered against 8 volumes of salt water (0.9%) then 4 volumes of water, and concentrated to a volume of about 100 mL. The percentages of BOC-putrescine and ethanolamine grafted, determined by proton RMΝ in D ₂ O, are 49 and 43%, respectively. Example 5 Synthesis of the Polymer (5)

Indices and groups: T = D, L-α-tocopherol, p = 5, q = 83, s = 12

According to a procedure similar to that used for the synthesis of the polymer (1), about 30O ml of a solution concentrated at 48 mg / g from 10 g of a poly (glutamic acid) of DP 100 grafted to 5% statistically with racemic α-tocopherol, 9.6 g of zsobutyl chloroformate, 7.7 mL of N-methyl-morpholine, 24.7 g of argininamide dihydrochloride and 7 mL of triethylamine. The percentage of grafted argininamide, determined by proton NMR in D ₂ O, is 83%.

Example 6 Synthesis of the Polymer (6)

Indices and groups: T = D, L-α-tocopherol, p = 11, q = 62, s = 147

According to a procedure similar to that used for the synthesis of the polymer (1), about 250 ml of a concentrated solution at 43 mg / g is obtained from 10 g of a poly (glutamic acid) of DP 220 grafted to 5% statistically with racemic α-tocopherol, 2.9 g of n-butyl chloroformate, 2.2 ml of N-methyl-morpholine, 4.93 g of argininamide dihydrochloride and 3.3 mL of triethylamine. The percentage of grafted argininamide, determined by proton NMR in D ₂ O, is 28%.

Example 7 Synthesis of the Polymer (7)

Indices and groups: T = D, L-α-tocopherol, p = 5, q = 40, r = 48, s = 7

According to a procedure similar to that used for the synthesis of the polymer (3), about 20O ml of a concentrated solution at 41 mg / g from 10 g of a poly (glutamic acid) of DP 100 grafted to 5% statistically with racemic α-tocopherol, 9.58 g of ώ-butyl chloroformate, 7.7 ml of N-methyl-morpholine, 8.22 g of argininamide dihydrochloride, 2.86 g of ethanolamine and 5.1 ml of triethylamine. The percentages of argininamide and ethanolamine grafted, determined by proton NMR in D ₂ O, are 40 and 48%, respectively.

Example 8 Synthesis of the Polymer (8)

Indices and groups: T = D, L-α-tocopherol, p = 11, q = 139, s = 70

According to a procedure similar to that used for the synthesis of the polymer (1), about 20O ml of a concentrated solution at 51 mg / g is obtained from 10 g of a poly (glutamic acid) of DP 220 grafted to 5% statistically with racemic α-tocopherol, 6.39 g of tert-butyl chloroformate, 5.1 ml of vV-methyl-morpholine, 13.16 g of argininamide dihydrochloride and 7.5 mL of triethylamine. The percentage of grafted argininamide, determined by proton NMR in D ₂ O, is 63%.

Example 9 Synthesis of the Polymer (9)

Indices and groups: T = D, L-α-tocopherol, p = 1, q = 121, s = 88

Five grams of a poly (glutamic acid) of DP 220 grafted at 5% randomly with racemic α-tocopherol are solubilized in 63 ml of NMP at 80 ^° C. This solution is cooled to 0 ^° C., and 2.38 mL of ώ-butyl chloroformate and then 2.02 mL of N-methyl-morpholine are added. This reaction mixture is stirred for 15 min at 0 ^° C. In parallel, 4.93 g of argininamide dihydrochloride are suspended in 62 ml of NMP and 2.8 ml of triethylamine are added. The suspension obtained is stirred for a few minutes at 20 ^° C. and then cooled to 0 ^° C. The milky suspension of activated polymer is then added to this suspension, and the reaction mixture is stirred for 2 h at 0 ^° C. and then 4 h at 20 ^° C. ⁰ C. After adding 1.04 mL of a solution of HCl 35% and 5O ml of water, the reaction mixture was poured dropwise in 50O mL of acidified water (pH = 3), maintaining the pH to 3 - 4 with a solution of 1N HCl. The solution obtained is diafiltered against 8 volumes of salt water (0.9%) then 4 volumes of water, and concentrated to a volume of approximately 250 mL. The percentage of grafted argininamide, determined by proton NMR in D ₂ O, is 55%. Example 10 Synthesis of the Polymer (10)

Indices and groups: T = D, L-α-tocopherol, p = 11, q = 88, r = 48, s = 73

Ten grams of a poly (glutamic acid) of DP 220 grafted at 5% randomly with racemic α-tocopherol are solubilized in 125 mL of NMP at 80 ^° C. This solution is cooled to 0 ^° C., and 5.6 ml of n-methyl-morpholine chloroformate and then 4.8 ml of N-methyl-morpholine are added. This reaction mixture is stirred for 15 min at 0 ^° C. In parallel, 7.4 g of argininamide dihydrochloride are suspended in 93 ml of NMP, then 4.7 ml of triethylamine and 1.2 ml of ethanolamine are added. The suspension obtained is stirred for a few minutes at 20 ^° C. and then cooled to 0 ^° C. The milky suspension of activated polymer is then added to this suspension, and the reaction mixture is stirred for 2 h at 0 ^° C. and then overnight at 20 ^° C. ⁰ C. After adding 2.07 mL of a solution of 35% HCl then 20O mL of water, the reaction mixture is poured dropwise into 670 mL of water acidified to pH = 3 with HCl, maintaining pH around 3 with 1N HCl solution. The solution obtained is diafiltered against 8 volumes of salt water (0.9%) and then 4 volumes of water, and concentrated to a volume of approximately 250 ml. . The percentages of grafted argininamide and ethanolamine, determined by proton NMR in D ₂ O, are 40 and 22%, respectively. Comparative Example 11: Cl Compound Not Functionalized by a Cationic Group

Comparative compound C1 is the precursor (in its anionic form) of the polyglutamate modified with a cationic group, ie DP20 polyglutamate grafted at 5% statistically with racemic alpha-tocopherol. This compound is obtained by the method described in application WO-A-03/104303.

Example 12: Insulin Association Study

An aqueous solution containing 10 mg of polymer per milliliter at pH 7.4 and 200 IU insulin (7.4 mg) is prepared. The solutions are incubated for 2 hours at room temperature and the free insulin is separated from the associated insulin by ultrafiltration (threshold at 100 kDa, 15 min at 10,000 ^° C. at 18 ^° C.). The free insulin recovered in the filtrate is then assayed by HPLC (High Performance Liquid Chromatography) and the amount of insulin associated is deduced. The results are given in Table 1 below.

TABLE 1

The results demonstrate that the polymers of the invention are capable of strongly associating insulin to give colloidal suspensions larger than 100 kDa and the levels of association with insulin are very high. In comparison with the polymer Cl, it is found that the presence of cationic charges does not decrease the level of insulin associated.

Example 13 Measurement of the viscosity (mPa / s) under shear of an aqueous solution at 29 mg / g with a gradient of speed of 10 s ^-1

The results demonstrate that the polymers of the invention are significantly less viscous than the reference C1 which does not contain pendant cationic groups.

Example 14: Solubility versus pH study

The results show that certain polymers of the invention (Examples 9 and 10) have pH-dependent solubility properties which, unlike the compound Cl, allow them to be formulated with an active ingredient at a moderately acidic pH (pH = 4). and form a deposit at physiological pH (pH around 7).

Example 15: Association Study of a Therapeutic RNA

Polymer / RNA combinations in aqueous solution are carried out by adding increasing amounts of polymers (1), (3) (two examples of polymers of the invention which are globally cationic at pH = 7.4) or Cl at fixed quantities. a therapeutic RNA of 1433 nucleotides. These mixtures are incubated for 2 h at 37 ^° C. and then analyzed by electrophoresis on 1% agarose gel under denaturing conditions (revelation of the RNAs with ethidium bromide). RNA alone serves as a positive control of integrity. RNA incubated with commercial RNAses serves as a control of degraded RNA. The results show that when the RNA studied was incubated with the polymers (1) or (3), the amount of RNA that migrates to the expected size in electrophoresis decreases gradually with the amount of polymer used for the combination and disappears. beyond a certain value without appearance of other bands. In contrast, in the mixture made with the compound Cl, the amount of RNA that migrates is unchanged, even in the presence of a large excess of polymer.

In a ^2nd time is added to the RNA the amount of polymer (1) or (3) to fully involve (conditions where the RNA is no longer visible to the expected size on gel) and allowed to these mixtures incubate for 2 h at 37 ^° C. After 2 h at 37 ^° C., increasing amounts of compound C1 are added to these mixtures and a new incubation is carried out for 16 h at 37 ^° C. The mixtures obtained are analyzed by electrophoresis. on 1% agarose gel under denaturing conditions (revelation of RNA ethidium bromide). The results show that the expected size is observed increasing amounts of RNA correlated with the amount of Cl polymer added to the RNA / polymer mixture (1) or (3).

These results show that certain polymers of the invention, globally cationic at pH = 7.4, make it possible to associate a model RNA of 1433 nucleotides and that this combination is reversible in the presence of a globally anionic polymer. In addition, the formulated RNA is not degraded.

Example 16: Study of the crossing of the cell membrane of a model oligonucleotide

Opti-MEM ^® medium without fetal calf serum is mixed with a Cy3-labeled 30-base RNA oligonucleotide at an amount of polymer (3) or (7) close to the minimum amount to associate the entire oligonucleotide. . This mixture is contacted on Huh-7 human hepatocarcinoma cells grown in 24-well plate at 25,000 cells / well. After 4 hours of incubation at 37 ^° C., 5% CO ₂ , D-MEM medium is added to 20% fetal calf serum (FCS) so that the final concentration of FCS is 10%. After 24 hours of incubation of the cells in the presence of oligonucleotide / polymer mixtures, the cells are washed, membrane-labeled with biotinylated concanavalin and then fixed for 3 min with 3.7% paraformaldehyde. After 2 washes with PBS, cells were incubated with DAPI (nuclear DNA) for 10 min, washed 3 times with PBS and then incubated with streptavidin labeled with Alexa Fluor ^® 488 which reveals the biotinylated concanavalin. The cells are observed by confocal microscopy. The localization of the cells is possible by observation of their membrane labeled with Alexa Fluor ^® 488 and their marked nucleus at the DAPI. The entry of the Cy3-labeled oligonucleotide into the cell is visualized by observation of Cy3 fluorescence (excitation at 550 nm, emission at 570 nm).

The results show that Cy3-labeled oligonucleotide is found in the cytoplasm of Huh-7 cells when the oligonucleotide has been incubated on the cells in the presence of polymers (3) or (7).

In comparison, when the oligonucleotide is incubated alone, or in the presence of the Cl polymer, on Huh-7 cells, the presence of the labeled oligonucleotide in the cells is not observed.

Claims

1. Polyamino acids, or their pharmaceutically acceptable salts, comprising glutamic residues, characterized in that some of the glutamic residues carry a pendant cationic group, which, if it is deprotonable, has a pKa greater than or equal to 7, said cationic groups being identical or different from each other, and in that other glutamic residues carry a hydrophobic group (GH) during, the hydrophobic groups (GH) being identical or different from each other, said cationic groups usable corresponding to the following formula:

X IW ₃ ⁺ Z

in which: X = O ₅ NH,

Y = independently an H or a CH ₃ ,

Z ^" = chloride, sulphate, phosphate or acetate,

L = linear (C ₂ -C ₆ ) alkylene and optionally substituted by a carboxyl or derivative functional group, said hydrophobic groups (GH) being chosen from the group comprising:

And the (poly) cyclic C 8 to C ₃₀ may optionally comprise at least one unsaturation and / or at least one heteroatom.

2. Polyamino acids according to claim 1, characterized in that the pendant cationic groups are grafted to the glutamic residues through an amide or ester bond.

3. Polyamino acids according to any one of the preceding claims, characterized in that they consist of functionalized homopolymers of L-glutamate or L-glutamic acid bound in the polyamino acid by their carboxyl alpha position.

4. Polyamino acids according to any one of the preceding claims, characterized in that the pendant cationic groups are chosen from the following group of radicals:

where Ra represents a hydroxy, alkoxy or alkylamino group, preferably a group -OMe, -OEt, -NH ₂ , -NHCH ₃ or -N (CH ₃ ) ₂ , and T represents a chloride, a sulfate, a phosphate or a acetate, preferably a chloride, or -NH- (CH ₂ ) ₄ -NH ₃ ⁺ , Z ^" ,

-NH- (CH ₂ ) ₄ -NH-C (= NH) -NH ₃ ⁺ , Z ^" , -O- (CH ₂ ) ₂ -N ⁺ (CH ₃ ) ₃ , Z ^" , -O- (CH ₂₎ ) ₂ -NH ₃ ^+, Z ^"wherein ^Z" represents a chloride, sulfate, phosphate or acetate, preferably chloride.

5. Polyamino acids according to any one of the preceding claims, characterized in that they comprise on average at least 3 hydrophobic groups (GH) per polymer chain.

6. Polyamino acids according to any one of the preceding claims, characterized in that still other glutamic residues carry a nonionizable group during, different from the hydrophobic groups (GH), said nonionizable groups being identical or different from each other. .

7. Polyamino acids according to claim 6, characterized in that said pendant nonionizable group is the hydroxyethylamino-radical.

8. Polyamino acids according to any one of the preceding claims, characterized in that still other glutamic residues carry a non-ionized group at neutral pH, different from hydrophobic groups (GH), said nonionized groups at neutral pH being identical or different from each other.

9. Polyaminocacids according to claim 9, characterized in that said nonionized group at neutral pH corresponds to the following formula:

where -R ¹⁰ is -H, -CO ₂ H, alkyl ester (preferably -COOMe or

-COOEt), -CH ₂ OH, -C (= O) -NH ₂ , -C (= O) -NH-CH ₃ or -C (= O) -N (CH ₃ ) ₂ .

10. Polyamino acids according to any one of the preceding claims, characterized in that they carry at least one graft of polyalkylene type (preferably ethylene) glycol bonded to a glutamate residue.

11. Polyamino acids according to any one of the preceding claims, characterized in that they correspond to the following formula (I):

in which :

A represents independently:

RNH- wherein R is H, C ₂ -C 10 linear alkyl, C ₃ -C 10 branched alkyl or benzyl, a terminal amino acid residue of formula:

H -NH - C- -COR '

in which

-R ⁷ is -OH, -OR ⁹ or -NHR ¹⁰ , and

R ⁸ , R ⁹ and R ¹⁰ independently represent H, linear C ₂ -C ₁₀ alkyl, branched C ₃ -C ₁₀ alkyl or benzyl; B is a direct bond, a divalent linking group, trivalent or tetravalent, preferably chosen from the following radicals: -O-, -NH-, -N _(C1 ₅ alkyl.) -, an amino acid residue ( preferably natural), diol, triol, diamine, triamine, aminoalcohol or hydroxyacid having 1 to 6 carbon atoms; D is H, linear C ₂ -C ₁₀ acyl, branched C ₃ -C ₁₀ acyl, or pyroglutamate; the hydrophobic groups GH each independently represent a radical chosen from: • linear or branched C ₈ to C ₃ o alkyls which may optionally comprise at least one unsaturation and / or at least one heteroatom (preferably O and / or N and / or S), or C ₈ -C ₃₀ alkylaryls or arylalkyls which may optionally comprise at least one unsaturation and / or at least one heteroatom (preferably O and / or N and / or S), or

O and / or N and / or S);

R ¹ represents a radical chosen from the group having the following formulas: -NH- (CH ₂ ) _W -NH ₃ ⁺ , Z ^" with w between 2 and 6, and preferably w is equal to 4,

-NH- (CH ₂ ) ₄ -NH-C (= NH) -NH ₃ ⁺ , Z 1 -O- (CH ₂ ) ₂ -NH ₃ ⁺ , Z ^" ,

-O- (CH ₂ ) ₂ -N ⁺ (CH ₃ ) ₃ , Z ^" , an amino acid residue or an amino acid derivative of formula:

in which :

X is an oxygen atom or a -NH-,

R is H, linear C ₂ -C ₁₀ alkyl, branched C ₃ -C ₁₀ alkyl or benzyl,

-R ¹³ is - (CH ₂ ) ₄ -NH ₃ ⁺ , Z ^" , - (CH ₂ ) ₃ -NH-C (= NH) -NH ₃ ⁺ , Z ^" , - (CH ₂ ) ₃ -NH ₃ ⁺ , Z; in which the counteranion Z ^" is a chloride, a sulfate, a phosphate or an acetate, preferably a chloride;

R ³ represents a hydroxyethylamino-, an alkylene glycol residue, a polyoxyalkylene glycol or a radical of formula:

where -R ¹⁰ represents -H, -CO ₂ H, an alkyl ester (preferably -COOMe or -COOEt), -CH ₂ OH, -C (= O) -NH ₂ , -Q = O) -NH -CH ₃ or -C (= O) -N (CH ₃ ) ₂ ;

• p, q, r and s are positive integers; (P) / (p + q + r + s) is defined as the molar grafting ratio of the hydrophobic groups GH and varies from 2 to 99 mol%, and preferably between 5 and 50%, provided that each copolymer chain has on average at least 3 hydrophobic grafts;

(P ^{+ c} 1 ^{+ r + s} ) ranges from 10 to 1000, preferably from 30 to 500;

• (r) / (p + q + r + s) varies from 0 to 98 mol%;

• (s) / (p + q + r + s) varies from 0 to 98 mol%; as well as their pharmaceutically acceptable salts.

12. A pharmaceutical, cosmetic, dietetic or phytosanitary composition comprising at least one polyamino acid comprising cationic group-modified glutamic residues according to any one of claims 1 to 11.

13. Composition according to claim 12, characterized in that it comprises at least one active ingredient.

14. A composition according to claim 13, characterized in that the active ingredient is associated with (x) polyamino acid (s) comprising glutamic residues modified (s) by a cationic group with one or more bonds other (s) that ( or chemical bond (s) covalent (s).

15. Composition according to any one of claims 12 to 14, characterized in that the active ingredient is selected from the group consisting of DNA, a DNA fragment, an RNA and an oligo RNA.

16. Composition according to any one of claims 12 to 15, characterized in that it is a colloidal suspension of nanoparticles and / or microparticles and / or polyamino acid micelles comprising glutamic residues modified with a cationic group, in a aqueous or oily phase.

17. Composition according to any one of claims 12 to 16, characterized in that the suspension is a colloidal solution of nanoparticles in an aqueous phase at acidic pH and which precipitates at physiological pH.

18. A process for the preparation of medicaments, in particular for oral, nasal, pulmonary, vaginal, ocular, subcutaneous, intravenous, intramuscular, intradermal, intraperitoneal or intracerebral administration, the active principles of these medicinal products being, in particular, proteins, glycoproteins, proteins bound to one or more polyalkylene glycol chains, peptides, polysaccharides, liposaccharides, oligonucleotides, polynucleotides and hydrophobic, hydrophilic or amphiphilic organic small molecules; and / or nutrients; and / or cosmetic or phytosanitary products; characterized in that it essentially consists in using at least one of the polyamino acids according to any one of claims 1 to 11 and / or the composition according to any one of claims 12 to 17.