FR3067247A1 - COMPOSITIONS IN THE FORM OF AN INJECTION AQUEOUS SOLUTION COMPRISING HUMAN GLUCAGON AND A CO-POLYAMINOACID - Google Patents

Abstract

The invention relates to a co-polyamino acid bearing carboxylate charges and hydrophobic radicals Hy.

Description

Compositions in the form of an aqueous solution for injection comprising human glucagon and a co-polyamino acid Human glucagon is a hyperglycemic hormone of short action which makes it possible to increase the glycemia, thus correcting a hypoglycemic level which may result from a excess insulin. It allows the release of glucose by stimulation of hepatic glycogenolysis, and has antagonistic properties of insulin (hypoglycemic). Human glucagon is normally secreted by alpha cells from the islets of Langerhans in the pancreas when hypoglycemia is detected.

Human glucagon is used for therapeutic purposes, such as the emergency treatment of severe hypoglycaemia, also called "rescue", but also in a diagnostic context when carrying out medical examinations, for example to inhibit the gastrointestinal motility. Other applications are also envisaged for human glucagon, in particular its use in a system of bi-hormonal regulation of the glycemia also called artificial pancreas and in the congenital hyperinsulinism which is a rare disease characterized by very high levels of insulin.

The clinical use of human glucagon has been limited because of some of its unfavorable properties for developing a stable pharmaceutical product for therapeutic purposes. In fact, human glucagon has very low solubility at physiological pH, high physical instability, because of its propensity to form fibrils over a wide range of pH. It is for this reason that the only commercial products based on human glucagon (Glucagen®, NOVO NORDISK and Glucagon for injection, ELI LILLY) are freeze-dried forms to be reconstituted extemporaneously. The work of Onoue et al. (Pharm. Res. 2004, 21 (7), 1274-83) have shown the potentially dangerous nature of these fibrils: the fibrillated human glucagon being cytotoxic in cultured mammalian cells.

In addition to its physical instability, human glucagon undergoes various types of chemical degradation. In aqueous solution, it degrades quickly to form several degradation products. At least 16 degradation products of human glucagon have been identified by Kirsh et al. (International Journal of Pharmaceutics, 2000, 203, 115-125). The chemical degradation of this human glucagon is therefore rapid and complex.

The poor chemical and physical stability of human glucagon in solution has led pharmaceutical companies such as NOVO NORDISK, ELI LILLY and more recently FRESENIUS KABI to market this human glucagon in the form of a lyophilisate to be reconstituted at acid pH (pH < 3) just before injection. Human glucagon in the form of lyophilisate is more stable, and the preparation of the formulation at acid pH just before use makes it possible to obtain a clear solution. However, once the product has been reconstituted, it must be used quickly because it undergoes extremely rapid chemical and physical degradation in the acid reconstitution buffer, with the appearance of human glucagon fibrils within 24 hours of reconstitution. This presentation of the product is however unsatisfactory because it requires very rapid use of the formulation. This instability not only makes use of the pump impossible, but it also has the disadvantage of leading to significant loss of product in diagnostic use. Indeed, a composition of this type is no longer usable a few hours after preparation that causes wastage.

Finally, even in the application of emergency treatment for severe hypoglycemic reactions, which may occur during insulin therapy in diabetic patients, the formulation to be reconstituted is also not ideal, since it involves long preparation and complicated, for example the GlucaGen® leaflet describes a 5-step process for injecting the recommended dose. Moreover, a study by the company LOCEMIA shows that very few people (around 10% of the participants) having to carry out the reconstitution in an emergency were able to deliver the adequate dose. Finally, the acid pH of human glucagon solutions can cause pain when injected into the patient.

[0008] There is therefore a need for a ready-to-use human glucagon solution. Today, the most advanced solutions from a clinical point of view to allow the delivery of human glucagon bypass the problem of stability of human glucagon in aqueous solution in different ways.

LOCEMIA has developed a spray of lyophilized human glucagon, currently being tested in a phase 3 clinical study, which is intended to be administered by the intranasal route. This spray is suitable for use called "rescue", that is to say in the case of severe hypoglycemia, because it is ready for use and therefore easy to use, unlike the solutions to be reconstituted. However, this product is not suitable for pump use or for use requiring precise control of the quantity of human glucagon delivered.

For its part, XERIS has developed a liquid formulation of human glucagon based on a polar aprotic solvent, such as DMSO, currently tested in clinical studies. However, if the injection of solution of organic solvents for use of the “rescue” type is possible, it is largely preferable to have an aqueous solution of human glucagon for chronic use. Compositions comprising an association with other peptides are envisaged, in particular amylin or a GLP-1 RA (Glucagon like peptide-1 receptor agonist).

Finally, faced with the difficulties in formulating human glucagon, analogs of human glucagon are being developed by large pharmaceutical companies, such as NOVO NORDISK, SANOFI OR ELI LILLY, in order to obtain formulations having stability compatible with pharmaceutical use.

However, these peptides whose primary sequence has been modified compared to the peptide of human origin may present a safety risk to patients.

There is therefore a major interest in a solution making it possible to improve the solubilization and the stability, both chemical and physical, of human glucagon in aqueous solution at a pH close to physiological pH, that is to say say between 6.0 and

8.0. This could make it possible to obtain a pharmaceutical product more easily usable by the patient in the event of an emergency, but also to open the field to new therapeutic applications of human glucagon, such as for example its use in an artificial bihormonal pancreas.

The prior art offers solutions to try to solve this problem.

Some documents suggest placing oneself at basic pH. for example

US2015291680 teaches the solubilization of human glucagon at 1 mg / ml by placing it at a pH between 8.8 and 9.4 and using ferulic acid or tetrahydrocurcumin. However, in addition to placing itself at basic pH, this solution has the drawback of leading to a fairly limited stability of human glucagon over time. The article by Jackson et al (Curr. Diab. Rep., 2012, 12, 705-710) proposes to formulate human glucagon at basic pH (around 10) in order to limit the formation of fibrils. However, this solution does not prevent rapid chemical degradation of human glucagon.

Application W02014096440 (NOVOZYME), on the contrary, envisages placing itself at a slightly acidic pH (around 5.5) in the presence of albumin and polysorbate, in order to improve stability by reducing the speed of fibrillation. However, this solution has a limited improvement in stability. Most of the solutions described in the prior art making it possible to obtain a clear solution of human glucagon and to prevent the aggregation, gelling or precipitation of human glucagon involve the use of known surfactants, detergents or solubilizing agents . For example, Matilainen et al (J. Pharm. Sci, 2008, 97, 2720-2729 and Eur.

J. Pharm. Sci., 2009, 36, 412-420) described the use of cyclodextrin in order to limit the rate of formation of human glucagon fibrils. However, the improvement made seems insufficient to envisage use as a pump.

Among the solutions proposed are hydrophilic surfactants:

- GB1202607 (NOVO NORDISK) describes the use of anionic or cationic detergents.

- US6384016 (NOVO NORDISK) and US2011097386 (BIODEL) use lysophospholipids (or lysolecithins).

- WO2015095389 (AEGIS) describes nonionic surfactants, such as dodecyl maltoside, for improving the bioavailability of therapeutic agents, in the case of delivery by application to the mucous membranes or the epidermis, and in particular in the case of ocular delivery , nasal, oral or nasolacrimal. This document describes that the presence of alkyl glycosides leads to an improvement in the absorption of human glucagon at the ocular level,

- Application WO2012059764 (ARECOR) describes cationic surfactants, and more specifically aromatic ammonium chlorides.

The surfactants indicated in the above documents may be too toxic or irritating for chronic use by the subcutaneous route. For example, lysophospholipids (or lysolecithins) are known to lyse red blood cells because of their hemolytic properties. During a subcutaneous injection, this can cause local tissue damage and pain at the injection site. In the case of a continuous injection by a pump, this can lead to pain and / or irritation at the needle insertion site. International application WO2011138802 (Sun Pharma) describes a ready-to-use solution of human glucagon in aqueous micellar solution at a pH between 5 and 7.5 in the presence of a pegylated lipid (pegylated distearoyl-phosphotidylethanolamine). However, Garay et al. (Expert Opin Drug Deliv (2012) 9, 1319-1323) teach that Poly Ethylene Glycol is both immunogenic and antigenic. This can be detrimental to patients with anti-PEG antibodies. Moreover, Ganson et al. (J. Allergy Clin. Immunol.

(2015) doi: 10.1016 / j.jaci.2015.10.034) describe that a clinical study involving pegnivacogin coupled with 40 kDa methoxypolyethylene glycol (mPEG) led to inflammatory responses from the first dose of pegnivacogin on 3 of 640 patients. Among these three patients two met the criteria for anaphylaxis and one presented an isolated dermal reaction, each event was considered serious, and one was even considered to be endangering the patient's life. These adverse events caused the termination of the clinical trial and raised the problem of the adverse effects of pegylated compounds.

Document W02013101749 (LATITUDE) describes nano-emulsions of human glucagon. However, it claims fairly modest performances in terms of chemical stability, that is to say that the composition comprises at least 75% of the initial concentration after 3-7 days at 37 ° C.

In addition, it should be noted that, to date, to the knowledge of the applicant, no pharmaceutical formulation comprising human glucagon in the form of an aqueous solution has been tested in a clinical study.

There therefore remains a need for a liquid aqueous formulation at a pH 5 close to the physiological pH of between 6.0 and 8.0 making it possible to dissolve and obtain good stability of human glucagon, both in terms of physical stability. than chemical stability. More particularly there is a need for such a formulation which can be used in a bihormonaie pump (insulin / human glucagon).

This need is all the more clear since Tan et al. (Diabetes, 2013, 62, 1131138) shows that combining human glucagon with a GLP-1 RA is an attractive proposition for the treatment of obesity and diabetes. However, being able to formulate human glucagon in a stable manner in an aqueous solution at a pH close to the physiological pH of between 6.0 and 8.0 makes it possible to be in more favorable conditions in order to be able to improve the stability of GLP-1 RA sensitive to acidic or basic conditions.

The invention thus relates to physically stable compositions in the form of an injectable aqueous solution, the pH of which is between 6.0 and 8.0, comprising at least:

a) human glucagon and

b) a co-polyamino acid carrying carboxyate charges and hydrophobic Hy radicals, said co-polyamino acid consisting of glutamic or aspartic units and said hydrophobic Hy radicals being of formula I below:

* - ^ GpR ^ -GpA ^ GpC) ^ra P Formula I in which

- GpR is a radical of formulas II or ΙΓ:

G

It HH H * _ _N _ _N _ _N _ * _π * _Qu _LL _R _ _ _N * _n,;

- GpA is a radical of formulas III or ΙΙΓ:

Ο HN- *

HN- * ni or * - ^ - A-N- * m ';

- GpC is a radical of formula IV:

- the * indicate the sites of attachment of the different groups;

- a is an integer equal to 0 or 1;

- b is an integer equal to 0 or 1;

p is an integer equal to 1 or 2 and o if p is equal to 1 then a is equal to 0 or to 1 and GpA is a radical of formula III 'and, o if p is equal to 2 then a is equal to 1, and GpA is a radical of formula III;

- c is an integer equal to 0 or 1, and if c is equal to 0 then d is equal to 1 or 2;

- d is an integer equal to 0, 1 or 2;

- r is an integer equal to 0 or to 1, and o if r is equal to 0 then the hydrophobic radical of formula I is linked to the copolyamino acid via a covalent bond between a carbonyl of the hydrophobic radical and a nitrogen atom in position N end of the copolyamino acid, thus forming an amide function resulting from the reaction of an amine function in the N terminal position of the co-polyamino acid precursor and an acid function carried by the precursor of the hydrophobic radical, and o if r is equal to 1 then the hydrophobic radical of formula I is linked to the copolyamino acid:

via a covalent bond between a nitrogen atom of the hydrophobic radical and a carbonyl of the co-polyamino acid, thus forming an amide function resulting from the reaction of an amine function of the precursor of the hydrophobic radical and an acid function carried by the precursor of the co -polyamino acid or via a covalent bond between a carbonyl of the hydrophobic radical and a nitrogen atom in the N-terminal position of the co-polyamino acid, thus forming an amide function resulting from the reaction of an acid function of the precursor of the hydrophobic radical and a function

Ί amine in the N terminal position carried by the precursor of the copolyamino acid;

- R is a radical chosen from the group consisting of:

o a divalent, linear or branched alkyl radical, comprising if GpR is a radical of formula II of 2 to 12 carbon atoms or if GpR is a radical of formula ΙΓ of 1 to 11 carbon atoms;

o a divalent, linear or branched alkyl radical, comprising if GpR is a radical of formula II of 2 to 11 carbon atoms or if GpR is a radical of formula ΙΓ of 1 to 11 carbon atoms, said alkyl radical carrying one or more -CONH2 functions, and o an ether or unsubstituted polyether radical comprising from 4 to 14 carbon atoms and from 1 to 5 oxygen atoms;

- A is a linear or branched alkyl radical comprising from 1 to 6 carbon atoms;

- B is a linear or branched alkyl radical, optionally comprising an aromatic ring, comprising from 1 to 9 carbon atoms;

- Cx is a linear or branched monovalent alkyl radical, in which x indicates the number of carbon atoms and:

o if p is equal to 1, x is between 11 and 25 (11 <x <25):

o if p is equal to 2, x is between 9 and 15 (9 <x <15), the ratio i between the number of hydrophobic radicals and Se number of glutamic or aspartic units being between 0 <i <0 , 5;

when several hydrophobic radicals are carried by a co-polyamino acid then they are identical or different, the degree of polymerization DP in glutamic or aspartic units is between 5 and 250;

the free acid functions being in the form of an alkaline cation salt chosen from the group consisting of Na ⁺ and K ⁺ .

In one embodiment, the composition is characterized in that the pH is between 6.6 and 7.8.

In one embodiment, the composition is characterized in that the pH is between 6.8 and 7.4.

In one embodiment, the composition is characterized in that said hydrophobic radicals are chosen from the hydrophobic radicals of formula I in which if p is equal to 1 and if x is less than or equal to 14 (x <14) then r = 0 or r = 1.

In one embodiment, the composition is characterized in that said hydrophobic radicals are chosen from hydrophobic radicals of formula I in which if p is equal to 1 and if x is between 15 and 16 (15 <x <16), then r = 1.

In one embodiment, the composition is characterized in that said hydrophobic radicals are chosen from the hydrophobic radicals of formula I in which if p is equal! at 1 and if x is greater than 17 (17 <x) then r = 1 and R is an ether or poiether radical.

In one embodiment, the composition is characterized in that said hydrophobic radicals are chosen from hydrophobic radicals of formula I in which if p is equal to 1 then x is between 17 and 25 (17 <x <25 ).

In one embodiment, its composition is characterized in that said hydrophobic radicals are chosen from among its hydrophobic radicals of formula I in which p - 1, represented by the following formula V:

^ GpR ^ -GpA ^ - GpC ^ra formula V

GpR, GpA, GpC, r and a have the definitions given above.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radica! of formula V in laqueile: r is equal to 1 (r = l) and a is equal to 0 (a = 0).

In one embodiment, the composition is characterized in that radica! hydrophobic is a radical of formula V in which r is equal to 1 (r = l) and a is equal to 1 (a = l).

In one embodiment, the composition is characterized in that the hydrophobic radical is a radica! of formula V in which GpR is a radical of formula II.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radica! of formula V in laqueile GpR is a radical of formula II in which R is a divalent linear alkyl radical comprising from 2 to 12 carbon atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in laqueile GpR is a radical of formula II in which R is a divalent alkyl radical comprising from 2 to 6 carbon atoms .

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II in which R is a divalent linear alkyl radical comprising from 2 to 6 atoms of carbon.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II in which R is a divalent alkyl radical comprising from 2 to 4 carbon atoms .

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II in which R is a divalent linear alkyl radical comprising from 2 to 4 atoms of carbon.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II in which R is a divalent alkyl radical comprising 2 carbon atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula ΙΓ.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II 'in which R is a divalent linear alkyl radical comprising from 1 to 11 atoms of carbon.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula ΙΓ in which R is a divalent alkyl radical comprising from 1 to 6 carbon atoms .

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II or ΙΓ, in which R is a divalent alkyl radical, comprising from 2 to 5 carbon atoms and carrying one or more amide functions (-CONH2).

In one embodiment, the composition is characterized in that Se hydrophobic radical is a radical of formula V in which GpR is a radical of formula ΙΓ or II, in which R is a divalent linear alkyl radical, comprising of 2 with 5 carbon atoms and carrying one or more amide functions (-CONH2).

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II or ΙΓ in which R is a radical chosen from the group consisting of radicals represented by the formulas below:

In one embodiment, the composition is characterized in that the radical R is linked to the co-polyamino acid via an amide function carried by the carbon in the delta or epsilon position (or in the 4 or 5 position) relative to the amide function (10 CONH ₂ ).

In one embodiment, the composition is characterized in that Se hydrophobic radical is a radical of formula V in which GpR is a radical of formula II or ΙΓ, in which R is a linear ether or unsubstituted polyether radical comprising from 4 to 14 carbon atoms and from 1 to 5 oxygen atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II or ΙΓ, in which R is an ether radical.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II or II ', in which R is an ether radical comprising from 4 to 6 carbon atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II or ΙΓ in which R is an ether radical represented by the formula

In one embodiment, its composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II or ΙΓ, in which R is a polyether radical.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which GpR is a radical of formula II or ΙΓ, in which R is a linear polyether radical comprising from 6 to 10 carbon atoms and 2 to 3 oxygen atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical of formula V in which GpR is a radical of formula II or ΙΓ, in which R is a polyether radical chosen from the group consisting of radicals

In one embodiment, the composition is characterized in that the hydrophobic radical of formula V in which GpR is a radical of formula II in which R is a polyether radical chosen from the group consisting of radicals

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which a is equal to 0 (a = 0) and r is equal to 0 (r = 0).

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which a is equal to 1 (a = 1) and the GpA radical of formula ΙΙΓ is chosen from the group consisting of the radicals represented pg r I es for mu les below:,

* * * * ★ ie _r ^J HjC ^ J i ch ₃ gh ₃ * * * * * * 4n ₃ ( h ₃ c ^CH 3 i * * * * η ₃ ο ^χ H ₃ cr ch ₃

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals of formulas IVa, IVb or IVc ci- after represented:

Formula IVa Vz O \ G 'V Formula IVb ^b o Formula IVc

In one embodiment, the composition is characterized in that Se hydrophobic radical is a radical of formula V in which the GpC radical is of formula IVa.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals of formulas IVa, IVb or IVc in which b is equal to 0, corresponding respectively to formulas IVd, IVe, and IVf below 15 represented: ....................

O O Y— N Formula IVd OO> - ^c 'ΐ N G Formula IVe

O 0 γ c _x Formula IVf

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in the aquile the GpC radical corresponds to ia formula IV or IVa in which b = 0, and corresponds to formula IVd.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radicai of formula IV in which b ~ 1 is chosen from the group consisting of radicals in which B is an amino acid residue chosen from the group consisting of radicals represented by the formulas below:

* * * * Y f h ₃ c ^ I ch ₃ CH3: * * * * * * X. ch ₃ h ₃ c * * v .. * * * ) h ₃ c ^x

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the radical GpC of formula IV or IVa in lesqueîîes b = 1, is chosen from the group consisting of radicals in which B is an amino acid residue selected from the group consisting of radicals represented by the formulas below: .................

* * ch ₃ * *

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting by linear alkyl radicals.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting by branched alkyl radicals.

In one embodiment, the composition is characterized in that Se hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting by alkyl radicals comprising between 11 and 14 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula

IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of radicals represented by the formulas below

* AT \ h ₃ x = ll * XH ₃ x = 13

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula 20 is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of alkyl radicals comprising between 15 and 16 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from Se 25 group consisting of the radicals represented by the formulas below:

x = 15 [00069] In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the

çh ₃ x = 16 * AT. Λ

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula

IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of alkyl radicals comprising between 17 and 25 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting by alkyl radicals comprising between 17 and 18 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting by the alkyl radicals represented by the formulas below:

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting by alkyl radicals comprising between 18 and 25 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula V in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting by the alkyl radicals represented by the formulas below:

x = 21 [00075] In one embodiment, the composition is characterized in that said hydrophobic radicals are chosen from the hydrophobic radicals of formula I in which a = 1 and p = 2, represented by the following formula VI:

© GpR V-GpA— (GpC) ^{r 2} Formula VI in which

GpR, GpA, GpC, r and a have the definitions given above.

In one embodiment, its composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula IL [00077] In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II in which R is a divalent linear alkyl radical comprising from 2 to 12 carbon atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II in which R is a divalent alkyl radical comprising from 2 to 6 carbon atoms .

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II in which R is a divalent linear alkyl radical comprising from 2 to 6 atoms of carbon.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II in which R is an alkyl radical comprising from 2 to 4 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II in which R is a divalent linear alkyl radical comprising from 2 to 4 atoms of carbon.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II in which R is a divalent linear alkyl radical comprising 2 carbon atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula ΙΓ.

In one embodiment, its composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula ΙΓ in which R is a divalent linear alkyl radical comprising from 1 to 11 atoms of carbon.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula ΙΓ in which R is a divalent alkyl radical comprising from 1 to 6 carbon atoms .

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II or II ', in which R is a divalent alkyl radical, comprising of 2 with 5 carbon atoms, and carrying one or more amide functions (-CONH2).

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II or ΙΓ, in which R is a divalent linear alkyl radical, comprising of 2 with 5 carbon atoms and carrying one or more amide functions (-CONH2). In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II or ΙΓ in which R is a radical chosen from the group consisting of Its radicals represented by the formulas below;

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the amine function of the GpR radical engaged in the formation of the amide function which binds said GpR radical to the co-polyamino acid is carried by a carbon in the delta or epsilon position (or in the 4 or 5 position) relative to the amide function (-CONH2), In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II or II ′, in which R is a linear ether or unsubstituted polyether radical comprising from 4 to 14 carbon atoms and from 1 to 5 oxygen atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II or ΙΓ in which R is an ether radical.

In one embodiment, the composition is characterized in that the ether radical R is a radical comprising from 4 to 6 carbon atoms.

In one embodiment, the composition is characterized in that the ether radical is * *.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II or II ', in which R is a polyether radical.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II or II ', in which R is a linear polyether radical comprising from 6 to 10 carbon atoms and 2 to 3 oxygen atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which GpR is a radical of formula II or IT in which R is a linear polyether radical chosen from the group

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpA radical of formula III is chosen from the group consisting of radicals of formulas Ilia and Illb below represented ^ _____

HN Ilia formula o .NH Illb Formula HN

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpA radical of formula

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula

IV is chosen from the group consisting of the radicals of formulas IVa, IVb and IVc below

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical is of formula IVa.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula IV is chosen from the group consisting of radicals of formulas IVa, IVb or IVc in which b is equal to 0, corresponding respectively to formulas IVd, IVe, and IVf below represented: _________

O 0 o Formula IVd O 0 · - {X ^c x J — N Formula IVe .GO Formula IVf

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the radicai GpC corresponds to formula IV or IVa in which b = 0, and corresponds to formula IVd.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula

IV is chosen from the group consisting of radicals in which Cx is chosen from Se group consisting of linear alkyl radicals comprising between 9 and 15 carbon atoms.

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula

IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of branched alkyl radicals comprising between 9 and 15 carbon atoms.

[000105] In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula

IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of alkyl radicals comprising 9 or 10 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula 20 is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of alkyl radicals comprising between 11 and 15 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of alkyl radicals comprising between 11 and 13 carbon atoms. In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting by the radicals represented by the formulas below:

* Λ ^^ ch ₃ x = 9 * x = 11 ★ A; c H 3 x = 13

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula

IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of alkyl radicals comprising 14 or 15 carbon atoms, [000110] In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical of formula 5 IV is chosen from the group consisting of radicals in which Cx is chosen from the group consisting of the radicals represented by the formulas below;

In one embodiment, the composition is characterized in that the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is chosen from the co-polyamino acids of formula VII below:

The

O D.

X

Hy formula VII in which, • D independently represents either a -CH2- group (aspartic unit) or a -CH2-CH2- group (glutamic unit), ® Hy is a hydrophobic radical chosen from the hydrophobic radicals of formulas I, V or VI, in which r = 1 and GpR is a radical of Formula II, • Ri is a hydrophobic radical chosen from the hydrophobic radicals of formulas I, V or VI in which r = 0 or r = 1 and GpR is a radical of

Formula II ', or a radical chosen from the group consisting of an H, a linear acyl group at C2 to CIO, a branched acyl group at C3 to CIO, a benzyl, a terminal "amino acid" unit and a pyroglutamate, • R2 is a hydrophobic radical chosen from its hydrophobic radicals of formulas I, V or VI in which r = 1 and GpR is a radical of Formula

II, or a radical -NR'R, R 'and R identical or different being chosen from the group consisting of H, linear or branched or cyclic C2 to CIO alkyls, benzyl and said R' and R alkyls which can form together one or more saturated, unsaturated and / or aromatic carbon rings and / or which may contain heteroatoms, chosen from the group consisting of O, N and S, * X represents an H or a cationic entity chosen from the group comprising metal cations;

• n + m represents the degree of polymerization DP of the co-polyamino acid, that is to say the average number of monomeric units per chain of copolyamino acid and 5 <n + m <250;

The co-polyamino acid carrying carboxylate charges and at least one hydrophobic radical of formula I can also be called "co-polyamino acid" in its present description.

The term “statistical co-polyamino acid” means a co-polyamino acid carrying carboxylate charges and at least one hydrophobic radical, a co-polyamino acid of formula VIIa.

In one embodiment, the composition is characterized in that the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is chosen from the co-polyamino acids of formulas VII, in which Ri = R'i and R2 = R '2, of the following formula Vlla:

R'1 ^Hy Formula Vlla in which,

- m, η, X, D and Hy have the definitions given above,

R'i is a radical chosen from the group consisting of an H, a linear acyl group at C2 to CIO, a branched acyl group at C3 to CIO, a benzyl, a terminal "amino acid" unit and a pyroglutamate,

- R'2 is a radical -NR'R, R 'and R identical or different being chosen from the group consisting of H, linear or branched or cyclic C2 to CIO alkyls, benzyl and said R' and R alkyls which may together form one or more saturated, unsaturated and / or aromatic carbon rings and / or which may contain heteroatoms, chosen from the group consisting of O, N and S.

The term “defined co-poiyamino acid” means a co-polyamino acid carrying carboxylate charges and at least one hydrophobic radical, a co-polyamino acid of formula Vllb.

In one embodiment, the composition is characterized in that the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is chosen from the co-polyamino acids of formula Vil in which n = 0 of formula Vllb below:

O

θ Formula Vllb in which m, X, D, Ri and R2 have the definitions given above and at least Ri or R2 is a hydrophobic radical of formula I, V or VI.

In one embodiment, its composition is characterized in that the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is chosen from the co-polyamino acids of formula VII in which n = 0 of formula Vllb and Ri or R2 is a hydrophobic radical of formula I, V or VI.

In one embodiment, the composition is characterized in that the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is chosen from the co-polyamino acids of formula Vllb in which Ri is a hydrophobic radical of formula I, V or VI in which r = 0 or r = 1 and GpR is of Formula II '.

In one embodiment, the composition is characterized in that the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is chosen from the co-polyamino acids of formulas Vllb in which R2 is a hydrophobic radical of formula I, V or VI in which r = 1 and GpR is of Formula IL [000120] In one embodiment, the composition is characterized in that Ri is a radical chosen from the group consisting of a linear C2 to C10 acyl group, a group branched acy in C3 to Cio, a benzyie, a terminal "amino acid" unit and a pyroglutamate.

In one embodiment, the composition is characterized in that Ri is a radical chosen from the group consisting of a linear C2 to Cio acyl group or a branched C3 to Cio acyl group.

In one embodiment, the composition is characterized in that the co-polyamino acid carrying carboxyate charges and hydrophobic radicals is chosen from the co-poiyamino acids of formulas VII, Vlla or Vllb in which group D is a group -CH2- (aspartic unit).

In one embodiment, the composition is characterized in that the co-polyamino acid carrying carboxy charges and hydrophobic radicals is chosen from the co-polyamino acids of formulas VII, Vlla or Vllb in which group D is a -CH2-CH2- group (glutamic unit).

In one embodiment, the composition is characterized in that the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.007 and 0.3.

In one embodiment, the composition is characterized in that the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.01 and 0.3.

In one embodiment, the composition is characterized in that the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.02 and 0.2.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula VI and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.007 and 0, 15.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula VI and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.01 and 0.1.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to ia formula VI and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.02 and 0.08.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula VI in which the Cx radical comprises between 9 and 10 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.03 and 0.15.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula VI in which the Cx radical comprises between 11 and 12 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.0.15 and 0.1.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula VI in which the Cx radical comprises between 11 and 12 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.02 and 0.08.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula VI in which the Cx radical comprises between 13 and 15 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.01 and 0.1.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula VI in which the Cx radical comprises between 13 and 15 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.01 and 0.06.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to its formula V and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.007 and 0, 3.

In one embodiment, its composition is characterized in that the hydrophobic radical corresponds to formula V and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.01 and 0.3.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula V and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.015 and 0, 2.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula V in which the Cx radical comprises between 11 and 14 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.1 and 0.2.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula V in which the Cx radical comprises between 15 and 16 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.04 and 0.15.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula V in which the Cx radical comprises between 17 and 18 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.02 and 0.06.

In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula V in which the Cx radical comprises between 19 and 25 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.01 and 0.06, In one embodiment, the composition is characterized in that the hydrophobic radical corresponds to formula V in which the Cx radical comprises between 19 and 25 carbon atoms and the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units is between 0.01 and 0.05.

In one embodiment, the composition according to the invention is characterized in that n + m is between 10 and 250.

In one embodiment, the composition according to the invention is characterized in that n + m is between 10 and 200.

In one embodiment, the composition according to the invention is characterized in that n + m is between 15 and 150.

In one embodiment, the composition according to the invention is characterized in that n + m is between 15 and 100.

In one embodiment, the composition according to the invention is characterized in that n + m is between 15 and 80.

In one embodiment, the composition according to the invention is characterized in that n + m is between 15 and 65.

In one embodiment, the composition according to the invention is characterized in that n + m is between 20 and 60, [000150] In one embodiment, the composition according to the invention is characterized in that n + m is between 20 and 50.

In one embodiment, the composition according to the invention is characterized in that n + m is between 20 and 40.

The invention also relates to said co-polyamino acids carrying carboxylate charges and hydrophobic radicals of formula I and the precursors of said hydrophobic radicals.

[000153] The co-polyamino acids carrying carboxylate charges and hydrophobic radicals of formula I are soluble in distilled water at a pH of between 6 and

8, at a temperature of 25 ° C. and at a concentration of less than 100 mg / ml.

The invention further relates to a method for preparing stable injectable compositions.

[000155] The term "soluble" is capable of making it possible to prepare a clear solution devoid of particles at a concentration of less than 100 mg / ml in distilled water at 25 ° C.

The term “solution” means a liquid composition devoid of visible particles, using the procedure in accordance with pharmacopoeias EP 8.0, in point 2.9.20, and US <790>.

The term “physically stable composition” is understood to mean compositions which, after a certain period of storage at a certain temperature, satisfy the criteria of visual inspection described in the European, American and international pharmacopoeia, that is to say, compositions which are clear and which contain no visible particles, but which are also colorless.

The term “chemically stable composition” is intended to mean compositions which, after storage for a certain time and at a certain temperature, have a minimum recovery of the active ingredients and comply with the specifications applicable to pharmaceutical products.

The term “aqueous injectable solution” is understood to mean water-based solutions which meet the conditions of the EP and US pharmacopoeias, and which are sufficiently liquid to be injected.

The term “co-polyamino acid consisting of glutamic or aspartic units” is understood to mean non-cyclic linear sequences of glutamic acid or aspartic acid units linked together by peptide bonds, said sequences having a terminal C part, corresponding to the carboxylic acid at one end, and an N-terminal part, corresponding to the amine at the other end of the chain.

The term "alkyl radical" means a carbon chain, linear or branched, which does not include a heteroatom.

[000162] The co-polyamino acid is a statistical or block co-polyamino acid. [000163] The co-polyamino acid is a statistical co-polyamino acid in the chain of glutamic and / or aspartic units.

In the formulas the * indicate the sites of attachment of the various elements represented.

In formulas I, V and VI, the * indicate the sites of attachment of hydrophobic radicals to the co-polyamino acid. The Hy radicals are attached to the copolyamino acid via amide functions, In Its formulas II and II ′, the * indicate, from left to right respectively, the attachment sites of GpR:

to the co-polyamino acid and to GpA if a - 1 or to GPC if a = 0.

In formulas III and HT, the * indicate, from left to right respectively, the GpA attachment sites:

to GpR if r = 1 or to the co-polyamino acid if r = 0 and to GpC.

In formula IV, the * indicates the GpC attachment site:

to GpA if a = 1, GpR if r = 1 and a = 0 or to the co-polyamino acid if r = 0 and a = 0.

All the connections between the different GpR, GpA and GpC groups are amide functions.

The radicals Hy, GpR, GpA, GpC, and D are each independently identical or different from one monomer unit to another.

When the co-polyamino acid comprises one or more aspartic unit (s), this (s) can (undergo) t undergo structural rearrangements.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acids can also comprise monomeric units of formula VIII and / or VIII ':

Formula VIII Formula VIII 'In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR responds to the formula II in which R is -CH2-CH2-, GpC corresponds to formula IVd in which x = 15 and Cx is [000174] In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1,

GpR corresponds to formula II in which R is -CH2-CH2-, GpC corresponds to formula IVd CH ₃

in which x = 16 and Cx is [000175] In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is, GpC corresponds to formula IVd in which 'CH, and Cx is [000176] In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is * _Λ , GpC corresponds to formula IVd in which x = 15 and Cx is ch ₃ [000177] In a mode of embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r - 1, a = 0, p - 1,

GpR corresponds to formula II in which R is formula IVd in which x

² , GpC responds to and Cx is [000178] In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to Its formula II in which R is _G ^ _c and Cx corresponds to Its formula IVd in which x [000179] In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is

O _t GpC corresponds to formula IVd in which x = 19 and Cx is [000180] In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1,

GpR corresponds to formula II in which R is -CH2-CH2-, GpC corresponds to formula IVa

In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r - 1, a - 0, p = 1, GpR corresponds to formula II in which R is -CH2-CH2-, GpC corresponds to formula IVa * *

In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II in which R is -CH2-CH2-, GpA corresponds to the formula Illb, GpC corresponds to the formula IVd in which x = 9 and Cx is

In one embodiment, the at least one hydrophobic radical of formula I is chosen from among its radicals of formula I in which, r = 1, a - 1, p = 2, GpR corresponds to formula II in which R is -CH2-CH2-, GpA corresponds to the formula Illb, GpC corresponds to the formula IVd in which x = 11 and Cx is

In one embodiment, the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r - 1, a - 1, p - 2, GpR corresponds to formula II in which R is -CH2-CH2-, GpA corresponds to the formula Illb, GpC corresponds to Its formula IVd in which x = 13 and Cx is [000185] In one embodiment, the at least one hydrophobic radical of formula I is chosen from radicals of formula I in which, r = 1, a = 1, p = 2,

GpR responds to the formula

AT

R is

Illb, GpC Cx is corresponds to Its formula IVd in it in which the, GpA corresponds to the formula which x = 13 and

In one embodiment, the co-polyamino acid carrying carboxyate charges and hydrophobic radicals is a co-polyamino acid of formula VIT or Vlla, in which DP = 23 +/- 5, i = 0.05 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is -CH2-CH2 -, GpC corresponds to the formula IVd in which x = 15 and Cx is [000187] The values of degree of polymerization DP and of ratio i are estimated by

NMR in D2O by comparing the integration of the signals from the hydrophobic groups to that of the signals from the main chain of the copolyamino acid.

In one embodiment, the co-polyamino acid carrying carboxyl charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 35 +/- 5, i = 0.05 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is -CH2-CH2 -, GpC corresponds to the formula IVd in which x = 15 and Cx is

In one embodiment, the co-polyamino acid carrying carboxyate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 35 +/- 5, i = 0.10 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is -CH2-CH2 -, GpC corresponds to formula IVd in which x = 15 and Cx is [000190] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or Vlla, in which DP = 35 +/- 5, i - 0.05 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is -CH2-CH2-, GpC corresponds to formula IVd in which x = 16 and Cx is

8.

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 23 +/- 5, i - 0.05 +/- 0.02 and Se at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is

Cx is, GpC responds to formula IVd in which x = 17 and a = 0, p = 1, GpR responds to formula II in, GpC responds to Sa which x = 19 and [000192] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in the laqueile DP = 22 +/- 5, i = 0.05 +/- 0.02 and the at least one radical 20 hydrophobic of formula I is chosen from the radicals of formula I in which, r - 1, which R is formula IVd in Cx is [000193] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 30 +/- 5, i - 0.10 +/- 0.03 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which , r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is

Cx is, GpC corresponds to the formula IVd in which x = 15 and

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 23 +/- 5, i = 0.07 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is

which x = 15 and Cx is, GpC corresponds to the formula IVd in [000195] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 23 +/- 5, i = 0.05 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1,

= 15 and Cx is a = 0, p = 1, GpR corresponds to Its formula II in which R is GpC corresponds to Its formula IVd in which x

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 26 +/- 5, i = 0.04 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r - 1, a = 0, p = 1, GpR corresponds to formula II in which R is -CH2-CH2 -, GpC responds to

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 35 +/- 5, i = 0.13 +/- 0.04 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p - 1, GpR corresponds to formula II in which R is -CH2-CH2 -, GpC responds to

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 23 +/- 5, i = 0.05 +/- 0.02 and Se at least one hydrophobic radical of formula I is chosen from Its radicals of formula I in which, r = 1, a = 0, p - 1, GpR corresponds to formula II in which R is

formula IVd in Cx is which x, GpC responds to Sa and

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or

Vllb, in which DP = 25 +/- 5, 0.033 <i <0.05 and the hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is -CH2-CH2-, GpC corresponds to formula IVd in which x = 15 and Cx is [000200] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or Vllb, in which DP - 30 +/- 5, 0.028 <i <0.04 and the at least one hydrophobic radical of formula I is chosen from Its radicals of formula I in which, r = 1, a = 0, p =

1, GpR corresponds to formula II in which R is ⁰ *, GpC corresponds to formula IVd in which x = 17 and Cx is [000201] In one embodiment, Se co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or

Vllb, in which DP = 45 +/- 10, 0.018 <i <0.028 and the at least one hydrophobic radical of formula i is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is, GpC corresponds to formula IVd in which x = 17 and

Cx is [000202] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or Vllb, in which DP = 60 +/- 10, 0.014 <i <0, 02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is, GpC corresponds to formula IVd in which x = 17 and

Cx is [000203] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or Vllb, in which DP - 25 +! - 5, 0.033 <i <0, 05 and the hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 0, p = 1, GpR corresponds to formula II in which R is corresponds to its formula IVd in which x = 19 and Cx is

GpC [000204] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 22 +/- 5, i ~ 0.05 + / - 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p - 2, GpR corresponds to its formula II in which R is -CH2- CH2-, GpA corresponds to the formula Illb, GpC corresponds to the formula IVd in which x = 11 and Cx is [000205] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or

Vlla, in which DP - 35 +/- 5, i = 0.05 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II and in which R is -CH2-CH2-, GpA corresponds to formula Illb, GpC corresponds to formula IVd in which x = 11 and Cx is [000206] In one mode of embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP = 65 +/- 5, i = 0.05 +/- 0.02 and the au at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II in which R is -CH2-CH2-, GpA corresponds to the formula Illb, GpC corresponds to formula IVd in which x = 11 and Cx is [000207] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or V lla, in which DP = 22 +/- 5, i = 0.04 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a - 1, p = 2, GpR corresponds to formula II in which R is -CH2-CH2-, GpA corresponds to formula Illb, GpC corresponds to formula IVd in which x - 13 and Cx is [000208] In a mode of embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIa, in which DP - 22 +/- 5, 0.03 +/- 0.01 and the at least one radical hydrophobic of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II in which R is

U, GpA corresponds to the formula IIIb, GpC corresponds to the formula IVd in which x = 13 and Cx is / ^GH 3 [000209] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co -polyamino acid of formula VII or Vlla, in which DP = 22 +/- 5, i = 0.07 +/- 0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r - 1, a = 1, p = 2, GpR corresponds to ia formula II in which R is θ, GpA corresponds to formula Illb, GpC corresponds to formula IVd in which x = 9 and Cx is, CH ₃ [000210 ] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or VIIb, in which DP - 27 +/- 5, 0.031 <i <0.045 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II in which R is -CH2- CH2-, GpA corresponds to the formula Illb, GpC corresponds to the formula IVd in which x = 11 and Cx is

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or

Vllb, in which DP = 22 +/- 5, 0.037 <i <0.055 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II in which R is -CH2-CH2-, GpA corresponds to formula IIIb, GpC corresponds to formula IVd in which x = 13 and Cx is [000212] In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or Vllb, in which DP = 22 +/- 5, 0.037 <i <0.055 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II in which R is

formula, GpA responds to formula Illb, GpC which x - 13 and Cx is responds

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or

Vllb, in which DP - 60 +/- 10, 0.014 <i <0.02 and the at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II in which R is -CH2-CH2-, GpA corresponds to formula IIIb, GpC corresponds to formula IVd in which x = 13 and Cx is

In one embodiment, the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is a co-polyamino acid of formula VII or Vllb, in which DP = 40 +/- 5, 0.022 <i <0.029 and the at at least one hydrophobic radical of formula I is chosen from the radicals of formula I in which, r = 1, a = 1, p = 2, GpR corresponds to formula II in which R is -CH2-CH2-, GpA corresponds to the formula Illb, GpC corresponds to formula IVd in which x = 13 and Cx is

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid comes from a polyamino acid obtained by polymerization.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is derived from a polyamino acid obtained by ring-opening polymerization of a derivative of N-carboxyanhydride of glutamic acid or of an N-carboxyanhydride derivative of aspartic acid.

In one embodiment, the composition according to the invention is characterized in that Se co-polyamino acid is derived from a polyamino acid obtained by polymerization of an N-carboxyanhydride derivative of glutamic acid or of a derivative of part-carboxyanhydride of aspartic acid as described in the review article Adv. Polym.

Sci. 2006, 202, 1-18 (Deming, T.J.).

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is derived from a polyamino acid obtained by polymerization of a derivative of N-carboxyanhydride of glutamic acid.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is derived from a polyamino acid obtained by polymerization of an N-carboxyanhydride derivative of glutamic acid chosen from the group consisting by methyl N-carboxyanhydride glutamate (GluOMe-NCA), Benzyl Ncarboxyanhydride glutamate (GluOBzl-NCA) and Se N-carboxyanhydride t-butyl glutamate (GluOtBu-NCA).

In one embodiment, the N-carboxyanhydride derivative of glutamic acid is N-carboxyanhydride L-methyl glutamate (L-GluOMe-NCA).

In one embodiment, the glutamic acid N-carboxyanhydride derivative is benzyl N-carboxyanhydride L-glutamate (L-GluOBzl-NCA). In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is derived from a polyamino acid obtained by polymerization of a derivative of N-carboxyanhydride of glutamic acid or of a derivative of N-carboxyanhydride of aspartic acid using as an initiator an organometallic complex of a transition metal as described in the publication Nature 1997, 390, 385-389 (Deming, TJ).

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is derived from a polyamino acid obtained by polymerization of a derivative of N-carboxyanhydride of glutamic acid or of a derived from

N-carboxyanhydride of aspartic acid using as initiator ammonia or a primary amine as described in patent FR 2,801,226 (Touraud, F.; et al.) And the references cited by this patent.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is derived from a polyamino acid obtained by polymerization of a derivative of N-carboxyanhydride of glutamic acid or of a derivative of N-carboxyanhydride of aspartic acid using as initiator hexamethyldisilazane as described in the publication J. Am. Chem. Soc. 2007, 129,

14114-14115 (Lu H.; et al.) Or a silylated amine as described in publication J.

Am. Chem. Soc. 2008, 130, 12562-12563 (Lu H.; et al.).

In one embodiment, the composition according to the invention is characterized in that the process for the synthesis of the polyamino acid obtained by polymerization of a derivative of N-carboxyanhydride of glutamic acid or of a derivative of N-carboxyanhydride of aspartic acid from which the co-polyamino acid is derived comprises a step of hydrolysis of ester functions.

In one embodiment, this step of hydrolysis of ester functions may consist of a hydrolysis in an acid medium or a hydrolysis in a basic medium or be carried out by hydrogenation.

In one embodiment, this step of hydrolysis of ester groups is a hydrolysis in an acid medium.

In one embodiment, this step of hydrolysis of ester groups is carried out by hydrogenation.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid comes from a polyamino acid obtained by depolymerization of a polyamino acid of higher molecular weight.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid comes from a poiyamino acid obtained by enzymatic depolymerization of a poiyamino acid of higher molecular weight. In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid comes from a poiyamino acid obtained by chemical depolymerization of a poiyamino acid of higher molecular weight. In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid comes from a poiyamino acid obtained by enzymatic and chemical depolymerization of a poiyamino acid of higher molecular weight.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is derived from a poiyamino acid obtained by depolymerization of a poiyamino acid of higher molecular weight chosen from the group consisting of polyglutamate sodium and sodium polyaspartate.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid comes from a poiyamino acid obtained by depolymerization of a sodium polyglutamate of higher molecular weight. In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is derived from a poiyamino acid obtained by depoiymerization of a sodium polyaspartate of higher molecular weight.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is obtained by grafting a hydrophobic group on an acid poly-L-glutamic acid or poly-L-aspartic acid using methods of amide bond formation well known to those of skill in the art.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is obtained by grafting a hydrophobic group on an acid poly-L-glutamic acid or poly-L-aspartic acid using the methods of amide bond formation used for peptide synthesis.

In one embodiment, the composition according to the invention is characterized in that the co-polyamino acid is obtained by grafting a hydrophobic group on a poly-L-glutamic acid or poly-L-aspartic acid as described in patent FR 2,840,614 (Chan, YP; et al.).

In one embodiment, the concentration of co-polyamino acid carrying carboxyate charges and hydrophobic radicals is at most 40 mg / ml. In one embodiment, the concentration of co-polyamino acid carrying carboxyate charges and hydrophobic radicals is at most 30 mg / ml.

In one embodiment, the concentration of co-polyamino acid carrying carboxylate charges and hydrophobic radicals is at most 20 mg / ml. In one embodiment, the concentration of co-polyamino acid carrying carboxylate charges and hydrophobic radicals is at most 10 mg / ml.

In one embodiment, the concentration of co-polyamino acid carrying carboxylate charges and hydrophobic radicals is at most 5 mg / ml. In one embodiment, the concentration of co-polyamino acid carrying carboxylate charges and hydrophobic radicals is at most 2.5 mg / ml. In one embodiment, the concentration of co-polyamino acid carrying carboxylate charges and hydrophobic radicals is at most 1 mg / ml. In one embodiment, the concentration of co-polyamino acid carrying carboxylate charges and hydrophobic radicals is at most 0.5 mg / ml.

In one embodiment, the co-polyamino acid mass ratio carrying carboxylate charges and hydrophobic radicals on glucagon is between 1.5 and 25.

In one embodiment, the co-polyamino acid mass ratio carrying carboxylate charges and hydrophobic radicals on glucagon is between 2 and 20.

In one embodiment, the co-polyamino acid mass ratio carrying carboxylate charges and hydrophobic radicals on glucagon is between 2.5 and 15.

In one embodiment, the co-polyamino acid mass ratio carrying carboxylate charges and hydrophobic radicals on glucagon is between 2 and 10.

In one embodiment, the co-polyamino acid mass ratio carrying carboxylate charges and hydrophobic radicals on glucagon is between 2 and 7.

Human glucagon is used in dosages which vary according to the applications.

In emergency treatment of hypoglycaemia Its recommended dosage is 1 mg intramuscularly or intravenously (0.5 g if the body mass is less than 25 kg). This administration is carried out with a solution of human glucagon at the concentration of 1 mg / ml.

In pumps, the daily dose envisaged is approximately 0.5 mg, the solutions can thus comprise from 0.25 mg / ml to 5 mg / ml of human glucagon.

[000255] According to one embodiment, the solutions can comprise from 0.5 mg / ml to 3 mg / ml of human glucagon.

In the treatment of obesity the daily dose envisaged is approximately 0.5 mg, the solutions can thus comprise from 0.25 mg / ml to 5 mg / ml of human glucagon.

In one embodiment, the concentration of human glucagon is between 0.25 and 5 mg / ml.

In one embodiment, the concentration of human glucagon is between 0.5 and 4 mg / ml.

In one embodiment, the concentration of human glucagon is between 0.75 and 3 mg / ml.

In one embodiment, the concentration of human glucagon is between 0.75 and 2.5 mg / ml.

In one embodiment, the concentration of human glucagon is between 0.75 and 2 mg / ml.

In one embodiment, the concentration of human glucagon is between 1 and 2 mg / ml.

20 [000263] In one embodiment, hydrophobic] / [human glucagon] is less than 20. the ratio molar [radical In one embodiment, hydrophobic] / [human glucagon] is less than 15. the ratio molar [radical [000265] In one embodiment, hydrophobic] / [human glucagon] is less than 10. the ratio molar [radical 25 [000266] In one embodiment, hydrophobic] / [human glucagon] is less than 5. the ratio molar [radical In one embodiment, hydrophobic] / [human glucagon] is less than 2.5. the ratio molar [radical 30 In one embodiment, hydrophobic] / [human glucagon] is less than 1.5 the ratio molar [radical

Human glucagon is a highly conserved polypeptide comprising a single chain of 29 amino acid residues having the following sequence H35 His-Ser-GIn-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys -Tyr-Leu-Asp-Ser-Arg-Arg-Ala-Phe-GInAsp-Val-Gln-Trp-Leu-Met-Asn-Thr-OH.

[000270] It can be obtained in various ways, by peptide synthesis by recombination.

Human glucagon is available from many sources. For example, it may be human glucagon produced by Bachem via peptide synthesis, in particular under the reference 407473.

In one embodiment, the compositions according to the invention further comprise a gastrointestinal hormone.

The term "gastrointestinal hormones" means the hormones chosen from the group consisting of GLP-1 RA for agonists of the human Glucagon receptor-Like Peptide-1 (Glucagon like peptide-1 receptor agonist) Glucagon like) and GIP (Glucose-dependent insulinotropic peptide), oxyntomodulin (a derivative of human proglucagon), peptide YY, S'amylin, cholecystokinin, pancreatic polypeptide (PP), ghreline and enterostatin, their analogs or derivatives and / or their pharmaceutically acceptable salts.

In one embodiment, its gastrointestinal hormones are analogs or derivatives of GLP-1 RA (Glucagon like peptide-1 receptor agonist) chosen from the group consisting of exenatide or Byetta® (ASTRA-ZENECA) , liraglutide or Victoza® (NOVO NORDISK), lixisenatide or Lyxumia® (SANOFI), albiglutide or

Tanzeum® (GSK) or dulaglutide or Trulicity® (ELI LILLY & CO), their analogs or derivatives and their pharmaceutically acceptable salts.

In one embodiment, the gastrointestinal hormone is pramlintide or Symlin® (ASTRA-ZENECA).

[000276] In one embodiment, the gastrointestinal hormone is exenatide or

Byetta® its analogs or derivatives and their pharmaceutically acceptable salts. In one embodiment, the gastrointestinal hormone is liraglutide or Victoza® its analogs or derivatives and their pharmaceutically acceptable salts. In one embodiment, the gastrointestinal hormone is lixisenatide or Lyxumia® its analogs or derivatives and their pharmaceutically acceptable salts. In one embodiment, the gastrointestinal hormone is albiglutide or Tanzeum® its analogs or derivatives and their pharmaceutically acceptable salts. In one embodiment, the gastrointestinal hormone is dulaglutide or Trulicity® its analogs or derivatives and their pharmaceutically acceptable salts.

In one embodiment, the gastrointestinal hormone is pramlintide or Symlin®, its analogs or derivatives and their pharmaceutically acceptable salts.

The term "analog" means, when used with reference to a peptide or a protein, a peptide or a protein, in which one or more constituent amino acid residues have been substituted with other residues d amino acids and / or in which one or more constituent amino acid residues have been deleted and / or in which one or more constitutive amino acid residues have been added. The percentage of homology accepted for the present definition of an analogue is 50%. The term "derivative" means, when used with reference to a peptide or a protein, a peptide or a protein or an analog chemically modified by a substituent which is not present in the peptide or the protein or the reference analogue, that is to say a peptide or a protein which has been modified by creation of covalent bonds, to introduce substituents.

In one embodiment, the substituent is chosen from the group consisting of fatty chains.

In one embodiment, the concentration of gastrointestinal hormone 15 is in the range of 0.01 to 10 mg / mL.

In one embodiment, the concentration of exenatide, its analogs or derivatives and their pharmaceutically acceptable salts is in the range of 0.04 to 0.5 mg / ml.

In one embodiment, the concentration of liraglutide, its analogs or derivatives and their pharmaceutically acceptable salts is in the range of 1 to 10 mg / ml.

In one embodiment, the concentration of lixisenatide, its analogs or derivatives and their pharmaceutically acceptable salts is in the range of 0.01 to 1 mg / ml.

In one embodiment, the concentration of pramlintide, its analogs or derivatives and their pharmaceutically acceptable salts is between

0.1 to 5 mg / mL.

In one embodiment, the compositions according to the invention are produced by mixing solutions of human glucagon obtained by reconstitution of lyophilisate and solutions of GLP-1 RA (Glucagon like peptide-1 receptor agonist) GLP1 RA, analog or derivative of GLP-1 RA, said solutions of GLP-1 RA being commercial or reconstituted from lyophilisate.

In one embodiment, the compositions according to the invention further comprise buffers.

In one embodiment, the compositions according to the invention comprise buffers at concentrations between 0 and 100 mM.

In one embodiment, the compositions according to the invention comprise buffers at concentrations between 15 and 50 mM. In one embodiment, the compositions according to the invention comprise a buffer chosen from the group consisting of a phosphate buffer, Tris (trishydroxymethylaminomethane) or sodium citrate.

In one embodiment, the buffer is sodium phosphate.

In one embodiment, the buffer is Tris (trishydroxymethylaminomethane).

In one embodiment, the buffer is sodium citrate.

In one embodiment, the compositions according to the invention further comprise preservatives.

In one embodiment, the preservatives are chosen from the group consisting of m-cresol and phenol, alone or as a mixture.

In one embodiment, the compositions according to the invention further comprise antioxidants.

[000301] In one embodiment, the antioxidants are chosen from methionine.

In one embodiment, the concentration of preservatives is between 10 and 50 mM.

In one embodiment, the concentration of preservatives is between 10 and 40 mM.

In one embodiment, the compositions according to the invention further comprise a surfactant.

[000305] In one embodiment, the surfactant is chosen from the group consisting of propylene glycol or polysorbate.

The compositions according to the invention can also comprise additives such as tonicity agents.

In one embodiment, the tonicity agents are chosen from the group consisting of sodium chloride, mannitol, sucrose, sobitol and glycerol.

The compositions according to the invention may further comprise all of its excipients in accordance with the pharmacopoeias and compatible with human glucagon and the GLP-1 RA used at the usual concentrations.

The invention also relates to a pharmaceutical formulation according to the invention, characterized in that it is obtained by drying and / or lyophilization. In the case of local and systemic releases, the modes of administration envisaged are by the intravenous, subcutaneous, intradermal or intramuscular route.

The transdermal, oral, nasal, vaginal, ocular, oral, pulmonary administration routes are also envisaged.

The invention also relates to single-dose formulations at pH between 6.6 and 7.8 comprising human glucagon.

The invention also relates to single-dose formulations at pH between 6.6 and 7.8 comprising human glucagon and a gastrointestinal hormone, as defined above.

In one embodiment the single-dose formulations further comprise a substituted co-polyamino acid as defined above.

In one embodiment, the formulations are in the form of an injectable solution. In one embodiment, GLP-1 RA, analog or derivative of GLP-1 RA is chosen from the group comprising exenatide (Byetta® ), liragîutide (Victoza®), lixisenatide (Lyxumia®), albiglutide (Tanzeum®), dulagîutide (Trulicity®) or one of their derivatives.

[000317] In one embodiment, the gastrointestinal hormone is exenatide.

In one embodiment, the gastrointestinal hormone is Liragîutide.

[000319] In one embodiment, the gastrointestinal hormone is lixisenatide. In one embodiment, the gastrointestinal hormone is albiglutide, [000321] In one embodiment, the gastrointestinal hormone is dulagutut.

Furthermore and just as importantly, the Applicant has been able to verify that human glucagon in the presence of a co-polyamino acid carrying carboxylate charges and at least one hydrophobic radical according to the invention retains its action as it either alone or in combination with a gastrointestinal hormone.

The preparation of a composition according to the invention has the advantage of being able to be carried out by simple mixing of a solution of human glucagon, a solution of GLP-1 RA, an analog or a derivative of GLP-1 RA, and of a copolyamino acid carrying carboxylate charges and of at least one hydrophobic radical according to the invention, in aqueous solution or in lyophilized form. If necessary, the pH of the preparation is adjusted to pH 7.

In one embodiment the mixture of human glucagon and substituted copolyamino acid is concentrated by ultrafiltration before mixing with GLP1 RA, an analog or derivative of GLP-1 RA in aqueous solution or in lyophilized form.

If necessary, the composition of the mixture is adjusted to excipients such as giycéroi, m-cresol, and polysorbate (Tween®) by adding concentrated solutions of these excipients within the mixture. If necessary, the pH of the preparation is adjusted to 7.

Description of Figure 1:

The median pharmacodynamic curves of the glycemia expressed by the difference in glucose compared to the basal level are shown in FIG. 1.

The curve representing the results obtained with the composition of Example CBle is represented by empty squares and the curve representing the results of its glucagen® composition is represented by solid circles.

Part A

AA .: Synthesis of hydrophobic molecules in which p = .1 [000328] The hydrophobic radicals are represented in the following table by the corresponding hydrophobic molecule before grafting on the co-polyamino acid.

No. Structure of the hydrophobic molecule before grafting on the copolyamino acid IAA ^H 2 ^{n /} \ NH q '^^' ^{C 3:31} p.m. AA2 η ₂ ι \ γ . J4H 1 0 - ^^ '^, ₃ Η ₂₆ h ₃ X ^CH ' AA3 H ₂ N ^x / AA4 / ° ^N X / ⁰ \ y ^ c ₁₅ H ₃₁ AA5 h ₂ n ^ % T nh ₂ '• nh .N X \

Table ΙΑ: list and structures of hydrophobic molecules synthesized according to the invention.

Example AAI: AAI molecule

Molecule Al: product obtained by the reaction between palmitoyl chloride and Lproline.

To a solution of L-proline (10.6 g, 92.1 mmol) in 1 N aqueous sodium hydroxide solution (230 mL; 230 mmol) is added dropwise a solution of palmitoyl chloride (90 minutes) 23.0 g, 83.7 mmol) in acetone (167 mL). After stirring for 14 h at room temperature, the heterogeneous mixture is cooled to 0 ° C., then filtered through a frit to give a white solid which is washed with water (2 x 100 mL) then diisopropyl ether (100 mL) . The solid is dried under reduced pressure. The solid is then dissolved at reflux in 200 ml of water and then 8 ml of a hydrochloric acid solution at 37% are added to obtain a pH = 1. The opalescent reaction medium is then cooled to 0 ° C. The precipitate obtained is filtered on a frit and then washed with water (5 x 50 mL) until filtrates of physiological pH between 6.0 and 8.0 are obtained, then they are dried in an oven at 50 ° C under empty overnight. The product is purified by recrystallization from diisopropyl ether. A white solid is obtained.

Yield: 22.7 g (77%).

Ψ NMR (CDCla, ppm): 0.88 (3H); 1.19-1.45 (24H); 1.58-1.74 (2H); 1.88-2.14 (3H); 2.15-2.54 (3H); 3.47 (1H); 3.58 (1H); 4.41 (0.1H); 4.61 (0.9H) 6.60-8.60 (1H).

Molecule A2: product obtained by reaction between the molecule Al and Bocethylenediamine.

To a solution of molecule Al (75.1 g, 212.4 mmol) in 1500 ml of chloroform are added successively and at room temperature N, Ndiisopropylethylamine (DIPEA) (68.8 g, 532.3 mmol ), 1-hydroxybenzotriazole (HOBt) (37.1 g, 274.6 mmol) then N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide (EDC) (53.1 g, 277.0 mmol). After 15 minutes of stirring at room temperature, a solution of Boc-ethylenediamine (Boc-ethylenediamine) (37.6 g, 234.7 mmol) in 35 ml of chloroform is added. After 18 h of stirring at room temperature, a solution of 0.1 N HCl (2.1 L), then a saturated NaCl solution (1 L) are added. The phases are separated and the organic phase is washed successively with a 0.1 N HCl / saturated NaCl solution (2.1 L / l L), a saturated NaCl solution (2 L), a saturated NaHCO ₃ solution ( 2 L), then a saturated NaCi solution (2 L). The organic phase is dried over anhydrous sodium sulfate, filtered and then concentrated under reduced pressure. The solid obtained is purified by trituration in diisopropyl ether (3 x 400 mL), to give a solid after drying under vacuum at 40 ° C.

Yield: 90.4 g (86%).

^T H NMR (CDCIa, ppm): 0.88 (3H); 1.20-1.37 (24H); 1.44 (9H); 1.54-1.70 (2H); 1.79-1.92 (1H); 1.92-2.04 (1H); 2.03-2.17 (1H); 2.17-2.44 (3H); 3.14-3.36 (4H); 3.43 (1H); 3.56 (1H); 4.29 (0.1 H); 4.51 (0.9 H); 4.82 (0.1H); 5.02 (0.9H); 6.84 (0.1H); 7.22 (0.9H).

AAI molecule To a solution of molecule A2 (20.1 g, 40.5 mmol) in 330 mL of dichloromethane is added dropwise and at 0 ° C a solution of 4 M hydrochloric acid in dioxane (100 mL, 400 mmol). After 3 h 30 min of stirring at room temperature, the solution is concentrated under reduced pressure. The residue is purified by flash chromatography (methane, dichloromethane) to give a white solid of molecule AAI in the form of the hydrochloride salt.

Yield: 16.3 g (93%).

^X H NMR (CDCl, ppm): 0.88 (3H); 1.07-1.40 (24H); 1.49-1.63 (2H); 1.77-2.18 (4H); 2.18-2.45 (2H); 3.14-3.32 (2H); 3.42-3.63 (2H); 3.63-3.84 (2H); 4.37 (O, 1H); 4.48 (0.9H); 6.81-8.81 (4H).

LC / MS (ESI): 396.5; (calculated ([M + H] ⁺ ): 396.4).

Example AA2: AA2 molecule

Molecule A3: 15-methvlhexadecan-l-ol.

In a three-necked flask under argon is introduced magnesium (9.46 g, 389 mmol) in shavings. The magnesium is covered with anhydrous THF (40 mL) and a few drops of l-bromo-3-methylbutane are added at room temperature to initiate the reaction. After the observation of an exotherm and a slight clouding of the medium, Se remains of l-bromo-3-methylbutane (53.87 g, 357 mmol) is added dropwise over 90 min while the temperature of the medium remains stable between 50 and 60 ° C. The reaction medium is then heated at 70 ° C for 2 h.

In a three-necked flask under argon, to a solution of CuCI (482 mg, 4.86 mmol) dissolved in the NMP (62 mL) at 0 ° C is added dropwise a solution of 12bromo-l-dodecanol ( 43 g, 162.1 mmol) in THF (60 mL). To this solution is then added dropwise the hot Organomagnesium solution freshly prepared so as to maintain the temperature of the medium below 20 ° C. The mixture is then stirred at room temperature for 16 h. The medium is cooled to 0 ° C and the reaction is stopped by adding an aqueous solution of HC! IN up to pH 1 and the medium is extracted with ethyl acetate. After washing the organic phase with a saturated NaCl solution and drying over NazSCU, the solution is filtered and concentrated in vacuo to give an oil. After purification by DCVC on silica gel (cyclohexane, ethyl acetate), an oil which crystallizes at room temperature is obtained.

Yield: 32.8 g (74%)

^X H NMR (CDCl, ppm): 0.87 (6H); 1.14 (2H); 1.20-1.35 (22H); 1.50-1.55 (3H); 3.64 (2H).

A4 molecule; 15-methylhexadecanoic acid.

To a solution of molecule A3 (20.65 g, 80.5 mmol) and tetrabutylammonium bromide (14.02 g, 42.5 mmol) in a mixture of acetic acid / dichlorethane / water (124/400 / 320 mL) at room temperature is added in small portions of potassium permanganate (38.2 g, 241.5 mmol). After stirring at reflux for 5 h and returning to ambient temperature, the medium is acidified to pH 1 by progressive addition of 5N HCl. NaïSCb (44.6 g, 354.3 mmol) is then added gradually until the medium becomes discolored. The aqueous phase is extracted with dichloromethane and the combined organic phases are dried over Na2SO4, filtered and concentrated in vacuo. After purification by chromatography on silica gel (cyclohexane, ethyl acetate, acetic acid), a white solid is obtained.

Yield: 19.1 g (quantitative)

Ψ NMR (CDCh, ppm): 0.87 (6H); 1.14 (2H); 1.22-1.38 (20H); 1.51 (1H); 1.63 (2H); 2.35 (2H).

Molecule_A5 j product obtained by reaction between molecule A4 and L-proline.

To a solution of molecule A4 (10 g, 37 mmol) in THF (360 mL) at

0 ° C are successively added dicyclohexyl carbodiimide (DCC) (8.01 g, 38.8 mmol) and N-hydroxysuccinimide (NHS) (4.47 g, 38.8 mmol). After 17 h of stirring at room temperature, the medium is cooled to 0 ° C for 20 min, filtered through a frit. L-proline (4 g, 37.7 mmol), triethylamine (34 mL) and water (30 mL) are added to the filtrate. After 20 h of stirring at room temperature, the medium is treated with an aqueous solution of HCl IN to pH 1. The aqueous phase is extracted with dichloromethane (2 x 125 mL). The combined organic phases are washed with an aqueous solution of IN HCl (2 x 100 mL), water (100 mL), then an aqueous solution saturated with NaCl (100 mL). After drying on NazSCL, the organic phase is filtered, concentrated under vacuum and the residue is purified by chromatography on silica gel (cyclohexane, ethyl acetate, acetic acid)

Yield: 9.2 g (72%)

Ψ NMR (CDCL, ppm): 0.86 (6H); 1.14 (2H); 1.22-1.38 (20H); 1.50 (1H); 1.67 (2H); 1.95-2.10 (3H); 2.34 (2H); 2.49 (1H); 3.47 (1H); 3.56 (1H); 4.61 (1H). LC / MS (ESI): 368.3; (calculated ([M + H] ⁺ ): 368.6).

Moléçuje _A6.j product obtained by reaction between Its molecule A5 and Bocethylenediamine.

To a solution of molecule A5 (9.22 g, 25.08 mmol) in a THF / DMF mixture (200/50 mL) are added triethylamine (TEA) (5.23 mL) and 2- (1H30 benzotriazol-1-yl) -1.3, 3,3-tetramethyluronium tetrafluoroborate (TBTU) at room temperature. After 10 min of stirring, Boc-ethylenediamine (4.42 g, 27.6 mmol) is added. After stirring at room temperature for 17 h, the mixture is diluted with water (300 mL) at 0 ° C and stirred cold for 20 min. The precipitate formed is filtered on a frit and the filtrate is extracted with ethyl acetate. The combined organic phases are washed with saturated NaHCO ₃ solution, dried over NazSCU, filtered, concentrated in vacuo and the residue is purified by flash chromatography (ethyl acetate, methanol).

Yield: 6.9 g (54%)

Ή NMR (CDCb, ppm): 0.86 (6H); 1.15 (2H); 1.22-1.38 (20H); 1.43 (9H); 1.50 (1H); 1.64 (4H); 1.85 (1H); 1.95 (1H); 2.10 (1H); 2.31 (2H); 3.20-3.35 (3H); 3.45 (1H); 3.56 (1H); 4.51 (1H); 5.05 (1H); 7.24 (1H).

LC / MS (ESI): 510.6; (calculated ([M + H] ⁺ ): 510.8).

Molecule AA2 To a solution of the molecule A6 (5.3 g, 10.40 mmol) in dichloromethane (50 mL) at 0 ° C is added a solution of 4N HCl in dioxane (13 mL). After 5 h of stirring at 0 ° C., the medium is concentrated under vacuum, taken up in water and lyophilized to give a white solid of molecule AA2 in the form of the hydrochloride salt.

Yield: 4.6 g (99%)

Ψ NMR (DzO, ppm): 0.91 (6H); 1.22 (2H); 1.22-1.50 (20H); 1.63 (3H); 1.98 (1H); 2.10 (2H); 2.26 (1H); 2.39 (1H); 2.43 (1H); 3.22 (2H); 3.45-3.60 (3H); 3.78 (1H); 4.42 (1H).

LC / MS (ESI): 410.4; (calculated ([M-CI] ⁺ ): 410.7).

Example AA3: AA3 molecule

M_Q! Écule_A.7 product obtained by the reaction between the molecule Al and ia Boctri (ethylene glycol) diamine.

By a process similar to that used for the preparation of the molecule A2 applied to the molecule Al (4.0 g, 11.3 mmol) and to Boc-tri (ethylene glycol) diamine (3.1 g, 12 , 4 mmol), a colorless oil is obtained after purification by flash chromatography (methanol, toluene).

Yield: 5.5 g (84%).

1 H NMR (CDCb, ppm): 0.88 (3H); 1.09-1.39 (24H); 1.44 (9H); 1.64 (2H); 1.79-2.01 (2H); 2.06-2.43 (4H); 3.23-3.68 (14H); 4.33 (0.2H); 4.56 (0.8H); 5.25 (1H); 6.49 (0.2H); 7.13-7.50 (0.8H).

AA3 molecule By a process similar to that used for the preparation of the AAI molecule applied to Its molecule A7 (5.5 g, 9.4 mmol), a white solid of molecule AA3 in the form of the hydrochloride salt is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 4.3 g (92%).

Ψ NMR (DMSO-d6, ppm): 0.85 (3H); 1.08-1.40 (24H); 1.40-1.52 (2H); 1.71-2.02 (4H); 2.02-2.31 (2H); 2.90-2.98 (2H); 3.15-3.47 (5H); 3.50-3.66 (7H); 4.24 (0.6H); 4.32 (0.4H); 7.83 (0.6H); 7.95 (3H); 8.17 (0.4H).

LC / MS (ESI): 484.6; (calculated ([M + H] ⁺ ): 484.4).

Example AA4: AA4 molecule

Molecule A8: product obtained by the reaction between the molecule Al and Boc-1-amino4,7,10-trioxa-13-tridecane amine.

By a process similar to that used for its preparation of the molecule A2 applied to the molecule Al (4.5 g, 12.7 mmol) and to Boc-l-amino-4,7,10-trioxa- 13 tridecane amine (4.5 g, 14.0 mmol), a yellow oil is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 7.7 g (92%).

^X H NMR (CDCl, ppm): 0.88 (3H); 1.22-1.37 (24H); 1.44 (9H); 1.59-1.67 (2H); 1.67-2.00 (6H); 2.06-2.45 (4H); 3.18-3.76 (18H); 4.28 (0.2H); 4.52 (0.8H); 4,695.04 (1H); 6.77 (0.2H); 7.20 (0.8H).

AA4 molecule By a process similar to that used for the preparation of the AAI molecule applied to the A8 molecule (7.7 g, 11.8 mmol), a yellow oil is obtained after purification by flash chromatography (methanol, dichloromethane ). Coevaporation with diisopropyl ether allows the molecule AA4 to be obtained in the form of the hydrochloride salt in the form of a white solid which is dried under vacuum at 50 ° C.

Yield: 5.4 g (76%).

Ψ NMR (CDCI ₃ , ppm): 0.88 (3H); 1.08-1.40 (24H); 1.49-1.65 (2H); 1.76-2.39 (10H); 3.07-3.28 (3H); 3.34-3.80 (15H); 4.34 (0.05H); 4.64 (0.95H); 7.35 (0.05H); 7.66-8.58 (3.95H).

LC / MS (ESI): 556.7; (calculated ([M + H] ⁺ ): 556.5).

Example AA5: molecule AA5 [000342] Molecule A9: product obtained by reaction between the molecule Al and the methyl ester of N-Boc-L-lysine.

By a process similar to that used for the preparation of the molecule A2 applied to the molecule Al (4 g, 11.3 mmol) and to the methyl ester of N-Boc-Lysine (3.2 g, 12.4 mmol), a colorless oil is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 4.9 g (73%).

Ψ NMR (CDCb, ppm): 0.88 (3H); 0.99-1.54 (37H); 1.54-1.75 (3H); 1.75-2.04 (3H); 2.04-2.41 (4H); 2.94-3.19 (2H); 3.19-3.81 (5H); 4.28-4.64 (2H); 4.94 (1H); 6.45 (0.1H); 7.36 (0.9H).

LC / MS (ESI): 596.7; (calculated ([M + H] ⁺ ): 596.5).

A10 molecule: product obtained by treatment of the A9 molecule with ammonia. To a suspension of molecule A9 (4.9 g, 8.2 mmol) in 10 ml of methanol are added 320 ml of a solution of 7 N ammonia in methanol. After 19 h of stirring at room temperature in a closed atmosphere, an additional 100 ml of ammonia solution are added. After 24 h of stirring at room temperature in a closed atmosphere, the reaction medium is concentrated under reduced pressure. The residue is purified by trituration in diisopropyl ether at reflux (100 ml), to give a white solid which is dried under vacuum at 50 ° C.

Yield: 4.1 g (85%).

Ψ NMR (CDCb, ppm): 0.88 (3H); 1.06-1.57 (37H); 1.57-1.79 (3H); 1.88-2.41 (7H); 3.09 (2H); 3.49 (1H); 3.62 (1H); 4.34 (1H); 4.51 (1H); 4.69-4.81 (1H); 5.43 (0.95H); 5.57 (0.05H); 6.25 (0.05H); 6.52 (0.95H); 6.83 (0.05H); 7.11 (0.95H).

AA5 molecule By a process similar to that used for the preparation of the molecule

AAI applied to the molecule A10 (388 mg, 0.67 mmol), a white solid of molecule AA5 in the form of the hydrochloride salt is obtained after purification by trituration in diisopropylether.

Yield: 292 mg (85%).

Ψ NMR (DMSO-d6, ppm): 0.85 (3H); 1.06-2.34 (38H); 2.61-2.81 (2H); 3.29-3.68 (2H); 4.05-4.17 (1.7H); 4.42 (0.3H); 7.00 (1H); 7.16 (0.71-1); 7.43 (0.3H); 7,738.04 (3.7H); 8.16 (0.3H).

LC / MS (ESI): 481.6; (calculated ([M + H] ⁺ ): 481.4).

Example AA6: AA6 molecule

Garlic molecule; product obtained by the reaction between stearoyl chloride and Lproline.

By a process similar to that used for the preparation of the molecule Al applied to L-proline (5.0 g, 43.4 mmol) and to stearoyl chloride (12.0 g, 39.6 mmol) , a white solid is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 5.37 g (36%)

1 H NMR (CDCb, ppm): 0.88 (3H); 1.26-1.37 (28H); 1.64-1.70 (2H); 1.88-2.10 (3H); 2.36 (2H); 2.54-2.58 (1H); 3.46 (1H); 3.56 (1H); 4.62 (1H).

LC / MS (ESI): 382.6; (calculated ([M + H] ⁺ ): 382.3).

Al2 molecule product obtained by reaction between the Ail molecule and Boctri (ethylene glycol) diamine.

By a process similar to that used for the preparation of the A6 molecule applied to the Ail molecule (33.81 g, 88.6 mmol) and to Boctri (ethylene glycol) diamine (26.4 g, 106.3 mmol) in THF using Sa DIPEA instead of Sa TEA, a white solid is obtained after purification by flash chromatography (ethyl acetate, methanol).

Yield: 43.3 g (80%)

^X H NMR (CDCl₃, ppm): 0.87 (3H); 1.24 (30H); 1.43 (9H); 1.61 (2H); 1.82 (1H); 1.96 (1H); 2.25-2.45 (2H); 3.25-3.65 (14H); 4.30 (0.15H); 4.53 (0.85H); 5.25 (1H); 6.43 (0.15H); 7.25 (0.85H).

LC / MS (ESI): 612.6; (calculated ([M + H] ⁺ ): 612.9).

AA6 molecule By a process similar to that used for the preparation of the AA2 molecule applied to the A12 molecule (43 g, 70.3 mmol), the residue obtained after concentration under vacuum is triturated in acetonitrile. The suspension is filtered and the solid washed with acetonitrile and then acetone. After drying under vacuum, a white solid of molecule AA6 in the form of the hydrochloride salt is obtained.

Yield: 31.2 g (81%)

Ψ NMR (DMSO-de, ppm): 0.85 (3H); 1.23 (28H); 1.45 (2H); 1.70-2.05 (4H); 2.13 (1H); 2.24 (1H); 2.95 (2H); 3.10-3.25 (2H); 3.30-3.65 (10H); 4.20-4.45 (1H); 7.85-8.25 (4H).

LC / MS (ESI): 512.4; (calculated ([M-CI] ⁺ ): 512.8).

Example AA7: AA7 molecule

Al3 molecule: product obtained by reaction between arachidic acid and L-proline. By a process similar to that used for the preparation of the molecule A5 applied to arachidic acid (15.51 g, 49.63 mmol) and to L-proline (6 g, 52.11 mmol) in using DIPEA instead of TEA, a white solid is obtained after purification by a chromatographic column on silica gel (cyclohexane, ethyl acetate, acetic acid).

Yield: 12.9 g (63%)

Ψ NMR (CDCH, ppm): 0.88 (3H); 1.28 (34H); 1.66 (2H); 1.95-2.15 (2H); 2.34 (2H); 2.45 (1H); 3.47 (1H); 3.56 (1H); 4.60 (1H).

LC / MS (ESI): 410.4; (calculated ([M + H] ⁺ ): 410.6).

Molecule A14 i product obtained by the reaction between the molecule A13 and Boc-1-amino4,7,10-trioxa-13-tridecane.

By a process similar to that used for its preparation of the molecule A12 applied to the molecule A13 (10.96 g, 26.75 mmol) and to Boc-l-amino-4,7,1056 trioxa-13 -tridecane (10.29 g, 32.11 mmol), a solid is obtained after purification by a chromatographic column on silica gel (cyclohexane, ethyl acetate, methanol). Yield: 14.2 g (75%)

^X H NMR (CDCl, ppm): 0.88 (3H); 1.24 (32H); 1.43 (9H); 1.61 (2H); 1.80 (1H); 1.96 (1H); 2.10-2.45 (4H); 3.20-3.75 (18H); 4.30 (0.20H); 4.55 (0.80H); 5.03 (1H); 6.75 (0.20H); 7.20 (0.80H).

LC / MS (ESI): 712.8; (calculated ([M + H] ⁺ ): 713.1).

AA7 molecule After a process similar to that used for the preparation of the AA2 molecule applied to the A14 molecule (14.25 g, 20.01 mmol), the residue obtained after vacuum concentration of the reaction medium is dissolved in the methanol and evaporated under reduced pressure, the operation being repeated 4 times to give a white solid of molecule AA7 in the form of the hydrochloride salt.

Yield: 12.7 g (98%)

¹ H NMR (DMSO-de, ppm): 0.85 (3H); 1.23 (32H); 1.45 (2H); 1.64 (2H); 1.70-2.05 (6H); 2.10-2.30 (2H); 2.82 (2H); 3.08 (2H); 3.30-3.60 (15H); 4.15-4.30 (1H); 7.73-8.13 (4H).

LC / MS (ESI): 612.7; (calculated ([M-CI] ⁺ ): 612.9).

Example AA8: AA8 molecule

Molecule A15: product obtained by the reaction between L-leucine and palmitoyl chloride.

By a process similar to that used for the preparation of its Al molecule applied to Sa L-leucine (15.0 g, 114.4 mmol) and palmitoyl chloride (34.5 g, 125 mmol), a white solid is obtained by trituration in diisopropyl ether. Yield: 13.0 g (31%)

Ψ NMR (CDCb, ppm): 0.88 (3H); 0.96 (6H); 1.16-1.35 (24H); 1.55-1.77 (5H); 2.23 (2H); 4.55-4.60 (1H); 5.88 (1H).

A16 molecule: product obtained by the reaction between the A15 molecule and the methyl ester of L-proline [000353] By a process similar to that used for the preparation of the A2 molecule applied to the A15 molecule (6.00 g, 16.2 mmol) and with the methyl ester of Lproline (3.23 g, 19.5 mmol), a slightly yellow oil is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 5.8 g (74%)

Ψ NMR (CDCI ₃ , ppm): 0.83-1.00 (9H); 1.18-1.32 (24H); 1.40-1.73 (5H); 1.84-2.33 (6H); 3.47-3.89 (2H); 3.70 (1.14H); 3.71 (1.21H); 3.74 (0.53H); 3.76 (0.12H); 4.40-4.56 (1H); 4.63-4.67 (0.04H); 4.84 (0.38); 4.90 (0.40); 5.06 (0.18); 5.99 (0.18H); 6.08-6.21 (0.82).

LC / MS (ESI): 481.6; (calculated ([M + H] ⁺ ): 481.4).

Molecule Al7: product obtained by the saponification of the methyl ester of the molecule A16.

To a solution of molecule A16 (5.8 g, 12.06 mmol) in 30 mL of methanol is added 1N sodium hydroxide (13.5 mL, 13.5 mmol). After 20 h of stirring at room temperature, the solution is diluted with water and then acidified with 20 ml of 1N hydrochloric acid at 0 ° C. The precipitate is filtered and then rinsed with water (50 ml) before being dissolved in 50 ml of dichloromethane. The organic phase is dried over Na2SO4, filtered and then concentrated under reduced pressure to give a colorless oil. Yield: 4.5 g (80%)

1 H-NMR (CDCl ₃ , ppm): 0.85-0.99 (9H); 1.14-1.41 (24H); 1.43-1.72 (5H); 1.87-2.47 (7H); 3.48-3.55 (0.6H); 3.56-3.62 (0.4H); 3.83-3.90 (0.4H); 3.90-3.96 (0.6H); 4.52-4.56 (0.6H); 4.56-4.59 (0.4H); 4.80-4.86 (0.4H); 4.86-4.91 (0.6H); 6.05 (0.4H); 6.11 (0.6H).

LC / MS (ESI): 467.6; (calculated ([M + H] ⁺ ): 467.4).

AÏS molecule: product obtained by the reaction between Boc-ethylenediamine and the A17 molecule.

By a process similar to that used for the preparation of the molecule A2 applied to the molecule A17 (4.5 g, 9.64 mmol) and to Boc-ethylenediamine (1.70 g, 10.61 mmol) , a colorless oil is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 2.0 g (34%)

Ψ NMR (CDCI ₃ , ppm): 0.83-0.99 (9H); 1.19-1.32 (24H); 1.44 (9H); 1.48-2.37 (14H); 3.09-3.99 (4H); 4.28-5.01 (2H); 5.64-6.04 (1H); 6.87-7.06 (1H).

LC / MS (ESI): 609.7; (calculated ([M + H] ⁺ ): 609.5).

AA8 molecule By a process similar to that used for the preparation of the AAI molecule applied to the A18 molecule (2 g, 3.28 mmol), a solid of molecule AA8 in the form of the hydrochloride salt is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 1.5 g (90%)

Ψ NMR (CDCI ₃ , ppm): 0.83-1.00 (9H); 1.18-1.32 (24H); 1.37-1.77 (5H); 1.93-2.41 (6H); 3.07-3.97 (6H); 4.44-4.77 (2H); 7.66-8.21 (2H).

LC / MS (ESI): 509.6; (calculated ([M + H] ⁺ ): 509.4).

Example AA9: AA9 molecule

Molecule A19: product obtained by the reaction between lauric acid and L-phenylalanine. By a process similar to that used for the preparation of the A5 molecule applied to lauric acid (8.10 g, 40.45 mmol) and to its L-phenylalanine (7 g, 42.38 mmol), a solid bianc is obtained.

Yield: 12.7 g (98%)

Ψ NMR (DMSO-de, ppm): 0.86 (3H); 1.10-1.30 (16H); 1.36 (2H); 2.02 (2H); 2.82 (1H); 3.05 (1H); 4.42 (1H); 7.15-7.30 (5H); 8.05 (1H); 12.61 (1H).

LC / MS (ESI): 348.2; (caiculate ([M + H] ⁺ ): 348.5).

A20 molecule: product obtained by the reaction between the A19 molecule and the hydrochloride salt of the methyl ester of L-proline.

By a process similar to that used for the preparation of the molecule A6 applied to the molecule A19 (9.98 g, 28.72 mmol) and to the hydrochloride salt of the methyl ester of L-proline (5, 23 g, 31.59 mm 3), a colorless oil is obtained after purification by a chromatographic column on silica gel (cyclohexane, ethyl acetate).

Yield: 5.75 g (44%)

1 H NMR (CDCb, ppm): 0.88 (3H); 1.10-1.30 (16H); 1.50-1.75 (3H); 1.80-2.02 (3H); 2.17 (2H); 2.65 (0.5H); 2.95 (1H); 3.05-3.20 (1.5H); 3.50-3.65 (1H); 3.75 (3H);

4.29 (0.5H); 4.46 (0.5H); 4.70 (0.1H); 4.95 (0.9H); 6.20-6.30 (1H); 7.15-7.30 (5H).

LC / MS (ESI): 459.2; (calculated ([M + H] ⁺ ): 459.6).

Molecule A2.1 product obtained by saponification of the molecule A20.

To a solution of molecule A20 (5.75 g, 12.54 mmol) in a mixture

THF / methanol / water (40/40/40 mL) at 0 ° C is added lithium hydroxide (LiOH) (600.49 mg, 25.07 mmol) then the mixture is stirred at room temperature for 20 h . After evaporation of the organic solvents under vacuum, the aqueous phase is diluted in water, acidified with an aqueous solution of IN HCl to pH 1. The product is then extracted with ethyl acetate. The combined organic phases are washed with a saturated aqueous NaCl solution, dried over Na2SC> 4, filtered and concentrated under reduced pressure to give a colorless oil.

Yield: 5.7 g (quantitative)

NMR (CDCb, ppm): 0.88 (3H); 1.10-1.30 (16H); 1.50-1.80 (3H); 1.67-2.02 (2H); 2.20 (2H); 2.25 (0.4H); 2.60 (0.6H); 2.85-3.10 (2.6H); 3.55-3.65 (1.4H); ; 4.35 (0.6H); 4.55 (0.4H); 4.94 (1H); 6.28 (0.4H); 6.38 (0.6H); 7.20-7.30 (5H).

LC / MS (ESI): 445.2; (calculated ([M + H] ⁺ ): 445.6).

Molecule A22: product obtained by reaction between ia Boc-ethylenediamine and the molecule A21.

By a process similar to that used for the preparation of the A6 molecule applied to the A21 molecule (5.67 g, 12.75 mmol) and to Boc-ethylenediamine (2.25 g, 14.03 mmol) , a colorless oil is obtained after purification by a chromatographic column on silica gel (dichloromethane, methanol).

Yield: 5.7 g (76%)

^Τ N NMR (CDCb, ppm): 0.88 (3H); 1.25 (16H); 1.43 (9H); 1.58 (2.6H); 1.75-1.95 (1.4H); 2.15-2.30 (3H); 2.64 (0.5H); 2.95-3.10 (2.5H); 3.20-3.40 (4H); 3.45 (0.5H); 3.55 (0.2H); 3.66 (1H); 4.44 (1H); 4.50 (0.2H); 4.60 (0.6H); 4.99 (0.7H); 5.54 (0.5H); 5.95 (0.2H); 6.17 (1H); 6.60 (0.5H); 7.07 (0.5H); 7.20-7.40 (5H). LC / MS (ESI): 587.4; (calculated ([M + H] ⁺ ): 587.8).

AA9 molecule After a process similar to that used for the preparation of the AA2 molecule applied to Its molecule A22 (5.66 g, 9.65 mmol), the residue obtained after concentration under vacuum of the reaction medium is dissolved in Se methanol and evaporated under reduced pressure, the operation being repeated 4 times to give a white foam of molecule AA9 in the form of the hydrochloride salt.

Yield: 4.9 g (97%)

^X NMR (DMSO-de, 120 ° C, ppm): 0.89 (3H); 1.26 (16H); 1.43 (2H); 1.68 (0.6H); 1.75-2.00 (3H); 2.05-2.25 (2.4H); 2.82-3.05 (5H); 3.38 (2H); 3.50-3.70 (1.4H); 4.25 (0.6H); 4.63 (0.41-1); 4.77 (0.6H); 7.25-7.50 (5H); 7.55-8.20 (4H).

LC / MS (ESI): 487.4; (calculated ([M-CI] ⁺ ): 487.7).

AB: Synthesis of co-polyamino acids

AB9

AB10

i = 0.08, DP (m + n) = 25

Hy =

HAS DRUNK

Ο

AB12 i = 0.05, DP (m + n) = 23

Table lb

AB18 i = 0.045, DP (m) = 22 AB19 i = 0.015, DP (m) = 65 AB20 i = 0.017, DP (m) = 60 R1 = CH ₃ -CO-

Table le: list of co-polyamino acids synthesized according to the invention.

Part AB: synthesis of co-polyamino acids

Example AB1: Co-polyamino acid AB1 - sodium poly-L-glutamate modified by the molecule AAI and having a number-average molar mass (Mn) of 2900 g / mol

Co-polyamino acid ASl-1: poly-L-glutamic acid of relative number-average molar mass (Mn) 3861 g / mol resulting from its polymerization of γ-benzyl-L-glutamate N10 carboxyanhydride initiated by hexylamine [000363] In a flask previously dried in an oven is placed under vacuum of ybenzyl-L-glutamate N-carboxyanhydride (89.9 g, 341 mmol) for 30 min, then anhydrous DMF (200 mL) is introduced. The mixture is then stirred under argon until complete dissolution, cooled to 4 ° C, then hexylamine (2.05 mL 15.5 mmol) is introduced quickly. The mixture is stirred between 4 ° C and room temperature for 2 days. The reaction medium is then heated at 65 ° C for 2 h, cooled to room temperature then poured dropwise into diisopropyl ether (3 L) with stirring. The white precipitate is recovered by filtration, washed with diisopropyl ether (2 x 200 mL) then dried under vacuum at 30 ° C to give a poly (gamma-benzyl-Lglutamic acid) (PBLG).

To a solution of PBLG (74.8 g) in trifluoroacetic acid (TFA, 340 mL) at 4 ° C is added dropwise a solution of hydrobromic acid (HBr) at 33% in acid acetic (240 mL, 1.37 mol). The mixture is stirred at room temperature for 2 h, then poured dropwise onto a 1: 1 (v / v) mixture of diisopropyl ether and water with stirring (4 L). After 2 h of stirring, the heterogeneous mixture is left to stand overnight. The white precipitate is recovered by filtration, washed with a 1: 1 (v / v) mixture of diisopropyl ether and water (340 mL) then with water (340 mL). The solid obtained is then dissolved in water (1.5 L) by adjusting the pH to 7 by adding a 10 N aqueous sodium hydroxide solution and then a 1 N aqueous sodium hydroxide solution. After solubilization, the theoretical concentration is adjusted to 20 g / L theoretical by adding water to obtain a final volume of 2.1 L.

The solution is filtered on a 0.45 μm filter and then purified by ultrafiltration against a 0.9% NaCl solution, then water until the conductimetry of the permeate is less than 50 pS / cm. The co-polyamino acid solution is then concentrated until a final volume of 1.8 L is obtained.

The aqueous solution is then acidified by adding 37% hydrochloric acid solution until reaching a pH of 2. After 4 h of stirring, the precipitate obtained is filtered, washed with water (2 x 340 mL) then dried under vacuum at 30 ° C to give a poly-L-glutamic acid of number average molar mass (Mn) 3861 g / mol relative to a polyoxyethylene (PEG) standard.

Co-polyamino acid AB1 [000368] The co-polyamino acid AB1-1 (10.0 g) is dissolved in DMF (700 mL) at 30 ° C-40 ° C and then cooled to 0 ° C. The AAI molecule in the form of the hydrochloride salt (1.64 g, 3.8 mmol) is suspended in DMF (23 mL) and triethylamine (0.39 g, 3.8 mmol) is then added and the mixture is slightly heated with stirring until complete dissolution. To the solution of co-polyamino acid at 0 ° C, N-methylmorpholine (NMM, 7.6 g, 75 mmol) in DMF (14 mL) and ethyl chloroformate (ECF, 8.2 g, 75 mmol) are added. After 10 min at 0 ° C, the solution containing the AAI molecule is added and the medium maintained at 30 ° C for 2 h. The reaction medium is poured dropwise onto 5.5 L of water containing 15% by mass sodium chloride and HCl (pH 2), then left to stand overnight. The precipitate is collected by filtration and dried under vacuum for approximately 30 min. The white solid obtained is taken up in water (500 mL) and the pH is adjusted to 7 by slow addition of an aqueous solution of NaOH IN. After filtration through a 0.45 μm filter, the clear solution obtained is purified by ultrafiltration against a 0.9% NaCl solution and then with water, until the conductimetry of the permeate is less than 50 pS / cm. After unloading, the solution is filtered through a 0.2 μm filter and stored at 2-8 ° C.

Dry extract: 24.9 mg / g

An average degree of polymerization (DP) of 23 is estimated by N NMR in D2O by comparing the integration of the signals from the grafted hydrophobic with that of the signals from the main chain.

According to NMR = 0.05

The calculated average molar mass of the co-polyamino acid AB1 is calculated on the basis of the molar masses of the radicals Ri and R2, the aspartate and / or glutamate residues (including an amide bond), the hydrophobic radical, the DS and the DP.

The calculated average molar mass of the co-polyamino acid AB1 is 3945 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 2900 g / mol.

Example AB2: Co-polyamino acid AB2 - sodium poly-L-glutamate modified by the AAI molecule and having a number-average molar mass (Mn) of 3700 g / mol [000369] By a process similar to that used for the preparation of the copolyaminoacidABl applied to the hydrochloride salt of the AAI molecule (1.64 g, 3.8 mmol) and to a poly-L-glutamic acid with relative Mn 5200 g / mol (10.0 g) obtained by a process similar to that used for the preparation of the co-polyamino acid ABl-1, a sodium poly-L-glutamate modified by the molecule AAI is obtained.

Dry extract: 14.1 mg / g DP (estimated according to Sa NMR Ψ): 35

According to His NMR ^: 1 = 0.05

The calculated average molar mass of the AB2 co-polyamino acid is 5972 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3700 g / mol.

Example AB3: Co-polyamino acid AB3 - sodium poly-L-glutamate modified by the molecule AAI and having an average molecular weight ers number (Mn) of 4900 g / mol [000370] By a process similar to that used for the preparation of the copolyamino acid AB1 applied to the hydrochloride salt of the AAI molecule (3.30 g, 7.6 mmol) and to a poly-L-glutamic acid of relative number average mass (Mn)

5200 g / mol (10.0 g) obtained by a process similar to that used for the preparation of the co-polyamino acid AB1-1, a sodium poly-L-glutamate modified by its molecule AAI is obtained.

Dry extract: 23.4 mg / g

PD (estimated from N NMR): 35

The calculated average molar mass of the AB3 co-polyamino acid is 6594 g / mol. According to the NMR ^: 1 = 0.10

Aqueous HPLC-SEC (PEG calibrator): Mn = 4900 g / mol.

Example AB4: Co-polyamino acid AB4 - sodium poly-L-glutamate modified by the molecule AA2 and having a number average molar mass (Mn) of 1800 g / mol [000371] By a process similar to that used for the preparation of the co10 polyamino acid AB1 applied to the hydrochloride salt of the molecule AA2 (1.09 g, 2.4 mmol) and to a poly-L-glutamic acid of average mass Mn = 5600 g / mol (6.3 g) obtained by a process similar to that used for the preparation of the co-polyamino acid AB1-1 but with a step of deprotection of the benzyl esters using trimethylsilane iodide according to the protocol described in the publication J. Am. Chem. Soc. 2000

122, 26-34 (Subramanian G., et al.), A sodium poly-L-glutamate modified by the molecule AA2 is obtained.

Dry extract: 21.5 mg / g DP (estimated from ^T H NMR): 35 From NMR ^: 1 = 0.052

The calculated average molar mass of the AB4 co-polyamino acid is 6022 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 1800 g / mol.

Example AB5: Co-polyamino acid AB5 - sodium poly-L-glutamate modified by the molecule AA6 and having a number-average molar mass (Mn) of

2600 g / mol [000372] By a process similar to that used for the preparation of copolyamino acid AB1 applied to the hydrochloride salt of the molecule AA6 (2.06 g, 3.8 mmol) and to a poly-L-glutamic acid ( 9.8 g) obtained by a process similar to that used for the preparation of the co-polyamino acid AB1-1, a sodium poly-L-glutamate modified with the molecule AA6 is obtained.

Solids content: 20.9 mg / g DP (estimated by NMR ^X H): 23 According to NMR: 1 = 0.05

The calculated average molar mass of the ABS co-polyamino acid is 4079 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 2600 g / mol.

Example AB6: Co-polyamino acid AB6 - sodium poly-L-glutamate modified by the molecule AA7 and having a number average molar mass (Mn) of 4000 g / mol [000373] A poly-L-glutamic acid of average mass Mn = 3500 g / mol and of 5 degree of polymerization 22 (10.0 g) obtained by a process similar to that used for the preparation of the co-polyamino acid AB1-1 is dissolved in DMF (420 mL) at 30 ° C 40 ° C then maintained at this temperature. In parallel, the hydrochloride salt of the molecule AA7 (1.47 g, 2.3 mmol) is suspended in DMF (12 mL) and triethylamine (0.23 g, 2.3 mmol) is added , then the mixture is slightly heated with stirring until complete dissolution. To the solution of co-polyamino acid in DMF, NMM (7.6 g, 75 mmol), the solution of AA7 then 2-hydroxypyridine / V-oxide (HOPO, 0.84 g, 7.5 mmol) are added successively. The reaction medium is then cooled to 0 ° C., then EDC (1.44 g, 7.5 mmol) is added and the medium is brought up to ambient temperature for 2 h. The reaction medium is filtered through a 0.2 mm woven filter and poured dropwise onto 3.5 L of water containing NaCl at 15% by mass and HCl (pH 2) with stirring. At the end of the addition, the pH is readjusted to 2 with a 37% HCl solution, and the suspension is left to stand overnight. The precipitate is collected by filtration, then rinsed with 100 ml of water. The white solid obtained is dissolved in 500 ml of water by slow addition of a 1N aqueous NaOH solution to pH 7 with stirring, then the solution is filtered through a 0.45 μm filter. The clear solution obtained is purified by ultrafiltration against a 0.9% NaCl solution and then with water, until the conductimetry of the permeate is less than 50 pS / cm. The solution is filtered through a 0.2 µm filter and stored at 2-8 ° C.

Dry extract: 21.6 mg / g

DP (estimated from H-NMR): 20 From NMR ’H: i = 0.025

The calculated average molar mass of the AB6 co-polyamino acid is 3369 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn - 4000 g / mol.

Example AB7: Co-polyamino acid AB7 - sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by the molecule AA7 and having a number-average molar mass (Mn) of 3300 g / mol [000374] Çq-polyaminpacid.e_AB7_sr _: _ poly-L-glutamic acid of average molar mass in number (Mn) relative 3600 g / mol and of DP 21 resulting from Its polymerization of γ-benzyl-L-glutamate N-carboxyanhydride initiated by hexylamine and capped at one of its ends by an acetyl group [000375] In a flask previously dried in an oven is placed under vacuum ybenzyl-L-glutamate N-carboxyanhydride (Glu (OBn) -NCA, 100.0 g, 380 mmol) for minutes and then anhydrous DMF (225 mL) is introduced. The mixture is then stirred under argon until complete dissolution, cooled to 4 ° C, then hexylamine (1.78 g, 17 mmol) is introduced quickly. The mixture is stirred between 4 ° C and room temperature for 2 days then precipitated in diisopropyl ether (3.4 L). The precipitate is recovered by filtration, washed twice with diisopropyl ether (225 mL) then dried to give a white solid which is dissolved in 450 mL of THF. To this solution are successively added DIPEA (31 mL, 176 mmol) and then acetic anhydride (17 mL, 176 mmol). After stirring overnight at room temperature, the solution is poured slowly into diisopropyl ether (3 L) with stirring. After 1 hour of agitation,

The precipitate is filtered, washed twice with diisopropyl ether (250 mL) then dried under vacuum at 30 ° C to give a poly (gamma-benzyl-L-glutamic acid) capped at one of its ends by an acetyl group.

To a solution of the above co-polyamino acid (72 g) in trifluoroacetic acid (TFA, 335 mL) at 4 ° C is added dropwise a solution of hydrobromic acid (HBr) at 33% in acetic acid (235 mL). The mixture is stirred at room temperature for 3 h 30, then poured dropwise onto a 1: 1 (v / v) mixture of diisopropyl ether and water with stirring (4 L). After 2 h of stirring, the heterogeneous mixture is left to stand overnight. The white precipitate is recovered by filtration, washed with a 1: 1 (v / v) mixture of diisopropyl ether and water (340 mL) then with water (340 mL).

The solid obtained is then dissolved in water (1.5 L) by adjusting the pH to 7 by adding a 10N aqueous sodium hydroxide solution and then an aqueous IN sodium hydroxide solution. After solubilization, the solution is diluted by adding water to obtain a final volume of 2.1 L. The solution is filtered through a 0.45 μm filter and then purified by ultrafiltration against a 0.9% NaCl solution, then l until the conductivity of the permeate is less than 50 pS / cm. The co-polyamino acid solution is then concentrated until a final volume of 1.8 L is obtained.

The aqueous solution is then acidified by adding 37% hydrochloric acid solution until reaching a pH of 2. After 4 h of stirring, the precipitate obtained is filtered, washed with water (330 mL ) then dried under vacuum at 30 ° C. to give a poly-L-glutamic acid of number average molar mass (Mn) 3600 g / mol relative to a polyoxyethylene (PEG) standard, and of average degree of polymerization 21.

Co-polyamino acid AB7: By a process similar to that used for the preparation of co-polyamino acid AB6 applied to the hydrochloride salt of the molecule AA7 (1.43 g, 2.2 mmol) and to the co-polyamino acid AB7- 1 (10.0 g), a sodium poly-Lgiutamate acid modified by the molecule AA7 is obtained.

Dry extract: 24.3 mg / g

DP (estimated from NMR: 21 Based on NMR ^: 1 = 0.03

The calculated average molar mass of the AB7 co-polyamino acid is 3677 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 3300 g / mol.

Example AB8: Co-polyamino acid AB8 - sodium poly-L-glutamate modified by the molecule AA7 and having a number-average molar mass (Mn) of 3600 g / mol [000380] Co-polyamino acid AB8-1: poly-L acid -glutamic acid of average molar mass in number (Mn) 3800 g / mol and of degree of polymerization 24 resulting from the polymerization of γ-methyl-L-glutamate N-carboxyanhydride initiated by ammonia [000381] By a process similar to that described in patent application FR-A-2 801 226 applied to γ-methyl-L-glutamic acid N-carboxyanhydride (25.0 g, 133.6 mmol) and to a 0.5N ammonia solution in ie dioxane (12.1 mL, 6.05 mmol), a poly-L-glutamic acid is obtained.

Co-polyamino acid AB8:

By a process similar to that used for the preparation of the copolyamino acid AB6 applied to the hydrochloride salt of the molecule AA7 (2.1 g, 3.24 mmol) and to the co-polyamino acid AB8-1 (14.3 g) , a sodium poly-L-glutamate modified by the molecule AA7 is obtained.

Dry extract: 25.2 mg / g DP (estimated according to NMR ^Τ Η): 24 According to ia NMR ^: 1 = 0.03

The calculated average molar mass of the AB8 co-polyamino acid is 4099 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3600 g / mol.

Example AB9: Co-polyamino acid AB9 - sodium poly-L-glutamate modified by the molecule AA3 and having a number-average molar mass (Mn) of

3200 g / mol [000383] By a process similar to that used for the preparation of copolyaminoacidABl applied to the hydrochloride salt of the molecule AA3 and to a poly-L-glutamic acid obtained by a process similar to that used for the preparation of co -polyamino acid AB1-1, a sodium poly-L-glutamate modified by the molecule AA3 is obtained.

Dry extract: 14.7 mg / g DP (estimated from Ψ NMR): 30 From NMR = 0.12

The calculated average molar mass of the AB9 co-polyamino acid is 6192 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3200 g / mol.

Example AB10: Co-polyamino acid ABlO - sodium poly-L-glutamate modified by the molecule AA4 and having a number-average molar mass (Mn) of

2630 g / mol [000384] By a process similar to that used for the preparation of copolyamino acid AB7 applied to the hydrochloride salt of the molecule AA4 and to a poly-L-glutamic acid obtained by a process similar to that used for the preparation of co -polyamino acid AB1-1, a sodium poly-L-glutamate modified by the molecule AA4 is obtained.

Dry extract: 18.3 mg / g DP (estimated according to ¹ H NMR): 25 Based on ^X H NMR: i = 0.08

The calculated average molar mass of the AB10 co-polyamino acid is 4870 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 2600 g / mol.

Example ABU: Co-polyamino acid ABU - sodium poly-L-glutamate modified by the molecule AA5 and having a number-average molar mass (Mn) of 2700 g / mol [000385] By a process similar to that used for the preparation of the copolyamino acid AB6 applied to the hydrochloride salt of the molecule AA5 and to a poly-L-glutamic acid obtained by a process similar to that used for the preparation of the co-polyamino acid AB1-1, a sodium poly-L-glutamate modified by the AA5 molecule is obtained.

Solids content: 20.2 mg / g DP (estimated by NMR ^X H): 23 According to the ^X H NMR: i = 0.05

The calculated average molar mass of the ABU co-polyamino acid is 4072 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 2700 g / mol.

Example AB12: Co-polyamino acid AB12 - sodium poly-L-glutamate modified by the molecule AA8 and having a number-average molar mass (Mn) of 3000 g / mol [000386] By a process similar to that used for the preparation of the copolyamino acid AB1 applied to the hydrochloride salt of the molecule AA8 and to a poly-L-glutamic acid obtained by a process similar to that used for the preparation of the co-polyamino acid AB1-1, a sodium poly-L-glutamate modified by the AA8 molecule is obtained.

Dry extract: 19.5 mg / g

PD (estimated from N NMR): 26

According to the NMR ^: 1 = 0.04

The calculated average molar mass of the AB12 co-polyamino acid is 4477 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3000 g / mol.

Example AB13: Co-polyamino acid AB13 - sodium poly-L-glutamate modified by the molecule AA9 and having a number-average molar mass (Mn) of 3300 g / mol [000387] By a process similar to that used for the preparation of the copolyamino acid AB6 applied to the hydrochloride salt of the molecule AA9 and to a poly-L-glutamic acid obtained by a process similar to that used for the preparation of the co-polyamino acid AB1-1 using isoamylamine as initiator in place of hexylamine, a sodium poly-L-glutamate modified by Its molecule AA9 is obtained. Dry extract: 22.3 mg / g

PD (estimated from N NMR): 35

According to NMR = 0.12

The calculated average molar mass of the AB13 co-polyamino acid is 7226 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3300 g / mol.

Example AB21: Co-polyamino acid AB21 - sodium poly-L-glutamate modified by the molecule AA7 and having a number-average molar mass (Mn) of 3400 g / mol [000388] By a process similar to that used for the preparation of the copolyamino acid AB6 applied to the hydrochloride salt of the molecule AA7 (2.44 g, 2.4 mmol) and to a poly-L-glutamic acid (10 g) obtained by a process similar to that used for the preparation of the co-polyamino acid AB1-1, a sodium poly-L-glutamate modified by the molecule AA7 is obtained.

Dry extract: 22.7 mg / g

DP (estimated from NMR: 22

According to the NMR ^: 1 = 0.056

The calculated average molar mass of the AB21 co-polyamino acid is 4090 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3400 g / mol.

Example AB14: Co-polyamino acid AB14 - sodium poly-L-glutamate modified at one of its ends by the molecule AAI and having a number-average molar mass (Mn) of 3400 g / mol [000389] A suitable container is introduced successively the hydrochloride salt of the AAI molecule (2.03 g, 4.70 mmol), chloroform (5 mL), 4Â molecular sieve (1.3 g), as well as Amberlite ion exchange resin IRN 150 (1.3 g). After 1 h of stirring on rollers, the medium is filtered and the resin is rinsed with chloroform. The mixture is evaporated and then co-evaporated with toluene. The residue is dissolved in anhydrous DMF (30 mL) to be used directly in the polymerization reaction.

In a flask previously dried in an oven, γ-benzyl-L-glutamate Ncarboxyanhydride (25.59 g, 97.2 mmol) is placed under vacuum for 30 min and then anhydrous DMF (140 mL) is introduced . The mixture is stirred under argon until complete solubilization, cooled to 4 ° C., then the solution of AAI molecule prepared as described above is introduced quickly. The mixture is stirred between 4 ° C and room temperature for 2 days, then heated to 65 ° C for 2 h. The reaction mixture is then cooled to room temperature then poured dropwise into diisopropyl ether (1.7 L) with stirring. The white precipitate is recovered by filtration, washed twice with diisopropyl ether (140 ml) and then dried under vacuum at

30 ° C to obtain a white solid. The solid is diluted in TFA (160 mL), and a 33% solution of hydrobromic acid (HBr) in acetic acid (62 mL, 354 mmol) is then added dropwise and at 0 ° C. The solution is stirred for 2 h at room temperature and is then poured dropwise onto a 1: 1 (v / v) mixture of diisopropyl ether / water and with stirring (1.9 L). After 2 h of stirring, the heterogeneous mixture is left to stand overnight. The white precipitate is recovered by filtration, washed successively with a 1: 1 mixture (v / v) of diisopropyl ether and water (280 ml) then with water (140 ml). The solid obtained is dissolved in water (530 ml) by adjusting the pH to 7 by adding a 10N aqueous sodium hydroxide solution and then a 1N aqueous sodium hydroxide solution. After solubilization, the theoretical concentration is adjusted to 20 theoretical g / L by adding water to obtain a final volume of 800 mL. The mixture is filtered on a 0.45 μm filter and is then purified by ultrafiltration against a 0.9% NaCl solution and then with water until the conductimetry of the permeate is less than 50 pS / cm. The co-polyamino acid solution is then concentrated to approximately 30 g / L theoretical and the pH is adjusted to 7.0. The aqueous solution is filtered through

0.2 µm and stored at 4 ° C.

Dry extract: 24.1 mg / g

DP (estimated by NMR ^) = 25 therefore i = 0.04

The calculated average molar mass of the AB14 co-polyamino acid is 3378 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3400 g / mol.

Example AB15: Co-polyamino acid AB15 - sodium poly-L-glutamate 5 modified at one of its ends by the molecule AA6 and having a number-average molar mass (Mn) 4100 g / mol [000391] By a process similar to that used for the preparation of copolyamino acid AB14 applied to the hydrochloride salt of the molecule AA6 (2.16 g, 3.94 mmol) and to 25.58 g (97.2 mmol) of γ-benzyl-L-glutamate N-carboxyanhydride , a sodium poly10 L-glutamate modified at one of its ends by the molecule AA6 is obtained. Dry extract: 45.5 mg / g DP (estimated by N NMR) = 30 therefore i = 0.033

The calculated average molar mass of the AB15 co-polyamino acid is 5005 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 4100 g / mol.

Example AB16: Co-polyamino acid AB16 - sodium poly-L-glutamate modified at one of its ends by the molecule AA6 and having a number-average molar mass (Mn) of 6500 g / mol [000392] By a process similar to that used for the preparation of the polyamino acid co20 AB14 applied to the hydrochloride salt of the molecule AA6 (2.39 g, 4.36 mmol) and to 50.0 g (189.9 mmol) of γ-benzyl-L-glutamate N- carboxyanhydride, a sodium polyL-glutamate modified at one of its ends by the molecule AA6 is obtained. Dry extract: 28.5 mg / g

DP (estimated by NMR ^) = 48 therefore i = 0.021

The calculated average molar mass of the AB16 co-polyamino acid is 7725 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 6500 g / mol.

Example AB17: Co-polyamino acid AB17 - sodium poly-L-glutamate modified at one of its ends by the molecule AA7 and having a number-average molar mass (Mn) of 3500 g / me [000393] By a process similar to that used for the preparation of copolyamino acid AB14 applied to the hydrochloride salt of the molecule AA7 (2.80 g, 4.32 mmol) and to 25.0 g (94.9 mmol) of y-benzyl-L-glutamate N-carboxyanhydride , a sodium polyL-glutamate modified at one of its ends by the molecule AA7 is obtained.

Dry extract: 25.2 mg / g

DP (estimated by NMR ^) = 26 therefore i = 0.038

The calculated average molar mass of AB17 co-polyamino acid is 4500 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn - 3500 g / mol.

Example AB18: Co-polyamino acid AB18 - sodium poly-L-glutamate modified at one of its ends by the molecule AA7 and having a number-average molar mass (Mn) of 3700 g / mol [000394] A poly-L-glutamate of sodium modified at one of its ends by the molecule AA7 is obtained by polymerization of γ-methyl N-carboxyanhydride of glutamic acid (25.0 g, 133.6 mmol) using the hydrochloride salt of the molecule AA7 (2, 80 g, 4.32 mmol) as initiator and by deprotecting the methyl esters by using a 37% hydrochloric acid solution according to the process described in patent application FR-A-2 801 226.

Dry extract: 44.3 mg / g

DP (estimated by N NMR) - 22 therefore i = 0.045

The calculated average molar mass of the AB18 co-polyamino acid is 3896 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn - 3700 g / mol.

Example AB19: Co-polyamino acid AB19 - sodium poly-L-glutamate modified at one of its ends by the molecule AA6 and having a number-average molar mass (Mn) of 10500 g / mol [000395] By a process similar to that used for the preparation of copolyamino acid AB14 applied to the hydrochloride salt of the molecule AA6 (1.64 g, 2.99 mmol) and to y-benzyl-L-glutamate N-carboxyanhydride (49.3 g, 187 mmol), a sodium polyL-glutamate modified at one of its ends by the molecule AA6 is obtained. Dry extract: 23.4 mg / g

DP (estimated by NMR Ψ) = 65 therefore i = 0.015

The calculated average molar mass of the AB19 co-polyamino acid is 10293 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn - 10500 g / mol.

Example AB20: Co-polyamino acid AB20 - sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified at one of its ends by the molecule AA6 and having a number-average molar mass (Mn) of 10,400 g / mol [000396] The hydrochloride salt of the molecule AA6 (0.545 g, 1.00 mmol), chloroform (10 mL), 4Â molecular sieve (3 g), as well as Amberlite IRN 150 ion exchange resin (3 g). After 1 h of stirring on rollers, the medium is filtered and the resin is rinsed with chloroform. The mixture is evaporated and then co-evaporated with toluene. The residue is dissolved in anhydrous DMF (10 mL) to be used directly in the polymerization reaction.

In a balloon previously dried in an oven, γ-benzyl-L-glutamate / Vcarboxyanhydride (17.0 g, 64.6 mmol) is placed under vacuum for 30 min and then anhydrous DMF (30 mL) is introduced. The mixture is stirred under argon until complete solubilization, cooled to 4 ° C., then the solution of molecule AA6 prepared as described above is introduced quickly. The mixture is stirred between 4 ° C and room temperature for 2 days, then precipitated in diisopropyl ether (0.6 L). The precipitate is recovered by filtration, washed twice with diisopropyl ether (40 ml) then dried to give a white solid which is dissolved in 80 ml of THF. To this solution are successively added DIPEA, (1.7 mL, 9.8 mmol) then acetic anhydride (0.9 mL, 9.5 mmol). After stirring overnight at room temperature, the solution is poured slowly into diisopropyl ether (480 mL) over a period of 30 min and with stirring. After 1 h of stirring, the precipitate is filtered, washed twice with diisopropyl ether (80 mL) then dried under vacuum at 30 ° C to give a poly (gamma-benzyl-L-glutamic acid) capped at one of its ends by an acetyl group and modified at the other of its ends by the molecule AA6 in the form of a white solid.

The solid is diluted in TFA (65 mL), and a solution of hydrobromic acid (HBr) at 33% in acetic acid (45 mL, 257.0 mmol) is then added dropwise and at 4 ° C. The solution is stirred for 2 h at room temperature and is then poured dropwise onto a 1: 1 (v / v) mixture of diisopropyl ether / water and with stirring (780 mL). After 2 h of stirring, the heterogeneous mixture is left to stand overnight. The white precipitate is recovered by filtration, washed successively with a 1: 1 (v / v) mixture of diisopropyl ether and water (70 mL) then with water (70 mL). The solid obtained is dissolved in water (300 ml) by adjusting the pH to 7 by adding a 10N aqueous sodium hydroxide solution and then a 1N aqueous sodium hydroxide solution. After solubilization, the theoretical concentration is adjusted to 20 g / L theoretical by adding water to obtain a final volume of 440 mL. The mixture is filtered on a 0.45 μm filter and is then purified by ultrafiltration against a 0.9% NaCl solution and then with water until the conductimetry of the permeate is less than 50 pS / cm. The co-polyamino acid solution is then concentrated to approximately 30 g / L theoretical and the pH is adjusted to 7.0. The aqueous solution is filtered through 0.2 μm and stored at 4 ° C. Dry extract: 21.5 mg / g

DP (estimated by NMR ^Χ Η) = 60 therefore i = 0.017

The calculated average molar mass of AB20 co-polyamino acid is 9619 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 10400 g / mol.

Part B:

BB: synthesis of the hydrophobic molecules in which p ~ 2 [000399] The radicals are represented in the following table by the corresponding hydrophobic molecule before grafting on Se co-polyamino acid.

No.

BAI

BA2

BA3

Structure of the hydrophobic molecule before grafting on the copolyamino acid

7 ~ T C ₉ H ₁₉

H, N

H, N

, 0 _Z ^z z • i i ....., "

CgH ₁₉

C _V1 H ⁿ 23

C ₁ p.m.

Table 1d: list of hydrophobic molecules synthesized according to the invention.

BA share: synthesis of hydrophobic molecules in which p = 2 BAI example: BAI molecule

Molecule Bl .: product obtained by the reaction between decanoic acid and L-proline. To a solution of decanoic acid (14.28 g, 82.91 mmol) in THF (520 mL) at 0 ° C are successively added dicyciohexyl carbodiimide (DCC) (16.29 g, 78.96 mmol) and N-hydroxysuccinimide (NHS) (9.09 g, 78.96 mmol). After 60 h of stirring at room temperature, the medium is cooled to 0 ° C for 20 min, filtered through a frit. L-proline (10 g, 86.86 mmol), diisopropylethylamine (DIPEA) (68.8 mL) and water (60 mL) are added to the filtrate. After 24 h of stirring at room temperature, the medium is diluted with water (300 ml). The aqueous phase is washed with ethyl acetate (2 x 250 mL), acidified to pH ~ 1 with an aqueous solution of IN HCl and then extracted with dichloromethane (3 x 150 mL). The combined organic phases are dried over Na2SO4, filtered, concentrated in vacuo and the residue is purified by chromatography on silica gel (cyclohexane, ethyl acetate). Yield: 14.6 g (69%)

Ψ NMR (CDCb, ppm): 0.87 (3H); 1.26 (12H); 1.65 (2H); 2.02 (3H); 2.34 (2H); 2.41 (1H); 3.48 (1H); 3.56 (1H); 4.58 (1H).

LC / MS (ESI): 270.2; (calculated ([M + H] ⁺ ): 270.4).

MolécuJe_B2j. product obtained by the reaction between the molecule B1 and L-lysine. By a process similar to that used for the preparation of the molecule B1 applied to the molecule B1 (14.57 g, 54.07 mmol) and to L-lysine (4.15 g, 28.39 mmol) , a yellow oil is obtained.

Yield: 16.4 g (93%)

Ψ NMR (CDCb, ppm): 0.88 (6H); 1.26 (24H); 1.35-1.65 (8H); 1.85-2.35 (12H); 2.53 (0.2H); 2.90 (0.8H); 3.45-3.75 (5H); 4.50-4.70 (3H); 7.82 (1H).

LC / MS (ESI): 649.6; (calculated ([M + H] ⁺ ): 649.9).

Molecule ............ B3 .............: product obtained by reaction between the molecule B2 and Bocethylenediamine.

To a solution of molecule B2 (16.4 g, 25.27 mmol) in THF (170 mL) are added DIPEA (8.80 mL) and 2- (1H-benzotriazol-l-yl ) -1,3,3,3-tetramethyluronium tetrafluoroborate (TBTU, 8.52 g, 26.54 mmol) at room temperature. After 30 min of stirring, Boc-ethylenediamine (4.45 g, 27.8 mmol) is added. After stirring at room temperature for 2 h, the solvent is evaporated off under reduced pressure and the residue is diluted with ethyl acetate (400 ml). The organic phase is washed with water (250 ml), a saturated aqueous solution of NaHCCb (250 ml), an aqueous solution of 1 N HCl (250 ml), a saturated aqueous solution of NaCl (250 ml) and is dried on NazSCU. After filtration and concentration under vacuum, the residue obtained is purified by chromatography on silica gel (ethyl acetate, methanol) to give a colorless oil.

Yield: 12.8 g (64%)

Ψ NMR (CDCb, ppm): 0.87 (6H); 1.25-1.60 (42H); 1.80-2.05 (4H); 2.15-2.45 (9H); 3.10-3.75 (10H); 4.30 (1H); 4.50 (2H); 5.50 (0.6H); 5.89 (0.2H); 6.15 (0.2H); 7.03 (1H); 7.47 (1H).

LC / MS (ESI): 791.8; (calculated ([M + H] ⁺ ): 792.1).

BAI molecule To a solution of the molecule B3 (12.78 g, 16.15 mmol) in dichloromethane (110 mL) at 5 ° C is added a solution of 4N HCl in dioxane (20.2 mL) . After 20 h of stirring at 5 ° C, the medium is concentrated in vacuo. The residue obtained is dissolved in methanol and evaporated in vacuo, this operation being repeated 4 times to give a white solid of BAI molecule in the form of the hydrochloride salt. Yield: 11.4 g (97%)

Ψ NMR (DMSO-de, ppm): 0.85 (6H); 1.25-1.50 (33H); 1.57 (1H); 1.70-2.40 (12H); 2.82 (2H); 3.00 (2H); 3.25-3.70 (6H); 4.05-4.50 (3H); 7.75-8.45 (6H).

LC / MS (ESI): 691.6; (calculated ([M-CI] ⁺ ): 692.0).

Example BA2: BA2 molecule

Molecule B4: product obtained by the reaction between isauric acid and L-proline. By a process similar to that used for the preparation of the molecule B1 applied to Iauric acid (31.83 g, 157.9 mmol) and to L-proline (20 g, 173.7 mmol), a yellow oil is obtained.

Yield: 34.3 g (73%)

Ψ NMR (CDCb, ppm): 0.87 (3H); 1.26 (16H); 1.70 (2H); 1.90-2.10 (3H); 2.35 (2H); 2.49 (1H); 3.48 (1H); 3.56 (1H); 4.60 (1H).

LC / MS (ESI): 298.2; (calculated ([M + H] ⁺ ): 298.4).

B5 molecule: product obtained by the reaction between the B4 molecule and L-lysine. By a process similar to that used for the preparation of the molecule B1 applied to the molecule B4 (33.72 g, 113.36 mmol) and to L-lysine (8.70 g, 59.51 mmol) , a white solid is obtained.

Yield: 26.2 g (66%)

NMR (CDCb, ppm): 0.88 (6H); 1.26 (32H); 1.35-1.65 (8H); 1.85-2.35 (15H); 2.87 (1H); 3.40-3.75 (5H); 4.50-4.75 (3H); 7.87 (1H).

LC / MS (ESI): 705.6; (calculated ([M + H] ⁺ ): 706.0).

B6 molecule: product obtained by reaction between Boc-ethylenediamine and the B5 molecule.

By a process similar to that used for the preparation of the B3 molecule applied to the B5 molecule (25.74 g, 36.51 mmol) and to Boc-ethylenediamine (6.43 g, 40.16 mmol) , a colorless oil is obtained.

Yield: 30.9 g (quantitative)

Ψ NMR (CDCb, ppm): 0.88 (6H); 1.35-1.65 (50H); 1.85-2.35 (13H); 3.05-3.75 (10H); 4.25-4.65 (3H); 5.50 (0.4H); 5.88 (0.2H); 6.16 (0.2H); 7.08 (1H); 7.26 (1H); 7.49 (0.2H)

LC / MS (ESI): 847.8; (calculated ([M + H] ⁺ ): 848.2).

BA2 molecule [000407] After a process similar to that used for the preparation of the BAI molecule applied to the B6 molecule (30.9 g, 36.47 mmol), the residue obtained after concentration in vacuo is dissolved in methane and evaporated under vacuum, this operation being repeated 4 times to give a white solid of BA2 molecule in the form of the hydrochloride salt after drying under reduced pressure.

Yield: 27.65 g (97%)

¹ H NMR (DMSO-de, ppm): 0.85 (6H); 1.10-2.40 (54H); 2.75-3.15 (4H); 3.25-3.60 (6H); 4.05-4.50 (3H); 7.50-8.50 (6H).

LC / MS (ESI): 747.6; (calculated ([M-CI] ⁺ ): 748.1).

Example BA3: BA3 molecule

B7 molecule. product obtained by the reaction between myristic acid and L-proline. By a process similar to that used for the preparation of the molecule B1 applied to myristic acid (18.93 g, 82.91 mmol) and to L-proline (10 g, 86.86 mmol), a yellowish oil is obtained.

Yield: 20 g (78%)

1 H NMR (CDCb, ppm): 0.88 (3H); 1.28 (20H); 1.70 (2H); 1.90-2.10 (3H); 2.36 (2H); 2.51 (1H); 3.47 (1H); 3.56 (1H); 4.61 (1H).

LC / MS (ESI): 326.2; (calculated ([M + H] ⁺ ): 326.6).

Molecule B8: product obtained by the reaction between molecule B7 and L-lysine [000409] By a process similar to that used for the preparation of molecule B1 applied to molecule B7 (20.02 g, 61.5 mmol) and with L-Sysine (4.72 g, 32.29 mmol), a white solid is obtained.

Yield: 12.3 g (53%)

Ψ NMR (DMSO-de, ppm): 0.85 (6H); 1.26 (40H); 1.35-1.50 (6H); 1.50-2.10 (10H); 2.10-2.25 (4H); 3.01 (2H); 3.31-3.55 (4H); 4.10-4.40 (3H); 7.68 (0.6H); 7.97 (1H); 8.27 (0.4H); 12.50 (1H).

LC / MS (ESI): 761.8; (calculated ([M + H] ⁺ ): 762.1).

Molecule B9: product obtained by the reaction between Sa Boc-ethylenediamine and the molecule B8.

By a process similar to that used for the preparation of the B3 molecule applied to the B8 molecule (12 g, 15.77 mmol) and to its Boc-ethylenediamine (3.03 g, 18.92 mmol), a colorless oil is obtained after purification by a chromatographic column on silica gel (ethyl acetate, methanol).

Yield: 12.5 g (88%)

NMR (DMSO-de, ppm): 0.85 (6H); 1.20-1.55 (55H); 1.50-2.25 (14H); 2.95-3.10 (6H); 3.31-3.55 (4H); 4.10-4.40 (3H); 6.74 (1H); 7.60-8.25 (3H).

LC / MS (ESI): 904.1; (calculated ([M + H] ⁺ ): 904.3).

BA3 molecule After a process similar to that used for the preparation of the BAI molecule applied to the B9 molecule (12.5 g, 13.84 mmol), the residue obtained after concentration in vacuo is dissolved in methanol and evaporated under vacuum, this operation being repeated 4 times to give a white solid of BA3 molecule in the form of the hydrochloride salt after drying under reduced pressure.

Yield: 9.2 g (79%)

1 H-NMR (DMSO-de, ppm): 0.85 (6H); 1.10-1.65 (48H); 1.70-2.35 (12H); 2.85 (2H); 3.01 (2H); 3.25-3.65 (6H); 4.10-4.50 (3H); 7.70-8.40 (6H).

LC / MS (ESI): 803.9; (calculated ([M-CI] ⁺ ): 804.2).

Example BA4: BA4 molecule

BIP molecule product obtained by the reaction between the B8 molecule and Boc-1-amino4,7,10-trioxa-13-tridecane.

By a process similar to that used for the preparation of the B3 molecule applied to the B8 molecule (29.80 g, 39.15 mmol) and to Boc-l-amino-4,7,10-trioxa13-tridecane (15.05 g, 46.96 mmol), a thick colorless oil is obtained. Yield: 25.3 g (61%)

1 H-NMR (DMSO-de, ppm): 0.85 (6H); 1.25-2.35 (75H); 2.85-3.20 (6H); 3.25-3.65 (16H); 4.10-4.45 (3H); 6.38 (0.1H); 6.72 (0.9H); 7.50-8.25 (3H).

LC / MS (ESI): 1064.2; (calculated ([M + H] ⁺ ): 1064.5).

BA4 molecule [000413] After a process similar to that used for the preparation of the BAI molecule applied to the B10 molecule (25.3 g, 23.8 mmol), the residue obtained after concentration in vacuo is dissolved in methanol and evaporated under vacuum, this operation being repeated 4 times to give a white solid of BA4 molecule in the form of the hydrochloride salt after drying under reduced pressure.

Yield: 20.02 g (84%)

Ψ NMR (DMSO-de, ppm): 0.85 (6H); 1.15-2.35 (66H); 2.80-3.20 (6H); 3.30-3.65 (16H); 4.10-4.45 (3H); 7.55-8.60 (6H).

LC / MS (ESI): 964.9; (calculated ([M-CI] ⁺ ): 964.6).

BD Synthèse.des copolyaminoaddes

O

BB2

Hy i = 0.047, DP (m_ + n) = 21

jy> θ11 ^Η 23

Ck ^ nh

O

NH

N H

O

C-i-iH

O

NOT

ⁿ 23

Hy =

O

We have

R <

u

Hy i - 0.049, DP (m + n) - 34

Hy = θ

Hy i = 0.042, DP (m + n) = 23

O

BB7

Hy =

Ο

BB1

Ο

Hy =

Ο

BB1

BB1

i ⁼ Q> Q3, DP (m + n) = 23,

BB1

f \ J _/ ^{C 1:27} PM

Cr NH

HyRI- CH3-COnh \ - ^ ⁿⁱ Î

,We have

i = 0.08, DP = 25

N x ^C 9 ^H 19

Ύ

CK NH

Hy

Hy ^:

C _g H ₁₉

Table the

Defined co-polyamino acids of formulas VII or Vllb

BB15

X

i = 0.042, DP (m) = 24

Part BB: synthesis of co-polyamino acids

Example BB1: Co-polyamino acid BB1 - sodium poly-L-glutamate modified by the molecule BA2 and having a number-average molar mass (Mn) of

2400 g / mol

Co-polyarnmoacid BB] -1 „poly-L-glutamic acid of relative number-average molar mass (Mn) 3860 g / mol resulting from the polymerization of γ-benzyl-L-glutamate Ncarboxyanhydride initiated by hexylamine [000414] In a flask previously dried in an oven is placed under vacuum of ybenzyl-L-glutamate N-carboxyanhydride (90.0 g, 342 mmol) for 30 min, then anhydrous DMF (465 mL) is introduced. The mixture is then stirred under argon until complete dissolution, cooled to 4 ° C, then hexylamine (1.8 mL 14 mmol) is introduced quickly. The mixture is stirred between 4 ° C and room temperature for 2 days. The reaction medium is then heated at 65 ° C for 4 h, cooled to room temperature then poured dropwise into cold diisopropyl ether (6 L) with stirring. The white precipitate is recovered by filtration, washed with diisopropyl ether (500 ml then 250 ml) then dried under vacuum at 30 ° C to give a poly (y-benzyl-L-glutamic acid) (PBLG).

To a solution of PBLG (42.1 g) in trifluoroacetic acid (TFA, 325 mL) at 4 ° C is added dropwise a solution of hydrobromic acid (HBr) at 33% in acid acetic (135 mL, 0.77 mol). The mixture is stirred at ambient temperature for 2 h, then poured dropwise onto a 1: 1 (v / v) mixture of diisopropyl ether and water with stirring (1.6 L). After 1 hour 30 minutes of stirring, the heterogeneous mixture is left to stand overnight. The white precipitate is recovered by filtration, washed with a 1: 1 (v / v) mixture of diisopropyl ether and water (200 mL).

The solid obtained is then solubilized in water (1 L) by adjusting the pH to 7 by adding a 10N aqueous sodium hydroxide solution and then a 1N aqueous sodium hydroxide solution. After solubilization, the theoretical concentration is adjusted to 25 g / L theoretical by adding water to obtain a final volume of 1.5 L.

The solution is filtered on a 0.45 μm filter and then purified by ultrafiltration against a 0.9% NaCl solution, then water until the conductimetry of the permeate is less than 50 pS / cm.

The aqueous solution is then acidified by adding 37% hydrochloric acid solution until reaching a pH of 2. After 4 h of stirring, the precipitate obtained is filtered and then dried under vacuum at 30 ° C to give a poly-L-glutamic acid with a number-average molar mass (Mn) 3860 g / mol relative to a polyoxyethylene (PEG) standard.

Co-polyamino acid BB1 [000419] Co-polyamino acid BB1-1 (10.0 g) is dissolved in DMF (700 mL) at 30-40 ° C and then cooled to 0 ° C. The hydrochloride salt of the molecule BA2 (2.95 g, 3.8 mmol) is suspended in DMF (45 ml) and triethylamine (0.39 g, 3.8 mmol) is then added to this. suspension then the mixture is slightly heated with stirring until complete dissolution. To the solution of co-polyamino acid at 0 ° C, N-methyl morphine (NMM, 7.6 g, 75 mmol) in DMF (14 mL) and ethyl chloroformate (ECF, 8.1 g, 75 mmol) are added. After 10 min at 0 ° C, the solution of BA2 molecule is added and the medium maintained at 30 ° C for 1 h. The reaction medium is poured dropwise onto 6 L of water containing sodium chloride at 15% by mass and HCl (pH 2), then left to stand overnight. The precipitate is collected by filtration, washed with sodium chloride solution at pH 2 (IL) and dried under vacuum for about 1 h. The white solid obtained is taken up in water (600 mL) and the pH is adjusted to 7 by adding Sent of a 1N aqueous NaOH solution. The volume is adjusted to 700 mL by adding water. After filtration through a 0.45 μm filter, the clear solution obtained is purified by ultrafiltration against a 0.9% NaCl solution and then with water, until the conductimetry of the permeate is less than 50 pS / cm. After unloading, its solution is filtered through a 0.2 μm filter and stored at 2-8 ° C.

Dry extract: 19.7 mg / g

DP (estimated according to NMR ^T H): 23 According to Sa NMR = 0.05

The calculated average molar mass of the co-polyamino acid BB1 is 4350 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 2400 g / mol.

Example BB2: co-polyamino acid BB2 - sodium poly-L-glutamate modified by the molecule BA2 and having a number-average molar mase (Mn) of 4900 g / mol [000420] A poly-L-glutamic acid of average molar mass in number (Mn)

4100 g / mol (5.0 g) obtained by a process similar to that used for the preparation of the co-polyamino acid BB1-1 is dissolved in DMF (205 mL) at 30-40 ° C and then maintained at this temperature. In parallel, the hydrochloride salt of the BA2 molecule (1.44 g, 1.84 mmol) is suspended in DMF (10 mL) and triethylamine (0.19 g, 1.84 mmol) is added , then the mixture is slightly heated with stirring until complete dissolution. To the solution of co-polyamino acid in ie DMF, to NMM (3.7 g, 36.7 mmol), the solution of foam BA2 then to 2-hydroxypyridine N-oxide (HOPO, 0.31 g, 2.76 mmol) are added successively. The reaction medium is then cooled to 0 ° C., then EDC (0.53 g, 2.76 mmol) is added and ie

100 medium rose to room temperature for 3 h. The reaction medium is poured dropwise onto 1.55 L of water containing NaCl at 15% by mass and HCl (pH 2) with stirring. At the end of the addition, the pH is readjusted to 2 with a 1N HCl solution, and the suspension is left to stand overnight. The precipitate is collected by filtration, then rinsed with 100 ml of water. The white solid obtained is dissolved in 200 ml of water by slow addition of a 1N aqueous NaOH solution to pH 7 with stirring, then the solution is filtered through a 0.45 μm filter. The clear solution obtained is purified by ultrafiltration against a 0.9% NaCl solution and then with water, until the conductimetry of the permeate is less than 50 pS / cm. The solution obtained is filtered through a 0.2 μm filter and stored at 2-8 ° C.

Solids content: 16.3 mg / g DP (estimated by NMR ^X H): 21 According to the ^X H NMR: i = 0.047

The calculated average molar mass of the co-polyamino acid BB2 is 3932 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 4900 g / mol.

Example BB3 Co-polyamino acid BB3 - sodium poly-L-glutamate modified by the molecule BA2 and having a number-average molar mass (Mn) of @ 400 g / mol

Co-poiyamino acid BB3-X: poly-L-glutamic acid of number-average molar mass (Mn) 17,500 g / mol resulting from the polymerization of y-methyl-L-glutamate Ncarboxyanhydride initiated by L-leucinamide [000421] An acid poly-L-glutamic mass average in number (Mn) 17500 g / mol relative to a standard polymethyl methacrylate (PMMA) is obtained by polymerization of y-methyl N-carboxyanhydride of glutamic acid using Lleucinamide as initiator and in performing a deprotection of the methyl esters by using a 37% hydrochloric acid solution according to the process described in patent application FR-A-2 801 226.

By a process similar to that used for the preparation of the polyamino acid co30 BB2 applied to the hydrochloride salt of the molecule BA2 (3.23 g, 4.1 mmol) and to the co-polyamino acid BB3-1 (11 g), a sodium poly-L-glutamate modified by Its molecule BA2 is obtained.

Solids content: 27.5 mg / g DP (estimated by NMR ^X H): 34

According to the NMR ^X H: li = 0.049

The calculated average molar mass of the co-polyamino acid BB3 is 6405 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 6400 g / mol.

101

Example BB4: co-polyamino acid BB4 - sodium poly-L-glutamate modified by the molecule BA2 and having a number-average molar mass (Mn) of 10500 g / mol [000423] By a process similar to that used for the preparation of the co5 polyamino acid BB2 applied to the hydrochloride salt of the molecule BA2 (5 g, 6.35 mmol) and to a poly-L-glutamic acid of average molar mass in number Mn =

10800 g / me (21.7 g) obtained by a process similar to that used for the preparation of the co-polyamino acid BB1-1, a sodium poly-L-glutamate modified by the molecule BA2 is obtained.

Dry extract: 28.2 mg / g

DP (estimated from ia NMR ^T H): 65 From NMR ^: 1 = 0.04

The calculated average molar mass of the co-polyamino acid BB4 is 11,721 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 10,500 g / mol.

Example BB5: Co-polyamino acid BB5 - sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by the molecule BA2 and having a number-average molar mass (Mn) of 3600 g / mol Ço.- .poly.amino acid BB5-1: poly-L-glutamic acid with Mn 3700 g / mol resulting from the polymerization of γ-benzyl-L-glutamate N-carboxyanhydride initiated by hexylamine and capped at one of its ends by a group acetyl [000424] In a flask previously dried in an oven is placed under vacuum ybenzyl-L-glutamate N-carboxyanhydride (100.0 g, 380 mmol) for 30 minutes and then anhydrous DMF (250 mL) is introduced. The mixture is then stirred under argon until complete dissolution, cooled to 4 ° C, then hexylamine (2.3 mL, 17 mmol) is introduced quickly. The mixture is stirred between 4 ° C and room temperature for 2 days then precipitated in diisopropyl ether (3.4 L). The precipitate is recovered by filtration, washed twice with diisopropyl ether (225 ml) then dried to give a white solid which is dissolved in 450 ml of THF. To this solution are successively added Ν, Ν-diisopropylethylamine (DIPEA, 31 mL, 176 mmol) and then acetic anhydride (17 mL, 176 mmol). After stirring overnight at room temperature, the solution is poured slowly into diisopropyl ether (3 L) over a period of 30 min and with stirring. After 1 h of stirring, the precipitate is filtered, washed twice with diisopropyl ether (200 ml) then dried under vacuum at 30 ° C to give a poly (y-benzyl-L-glutamic acid) capped at one of its ends by an acetyl group. To a solution of the capped co-polyamino acid (72 g) in trifluoroacetic acid (TFA, 335 mL) at 4 ° C is added dropwise a 33% hydrobromic acid (HBr) solution in the acetic acid (235 mL, 1.34 mol). The mixture is

102 stirred at room temperature for 3 h 30, then poured dropwise onto a 1: 1 (v / v) mixture of diisopropyl ether and water with stirring (4 L). After 2 h of stirring, the heterogeneous mixture is left to stand overnight. The white precipitate is recovered by filtration, washed with a 1: 1 (v / v) mixture of diisopropyl ether and water (340 mL) then with water (340 mL). The solid obtained is then dissolved in water (1.5 L) by adjusting the pH to 7 by adding a 10N aqueous sodium hydroxide solution and then a 1N aqueous sodium hydroxide solution. After solubilization, the theoretical concentration is adjusted to 20 g / L theoretical by adding water to obtain a final volume of 2.1 L. The solution is filtered on a 0.45 μm filter and then purified by ultrafiltration against a 0.9% NaCl solution, then l until the conductivity of the permeate is less than 50 pS / cm.

The co-polyamino acid solution is then concentrated until a final volume of 1.8 L is obtained. The aqueous solution is then acidified by adding 37% hydrochloric acid solution until reaching a pH of 2. After 4 h stirring, the precipitate obtained is filtered, washed with water (330 mL) and then dried under vacuum at 30 ° C to give a poly-L-glutamic acid of number average molar mass (Mn) 3700 g / mol compared to a polyoxyethylene (PEG) standard.

Co-polyamino acid BBS By a process similar to that used for the preparation of co20 polyamino acid BB2 applied to the hydrochloride salt of the molecule BA2 (6.92 g, 8.8 mmol) and to co-polyamino acid BB5-1 (30 , 0 g), a sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by the molecule BA2 is obtained. Dry extract: 29.4 mg / g

PD (estimated from N NMR): 23

According to the NMR ^: 1 = 0.042

The calculated average molar mass of the co-polyamino acid BB5 is 4302 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3600 g / mol.

Example BB6: Co-polyamino acid BB6 - sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by the molecule BA2 and having a number-average molar mass (Mn) of 4100 g / mol [000427] By a process similar to that used for the preparation of the copolyamino acid BB2 applied to the hydrochloride salt of the molecule BA2 (5.8 g, 7.4 mmol) and to a poly-L-glutamic acid of average molar mass in number Mn = 3800 g / mol (25 g) obtained by a process similar to that used for the preparation of copolyamino acid BB5-1 using ammonia in place of hexylamine, a sodium poly-Lglutamate capped at one of its ends by a acetyl group and modified by the molecule BA2 is obtained.

103

Solids content: 27.6 mg / g DP (estimated by NMR ^X H): 24 According to the ^X H NMR: i = 0.04

The calculated average molar mass of the co-polyamino acid BB6 is 4387 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 4100 g / mol.

Example BB7: Co-polyamino acid BB7 - sodium poly-L-glutamate modified by the molecule BA2 and having a number-average molar mass (Mn) of 4200 g / mol [000428] By a process similar to that used for the preparation of the copolyamino acid BB2 applied to the hydrochloride salt of the molecule BA2 (7.07 g, 9.0 mmol) and to a poly-L-glutamic acid of average molar mass in number Mn ~ 3600 g / mol (30.0 g) obtained by a process similar to that used for the preparation of the copolyamino acid BB1-1, a sodium poly-L-glutamate modified by the molecule BA2 is obtained.

Solids content: 28.3 mg / g DP (estimated by NMR ^X H): 22 According to the ^X H NMR: i == 0.042

The calculated average molar mass of the co-polyamino acid BB7 is 4039 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 4200 g / mol.

BBS example; co-polyamino acid BBS - sodium poly-L-glutamate modified by the molecule 8A2 and having a number-average molar mass (Mn) of 5200 g / mol [000429] By a process similar to that used for its preparation of the copolyamino acid BB2 applied with the hydrochloride salt of the molecule BA2 (0.85 g, 1.1 mmol) and with a poly-L-glutamic acid of average molar mass in number Mn = 4100 g / mol (5.0 g) obtained by a similar process to that used for the preparation of the copolyamino acid BB1-1, a sodium poly-L-glutamate modified by its molecule BA2 is obtained.

Solids content: 28.6 mg / g DP (estimated by NMR ^X H): 21 According to the ^X H NMR: i = 0.026

The calculated average molar mass of the co-polyamino acid BB8 is 3620 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn - 5200 g / mol.

104 f

/

I ί

ï

Example BB9: co-polyamino acid BB9 - sodium poly-L-glutamate modified by the molecule BA3 and having a number-average molar mass (Mn) of

4700 g / mol [000430] By a process similar to that used for the preparation of co5 polyamino acid BB2 applied to the hydrochloride salt of the molecule BA3 (3.05 g, 3.6 mmol) and to a poly-L-glutamic acid of number-average molar mass Mn = 4100 g / mol (10.0 g) obtained by a process similar to that used for the preparation of copolyamino acid BB1-1, a sodium poly-L-glutamate modified by the molecule BA3 is obtained.

Dry extract: 28.6 mg / g

DP (estimated from NMR: 26 Based on NMR ^: 1 = 0.05

The calculated average molar mass of the co-polyamino acid BB9 is 4982 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 4700 g / mol.

Example BB1O: co-polyamino acid BB10 - sodium poly-L-glutamate modified by the molecule BA3 and having a number-average molar mass (Mn) of 4200 g / mol [000431] By a process similar to that used for the preparation of the co20 polyamino acid BB2 applied to the hydrochloride salt of the molecule BA3 (1.90 g, 2.3 mmol) and to a poly-L-glutamic acid of average molar mass in number Mn = 3500 g / mol (10.0 g) obtained by a process similar to that used for the preparation of the copolyamino acid BB1-1, a sodium poly-L-glutamate modified by the molecule BA3 is obtained.

Dry extract: 25.9 mg / g

DP (estimated by NMR ^X H): 22 According to NMR: 1 = 0.029

The calculated average molar mass of the co-polyamino acid BB10 is 3872 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 4200 g / mol.

Example BBli: co-polyamino acid BB11 - sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by the molecule BA4 and having a number-average molar mass (Mn) of 3900 g / mol [000432] By a process similar to that used for the preparation of the polyamino acid co35 BB2 applied to the hydrochloride salt of the molecule BA4 (2.21 g, 2.2 mmol) and to a poly-L-glutamic acid of average mass in number Mn = 3700 g / mol (10 g) obtained by a process similar to that used for the preparation of co1

105 polyamino acid BB5-1, a sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by the molecule BA4 is obtained.

Dry extract: 28.1 mg / g

DP (estimated from ^X H NMR): 22 5 Based on ^X H NMR: i = 0.032

The calculated average molar mass of the co-polyamino acid BB11 is 4118 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3900 g / mol.

Example BB12: co-polyamino acid BB12 - sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by Its molecule BA3 and having a number-average molar mass (Mn) of 3900 g / mol [000433] By a process similar to that used for the preparation of the copolyamino acid BB2 applied to the hydrochloride salt of the molecule BA3 (1.9 g, 2.3 mmol) and to a poly-L-glutamic acid of average mass in number Mn = 3600 g / mol (10

g) obtained by a process similar to that used for the preparation of copolyamino acid BB5-1, a sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by the molecule BA3 is obtained.

Solids content: 26.7 mg / g DP (estimated by NMR ^X H): 23

According to the NMR ^X H: i = 0.03

The calculated average molar mass of the co-polyamino acid BB12 is 4145 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3900 g / mol.

Example BB13: co-polyamino acid BB13 - sodium poly-L-glutamate modified by the molecule BAI and having a number-average molar mass (Mn) of 2800 g / mol [000434] By a process similar to that used for the preparation of the copolyamino acid BB1 applied to the hydrochloride salt of the BAI molecule (3.65 g, 5 mmol) and to a poly-L-glutamic acid of average molar mass in number Mn =

3600 g / mol (10 g) obtained by a process similar to that used for the preparation of the co-polyamino acid BB1-1, a sodium poly-L-glutamate modified with the BAI molecule is obtained.

Dry extract: 25.6 mg / g

DP (estimated by NMR ^X H): 25

According to the NMR ^X H: i = 0.08

The calculated average molar mass of the co-polyamino acid BB13 is 5253 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 2800 g / mol.

106

Example BB14: co-polyamino acid BB14 - sodium poly-L-glutamate modified at one of its ends by the molecule BA2 and having a number-average molar mass (Mn) of 4020 g / mol [000435] A suitable container is introduced successively the hydrochloride salt of the BA2 molecule (2.12 g, 2.70 mmol), chloroform (40 mL), 4Â molecular sieve (1.5 g), as well as the ion exchange resin Amberlite IRN 150 (1.5 g). After 1 h of stirring on rollers, the medium is filtered and the resin is rinsed with chloroform. The mixture is evaporated and then co-evaporated with toluene. The residue is dissolved in anhydrous DMF (20 mL) to be used directly in the polymerization reaction.

In a balloon previously dried in an oven, γ-benzyl-L-glutamate Ncarboxyanhydride (18 g, 68.42 mmol) is placed under vacuum for 30 min and then anhydrous DMF (100 mL) is introduced. The mixture is stirred under argon until complete solubilization, cooled to 4 ° C., then Its solution of BA2 molecule prepared as described above is introduced quickly. The mixture is stirred between 4 ° C and room temperature for 2 days, then heated to 65 ° C for 2 h. The reaction mixture is then cooled to room temperature then poured dropwise into diisopropyl ether (1.2 L) with stirring. The white precipitate is recovered by filtration, washed twice with diisopropyl ether (100 ml) then dried under vacuum at 30 ° C to obtain a white solid. The solid is diluted in TFA (105 mL), and a 33% solution of hydrobromic acid (HBr) in acetic acid (38 mL, 220 mmol) is then added dropwise and at 0 ° C. The solution is stirred for 2 h at room temperature and is then poured dropwise onto a 1: 1 (v / v) mixture of diisopropyl ether / water and with stirring (600 mL). After 2 h of stirring, the heterogeneous mixture is left to stand overnight. The white precipitate is recovered by filtration, washed successively with a 1: 1 mixture (v / v) of diisopropyl ether and water (200 ml) then with water (100 ml). The solid obtained is dissolved in water (450 mL) by adjusting the pH to 7 by adding a 10N aqueous sodium hydroxide solution and then a 1N aqueous sodium hydroxide solution. The mixture is filtered on a 0.45 μm filter. then is purified by ultrafiltration against a 0.9% NaCl solution and then water until the conductimetry of the permeate is less than 50 pS / cm. The co-polyamino acid solution is then concentrated to approximately 30 g / L theoretical and the pH is adjusted to 7.0. The aqueous solution is filtered through 0.2 μm and stored at 4 ° C.

Dry extract: 22.3 mg / g

DP (estimated by NMR = 29 therefore i = 0.034

The calculated average molar mass of the co-polyamino acid BB14 is 5089 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 4020 g / mol.

107

Example BB15: co-polyamino acid BB1S - sodium poly-L-glutamate modified at one of its ends by the molecule BA3 and having a number-average molar mass (Mn) of 3610 g / mol [000437] By a process similar to that used for the preparation of the copolyamino acid BB14 applied to the hydrochloride salt of the molecule BA3 (3.62 g, 4.32 mmol) and to 25.0 g (94.97 mmol) of γ-benzyl-L-glutamate N-carboxyanhydride, a sodium polyL-glutamate modified at one of its ends by the molecule BA3 is obtained. Dry extract: 26.5 mg / g

DP (estimated by NMR Ψ) = 24 therefore i = 0.042

The calculated average molar mass of the co-polyamino acid BB15 is 4390 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3610 g / mol.

Example BBÎ6: co-polyamino acid BB16 - sodium poly-L-glutamate modified at one of its ends by the molecule BA4 and having a number-average molar mass (Mn) of 3300 g / mol [000438] By a process similar to that used for the preparation of the copolyamino acid BB14 applied to the hydrochloride salt of the molecule BA4 (5.70 g, 5.70 mmol) and to 29.99 g (113.9 mmol) of γ-benzyl-L-glutamate N-carboxyanhydride, a sodium poly-L-glutamate modified at one of its ends by its molecule BA4 is obtained.

Dry extract: 32.3 mg / g

DP (estimated by NMR Ψ) = 23 therefore i = 0.043

The calculated average molar mass of co-poSyamino acid BB16 is 4399 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn = 3300 g / mol.

Example BB17: co-polyamino acid BB17 - sodium poly-L-glutamate modified at one of its ends by the molecule BA3 and having a number-average molar mass of 10730 g / mol [000439] By a process similar to that used for the preparation of the copolyamino acid BB14 applied to the hydrochloride salt of the molecule BA3 (2.51 g, 3 mmol) and to 52.7 g (200 mmol) of γ-benzyl-L-glutamate N-carboxyanhydride, a modified poly-Lglutamate at one of its ends by the BA3 molecule is obtained. Dry extract: 24.5 mg / g

DP (estimated by 1 H NMR) - 65 so i = 0.015

The calculated average molar mass of the co-polyamino acid BB17 is 10585 g / mol.

Aqueous HPLC-SEC (PEG calibrator): Mn - 10700 g / mol.

108

Example BB18: co-polyamino acid BB18 - sodium poly-L-glutamate modified at one of its ends by the molecule BA3 and having a number-average molar mass of 6600 g / mol [000440] By a process similar to that used for the preparation of co5 polyamino acid BB14 applied to the hydrochloride salt of the molecule BAS (2.51 g, 3 mmol) and to 31.6 g (120 mmol) of γ-benzyl-L-glutamate N-carboxyanhydride, a sodium poly-Lglutamate modified at one of its ends by the BA3 molecule is obtained.

Dry extract: 27.3 mg / g

DP (estimated by NMR Ψ) = 40 therefore i = 0.025

The calculated average molar mass of the co-polyamino acid BB18 is 6889 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 6600 g / mol.

Part C:

The glucagon used is human glucagon resulting from a process of peptide synthesis. It comes from the company Bachem (reference 407473).

Example C1: Glucagon solution at 2 mg / ml [000441] Glucagon (80 mg) powder is introduced into a 45 ml faecal tube.

A 0.01 N aqueous hydrochloric acid solution (40 ml) is added. The glucagon powder is mixed by repeated inversions of the tube until the glucagon is completely dissolved. The 2 mg / ml glucagon solution is then filtered through a membrane (0.22 μm).

Example C2: Glucagon solution at 4 mg / ml [000442] Glucagon (160 mg) powder is introduced into a 45 ml falcon tube. A 0.003 N aqueous hydrochloric acid solution (40 ml) is added. The glucagon powder is mixed by repeated inversions of the tube until the glucagon is completely dissolved. The glucagon solution at 4 mg / ml is then filtered through a membrane (0.22 μm).

Example C3: Glucagon solution at 6 mg / ml [000443] Glucagon (240 mg) powder is introduced into a falcon tube of 45 ml. A 0.003 N aqueous hydrochloric acid solution (40 ml) is added. The glucagon powder is mixed by repeated inversions of the tube until the glucagon is completely dissolved. The glucagon solution at 6 mg / ml is then filtered through a membrane (0.22 μm).

109

Example C4: Glucagon solution at 10 mg / ml [000444] Glucagon (400 mg) powder is introduced into a 45 ml falcon tube. A 0.003 N aqueous hydrochloric acid solution (40 ml) is added. The glucagon powder is mixed by repeated inversions of the tube until the glucagon is completely dissolved. The 10 mg / ml glucagon solution is then filtered through a membrane (0.22 μm).

Tests were first carried out in order to check whether the copolyamino acids make it possible to dissolve the glucagon, and its minimum concentration of co-polyamino acid necessary to dissolve the glucagon was determined.

Example CAI: Compositions of AB16 co-polyamino acid at variable concentrations and glucagon at 1 mg / ml.

X mg of AB16 co-polyamino acid are weighed precisely, and added to 2 ml of a 10 mM phosphate buffer solution comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm), [000447] 2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above in order to lead to a composition comprising X mg / ml of co-polyamino acid and 1 mg / ml of glucagon.

[000448] A visual inspection is carried out to determine whether or not a clear solution is obtained. The result of the minimum concentration is presented in Table 5.

Example CA2: Compositions of AB21 co-polyamino acid at variable concentrations and glucagon at 1 mg / ml.

As described in the CAI example, compositions comprising X mg / ml of AB21 co-polyamino acid and 1 mg / ml of glucagon are prepared.

A visual inspection is carried out to determine whether or not a clear solution is obtained. The result of the minimum concentration is presented in Table 5.

Example CA3: Compositions of co-polyamino acid BB14 at variable concentrations and glucagon at 1 mg / ml.

In the same manner as described in the CAI example, compositions comprising X mg / ml of co-polyamino acid BB14 and 1 mg / ml of glucagon are prepared.

Visual inspection is carried out to determine whether or not a clear solution is obtained. The result of the minimum concentration is presented in Table

5.

Example CA4: Compositions of co-polyamino acid BB2 at variable concentrations and glucagon at 1 mg / ml.

In the same way as described in the CAI example, compositions comprising X mg / ml of co-polyamino acid BB2 and 1 mg / ml of glucagon are prepared.

A visual inspection is carried out to determine whether or not a clear solution is obtained. The result of the minimum concentration is presented in Table

5.

Example CA5: Compositions of co-polyamino acid BB15 at variable concentrations and glucagon at 1 mg / ml.

In the same manner as described in the CAI example, compositions comprising X mg / ml of co-polyamino acid BB15 and 1 mg / ml of glucagon are prepared.

A visual inspection is carried out to determine whether or not a clear solution is obtained. The result of the minimum concentration is presented in Table 5.

Example CA6: Compositions of co-polyamino acid BB9 at variable concentrations and glucagon at 1 mg / ml.

In the same manner as described in the CAI example, compositions comprising X mg / ml of co-polyamino acids BB9 and 1 mg / ml of glucagon are prepared.

A visual inspection is carried out to determine whether or not a clear solution is obtained. The result of the minimum concentration is presented in Table 5.

111

Example Co- polyamino Minimum concentration of copolyamino acid (in mg / ml) for the solubilization of human glucagon (1 mg / ml) CAI AB16 <0.82 CA2 AB21 <1.25 CA3 BB14 <0.82 CA4 BB2 <0.82 CASE BB15 <1.25 CA6 BB9 <1.25

Table 5: Minimum concentration of co-polyamino acid (in mg / ml) for the solubilization of human glucagon (1 mg / ml).

Example CA7: Solution of co-polyamino acid BB15 at 4.4 mg / ml and glucagon at 2 mg / ml [00010] 17.2 mg of co-polyamino acid BB15 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example C2 are mixed with 2 ml of the co-polyamino acid solution BB15 as prepared above. We obtain a clear solution.

Example CA8: Solution of co-polyamino acid BB15 at 4.4 mg / ml and glucagon at 3 mg / ml [00013] 2 ml of a glucagon solution as prepared in Example C3 are mixed with 2 ml of the BB15 co-polyamino acid solution as prepared in Example CA7.

We obtain a clear solution.

Example CA9: Solution of co-polyamino acid BB15 at 4.4 mg / ml and glucagon at 5 mg / ml [00015] 2 ml of a glucagon solution as prepared in Example C4 are mixed with 2 ml of the BB15 co-polyamino acid solution as prepared in Example CA7.

We obtain a clear solution.

112 [00017] Tests were carried out in order to verify whether the co-polyamino acids made it possible to stabilize the glucagon, then to determine a minimum concentration of co-polyamino acid necessary to stabilize the glucagon.

Example CB1: Solution of co-polyamino acid BB15 at variable concentrations and glucagon at 1 mg / ml [00018] 4X mg of co-polyamino acid BB15 are weighed precisely, and added to 2 ml of a 10 mM phosphate buffer solution comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above in order to lead to a composition comprising X mg / ml of co -polyamino acid and 1 mg / ml of glucagon.

Then three samples of 1 ml each of these solutions are prepared and placed under static conditions at 37 ° C.

A visual inspection is carried out at 7 days, 14 days, and 2 days, see Table 6

Example Co-concentration polyamino acid BB15 (Mg / ml) Stable at 7 days Stable at 14 d Stable at 21 d cblA 0.4 no no no cblB 0.8 no no no CBLC 1.2 no no no cblD 2.5 no no no cable 3.8 Yes Yes Yes cblF 5.1 Yes Yes OR! CBlg 1 ........................................ Λ4 Yes Yes Yes

Table 6

Concentration range to determine the minimum hydrophobic / glucagon radical molar ratio

Concentration ranges were carried out with other co-polyamino acids and led to the following stable solutions being obtained.

113

Example CB2: Solution of co-polyamino acid AB1 at 15 mg / ml and glucagon at 1 mg / ml [00022] 60 mg of co-polyamino acid AB1 are weighed precisely, and added to 2 ml of a phosphate buffer solution at 10 mM comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution AB1 as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB3: Solution of ABS co-polyamino acid at 10 mg / ml and of glucagon at 1 mg / ml [00024] 40 mg of co-polyamino acid AB5 are weighed precisely, and added to 2 ml of a phosphate buffer solution at 10 mM comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB4: Solution of co-polyamino acid AB7 at 8.6 mg / ml and glucagon at 1 mg / ml [00026] 34.4 mg of co-polyamino acid AB7 are weighed precisely and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB5: Solution of co-polyamino acid AB15 at 14 mg / ml and glucagon at 1 mg / ml [00028] 56 mg of co-polyamino acid AB15 are weighed precisely and added to 2 ml of a phosphate buffer solution at 10 mM comprising m-cresol (46 mM),

114 glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of its co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB6: Solution of co-polyamino acid AB16 at 16.2 mg / ml and of glucagon at 1 mg / ml [00030] 64.8 mg of co-polyamino acid AB16 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB7: Solution of co-polyamino acid AB17 at 6.4 mg / ml and glucagon at 1 mg / ml [00032] 25.6 mg of co-polyamino acid AB17 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB8: Solution of co-polyamino acid BB2 at 8 mg / ml and of glucagon at 1 mg / ml [00034] 32 mg of co-polyamino acid BB2 are weighed precisely, and added to 2 ml of a phosphate buffer solution at 10 mM comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then

115 three I ml samples each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB9: Solution of co-polyamino acid BBS at 9 mg / ml and of glucagon at 15 mg / ml [00036] 36 mg of co-polyamino acid BBS are weighed precisely, and added to 2 ml of a phosphate buffer solution with 10 mM comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB10: Solution of co-polyamino acid BB7 at 15.4 mg / ml and of glucagon at 1 mg / ml [00038] 61.6 mg of co-polyamino acid BB7 are weighed precisely and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB11: Solution of co-polyamino acid BB8 at 7.6 mg / ml and of glucagon at 1 mg / ml [00040] 30.4 mg of co-polyamino acid BB8 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

116

Example CB12: Solution of co-polyamino acid BB9 at 4.3 mg / ml and of glucagon at 1 mg / ml [00042] 17.2 mg of co-polyamino acid BB9 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB13: Solution of co-polyamino acid BB11 at 5.9 mg / ml and of glucagon at 1 mg / ml [00044] 23.6 mg of co-polyamino acid BB11 are weighed precisely and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB14: Solution of co-polyamino acid BB11 at 8.6 mg / ml and of glucagon at 1 mg / ml [00046] 34.4 mg of co-polyamino acid BB11 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until the co-polyamino acid is dissolved, then its solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB15: Solution of co-polyamino acid BB14 at 9.1 mg / ml and of glucagon at 1 mg / ml [00048] 36.4 mg of co-polyamino acid BB14 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM),

117 glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB16: Solution of co-polyamino acid BB16 at 3.8 mg / ml and of glucagon at 1 mg / ml [00050] 15.2 mg of co-polyamino acid BB16 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB17: Solution of co-polyamino acid BB16 at 6.3 mg / ml and glucagon at 1 mg / ml [00052] 25.2 mg of co-polyamino acid BB16 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB18: Solution of co-polyamino acid BB15 at 4.4 mg / ml and glucagon at 2 mg / ml [00054] 17.6 mg of co-polyamino acid BB15 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example C2 are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then

118 three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB19: Solution of co-polyamino acid BB15 at 8.8 mg / ml and glucagon at 25 mg / ml [00056] 35.4 mg of co-polyamino acid BB15 are weighed precisely, and added to 2 ml of a solution 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example C2 are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB20: Solution of co-polyamino acid BB15 at 6.3 mg / ml, glucagon at 1 mg / ml and L-methionine at 0.1 mg / ml [00058] A solution of co-polyamino acid BB15 at 12.6 mg / ml is prepared by dissolving 46.5 mg of co-polyamino acid lyophilizate BB15 with 1.6 ml of water, 1.3 ml of 126.7 mM m-cresol, 358 µL of 4.9 M glycerol , 360 µL of a 100 mM phosphate buffer solution and 75 µL of 9.8 mg / mL L-methionine. The solution is filtered on a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example C2 are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB21: Solution of co-polyamino acid BB15 at 6.3 mg / ml, glucagon at 1 mg / ml and L-methionine at 1 mg / ml [00060] A solution of co-polyamino acid BB15 at 12.6 mg / ml is prepared by dissolving 46.5 mg of co-polyamino acid lyophilizate BB15 with 937 pL of water, 1.3 mL of 126.7 mM m-cresol, 358 pL of 4.9 M glycerol, 360 pL d '' a 100 mM phosphate buffer solution and 733 µL of L-methionine at 9.8 mg / mL. The solution is filtered on a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example C2 are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of Mi each of this solution are prepared and placed under static conditions at 37 ° C.

119

Example CB22: Solution of co-polyamino acid BB15 at 6.3 mg / ml, glucagon at 1 mg / ml and exenatide at 0.1 mg / ml [00062] 14 mg of exenatide (Bachem; Pdt N ° - 4044219) are introduced into an Eppendorf tube and then 1.4 ml of water is added. The powder is mixed by repeated inversions and the 10 mg / ml exenatide solution is filtered through a membrane (0.22 μm). A solution of co-polyamino acid BB15 at 12.6 mg / ml is prepared by dissolving 28.3 mg of lyophilisate of co-polyamino acid BB15 with 937 pL of water, 817 pL of m-cresoi at 126.7 mM , 224 µL of 4.9 M glycerol, 225 µL of 100 mM phosphate buffer solution and 45 µL of 10 mg / mL exenatide. The solution is filtered on a membrane (0.22 μm).

The final solution is prepared by mixing 2 ml of a glucagon solution as prepared in Example C1 and 2 ml of the BB15 solution at 12.6 mg / ml as prepared above. The mixture is homogenized manually and contains 1 mg / mL of glucagon, 0.1 mg / mL of exenatide, 6.3 mg / mL of BB15, 5 mM phosphate buffer, 23 mM m-cresol and 249 mM glycerol.

Three I ml samples each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB23: Solution of co-polyamino acid BB15 at 6.3 mg / ml, glucagon at 1 mg / ml and exenatide at 0.25 mg / ml [00066] A solution of exenatide at 10 mg / ml is obtained in the same way as described in example CB22.

A solution of co-polyamino acid BB15 at 12.6 mg / ml is prepared by dissolving 28.4 mg of lyophilisate of co-polyamino acid BB15 with 872 pL of water, 817 pL of m-cresol at 126.7 mM, 224 µL of 4.9 M glycerol, 225 µL of 100 mM phosphate buffer solution and 113 µL of 10 mg / mL exenatide. The solution is filtered on a membrane (0.22 μm).

The final solution is prepared by mixing 2 ml of a glucagon solution as prepared in Example C1 and 2 ml of the BB15 solution at 12.6 mg / ml as prepared above. The mixture is homogenized manually and contains 1 mg / mL of glucagon, 0.25 mg / mL of exenatide, 6.3 mg / mL of BB15, 5 mM phosphate buffer, 23 mM m-cresol and 249 mM glycerol.

Three I ml samples each of this solution are prepared and placed under static conditions at 37 ° C.

120

Example CB24: Solution of co-polyamino acid BB15 at 6.3 mg / ml, glucagon at 1 mg / ml and exenatide at 0.5 mg / ml [00070] A solution of co-polyamino acid BB15 at 12.6 mg / ml is prepared by dissolving 28.2 mg of lyophilisate of co-polyamino acid BB15 with 750 pL of water, 817 pL of m-cresol at 126.7 mM, 224 pL of Glycerol at 4.9 M, 225 pL of a 100 mM phosphate buffer solution and 226 μL of exenatide at 10 mg / ml, as prepared in the example above. The solution is filtered on a membrane (0.22 μm).

The final solution is prepared by mixing 2 ml of a glucagon solution as prepared in Example Cl and 2 ml of the co-polyamino acid BB15 solution at 12.6 mg / ml as prepared above . The mixture is homogenized manually and contains 1 mg / ml of glucagon, 0.5 mg / ml of exenatide, 6.3 mg / ml of co-polyamino acid BB15, 5 mM of phosphate buffer, 23 mM m-cresol and 249 mM of glycerol.

Three I ml samples each of this solution are prepared and placed under static conditions at 37 ° C.

Example CB25: Solution of co-polyamino acid AB21 at 8.6 mg / ml and glucagon at 1 mg / ml [00073] 34.4 mg of co-polyamino acid AB21 are weighed precisely, and added to 2 ml of a solution of 10 mM phosphate buffer comprising m-cresol (46 mM), glycerol (548 mM). The composition is stirred until dissolution of the co-polyamino acid, then the solution is filtered through a membrane (0.22 μm).

2 ml of a glucagon solution as prepared in Example Cl are mixed with 2 ml of the co-polyamino acid solution as prepared above. Then three samples of I ml each of this solution are prepared and placed under static conditions at 37 ° C.

Part D: STABILITY

Example DI: Physical Stability of Co-Polyamino Acid / Glucagon Compositions Visual inspection of the samples placed under static conditions at 37 ° C. is carried out at 0, 7, 14 and 21 days at 37 ° C. in order to detect the appearance visible particles or turbidity. This inspection is carried out according to the recommendations of the European Pharmacopoeia (EP 2.9.20): the vials are subjected to lighting of at least 2000 Lux and are observed in front of a white background and a black background. When particles are visible in at least 2 of the 3 samples, the composition is estimated to be unstable. Stable therefore means that at the date of the inspection at least 2 samples were free of particles.

The results of the visual inspections are shown in Table 7 below.

Example Co- polyamino fmg / ml) glucagon (Mg / ml) exenatide (Mg / ml) L-methionine (Mg / ml) Stable at 7 days Stable at 14 d Stable at 21 d cable BB15 (3.8) 1 0 0 Yes Yes Yes CB2 AB1 (15) 1 0 0 Yes no no CB3 ABS (10) 1 0 0 Yes OR! Yes CB4 AB7 (8.6) 1 0 0 Yes Yes Yes CBS AB15 (14) 1 0 0 Yes Yes Yes CB6 AB16 (16.2) 1 0 0 Yes Yes Yes CB7 AB17 (6.4) 1 0 0 Yes Yes Yes CB8 BB2 (8) 1 0 0 Yes OR! Yes CB9 BB5 (9) 1 0 0 Yes Yes no CB10 BB7 (15.4) 1 0 0 Yes Yes CB11 BB8 (7.6) 1 0 0 Yes no no CB12 BB9 (4.3) 1 0 0 Yes Yes Yes CB13 BB11 (5.9) 1 0 0 Yes Yes no CB14 BB11 (8.6) 1 0 0 Yes Yes Yes CB15 BB14 (9.1) 1 0 0 Yes Yes no CB16 BB16 (3.8) 1 0 0 Yes Yes no CB17 BB16 (6.3) 1 0 0 Yes Yes Yes CB18 BB15 (4.4) 2 0 0 Yes Yes no CB19 BB15 (8.6) 2 0 0 Yes Yes Yes CB20 BB15 (6.3) 1 0 0.1 OR! Yes * CB21 BB15 (6.3) 1 0 1 Yes Yes CB22 BB15 (6.3) 1 0.1 0 Yes Yes CB23 BB15 (6.3) 1 0.25 0 Yes Yes CB24 BB15 (6.3) 1 0.5 0 Yes Yes CB25 AB21 (8.6) 0 0 0 Yes Yes Yes

"-" means not observed

Table 7: Results of visual inspections of compositions comprising a copolyamino acid and glucagon

122

Example D2: Chemical Stability of Co-Polyamino Acid / Glucagon Compositions An RP-HPLC method adapted from USP directives was used to determine the concentration of glucagon and its degradation products. This method was used to evaluate the chemical stability of the glucagon of the compositions.

The HPLC conditions are as follows, and the HPLC gradient is summarized below: Column: 4.6 x 150 mm, C-18

Mobile phase A: Solution S / Acetonitrile 80/20 (v / v), solution S being a solution of potassium dihydrogen phosphate 150 mm in water, adjusted to pH 2.7 with an 85% phosphoric acid solution

- Mobile phase B: water / acetonitrile 60/40 (v / v)

Mobile phase C: water / acetonitrile 10/90 (v / v)

Column temperature: 45 ° C

Detection: UV 210 nm

Autosampler temperature: 4 ° C. The recovery was measured on samples at 7, 14 and 21 days at 37 ° C. under static conditions. The chemical stability data, that is to say recovery of glucagon obtained by RP-HPLC are presented in Table 8 below.

Example Co- polyamino (Mg / ml) glucagon (Mg / ml) exenatide (Mg / ml) L- methionine (Mg / ml Overlap rancid at 7d Overlap rancid at 14 d Overlap rancid at 21 d cable BB15 (3.8) 1 0 0 > 95 > 85 CB4 AB7 (8.6) 1 0 0 > 95 > 90 > 85 CB5 AB15 (14) 1 0 0 > 90 > 90 CB7 AB17 (6.4) 1 0 0 > 95 > 90 > 90 CB8 BB2 (8) 1 0 0 > 90 > 90 CB10 BB7 (15.4) 1 0 0 > 95 > 90 CB14 BB11 (8.6) 1 0 0 > 95 > 90 > 90 CB15 BB14 (9.1) 1 0 0 > 90 CB17 BB16 (6.3) 1 0 0 > 95 > 90 > 90 CB18 BB15 (4.4) 2 i 0 0 > 95 > 90 > 85 CB19 BB15 (8.6) 2 0 0 > 95 > 90 > 90 CB20 BB15 (6.3) 1 0 0.1 > 95 > 90 . J

123

CB21 BB15 (6.3) 1 0 1 > 95 > 90 CB23 BB15 (6.3) 1 0.25 0 > 95 > 90 CB24 BB15 (6.3) 1 0.5 0 > 95 > 90 " CB25 AB21 (8.6) 0 0 0 ; > 90 > 85

"-" means not measured

Table 8: Measures of recovery of compositions comprising a co-polyamino acid and glucagon

Part F: Pharmacodynamic studies in pigs Studies have been conducted with the aim of evaluating the pharmacodynamics of a composition of co-polyamino acid BB15 and glucagon (example CBle) at a dose of 2pg / kg in the pig.

The hyperglycemic effects of this composition in Example CBle were compared with an injection of a glucagon solution (Glucagen®, NOVO NORDISK) at 2pg / kg.

Twelve animals which have been fasted for about 5.5 hours were injected into the side at a dose of 2pg / kg using a Junior Star® pen. In order to overcome regulatory effects of its glycaemia by the secretion of insulin, in response to the hyperglycemic effect induced by the injection of glucagon, 30 minutes before the injection, 44 pg / kg of octreotide is administered to pigs subcutaneously. Three blood samples are taken in the hour preceding the injection (T-40min, T20min and T-10min), in order to determine the basal level of glucose and glucagon. Blood samples are then taken for 3 hours after administration. Blood sugar is determined using a glucometer.

The median pharmacodynamic curves of the glycemia expressed by the difference in glucose relative to the basal level are shown in FIG. 1. The curve representing the results obtained with the composition of Example CBle is represented by empty squares and the curve representing the results of the composition of glucagen® is represented by solid circles.

The pharmacodynamic results obtained from the administration of the formulation of the example CBle and of Glucagen® show a hyperglycemic activity quickly after injection with a maximum blood sugar level reached 30 minutes after injection. These pharmacodynamic profiles show that the formulation of Example CBle and the commercial human glucagon (Glucagen®) have similar pharmacodynamic properties.

Claims (9)

1. Co-polyamino acid carrying carboxylate charges and hydrophobic Hy radicals consisting of glutamic or aspartic units characterized in that said hydrophobic Hy radicals are chosen from Its radicals of formula I below: * - ^ GpR GpC) P Formula I in which GpR is a radicai of formulas II or II ': H R-N- * n '; GpA is a radical of formulas III or III ': he A O 'Il H HN— * m or * - ^l AN- 'mGpC is a radical of formula IV: O - the * indicate the sites of attachment of the different groups linked by amide functions; - a is an integer equal to 0 or 1; - b is an integer equal to 0 or 1; p is an integer equal to 1 or 2 and o if p is equal to 1 then a is equal to 0 or 1 and GpA is a radical of formula III 'and, o if p is equal to 2 then a is equal to 1, and GpA is a radical of formula III; 125 c is an integer equal to 0 or 1, and if c is equal to O then d is equal to 1 or 2; d is an integer equal to 0, 1 or 2; r is an integer equal to 0 or 1, and o if r is equal to 0 then the hydrophobic radical of formula i is linked to the copolyamino acid via a covalent bond between a carbonyl of the hydrophobic radical and a nitrogen atom in the N terminal position of the copolyamino acid, thus forming an amide function resulting from the reaction of an amine function in the N-terminal position of the co-polyamino acid precursor and an acid function carried by the precursor of the hydrophobic radical, and o if r is equal to 1 then Se radical hydrophobic of formula I is linked to the copolyamino acid: • via a covalent bond between a nitrogen atom of the hydrophobic radical and a carbonyl of the co-polyamino acid, thus forming an amide function resulting from the reaction of an amine function of the precursor of the hydrophobic radical and an acid function carried by the precursor of the co-polyamino acid or • via a covalent bond between a carbonyl of the hydrophobic radical and a nitrogen atom in position N terminai of the co-poiyamino acid, thus forming an amide function resulting from the reaction of an acid function of the precursor of the hydrophobic radical and an amine function in the N-terminal position carried by the precursor of the copolyamino acid; R is a radical chosen from the group consisting of: o a divalent, linear or branched alkyl radical, comprising if GpR is a radical of formula II of 2 to 12 carbon atoms or if GpR is a radical of formula IT of 1 to 11 carbon atoms; o a divalent, linear or branched alkyl radical, comprising if GpR is a radical of formula II of 2 to 11 carbon atoms or if GpR is a radical of formula IT of 1 to 11 carbon atoms, said alkyl radical bearing one or more -CONH2 functions, and o an unsubstituted ether or polyether radical comprising from 4 to 14 carbon atoms and from 1 to 5 oxygen atoms; A is a linear or branched alkyl radical comprising from 1 to 6 carbon atoms; B is a linear or branched alkyl radical, optionally comprising an aromatic ring, comprising from 1 to 9 carbon atoms; Cx is a linear or branched monovalent aikyl radical, in which x indicates the number of carbon atoms and: 126 o if p is equal to 1, x is between 11 and 25 (11 <x <25): o if p is equal to 2, x is between 9 and 15 (9 <x <15), the ratio i between the number of hydrophobic radicals and the number of glutamic or aspartic units being between 0 <i <0.5; when several hydrophobic radicals are carried by a co-polyamino acid then they are identical or different, the degree of polymerization DP in glutamic or aspartic units is between 10 and 250; the free acid functions being in the form of an alkaline cation salt chosen from the group consisting of Na ⁺ and K ⁺ . 2. Co-polyamino acid according to claim 1, characterized in that said hydrophobic radicals are chosen from hydrophobic radicals of formula I in which p = 1, represented by the following formula V: ^ GpR ^ GpA) —GpC formula V GpR, GpA, GpC, r and a have the definitions given above. 3. Co-polyamino acid according to claim 1, characterized in that said hydrophobic radicals are chosen from hydrophobic radicals of formula I in which a ~ 1 and p - 2, represented by the following formula VI: GpR —GpA — GpC) ² Formula VI in which GpR, GpA, GpC, r and a have the definitions given above. 4. Co-polyamino acid according to any one of the preceding claims, characterized in that it is chosen from the co-polyamino acids of formula VII below: 127 Q ΟΧ m We Hy formula VII in which, • D represents, independently, either a group -CH2- (aspartic unit) or a group -CH2-CH2- (glutamic unit), • Hy is a hydrophobic radical chosen from the hydrophobic radicals of formulas I, V or VI, in which r = 1 and GpR is a radical of Formula II, • Ri is a hydrophobic radical chosen from the hydrophobic radicals of formulas I, V or VI in which r = 0 or r = 1 and GpR is a radical of Formula II ′, or a radical chosen from the group consisting of an H, a linear C2 to CIO acyl group, a branched C3 to CIO acyl group, a benzyl, a terminal "amino acid" unit and a pyroglutamate, • R2 is a hydrophobic radical chosen from hydrophobic radicals of formulas I, V or VI in which r = 1 and GpR is a radical of Formula II, or an identical or different radical -NR'R, R 'and R being chosen from group consisting of H, linear or branched or cyclic C2 to CIO alkyl, Se b enzyl and said R 'and R alkyl which can together form one or more saturated, unsaturated and / or aromatic carbon rings and / or which may contain heteroatoms, chosen from the group consisting of O, N and S, • X represents an H or a cationic entity chosen from the group comprising metallic cations; • n + m represents the degree of polymerization DP of the co-polyamino acid, that is to say the average number of monomeric units per chain of copolyamino acid and 5 <n + m <250; 128 5. Co-polyamino acid according to claim 4, characterized in that it is chosen from co-polyamino acids of formulas VII, in which Ri - R'i and R2 = R'2, of formula Vlla below: in which, - m, η, X, D and Hy have the definitions given above, R'i is a radical chosen from the group consisting of an H, a linear acyl group at C2 to CIO, a branched acyl group at C3 to CIO, a benzyl, a terminal "amino acid" unit and a pyroglutamate, - R'2 is a radical -NR'R, R 'and R identical or different being chosen from the group consisting of H, linear or branched or cyclic C2 to CIO alkyls, benzyl and said R' and R alkyls which may together form one or more saturated, unsaturated and / or aromatic carbon rings and / or which may contain heteroatoms, chosen from the group consisting of O, N and S. 6. Co-polyamino acid according to claim 4, characterized in that it is chosen from among its co-polyamino acids of formula VII in which n = 0 of formula Vllb below: R ₁ in which m, X, D, Ri and R2 have the definitions given above and at least Ri or R2 is a hydrophobic radical of formula I, V or VI. 129 7. Co-polyamino acid according to claim 6, characterized in that it is chosen from co-polyamino acids of formula Vllb in which R.2 is a radical 5 hydrophobic of formula I, V or VI in which r = 1 and GpR is of Formula ΙΓ. 8. Co-polyamino acid according to any one of claims 4 to 7, characterized in that it is chosen from among its co-polyamino acids of formulas VII, Vlla or Vllb in which the at least one co-polyamino acid is chosen from co- polyamino acids in 10 which group D is a group -CFk- (aspartic unit). 9. Co-polyamino acid according to any one of claims 4 to 7, characterized in that it is chosen from co-polyamino acids of formulas VII, Vlla or Vllb in which the at least one co-polyamino acid is chosen from co -polyamino acids in Which group D is a group -CH2-CH2- (glutamic unit).