FR3061023A1 - COMPOSITIONS IN THE FORM OF AQUEOUS INJECTABLE SOLUTION COMPRISING AMYLINE, AMYLINE RECEPTOR AGONIST OR AMYLINE ANALOGUE AND CO-POLYAMINOACID - Google Patents

Abstract

The invention relates to a composition in the form of an injectable aqueous solution, the pH of which is between 6.0 and 8.0, comprising at least: a) amylin, an amylin receptor agonist or a amylin analogue; b) a co-polyamino acid bearing carboxylate charges and hydrophobic radicals Hy, said co-polyamino acid being constituted by glutamic or aspartic units and said hydrophobic radicals Hy being of formula I below: characterized in that the composition does not comprise any a basal insulin whose isoelectric point pI is between 5.8 and 8.5. It also relates to a composition characterized in that it further comprises prandial insulin.

Description

Holder (s):

ADOCIA Limited company.

O Extension request (s):

® Agent (s): TRIPOZ CABINET.

FR 3 061 023 - A1 ® COMPOSITIONS IN THE FORM OF AN AQUEOUS INJECTION SOLUTION COMPRISING AMYLINE, AN AMYLINE RECEPTOR AGONIST OR AN AMYLINE ANALOG AND A CO-POLYAMINOACID.

(® The invention relates to a composition in the form of an injectable aqueous solution, the pH of which is between 6.0 and 8.0, comprising at least:

a) amylin, an amylin receptor agonist or an amylin analog;

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

®GpR) -f-GpA) - (Gpc) ^ra P Formula I characterized in that the composition does not comprise a basal insulin whose isoelectric point pl is between 5.8 and 8.5.

It also relates to a composition characterized in that it further comprises a mealtime insulin.

COMPOSITIONS IN THE FORM OF AN AQUEOUS SOLUTION

INJECTABLE COMPRISING AMYLINE, AN AGONIST IN

AMYLINE RECEPTOR OR AN AMYLINE ANALOG AND A

CO-POLYAMINOACID [0001] The invention relates to its therapies by injection of amylin, of amylin receptor agonist or of amylin analog to treat diabetes.

The invention relates to a composition in the form of an injectable aqueous solution, the pH of which is between 6.0 and 8.0, comprising at least amylin, an agonist at the amylin receptor or a analog of amylin and a co-polyamino acid carrying carboxylate charges and hydrophobic radicals according to the invention and compositions further comprising insulin (excluding basal insulins whose isoelectric point pi is between 5.8 and 8 , 5). The invention also relates to pharmaceutical formulations comprising the compositions according to the invention. Finally, the invention also relates to a use of the co-polyamino acids carrying carboxylate charges and of hydrophobic radicals according to the invention for stabilizing compositions of amylin, of agonist at the amylin receptor or analog of amylin as well as compositions of amylin, amylin receptor agonist or amylin analog further comprising an insulin.

Type 1 diabetes is an autoimmune disease leading to the destruction of beta cells in the pancreas. These cells are known to produce insulin, the main role of which is to regulate the use of glucose in peripheral tissues (Gerich 1993 Control of glycaemia). Consequently, patients with type 1 diabetes suffer from chronic hyperglycemia and must administer exogenous insulin in order to limit this hyperglycemia. Insulin therapy has drastically changed the life expectancy of these patients. However, the glycemic control provided by exogenous insulin is not optimal, especially after taking a meal. This is related to the fact that these patients produce glucagon after taking a meal, which leads to the destocking of part of the glucose stored in the liver, which is not the case in the healthy person. This glucagon-mediated glucose production exacerbates the problem of regulating blood sugar in these patients.

It has been shown that amylin, another hormone produced by beta cells in the pancreas and therefore also deficient in type 1 diabetic patients, plays a key role in the regulation of postprandial blood sugar. Amylin, also known as the "islet amyloid polypeptide" or IAPP, is a 37 amino acid peptide that is co-stored and co-secreted with insulin (Schmitz 2004 Amylin Agonists). This peptide is described to block the production of gîucagon by the alpha cells of the pancreas. Thus, insulin and amylin have complementary and synergistic roles, since insulin makes it possible to reduce the concentration of glucose in the blood whereas amyline makes it possible to reduce the entry of endogenous glucose into the blood by inhibiting the production (secretion) of endogenous gîucagon.

This problem of regulating postprandial glycemia is fairly similar for patients with type 2 diabetes treated with insulin insofar as their disease has led to a very significant loss of their beta cell mass and therefore, their ability to produce insulin and amylin.

Human amylin has properties which are not compatible with pharmaceutical requirements in terms of solubility and stability (Goldsbury CS, Cooper GJ, Goldie KN, Muller SA, Saafi EL, Gruijters WT, Misur MP, Engel A , Aebi U, Kistler J: Polymorphie fibrillar assembîy of human amylin. J Struct Biol 119: 17-27, 1997). Amylin is known to form amyloid fibers which lead to the formation of plaques which are insoluble in water. Although being the natural hormone, it was necessary to develop an analog in order to solve these solubility problems.

The physico-chemical properties of amylin thus make it impossible to use: amylin is only stable for about fifteen minutes at acidic pH, and less than a minute at neutral pH.

The company Amylin has developed an analog of amylin, pramlintide, to overcome the lack of stability of human amylin. This product, marketed under the name of Symlin, was approved in 2005 by the FDA for the treatment of type 1 and type 2 diabetics. It must be administered subcutaneously three times a day, within one hour before a meal. improve post-meal blood sugar control. This peptide is formulated at acidic pH and is described to fibrillate when the pH of the solution is higher than 5.5. Variants of analogs are described in US Patent 5,686,411.

This analog is thus not satisfactory in terms of stability when a formulation at neutral pH is envisaged.

To date, there is no way to stabilize human amylin to make it a pharmaceutical product. However, it would be advantageous for patients to have access to the human form of this physiological hormone. It would also be advantageous to be able to formulate a neutral pH amylin receptor analog or agonist.

In addition, there would be an advantage in being able to mix amyline, an analogue of amylin, or an amylin receptor agonist, with a mealtime insulin in aqueous solution since these two products are to be administered before the meal. This would also mimic physiology since these two hormones are co-secreted by beta cells in response to a meal to improve control of postprandial blood sugar.

However, taking into account the fact that the prandial insulin solutions have a pH close to neutral for reasons of chemical stability, it is not possible to obtain an aqueous solution meeting the pharmaceutical requirements in terms of solubility and stability.

For this reason, patent application US2016 / 001002 from the company ROCHE describes a pump containing two separate reservoirs in order to make possible the co-administration of these two hormones with a single medical device. However, this patent does not solve the problem of mixing these two hormones in solution which would make it possible to administer them with conventional pumps already on the market which contain only one reservoir.

Patent application W02013067022 from the company XERIS provides a solution to the problem of the stability of amylin and its compatibility with insulin by using an organic solvent instead of water. The absence of water seems to solve the stability problems but the use of an organic solvent poses safety problems of chronic use for diabetic patients and also problems of compatibility with the usual medical devices, at the level of the tubes. , seals and plasticizers used.

Patent application WO2007104786 from the company NOVO NORDISK describes a method for stabilizing a solution of pramlintide, which is an analogue of amylin, and of insulin by the addition of a phospholipid, derived from glycerophosphoglycerol, in particular dimyristoyl glycerophosphoglycerol (DMPG). However, this solution requires the use of large quantities of DMPG which can pose a problem of local tolerance.

To the knowledge of the applicant, there is no satisfactory means which makes it possible to combine in aqueous solution a mealtime insulin and human amylin, an amylin receptor agonist or an analog of amylin in order to be able to be administered with conventional devices.

The acid formulation pH and rapid fibrillation are obstacles to obtaining a pharmaceutical formulation with neutral pH based on amylin and pramlintide, but also a brake in combining amylin or pramlintide with other pharmaceutical ingredients. active, in particular peptides or proteins.

The Applicant has noticed that, surprisingly, the co-polyamino acids according to the invention stabilize compositions of amylin, of agonist at the amylin receptor or of amylal analog at a pH of between 6 and 8. In fact, compositions comprising amylin, an amylin receptor agonist or an amylin analog in combination with a co-polyamino acid according to the invention exhibit increased stability over time, which is of great interest for pharmaceutical development.

The Applicant has also found that the co-polyamino acids according to the invention also make it possible to obtain a composition comprising prandial insulin and amylin, agonist at the amylin receptor or amylin analog, said composition being clear and having improved fibrillation stability.

A conventional method for measuring the stability of proteins or peptides consists in measuring the formation of fibrils using Thioflavin T, also called ThT. This method makes it possible to measure, under temperature and stirring conditions which allow the phenomenon to accelerate, the lag time before the formation of fibrils by measuring the increase in fluorescence. The compositions according to the invention have a latency time before the formation of fibrils significantly greater than that of amylin, of an agonist at the amylin receptor or of an amylin analog at the pH of interest.

The compositions according to the invention have physical, and possibly chemical, stability, satisfactory at the desired pH.

In one embodiment, the invention relates to a composition in the form of an injectable aqueous solution, the pH of which is between 6.0 and 8.0, comprising at least:

a) amylin, an amylin receptor agonist or an amylin analog;

b) a co-polyamino acid carrying carboxylate 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:

* - (g _p r) - (- gpa) - (gpc) ^ra P Formula I in which

GpR is a radical of formulas II or ΙΓ:

HH _n or *

- GpA is a radical of formulas III or ΙΙΓ:

O HN— *

-HE/

HN— * neither or *

H

A-N- * ni ';

GpC is a radical of formula IV:

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 1 and GpA is a radical of formula ΙΙΓ 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 terminal of the co-polyamino 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 co-polyamino 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 an amine function in the N-terminal position carried by the precursor of the co-polyamino 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 -CONH ₂ 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 5 carbon atoms;

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

- C _x 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 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 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 ⁺ ;

characterized in that the composition does not include a basal insulin whose isoelectric point pi is between 5.8 and 8.5.

In one embodiment, Hy comprises more than 30 carbon atoms.

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 7.0 and 7.8.

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

The compositions in the form of an injectable aqueous solution according to the invention are clear solutions. The term “clear solution” is understood to mean compositions which satisfy the criteria described in the American and European pharmacopoeias concerning injectable solutions. In the US Pharmacopoeia, the solutions are defined in the section <1151> referring to the injection (<1>) (referring to <788> according to USP 35 and specified in <788> according to USP 35 and in <787> , <788> and <790> USP 38 (from ¹ August 2014), according to USP 38). In the European Pharmacopoeia, injectable solutions must meet the criteria given in sections 2.9.19 and 2.9.20.

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 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 the 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 hydrophobic radicals of formula I in which if p is equal to 1 and if x is greater than 17 (17 <x) then r = 1 and R is an ether or polyether 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, the composition is characterized in that said hydrophobic radicals are chosen from the hydrophobic radicals of formula I in which p = 1, represented by the following formula V:

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 radical of formula V in which: r is equal to 1 (r = l) and a is equal to 0 (a = 0).

In one embodiment, the composition is characterized in that the hydrophobic radical 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 radical 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 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 12 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 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 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 V in which GpR is a radical of formula ΙΓ in which R is a divalent alkyl radical comprising from 1 to β 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 (-CONH ₂ ).

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 11, in which R is a divalent linear alkyl radical, comprising of 2 to 5 carbon atoms and carrying one or more amide functions (-CONH ₂ ).

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:

ίο [00047] 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 (CONH ₂ ).

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 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 ΙΓ, in which R is an ether radical comprising from 4 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 or ΙΓ in which R is an ether radical represented by the formula

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 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 II ', in which R is a polyether radical chosen from the group consisting of

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 by the formulas below:

★ * .. * * ^H 3 ^C \ j ch ₃ * * * * * * ^ ch ₃ h ₃ c * * * * 'V HjcZ h, cXh 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 the radicals of formulas IVa, IVb or IVc below represented:

* 0 Z ^k, ' ^NH V Formula IVb O Formula IVc

In one embodiment, the composition is characterized in that the 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 represented below: ______________________________________________

O O -Λ Vo. O Formula IVd ί r Formula IVe 0 0 Formula IVf

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

* * * * ch ₃ ch ₃ ★ * * * * * X b ^ ^Àx CH ₃ ch ₃ h ₃ c ^ * * Y * * * * h ₃ Y h ₃ cXXh 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 or IVa in which 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

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 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 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 by your radicals represented by the formulas below:

* = 'CH ₃ x = ll  * 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 IV is chosen from the group consisting of radicals in which Cx is chosen from the 1st group consisting by 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 the group consisting of 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

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 of 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 formed by alkyl radicals represented by the formu are below: x = 17

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 compound radicals the formulas below

In one embodiment, the composition is characterized in that the 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:

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

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 radical of formula VI in which GpR is a radical of formula II.

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 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 Π 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, 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 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 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 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 linear alkyl radical, comprising of 2 to 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

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 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 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 [00095] 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 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 ΙΓ, in which R is a linear polyether radical comprising from 6 to atoms carbon 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 ΙΓ 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 the radicals of formulas Ilia and Illb below represented:

Θ: HN Ilia formula O HN S Illb Formula

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

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 and IVc ci -after represented:

* ~ Y ° / 0 \ θ 'V Formula IVa ‘Y ° / o \ Formula IVb O / 0 \ ΥΛΎ M Formula IVc

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: ____________________

0 0 * —kk ~ C _x O Formula IVd 0 0 \ - _N ^z Formula IVe 0 0 : Formula IVf

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula VI in which the GpC radical 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 the group consisting by 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 by branched 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 by 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 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 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 by 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:

* x = 9 x - 11 ★ ch ₃ x = 13

In one embodiment, the composition is characterized in that the hydrophobic radical is a radical of formula Vï 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 14 or 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 by 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:

formula VII in which, • D represents, independently, either a group -CH ₂ - (aspartic unit) or a group -CH ₂ -CH ₂ - (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 hydrophobic radicals of formulas I, V or VI in which r = 0 or r = l and GpR is a radical of Formula ΙΓ, or a radical chosen from the group consisting of an H, a linear C2 to C10 acyl group, a branched C3 to C10 acyl group, a benzyl, a terminal "amino acid" unit and a pyroglutamate, • R ₂ 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, or an identical or different radical -NR'R, R 'and R being chosen in the group consisting of H, linear or branched or cyclic C2 to CIO alkyls, l e benzyl and said R ′ and R alkyl 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, • 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 co-polyamino 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 the 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 Vlla.

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 R ₂ = R ' ₂ , of the following formula Vlla:

D

OX

O

R '

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 C2 to C10 acyl group, a branched C3 to C10 acyl group, a benzyîe, 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 C10 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.

Called "defined co-polyamino acid" 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 VII in which n = 0 of formula Vllb below:

in which m, X, D, Ri and R ₂ have the definitions given above and at least Ri or R ₂ 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 VII in which n = 0 of formula Vllb and Ri or R ₂ 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 ΙΓ.

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 R ₂ is a hydrophobic radical of formula I, V or VI in which r = 1 and GpR is of Formula II.

[000121] In one embodiment, the composition is characterized in that Ri is a radical selected from the group consisting of linear acyl, C ₂ -C ₁₀ acyl group branched C ₃ to Ci _0, benzyl , a terminal "amino acid" unit and a pyroglutamate.

In one embodiment, the composition is characterized in that R! is a radical chosen from the group consisting of a linear C ₂ to C ₁₀ acyl group or a branched C ₃ to C ₁₀ acyl group.

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, Vlla or Vllb in which group D is a group -CH ₂ - (aspartic unit).

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, Vlla or Vllb in which group D is a group -CH ₂ -CH ₂ - (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 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 0.015 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 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, 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 0.01 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 and 14 carbon atoms and the ratio i between the number of hydrophobic radicals and the number d '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 a mode characterized in [000145] In characterized in that n + a mode [000146] In characterized in that n + a mode [000147] In characterized in that n + a mode [000148] In characterized in that n + a mode that embodiment, the composition m is between 10 and 250.

of embodiment, the composition m is between 10 and 200.

of embodiment, the composition m is between 15 and 150.

of embodiment, the composition m is between 15 and 100.

the composition is between 15 and 80.

In one embodiment, the composition i is between 15 and 65.

of embodiment, the composition m is between 20 and 60.

embodiment, the composition is between 20 and 50.

the composition is between 20 and 40.

Characterized

according to the invention East according to the invention East according to the invention East according to the invention East according to the invention East according to the invention East according to the invention East according to the invention East according to the invention East

in characterized in [000151] in characterized in [000152] characterized in that a mode what n + a mode what

In this one mode [000153] Amylin, or islet amyloid polypeptide (IAPP), is a peptide hormone with 37 residues. It is co-secreted with insulin from pancreatic beta cells in the ratio of about 100: 1. Amylin plays a role in glycemic regulation by stopping the secretion of endogenous glucagon and slowing gastric emptying and promoting satiety, thereby reducing postprandial glycemic excursions in blood glucose levels.

IAPP is treated from a coding sequence of 89 residues. The amyloid Proislet polypeptide (proIAPP, proamyline, proislet protein) is produced in pancreatic beta cells (beta cells) as a pro-peptide of RSO 67 amino acids, 7404 Dalton, and undergoes post-translational modifications including cleavage of protease to produce amylin.

In the present application, the amylin as mentioned refers to the compounds described in US patents 5,124,314 and US 5,234,906.

The term “analog” is understood to mean, when used with reference to a peptide or a protein, a peptide or a protein, in which one or more amino acid residues constituting the primary sequence have been substituted with d other amino acid residues 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%. In the case of amylin, an analog may for example be derived from the primary amino acid sequence of amylin by substituting one or more natural or non-natural or peptidomimetic amino acids.

The term “derivative” is understood to mean, 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 analog, that is to say a peptide or a protein which has been modified by creation of covalent bonds, to introduce substituents of the non-amino acid type.

An agonist at the amylin receptor refers to a compound which mimics one or more characteristics of the activity of amylin.

[000159] Amylin derivatives are described in the article Yan et al., PNAS, vol.

103, no 7, p 2046-2051, 2006.

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

Analogs of amylin are described in US Patents 5,686,411, US

6,114,304 or even US 6,410,511.

In one embodiment, the composition is characterized in that the amylin analog is pramlintide (Symlin®) sold by the company

ASTRAZENECA AB.

In one embodiment, the molar ratios of co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog are between

1.5 and 75.

In one embodiment, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog molar ratios are between 1.8 and 50.

In one embodiment, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog molar ratios are between and 35.

In one embodiment, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog molar ratios are between

2.5 and 30.

In one embodiment, the molar ratios of co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog are between and 30.

In one embodiment, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog molar ratios are between

3.5 and 30.

In one embodiment, the molar ratios of co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog are between and 30.

In one embodiment, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analogue molar ratios are between and 30.

In one embodiment, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog molar ratios are between 7 and 30.

In one embodiment, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analogue molar ratios are between 9 and 30.

In one embodiment, the co-polyamino acid / amylin molar ratios are between 3 and 75.

In one embodiment, the co-polyamino acid / amylin molar ratios are between 7 and 50.

In one embodiment, the co-polyamino acid / amylin molar ratios are between 10 and 30.

In one embodiment, the co-polyamino acid / amylin molar ratios are between 15 and 30 [000177] In one embodiment, pramlintide is between 1.5 and 75.

In one embodiment, pramlintide are between 2 and 50.

[000179] In one embodiment, pramlintide are between 3 and 30.

In one embodiment, pramlintide are between 4 and 30.

In one embodiment, pramlintide are between 5 and 30.

In one embodiment, pramlintide are between 8 and 30.

In one embodiment, the co-polyamino acid molar ratios / the co-polyamino acid molar ratios / the co-polyamino acid molar ratios / the co-polyamino acid molar ratios / the co-polyamino acid molar ratios / the co-polyamino acid molar ratios / the co-polyamino acid / pramlintide molar ratios are between 10 and 30.

In one embodiment, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between

1.5 and 150.

In one embodiment, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between 1.8 and 100.

In one embodiment, the hydrophobic radical molar ratios Hy! amylin, amylin receptor agonist or amylin analog are between and 70.

In one embodiment, the hydrophobic Hy / amylin radical molar ratios, agonist at the amylin receptor or amylin analog are between

2.5 and 60.

In one embodiment, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between and 60.

In one embodiment, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between

3.5 and 60.

In one embodiment, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between and 60.

In one embodiment, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between and 60.

In one embodiment, the hydrophobic Hy / amylin radical molar ratios, agonist at the amylin receptor or amylin analog are between 7 and 60.

In one embodiment, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between 9 and 60.

an embodiment, [000194] In amyline are between 5 and 60. In one embodiment, amyline are between 10 and 60.

In one embodiment, amylin are between 15 and 60.

In one embodiment, pramlintide are between 1.5 and 60.

In one embodiment, pramlintide are between 2 and 60.

In one embodiment, pramlintide are between 3 and 60.

In one embodiment, pramlintide are between 4 and 60.

In one embodiment, pramlintide are between 5 and 60.

[000202] In one embodiment, pramlintide are between 8 and 60.

[000203] In one embodiment, pramlintide are between 10 and 60.

les les ratios ratios ratios molaires molaires molaires radical radical radical the molar ratios radical les les les les les ratios ratios ratios ratios ratios ratios molar molar molar molar molar molar molar radical radical radical radical radical hydrophobic Hy / hydrophobic Hy / hydrophobic Hy / hydrophobic Hy / hydrophobic Hy / hydrophobic Hy / hydrophobic Hy / hydrophobic Hy / hydrophobic Hy / hydrophobic Hy / [000204] In one embodiment, the mass ratios co-polyamino acid / amylin, agonist at the amylin receptor or amylin analogue are between 1.0 and 70.

In a mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amyli don't or analog of amylin are between 1.2 and 45. In one mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amyli don't or analog of amylin are between 1.3 and 30. In a mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amylin or analog of amylin are between 1.7 and 27. In one mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amylin or analog of amylin are between 2.0 and 27. In a mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amylin or analog of amylin are between 2.3 and 27. In one mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amylin or analog of amylin are between 2.7 and 27. In one mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amylin or analog of amylin are between 3.3 and 27. In one mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amylin or analog of amylin are between 4.7 and 27. In one mode of production, the ratios mass co- polyamino acid / amylin, agonist at receiver of amylin or analog of amylin

are between 6.0 and 27.

In one embodiment, the ratios mass co- polyamino acid / amyline are between 2.0 and 67. In one embodiment, polyamino acid / amylin are between 4.7 and 27. the ratios mass co- In one embodiment, polyamino acid / amylin are between 6.7 and 27. the ratios mass co- In one embodiment, polyamino acid / amylin are between 10 and 27. the ratios mass co-

In one embodiment, the co-polyamino acid / pramlintide mass ratios are between 1.0 and 67.

In one embodiment, the co-polyamino acid / pramlintide mass ratios are between 1.3 and 45.

In one embodiment, the co-polyamino acid / pramlintide mass ratios are between 2.7 and 27.

In one embodiment, the co-polyamino acid / pramlintide mass ratios are between 3.3 and 27.

In one embodiment, the co-polyamino acid / pramlintide mass ratios are between 5.3 and 27.

[000223] In one embodiment, the co-polyamino acid / pramlintide mass ratios are between 6.7 and 27.

In one embodiment, the composition is characterized in that it also comprises insulin.

In one embodiment, the composition is characterized in that the insulin is a mealtime insulin. Prandial insulins are soluble at pH 7.

By mealtime insulin is meant a so-called rapid or "regular" insulin.

The so-called rapid meal insulins are insulins which must meet the needs caused by the ingestion of proteins and carbohydrates during a meal, they must act in less than 30 minutes.

In one embodiment, the so-called “regular” mealtime insulin is human insulin.

In one embodiment, the mealtime insulin is a recombinant human insulin as described in the European Pharmacopoeia and the American Pharmacopoeia.

Human insulin is for example marketed under the brands Humulin® (ELI LILLY) and Novolin® (NOVO NORDISK).

[000231] The so-called fast acting prandial insulins are insulins which are obtained by recombination and whose primary sequence has been modified to reduce their action time.

In one embodiment, the so-called fast acting mealtime insulins are chosen from the group comprising insulin lispro (Humalog®), insulin glulisine (Apidra®) and insulin aspart (NovoLog®) .

[000233] In one embodiment, the meal insulin is insulin lispro.

In one embodiment, the meal insulin is insulin glulisine.

In one embodiment, the meal insulin is insulin aspart.

The units recommended by the pharmacopoeias for insulins are presented in the table below with their correspondences in mg:

Insulin Pharmacopoeia EP 8.0 (2014) US Pharmacopoeia - USP38 (2015) Aspart 1U = 0.0350 mg insulin aspart 1 USP = 0.0350 mg insulin aspart Lispro 1U = 0.0347 mg insulin lispro 1 USP = 0.0347 mq insulin lispro Human 1IU = 0.0347 mg of human insulin 1 USP = 0.0347 mg human insulin

In the case of insulin gulisine, 100U = 3.49 mg of insulin glulisine (according to Annex 1 - Summary of product characteristics relating to ADIPRA®).

However in the following text U is systematically used interchangeably for the quantities and concentrations of all insulins. The respective values corresponding in mg are those given above for values expressed in U, UI or USP.

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is between 240 and 3000 μΜ (40 to 500 U / mL).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is between 600 and 3000 μΜ (100 to 500 U / ml).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is between 600 and 2400 μΜ (100 to 400 U / mL).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is between 600 and 1800 μΜ (100 to 300 U / mL).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is between 600 and 1200 μΜ (100 to 200 U / ml).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is 600 μΜ (100 U / ml).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is 1200 μΜ (200 U / ml).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is 1800 μΜ (300 U / ml).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is 2400 μΜ (400

U / mL).

In one embodiment, it relates to a pharmaceutical formulation characterized in that the insulin concentration is 3000 pM (500 U / ml).

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog receptor molar ratios are between 1.5 and 75.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog molar ratios are between 1.8 and 50.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 2 and 35.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 2.5 and 30.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 3 and 30.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog receptor molar ratios are between 3.5 and 30.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 4 and 30.

In one embodiment comprising prandial insulin, the molar ratios of co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog are between 5 and 30.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 7 and 30.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 9 and 30.

In an embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios are between 5 and 75.

In an embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios are between 10 and 50.

In an embodiment comprising prandial insulin, the co-polyamino acid / amylin molar ratios are between 15 and 30.

In one embodiment comprising prandial insulin, the co-polyamino acid / pramlintide molar ratios are between 1.5 and 75.

[000263] In an embodiment comprising prandial insulin, the co-polyamino acid / pramlintide molar ratios are between 2 and 50.

In one embodiment comprising prandial insulin, the co-polyamino acid / pramlintide molar ratios are between 3 and 30.

[000265] In an embodiment comprising prandial insulin, the co-polyamino acid / pramlintide molar ratios are between 4 and 30.

[000266] In an embodiment comprising prandial insulin, the co-polyamino acid / pramlintide molar ratios are between 5 and 30.

In one embodiment comprising prandial insulin, the co-polyamino acid / pramlintide molar ratios are between 8 and 30.

[000268] In an embodiment comprising prandial insulin, the co-polyamino acid / pramlintide molar ratios are between 10 and 30.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 1.5 and 150.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 1.8 and 100.

In one embodiment comprising prandial insulin, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between 2 and 70.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 2.5 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 3 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin radical ratios, agonist at the amylin receptor or amylin analog are between 3.5 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 4 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical radicals Hy / amylin, agonist at the amylin receptor or amylin analog are between 5 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 7 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios, agonist at the amylin receptor or amylin analog are between 9 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios are between 5 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios are between 10 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / amylin molar ratios are between 15 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / pramlintide molar ratios are between 1.5 and 60.

[000283] In an embodiment comprising prandial insulin, the hydrophobic radical Hy / pramlintide molar ratios are between 2 and 60.

[000284] In an embodiment comprising prandial insulin, the hydrophobic radical Hy / pramlintide molar ratios are between 3 and 60.

[000285] In an embodiment comprising prandial insulin, the hydrophobic radical Hy / pramlintide molar ratios are between 4 and 60.

[000286] In an embodiment comprising prandial insulin, the hydrophobic radical Hy / pramlintide molar ratios are between 5 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / pramlintide molar ratios are between 8 and 60.

In one embodiment comprising prandial insulin, the hydrophobic radical Hy / pramlintide molar ratios are between 10 and 60.

In one embodiment comprising prandial insulin, the mass ratios of co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog are between 1.0 and 70.

In one embodiment comprising prandial insulin, the mass ratios of co-polyamino acid / amylin, agonist at the receptor for aminyl or amylin analog are between 1.2 and 45.

[000291] In an embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist at the amine receptor or amylin analog mass ratios are between 1.3 and 30.

In an embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist at the aminin receptor or amylin analog mass ratios are between 1.7 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist at the aminin receptor or amylin analog mass ratios are between 2.0 and 27.

In one embodiment comprising prandial insulin, the mass ratios of co-polyamino acid / amylin, agonist at the amine receptor or amylin analog are between 2.3 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist, receptor agonist or amylin analog ratios are between 2.7 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist at the aminin receptor or amylin analog mass ratios are between 3.3 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin, agonist at the amine receptor or amylin analog mass ratios are between 4.7 and 27.

In an embodiment comprising prandial insulin, the mass ratios of co-polyamino acid / amylin, agonist at the amine receptor or amylin analog are between 6.0 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin mass ratios are between 3.3 and 67.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin mass ratios are between 6.6 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / amylin mass ratios are between 10 and 27.

In an embodiment comprising prandial insulin, the co-polyamino acid / pramlintide mass ratios are between 1.0 and 67.

In one embodiment comprising prandial insulin, the co-polyamino acid / pramlintide mass ratios are between 1.2 and 45.

In an embodiment comprising prandial insulin, the co-polyamino acid / pramiintide mass ratios are between 1.3 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / pramiintide mass ratios are between 1.7 and 27.

In an embodiment comprising prandial insulin, the co-polyamino acid / pramiintide mass ratios are between 2.0 and 27.

In an embodiment comprising prandial insulin, the co-polyamino acid / pramiintide mass ratios are between 2.3 and 27.

In an embodiment comprising prandial insulin, the co-polyamino acid / pramiintide mass ratios are between 2.7 and 27.

In an embodiment comprising prandial insulin, the co-polyamino acid / pramiintide mass ratios are between 3.3 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / pramiintide mass ratios are between 4.7 and 27.

In one embodiment comprising prandial insulin, the co-polyamino acid / pramiintide mass ratios are between 6.0 and 27.

In addition, it is particularly advantageous to combine amylin, an agonist at the amylin receptor or an amylin analog, in combination or not with a mealtime insulin, with GLP-1, GLP analogs -1, GLP-1 receptor agonists, these are commonly called GLP-1 RA. This allows in particular to potentiate the effect of insulin and is recommended in certain types of diabetes treatment.

In one embodiment, GLP-1, analogs of GLP-1, or GLP-1 RA are said to be "fast". The term "rapid" means GLP-1, analogs of GLP-1, or GLP-1 RA, the apparent elimination half-life after subcutaneous injection in humans is less than 8 hours, in particular less 5 hours, preferably less than 4 hours or even less than 3 hours, such as exenatide and iixisenatide.

In one embodiment, GLP-1, analogs of GLP-1, or

GLP-1 RA are chosen from the group consisting of exenatide or Byetta® (ASTRAZENECA), Iixisenatide or Lyxumia® (SANOFI), their analogs or derivatives and their pharmaceutically acceptable salts.

In one embodiment, GLP-1, analog of GLP-1, or GLP-1 RA is exenatide or Byetta®, its analogs or derivatives and their pharmaceutically acceptable salts.

In one embodiment, GLP-1, analog of GLP-1, or GLP-1 RA is lixisenatide or Lyxumia®, its analogs or derivatives and their pharmaceutically acceptable salts.

In one embodiment, the concentration of exenatide, its analogs or derivatives and their pharmaceutically acceptable salts is in the range of 0.01 to 1.0 mg per 100 U of insulin.

In one embodiment, the concentration of exenatide, its analogs or derivatives and their pharmaceutically acceptable salts is 0.01 to 0.5 mg per 100 U of insulin.

In one embodiment, the concentration of exenatide, its analogs or derivatives and their pharmaceutically acceptable salts is 0.02 to 0.4 mg per 100 U of insulin.

In one embodiment, the concentration of exenatide, its analogs or derivatives and their pharmaceutically acceptable salts is 0.03 to 0.3 mg per 100 U of insulin.

In one embodiment, the concentration of exenatide, its analogs or derivatives and their pharmaceutically acceptable salts is 0.04 to 0.2 mg per 100 U of insulin.

In one embodiment, the concentration of exenatide, its analogs or derivatives and their pharmaceutically acceptable salts is 0.04 to 0.15 mg per 100 U of insulin.

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 per 100 U of insulin.

In one embodiment, the concentration of lixisenatide, its analogs or derivatives and their pharmaceutically acceptable salts is 0.01 to 0.5 mg per 100 U of insulin.

In one embodiment, the concentration of lixisenatide, its analogs or derivatives and their pharmaceutically acceptable salts is 0.02 to 0.4 mg per 100 U of insulin.

In one embodiment, the concentration of lixisenatide, its analogs or derivatives and their pharmaceutically acceptable salts is from 0.03 to 0.3 mg for

100 U of insulin.

In one embodiment, the concentration of lixisenatide, its analogs or derivatives and their pharmaceutically acceptable salts is from 0.04 to 0.2 mg for

100 U of insulin.

In one embodiment, the concentration of lixisenatide, its analogs or derivatives and their pharmaceutically acceptable salts is from 0.04 to 0.15 mg per 100 U of insulin.

In one embodiment, the compositions according to the invention are produced by mixing amylin solutions and commercial solutions of GLP-1, of GLP-1 analog or of agonist of GLP receptors- 1 RA in volume ratios in the range 10/90 to 90/10 in the presence of a co-polyamino acid.

In one embodiment, the compositions according to the invention also comprise zinc salts at a concentration of between 0 and 500 μΜ per 100 U of insuiine.

In one embodiment, the compositions according to the invention further comprise zinc salts at a concentration of between 0 and 400 μΜ per 100 U of insuiine.

In one embodiment, Its compositions according to the invention also comprise zinc salts at a concentration between 0 and 300 μΜ per 100 U of insulin.

In one embodiment, the compositions according to the invention further comprise zinc seis at a concentration of between 0 and 200 μΜ per 100 U of insulin.

In one embodiment, the compositions according to the invention further comprise zinc seis at a concentration of between 0 and 100 μM per 100 U of insulin.

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, ie Tris (trishydroxymethylaminomethane) and sodium citrate.

In one embodiment, the buffer is sodium phosphate.

[000340] In one embodiment, the buffer is ie 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 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 also comprise a surfactant.

In one embodiment, the surfactant is chosen from the group consisting of propylene glycol and 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 glycerin, sodium chloride, mannitol and glycine.

The compositions according to the invention may also comprise all the excipients in accordance with the pharmacopoeias and compatible with the insulins 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.

[000353] The transdermal, oral, nasal, vaginal, ocular, oral, pulmonary administration routes are also envisaged.

The invention also relates to a pump, implantable or transportable, comprising a composition according to the invention.

The invention also relates to the use of a composition according to the invention intended to be placed in a pump, implantable or transportable.

The invention also relates to single-dose formulations at pH between 6.0 and 8.0 comprising amylin, an agonist at the amylin receptor or an amylin analog and a co-polyamino acid according to the invention.

The invention also relates to single-dose formulations at pH between 6.0 and 8.0 comprising amylin, an agonist at the amylin receptor or amylin analog, a co-polyamino acid according to the invention and a GLP-1, an analog of GLP-1 or a GLP-1 RA, as defined above.

The invention also relates to single-dose formulations at pH between 6.6 and 7.8 comprising amylin, an agonist at the amylin receptor or an amine analog and a co-polyamino acid according to the invention.

The invention also relates to single-dose formulations at pH between 6.6 and 7.8 comprising amylin, an agonist at the amylin receptor or an amine analog, a co-polyamino acid according to the invention and a mealtime insulin, as defined above.

The invention also relates to single-dose formulations at pH between 6.6 and 7.6 comprising amylin, an agonist at the amylin receptor or an amine analog and a co-polyamino acid according to the invention.

The invention also relates to single-dose formulations at pH between 6.6 and 7.6 comprising amylin, an agonist at the amylin receptor or an amine analog, a co-polyamino acid according to the invention and a mealtime insulin, as defined above.

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

In one embodiment, the formulations are in the form of an injectable solution.

The preparation of a composition according to the invention has the advantage of being able to be carried out by simple mixing of an aqueous solution of amylin, of an agonist at the amylin receptor or of an analogue of amyîine, and a co-polyamino acid carrying carboxylate charges and 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 between 6 and 8.

The preparation of a composition according to the invention has the advantage of being able to be carried out by simple mixing of an aqueous solution of amylin, of an agonist at the amylin receptor or of an analogue of amyîine, prandial insulin, and a co-polyamino acid carrying carboxylate charges and 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 between 6 and 8.

In one embodiment, the mixture of mealtime insulin and copolyamino acid is concentrated by ultrafiltration.

If necessary, the composition of the mixture is adjusted to excipients such as glycerin, m-cresol, zinc chloride, and polysorbate (Tween®) by adding concentrated solutions of these excipients within the mixture. If necessary, the pH of the preparation is adjusted to pH between 6 and 8.

In one embodiment, the compositions are characterized in that said compositions have a stability measured by ThT greater than that of a reference composition comprising amylin, an agonist at the amylin receptor or the like of amylin but not comprising a co-polyamino acid carrying carboxylate charges and hydrophobic Hy radicals.

In one embodiment, the compositions are characterized in that said compositions have a stability measured by ThT greater than that of a reference composition comprising amylin, an agonist at the amylin receptor or the like of amylin in combination with an insulin but not comprising a co-polyamino acid carrying carboxylate charges and hydrophobic radicals Hy.

In one embodiment, the compositions are characterized in that said compositions have a stability measured by ThT greater than that of a reference composition comprising amylin, an agonist at the amylin receptor or the like amylin in combination with GLP-1, a GLP-1 analog or a GLP-1 receptor agonist, but not comprising a co-polyamino acid carrying carboxylate charges and hydrophobic Hy radicals.

In one embodiment, the compositions are characterized in that said compositions have a stability measured by ThT greater than that of a reference composition comprising amylin, an agonist at the amylin receptor or the like amylin in combination with insulin and GLP-1, a GLP-1 analogue or a GLP-1 receptor agonist, but not comprising a co-polyamino acid carrying carboxylate charges and hydrophobic Hy radicals.

The invention also relates to the use of a co-polyamino acid carrying carboxylate charges and hydrophobic Hy radicals to stabilize a composition comprising amylin, an amylin receptor agonist or an amylin analog.

The invention also relates to the use of a co-polyamino acid carrying carboxylate charges and hydrophobic Hy radicals for stabilizing a composition comprising amylin, an amylin receptor agonist or an amylin analog and a mealtime insulin, and optionally a GLP-1, a GLP-1 analog or a GLP-1 receptor agonist.

The invention relates to a composition stabilization method comprising amylin, an amylin receptor agonist or an amylin analog or a composition stabilization method comprising amylin, a receptor agonist amylin or an amylin analog and a mealtime insulin, and optionally a GLP-1, a GLP-1 analog or a GLP-1 receptor agonist.

The following examples illustrate, without limitation, the invention.

Part A

AA_L_Sÿ_nth.èse of hydrophobic molecules, in which p = 1 [000376] 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 co-polyamino acid AAI ° cT θ ¹⁵ ” ³¹ AA2 H ₂ hT -γ N ° ₀ / ^ CuH ₂₆ Î AA3 AA4 ZL / 'Z A''.Z;^0; T 1: AA5 o <^^ nh ₂ Ai

AA6 ⁰ qÎ ^^ Ci7H35 AA7 - \ / ^u 19 ^m 39 AA8 CH o L z ^NK Ύ ^{15 31} ° '\ I AA9 h _? n ^ X / ^nh _x X / II ^N ο Υ _Υ ^ζΝΗ γ ^{0, ιΗ21}

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 L-proline.

A solution of L-proline (10.6 g, 92.1 mmol) in 1 N aqueous sodium hydroxide (230 mL; 230 mmol) is added dropwise for 90 minutes a solution of palmitoyl chloride ( 23.0 g, 83.7 mmol) in acetone (157 mL).

After 14 h of stirring 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 37% hydrochloric acid solution 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%).

^X H NMR (CDCl _3, 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 (1 H); 4.41 (0.1H); 4.61 (0.9η) 6.60-8.60 (1H).

Molecule A2: product obtained by reaction between the molecule Al and Boc-ethylenediamine.

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, N-diisopropylethylamine (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 0.1 N HCl solution (2.1 L), then a saturated NaCl solution (1 L) are added. The phases are separated then the organic phase is washed successively with a 0.1 N HCl solution / saturated NaCl (2.1 L / l L), a saturated NaCl solution (2 L), a saturated NaHCO ₃ solution ( 2 L), then a saturated NaCl 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%).

^X NMR (CDCl3, 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 [000379] 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 (methanol, dichloromethane) to give a white solid of molecule AAI in the form of the hydrochloride salt.

Yield: 16.3 g (93%).

NMR (CDCI ₃ , 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-methylhexadecan-1-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 observing an exotherm and a slight clouding of the medium, the rest of the 1-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 environment reacts! is then heated at 70 ° C for 2 h.

In a three-necked flask under argon, to a CuCI solution (482 mg, 4.86 mmol) dissolved in the NMP (62 mL) at 0 ° C. is added dropwise a solution of 12-bromo-l- dodecanol (43 g, 162.1 mmol) in THF (50 mL). To this solution is then added dropwise the solution of the hot organomagnesium 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 HCl IN to pH 1 and the medium is extracted with ethyl acetate. After washing the organic phase with a saturated NaCl solution and drying over Na ₂ SO ₄ , 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 _3, 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 room temperature, the medium is acidified to pH 1 by progressive addition of 5N HCl. Na ₂ SO ₃ (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 Na ₂ SO ₄ , 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)

^X H NMR (CDCl _3, ppm): 0.87 (6H); 1.14 (2H); 1.22-1.38 (20H); 1.51 (1H); 1.63 (2H); 2.35 (2H).

Molecule A5: product obtained by reaction between the 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 over Na ₂ SO ₄ , 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%)

^X NMR (CDCl3, 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).

A6 molecule: product obtained by reaction between the A5 molecule and Boc-ethylenediamine.

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- (1 H -benzotriazol-1-yl) -1,3,3-tetramethyluronium tetrafluoroborate (TBTLJ) 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 Na ₂ SO ₄ , filtered, concentrated in vacuo and the residue is purified by flash chromatography (ethyl acetate, methanol).

Yield: 6.9 g (54%)

^X H NMR (CDCl _3, 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).

AA2 molecule [000385] 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 hours 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%)

NMR ^X H (D ₂ O, 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

Molecule A7: product obtained by the reaction between the molecule Al and Boc-tri (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%).

NMR (CDCI3, 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 molecule

AAI applied to the 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%).

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-amino-4,7,10-trioxa-13-tridecane amine.

By a process similar to that used for the 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%).

NMR (CDCI ₃ , 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); 4.69-5.04 (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 makes it possible to obtain the molecule AA4 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%).

^T NMR (CDCl3, 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: AA5 molecule [000390] 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-Llysine (3.2 g, 12.4 mmol), a colorless oil is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 4.9 g (73%).

NMR (CDCI ₃ , 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 (O, 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%).

^X NMR (CDCl3, 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 AAI molecule applied to the A10 molecule (388 mg, 0.67 mmol), a white solid of AA5 molecule in the form of the hydrochloride salt is obtained after purification. by trituration in diisopropyl ether.

Yield: 292 mg (85%).

^X H 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 (0.7H); 7.43 (0.3H); 7.73-8.04 (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 L-proline.

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%)

^X H NMR (CDCl _3, 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).

Molecule A12: product obtained by reaction between the molecule Ail and Boc-tri (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 Boc-tri (ethylene glycol) diamine (26.4 g, 106 , 3 mmol) in THF using DIPEA instead of TEA, a white solid is obtained after purification by flash chromatography (ethyl acetate, methanol).

Yield: 43.3 g (80%)

N ^N NMR (CDCI ₃ , 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%)

^X 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

Molecule A13j. 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) using DIPEA instead of TEA, a white solid is obtained after purification by chromatographic column on silica gel (cyclohexane, ethyl acetate, acetic acid).

Yield: 12.9 g (63%)

^X H NMR (CDCl _3, 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).

A14 molecule: product obtained by the reaction between the A13 molecule and Boc-1amino-4,7,10-trioxa-13-tridecane.

By a process similar to that used for the preparation of the molecule A12 applied to the molecule A13 (10.96 g, 26.75 mmol) and to Boc-l-amino-4,7,10-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 NMR (CDCl3, ppm): 0.88 (3H); 1.24 (32H); 1.43 (9H); 1.51 (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 concentration in vacuo 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%)

^X 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 the molecule

Al applied to L-leucine (15.0 g, 114.4 mmol) and to palmitoyl chloride (34.5 g, 125 mmol), a white solid is obtained by trituration in diisopropyl ether.

Yield: 13.0 g (31%)

^X NMR (CDCl3, ppm): 0.88 (3H); 0.96 (6H); 1.15-1.35 (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 [000401] 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 L-proline (3.23 g, 19.5 mmol), a slightly yellow oil is obtained after purification by flash chromatography (methanol, dichloromethane).

Yield: 5.8 g (74%)

^X H NMR (CDCl _3, ppm): 0.83 to 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): 481.5; (calculated ([M + H] ⁺ ): 481.4).

A17 molecule: product obtained by the saponification of the methyl ester of the A16 molecule.

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 Na ₂ SO ₄ , filtered and then concentrated under reduced pressure to give a colorless oil.

Yield: 4.5 g (80%)

NMR (CDCI3, 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 (0.41-1); 4.86-4.91 (0.6H);

6.05 (0.4H); 6.11 (0.6H).

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

Molecule A18.:. product obtained by the reaction between Boc-ethylenediamine and the molecule A17.

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%)

NMR (CDCI3, 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%)

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.j product obtained by the reaction between lauric acid and L-phenylalanine.

By a process similar to that used for the preparation of the molecule A5 applied to lauric acid (8.10 g, 40.45 mmol) and to L-phenylalanine (7 g, 42.38 mmol), a white solid is obtained.

Yield: 12.7 g (98%)

^X H NMR (DMSO-d _6, 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; (calculated ([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 mmol), a colorless oil is obtained after purification by a chromatographic column on silica gel (cyclohexane, ethyl acetate).

Yield: 5.75 g (44%)

N ^N NMR (CDCI ₃ , 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); 7,157.30 (5H).

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

A21 molecule: product obtained by saponification of the A20 molecule.

Lithium hydroxide (LiOH) is added to a solution of molecule A20 (5.75 g, 12.54 mmol) in a THF / methanol / water mixture (40/40/40 ml) at 0 ° C ) (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 Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a colorless oil.

Yield: 5.7 g (quantitative)

NMR (CDCI ₃ , 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 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%)

^X NMR (CDCl3, 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 the A22 molecule (5.66 g, 9.65 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 foam of molecule AA9 in the form of the hydrochloride salt.

Yield: 4.9 g (97%)

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 (0.4H); 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

AB5

AB6

Ri == H or pyroglutamate

.0

Ri = H or pyroglutamate

AB7

AB8

Hy i = 0.03, DP (m + n) = 21 Hy =

R1 = CH ₃ -CO-, H or pyroglutamate

We have'

i = 0.03, DP (m + n) - 24

Hy =

Ri = H or pyroglutamate

H ^ O i = 0.056, DP (m + n) = 22

Hy =

_____ Ri = H or pyroglutamate

Table lb

Defined co-polyamino acids of formula VII or Vllb:

example number co-polyamino acids carrying carboxylate charges and hydrophobic radicals AB14 0¾ We have ; ° X \ Z VS ) \ ^ C ₁₅ H ₃₁ H / ^R N '' NT <.N H H L ^J m 0 i = 0.04, DP (m) = = 25 R, = H or pyroglutamate

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 AB1-1: poly-L-glutamic acid of number-average molar mass (Mn) relative 3861 g / mol resulting from the polymerization of y-benzyl-L-glutamate N-carboxyanhydride initiated by hexylamine [000411] In a flask previously dried in an oven is placed under vacuum y-benzyl-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-L-glutamic 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 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 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 [000416] 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. With 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 about 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 ² H NMR in D ₂ O by comparing the integration of the signals coming from the grafted hydrophobic with that of the signals coming from the main chain.

According to the NMR ^: 1 = 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 R _x and R ₂ , of the aspartate and / or glutamate residues (including an amide bond), of the hydrophobic radical, of DS and of 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 molecule AAI and having a number-average molar mass (Mn) of 3700 g / mol [000417] By a process similar to that used for the preparation of the co-polyamino acid ABl applied to the hydrochloride salt of the molecule AAI (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 similar process 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 by NMR ^X H): 35

According to NMR = 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 its molecule AAI and having a number-average molar mass (Mn) of 4900 g / mol [000418] By a process similar to that used for the preparation of the co-polyamino acid AB1 applied to the hydrochloride salt of the molecule AAI (3.30 g, 7.6 mmol) and to a poly-L-glutamic acid of average mass in number (Mn) relative 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 with the molecule AAI is obtained.

Dry extract: 23.4 mg / g

DP (estimated according to ⁴ H 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 [000419] By a process similar to that used for the preparation of the co-polyamino acid AB1 applied to the hydrochloride salt of the molecule AA2 (1.09 g, 2.4 mmol) and to a poly-L- acid. glutamic 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 NMR: 35

According to the NMR ^X H: i = 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 2,600 g / mol [000420] By a process similar to that used for the preparation of the co-polyamino 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 from co-poiyamino acid AB1-1, a sodium poly-L-glutamate modified by the molecule AA6 is obtained.

Dry extract: 20.9 mg / g

DP (estimated by NMR ^X H): 23

According to the NMR ^X H: i = 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 [000421] A poly-L-giutamic acid of average mass Mn = 3500 g / mol and 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 by NMR ^X H): 20

According to the NMR ^X 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 acety group and modified by the molecule AA7 and having a number-average molar mass (Mn) of 3300 g / mol [000422] Co-polyamLnoaçicLe__AB7-.l_ poly-L-glutamic acid of average molar mass in number (Mn) relative 3600 g / mol and of DP 21 resulting from the polymerization of γ-benzyl-L-glutamate N-carboxyanhydride initiated by hexylamine and capped at one of its ends by an acety group [000423] In a flask previously dried in an oven is placed under vacuum the y-benzyl-L-glutamate N-carboxyanhydride (Glu (OBn) -NCA, 100, 0 g, 380 mm 3) for 30 minutes 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 h of stirring, 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 min, 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 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 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-L-glutamate acid modified with the molecule AA7 is obtained.

Dry extract: 24.3 mg / g

DP (estimated by NMR ^X H): 21

According to the NMR ^X H: i = 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 [000428] Çojjolyaminoacid AB8-1. 1 poly-L-glutamic acid of number-average molar mass (Mn) 3800 g / mol and of degree of polymerization 24 resulting from the polymerization of γ-methyl-L-glutamate N-carboxyanhydride initiated by ammonia [000429] 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 an ammonia solution 0 , 5N in 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 co-polyamino 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 from NMR * H): 24

According to the NMR ^X H: i = 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 [000431] By a process similar to that used for the preparation of co -polyaminoacidABl 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 the co-polyamino acid AB1-1, a sodium poly-L-glutamate modified by the AA3 molecule is obtained.

Dry extract: 14.7 mg / g

DP (estimated from NMR ^): 30

According to the NMR ^: 1 = 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 AB1O: Co-polyamino acid ABlO - sodium poly-L-glutamate modified by the molecule AA4 and having a number-average molar mass (Mn) of 2600 g / mol [000432] By a process similar to that used for the preparation of co -polyamino 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 the co-polyamino acid AB1-1, a sodium poly-L-glutamate modified by the AA4 molecule is obtained.

Dry extract: 18.3 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 AB10 co-polyamino acid is 4870 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 2600 g / mol.

ABU example; 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 [000433] By a process similar to that used for the preparation of the co-polyamino 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 molecule AA5 is obtained.

Dry extract: 20.2 mg / g

DP (estimated by NMR ^X H): 23

According to the NMR ^X H: 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 [000434] By a process similar to that used for the preparation of the co-polyamino 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 co-polyamino acid AB1-1, a modified sodium poly-L-glutamate by the molecule AA8 is obtained.

Dry extract: 19.5 mg / g

DP (estimated by NMR ^X H): 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 [000435] By a process similar to that used for the preparation of the co-polyamino 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 co-polyamino acid AB1-1 using isoamylamine as initiator in place of hexylamine, a sodium poly-L-glutamate modified by the molecule AA9 is obtained. Dry extract: 22.3 mg / g

DP (estimated from NMR ^Χ Η): 35

According to the NMR ^: 1 = 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 [000436] By a process similar to that used for the preparation of the co-polyamino 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 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 ¹ H 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 [000437] 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 the ion exchange resin Amberlite 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, y-benzyl-L-glutamate / V-carboxyanhydride (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) 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 (v / v) mixture 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 ¹ H 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 modified at one of its ends by the molecule AA6 and having a number-average molar mass (Mn) 4100 g / mol [000439] By a process similar to that used for the preparation of the co-polyamino 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 poly-L-glutamate modified at one of its ends by the molecule AA6 is obtained.

Dry extract: 45.5 mg / g

DP (estimated by NMR ^X H) = 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 [000440] By a process similar to that used for the preparation of the co-polyamino acid 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 poly-L-glutamate modified at one of its ends by the molecule AA6 is obtained.

Dry extract: 28.5 mg / g

DP (estimated by ¹ H 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 / mol [000441] By a process similar to that used for the preparation of the co-polyamino 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 poly-L-glutamate modified at one of its ends by the molecule AA7 is obtained.

Dry extract: 25.2 mg / g

DP (estimated by NMR ^T H) - 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 [000442] 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 NMR * Η) - 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 [000443] By a process similar to that used for the preparation of the co-polyamino acid AB14 applied to the hydrochloride salt of the molecule AA6 (1.64 g, 2.99 mmol) and to γ-benzyl-L-glutamate N-carboxyanhydride (49.3 g, 187 mmol) , a sodium poly-L-glutamate modified at one of its ends by the molecule AA6 is obtained.

Dry extract: 23.4 mg / g

DP (estimated by ¹ H NMR) = 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 = 10,500 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 [000444] In a suitable container are successively introduced 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, y-benzyl-L-glutamate / V-carboxyanhydride (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 ^X H) = 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 The 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 co-polyamino acid BAI (N x ^C 9 ^H 19 YY D 0 O ^ NH \ Q , O AT \ Η ₂ Ν ^^ ^Ν γΥ, Λ ₀ ^C ü ^H 19 O BA2 / N. z ^C H ^H 23 Y Ίί _Y 0 O '^' NH k _y O Y h ₂ n ^χχ ^ ^ΝΗ \ γ \ ι ^z Tri NH ^x -q ^C 11 ^H 23 0 BA3 y3 <^ / ^C 13 ^H 27 ⁰ O ^^ NH ..... s çv U ri X ^HîN j <xtrih _o ^{C 1:27} PM 0

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

Part BA: synthesis of hydrophobic molecules in which p = 2

BAI example: BAI molecule

Molecule B1: product obtained by the reaction between decanoic acid and L-proline. Dicyclohexyl carbodiimide (DCC) (16.29 g, 78.96) are successively added to a solution of decanoic acid (14.28 g, 82.91 mmol) in THF (520 mL) at 0 ° C 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-proin (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 Na ₂ SO ₄ , filtered, concentrated in vacuo and the residue is purified by chromatography on silica gel (cyclohexane, ethyl acetate).

Yield: 14.6 g (69%)

^X H NMR (CDCl _3, 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).

Molecule B2: product obtained by the reaction between the molecule B1 and L-lysine. By a process similar to that used for the preparation of the B1 molecule applied to the B1 molecule (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%)

^X NMR (CDCl3, 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 molecule B2 and Boc-ethylenediamine.

To a solution of molecule B2 (16.4 g, 25.27 mmol) in THF (170 mL) are added D1PEA (8.80 mL) and 2- (1H-benzotriazol-1-yl ) -1,3,3-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 NaHCO ₃ (250 ml), an aqueous solution of 1 N HCl (250 ml), a saturated aqueous solution of NaCl (250 ml) and is dried over Na ₂ SO ₄ . 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 (54%)

^X H NMR (CDCl _3, 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 [000451] 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%)

^X NMR (DMSO-d₆, 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: BAS molecule

Molecule B4: product obtained by the reaction between lauric acid and L-proline.

By a process similar to that used for the preparation of the molecule B1 applied to lauric 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%)

^X H NMR (CDCl _3, 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_i 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 B1 molecule applied to the B4 molecule (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%)

¹ H NMR (CDCl3, 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)

^X NMR (CDCl3, 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 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 methanol 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%)

^X H NMR (DMSO-ds, 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

Molecule 87: 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 L-proline (10 g, 86.86 mmol), a yellowish oil is obtained.

Yield: 20 g (78%)

NMR (CDCI ₃ , 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 the molecule B7 and L-lysine [000457] By a process similar to that used for the preparation of the molecule B1 applied to the molecule B7 (20.02 g, 61.5 mmol) and with L-isine (4.72 g, 32.29 mmol), a white solid is obtained.

Yield: 12.3 g (53%)

^Τ N 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).

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

By a process similar to that used for the preparation of the molecule B3 applied to the molecule B8 (12 g, 15.77 mmol) and to 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-d ₅ , 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%)

NMR (DMSO-d ₆ , 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-amino-4,7,10-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-trioxa-13 -tridecane (15.05 g, 46.96 mmol), a thick colorless oil is obtained.

Yield: 25.3 g (61%)

^X H NMR (DMSO-d _6, 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 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%)

NMR (DMSO-d ₆ , 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).

BB_ ± _Synthèse_cLe_s_ copolya minoacides.

Çpolyaminoacids staistaiques delormulgs VII or Vlla

n ° hydrophobic co-polyamino acids carriers of carboxylate charges and of radicals BB1 J We have 0¾ Y 2 0 H Ri _x .NOT \ .x \ NOT N x H H not m 0 Hy- ^ k i = 0.05, DP (m + n) = 23 ,NOT Y ^11:23 a.m. 0 0 ^ ' NH 1 , Ν- ^ ζζ ★ r Λ Nh-XÆA 'NH x ^ 11 ^ 23 Ο Hy = 0 Ri - H or pyroglutamate

BB4

/ ç 0 ^ λ ,NOT OR Y 0 Ί _H 11 ^m 23 ^x nh \ ^ ⁿ 1 J NH ^ ^C 11 ^H 23 0 Hy = Rl = CH3-CO-, H or pyroglutamate BB7 We have 0¾ VS ) H Ri _x Y H "NOT H m not 0 Hy ^ X i = 0.042, DP f m n) = 22 11 ^ 23 1 1 cX NH ^X 'NH ^^ ^N t A. w θ11 ^Η 23 0 Hy = & Rj = H or pyroglutamate

BB8

H <

R i II or pyrogl utamate

BB9

O

i = 0.05, DP (m + n) = 26

VY ⁰ O ^ ^X NH ^1:27 p.m. ^V nh ^ / ^nh _v A Çk ^C 13 ^H 27 NH \ q He II 0 H or pyroglutamate BB10 0¾ We have / VS ^R N H • H not 0 ^J n Ί i = 0.029, DP (m + n) = 22 V 0 NH ^1:27 p.m. ' ^Χ ΝΗ''Χ / ^ΝΗ _γ Λ (( 'NH c ₁ p.m. _Z7 He II & H or pyroglutamate

BB11

Rl = CH3-CO-, H or pyroglutamate

BB12

Hy ^ O

L = 0.03, DP (m + n) = 23_________

Y : Λ Y 0 Ί ^1:27 p.m. (( ; Λ. At NH K C13H27 V 0 Hy = Ô Rl = CH3-CO-, H or pyroglutamate BB13 ,We have 0¾ VS ) H ^R 1X H ,NOT H L J ^J m not VS Hy ^ k i = 0.08, DP = = 25 r ~~ vs U ^S NH • x v Γ 4 0 * Y s' \ Sîh- ^ ⁿⁱ 1 Y CgH ₁₉ he N H Λ D Hy = 0 Ri = H or pyroglutamate

Table the

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

Çojsolya m i noacjdeJBBUl. 1 poly-L-glutamic acid of average molar mass in number (Mn) relative 3860 g / mol resulting from the polymerization of y-benzyl-L-glutamate N-carboxyanhydride initiated by hexiamiamine [000462] In a flask previously dried at the oven is placed under vacuum of y-benzyl-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 hexiamiamine (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 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 poiy-L-glutamic acid with an average mass by number (Mn) 3860 g / mol relative to a polyoxyethylene (PEG) standard.

Co-polyamino acid BB1 [000467] The 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. With 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, 8.1 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 β 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 slow addition of a 1N aqueous NaOH solution. The volume is adjusted to 700 ml by addition of 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, the solution is filtered through a 0.2 μm filter and stored at 2-8 ° C.

Dry extract: 19.7 mg / g

DP (estimated from NMR: 23

According to the NMR ^: 1 = 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 Its molecule BA2 and having a number-average molar mase (Mn) of 4900 g / mol [000468] 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. With the solution of co-polyamino acid in DMF, NMM (3.7 g, 36.7 mmol), the solution of molecule BA2 then 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 the medium is brought up to ambient 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.

Dry extract: 16.3 mg / g

DP (estimated by NMR ^X H): 21

According to the NMR ^X H: 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 6400 g / mol

Co-polyamino acid BB3-1: poly-L-glutamic acid of number-average molar mass (Mn) 17,500 g / mol resulting from the polymerization of γ-methyl-L-glutamate N-carboxyanhydride initiated by L-leucinamide [000469] A poly-L-glutamic acid of average mass in number (Mn) 17500 g / mol relative to a standard polymethyl methacrylate (PMMA) is obtained by polymerization of γ-methyl N-carboxyanhydride of glutamic acid using L-leucinamide 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.

By a process similar to that used for the preparation of the co-polyamino acid 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 the molecule BA2 is obtained.

Dry extract: 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.

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 [000471] By a process similar to that used for the preparation of the co-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 number-average molar mass Mn = 10,800 g / mol (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 got.

Dry extract: 28.2 mg / g

DP (estimated by NMR ^X H): 55

According to the NMR ^X H: i = 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 Co-polyamino 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 an acetyl group [000472] In a balloon beforehand dried in an oven is placed under vacuum of y-benzyl-L-glutamate N-carboxyanhydride (100.0 g, 380 mmol) for 30 minutes 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, 175 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.

100 [000473] A solution of co-poiyamino acid cappé (72 g) in trifluoroacetic acid (TFA, 335 mL) at 4 ° C is added dropwise a solution of hydrobromic acid (HBr) at 33% in l 'acetic acid (235 mL, 1.34 mol). The mixture is stirred at room temperature for 3 h 30 min, then added dropwise to 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 addition of 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, ie 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) 3700 g / mol compared to a polyoxyethylene (PEG) standard.

Co-polyamino acid BB5 By a process similar to that used for the preparation of co-polyamino acid BB2 applied to the hydrochloride of the molecule BA2 (6.92 g, 8.8 mmoi) and to the co-poiyamino 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 NMR ’Ή): 23

According to ¹ H NMR: i = 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.

ιοί

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 [000475] By a process similar to that used for the preparation of co-polyamino 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 co-polyamino acid BB5-1 using ammonia instead of hexylamine, a sodium poly-L-glutamate capped at one of its ends by an acetyl group and modified by the BA2 molecule is obtained.

Dry extract: 27.5 mg / g

DP (estimated by NMR ^X H): 24

According to ¹ H NMR: i = 0.04

The calculated average molar mass of the co-polyamino acid BB5 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 [000476] By a process similar to that used for the preparation of the co-polyamino 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 / me (30.0 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: 28.3 mg / g

DP (estimated from NMR ^): 22

According to NMR = 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.

Example BB8: co-polyamino acid BB8 - sodium poly-L-glutamate modified by the molecule BA2 and having a number-average molar mass (Mn) of 5200 g / mol [000477] By a process similar to that used for the preparation of the co-polyamino acid BB2 applied to the hydrochloride salt of the molecule BA2 (0.85 g, 1.1 mmol) and to a poly-L-glutamic acid with an average molar mass in number Mn = 4100 g / mol (5.0 g) obtained by a process similar to that used for the preparation

102 of co-polyamino acid BB1-1, a sodium poly-L-glutamate modified by the molecule

BA2 is obtained.

Dry extract: 28.6 mg / g

PD (estimated from NMR ^Χ Η): 21

From NMR = 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.

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 [000478] By a process similar to that used for the preparation of the co-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 average molar mass in number Mn = 4100 g / mol (10.0 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 BA3 is obtained.

Dry extract: 28.6 mg / g

DP (estimated from NMR: 26

According to the 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 BB1O - sodium poly-L-glutamate modified by the molecule BA3 and having a number-average molar mass (Mn) of 4200 g / mol [000479] By a process similar to that used for the preparation of the co-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 co-polyamino 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 the NMR: i = 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.

103

Example BB11: co-polyamino acid BB11 - sodium poly-L-glutamate capped at one of its ends by an acety group and modified by the molecule BA4 and having a number-average molar mass (Mn) of 3900 g / mol [000480] By a process similar to that used for the preparation of co-polyamino acid 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 co-polyamino acid BB5-1, a sodium poly-L-glutamate capped at one of its ends by an acety group and modified by the molecule BA4 is obtained.

Dry extract: 28.1 mg / g

DP (estimated by NMR ^X H): 22

According to the NMR ^X H: 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 acety group and modified by the molecule BA3 and having a number-average molar mass (Mn) of 3900 g / mol [000481] By a process similar to that used for the preparation of the co-polyamino acid BB2 applied to the se! hydrochloride 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 ia preparation of co-polyamino acid BB5-1, a sodium poly-L-glutamate capped at one of its ends by an acety group and modified by the molecule BA3 is obtained.

Dry extract: 25.7 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 [000482] 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 poiy-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

104 of co-polyamino acid BB1-1, a sodium poly-L-glutamate modified by the molecule

BAI is obtained.

Dry extract: 25.6 mg / g

DP (estimated from NMR ^): 25

According to the NMR ^: 1 = 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.

Example BB14: co-polyamino acid BB14 - sodium poiy-L-glutamate modified at one of its ends by the molecule BA2 and having a number-average molar mass (Mn) of 4020 g / mol [000483] 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 flask previously dried in an oven, y-benzyl-L-glutamate / V-carboxyanhydride (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 the 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 solution of hydrobromic acid (HBr) at 33% 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 then is 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

105 that the conductivity of the permeate is less than 50 pS / cm. The copolyamino acid solution is then concentrated to around 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 ¹ H 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.

Example BB15: co-polyamino acid BB15 - sodium poly-L-glutamate modified at one of its ends by the molecule BA3 and having a number-average molar mass (Mn) of 3389 g / mol [000485] By a process similar to that used for the preparation of the co-polyamino 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 poly-L-glutamate modified at one of its ends by the molecule BA3 is obtained.

Dry extract: 30.4 mg / g

DP (estimated by NMR ^X H) = 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 - 3389 g / mol.

Example BB16: 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 [000486] By a process similar to that used for the preparation of the co-polyamino 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 the molecule BA4 is obtained.

Dry extract: 32.3 mg / g

DP (estimated by NMR ⁴ Η) = 23 therefore i - 0.043

The calculated average molar mass of the co-polyamino acid BB16 is 4399 g / mol. Aqueous HPLC-SEC (PEG calibrator): Mn = 3300 g / mol.

106

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 10700 g / mol [000487] By a process similar to that used for the preparation of co -polyamino 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 sodium poly-L-glutamate modified at one of its ends by the BA3 molecule is obtained.

Dry extract: 24.5 mg / g

DP (estimated by ¹ H NMR) = 65 therefore 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.

Example BB18: 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 of 6600 g / mol [000488] By a process similar to that used for the preparation of the co-polyamino acid BB14 applied to the hydrochloride salt of the molecule BA3 (2.51 g, 3 mmol) and to 31.6 g (120 mmol) of γ-benzyl-L-glutamate N-carboxyanhydride, a poly-L- sodium glutamate modified at one of its ends by the molecule BA3 is obtained.

Dry extract: 27.3 mg / g

DP (estimated by NMR ’Ή) = 40 so 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.

107

B. COMPOSITIONS

Example BV1: Preparation of a 0.6 mg / ml human amylin solution containing m-cresol (29 mM), glycerin (174 mM) at pH 7.4.

A concentrated human amylin solution at 3 mg / ml is prepared by dissolving human amylin in the form of a powder purchased from AmbioPharm. This solution is added to a concentrated solution of excipients (m-cresol, glycerin) so as to obtain the intended final composition. The final pH is adjusted to 7.4 by adding NaOH / HCl.

Example BV2: Preparation of a 0.6 mg / mL human amylin solution containing co-polyamino acid BB15, m-cresol (29 mM) and glycerine (174 mM) at pH 7.4.

[000490] A concentrated solution of co-polyamino acid BB15 and of excipients is prepared by adding concentrated solutions of excipients (m-cresol, glycerin) to a concentrated solution of co-polyamino acid BB15.

The concentrated human amylin solution at 3 mg / MI BV1 is added to this concentrated solution of co-polyamino acid BB15 and of excipients so as to obtain the final compositions BV5 to BV11 (Table 1). The final pH is adjusted to 7.4 by adding NaOH / HCl.

Solution BB15 / human amylin ratio Co-polyamino acid BB15 concentration Visual aspect of the solution mol / mol mg / mL mM BV1 • ·· • Crystal clear BV5 5 3 0.73 Crystal clear BV6 6 3.6 0.88 Crystal clear BV7 7 ... 4.2 1.03 Crystal clear BV8 8 4.8 1.17 Crystal clear BV9 9 ........................ 5.4 ................ 1.32 Crystal clear BV10 10 6 1.47 Crystal clear BV11 17 10.5 2.57 Crystal clear

Table 1: Compositions and visual appearance of solutions of human amylin at pH 7.4 at different concentrations of co-polyamino acid BB15.

Example BY1: Preparation of a 0.9 mg / ml pramlintide solution containing m-cresol (29 mM) and glycerin (174 mM) at pH 7.4.

A concentrated pramlintide solution at 5 mg / mL is prepared by dissolving pramlintide in the form of a powder purchased from Hybio. This solution

108 is added to a concentrated solution of excipients (m-cresol, glycerin) so as to obtain the intended final composition. The final pH is adjusted to 7.4 by adding NaOH / HCl.

Example BW1: Preparation of a 0.4 mg / mL pramlintide solution containing phenol (30 mM), glycerin (174 mM) and glycylglycine (8 mM) at pH 7.4.

By a process similar to that used in Example BY1, a 0.4 mg / ml pramlintide solution containing phenol (30 mM), ia glycerin (174 mM) and ia giycyigiycin (8 mM) at pH 7.4 is obtained.

Example BYO: Preparation of a 0.9 mg / mL pramlintide solution containing co-polyamino acid BB15, m-cresol (29 mM) and glycerin (174 mM) at pH 7.4.

A concentrated solution of co-polyamino acid BB15 and of excipients is prepared by adding concentrated solutions of excipients (m-cresol, glycerin) to a concentrated solution of co-polyamino acid BB15.

A concentrated pramlintide solution at 5 mg / ml is added to this concentrated solution of co-polyamino acid BB15 and of excipients so as to obtain the final compositions BY2 to BY8 (Table 3). The final pH is adjusted to 7.4 by adding NaOH / HCl.

Solution Ratio BB15 / pramlintide Co-polyamino acid BB15 concentration Visual aspect of the solution mol / mol mg / mL mM BY1 - - - Crystal clear BY2 2 1.8 0.44 Crystal clear BY3 3 2.7 0.66 Crystal clear BY4 4 3.6 0.88 Crystal clear BY5 5 4.5 1.10 Crystal clear BY6 6 5.4 1.32 Crystal clear BY7 10 9 2.20 Crystal clear

Table 3: Compositions and visual appearance of pramlintide solutions at pH 7.4 at different concentrations of co-polyamino acid BB15.

Example BWO: Preparation of a pramlintide solution at 0.4 mg / ml containing co-polyamino acid BB15, phenol (30 mM), glycerin (174 mM) and glycylglycine (8 mM) at pH 7, 4.

By a protocol similar to that used in the example BYO, from a pramlintide solution at 0.4 mg / mL BW1, pramlintide solutions at 0.4 mg / mL containing co-polyamino acid BB15 , phenol (30 mM), glycerin (174 mM) and glycylglycine (8 mM) at pH 7.4 BW2 and BW3 are obtained.

109

Solution Co-polyamino acid BB15 concentration BB15 / pramlintlde ratio Visual aspect of the solution mg / mL mM mol / mol BW2 2.4 0.59 6 Crystal clear BW3 4 0.98 10 Crystal clear

Table 4: Compositions and visual appearance of solutions of pramlintide at 0.4 mg / ml at pH 7.4 at different concentrations of co-polyamino acid BB15.

Example BPO: Preparation of a pramlintide solution at 0.9 mg / ml containing various co-polyamino acids of the invention, m-cresol (29 mM) and glycerin (174 mM) at pH 7.4.

By a protocol similar to that described in Example ΒΥ0, solutions of pramlintide at 0.9 mg / mL containing different co-polyamino acids of the invention, 10 m-cresol (29 mM) and glycerin (174 mM) at pH 7.4 BP2 to BP12 are obtained.

Solut ion Co-polya mino acid Concentration in co-polyamino acid Ratio co- poiyamino acid / pr amlintide Visual aspect of the solution mg / m L mM mol / mol BP2 BB15 5 10 1.22 2.45 5.4 10.8 Crystal clear Crystal clear BP3 BB14 5 10 0.98 1.96 4.3 8.6 Crystal clear Crystal clear BP4 AB17 5 10 1.11 1.22 4.9 9.8 Crystal clear Crystal clear BP5 AB15 5 10 0.99 1.99 4.4 8.8 Crystal clear Crystal clear BP6 AB14 5 10 1.48 2.96 6.5 13 Crystal clear Crystal clear BP10 BB18 5 0.72 3.2 Crystal clear 10 1.44 6.3 Crystal clear BP11 BB9 5 10 1 2.01 4.4 8.8 Crystal clear Crystal clear BP12 BB2 5 10 1.27 2.54 5.6 11.2 Crystal clear Crystal clear

Table 8: Compositions and visual appearance of solutions of pramlintide at 0.9 mg / ml at pH 7.4 in the presence of different co-polyamino acids.

110

Example BX1: Preparation of a 0.6 mg / mL human amylin solution and 100 IU / mL human insulin containing m-cresol (29 mM), glycerin (174 mM) and zinc (229 pM) at pH 7.4.

The concentrated human amylin solution at 3 mg / mL BV1 is added to a concentrated solution of excipients (m-cresol, glycerin). A 500 IU / mL human insulin solution is prepared by dissolving human insulin in the form of a powder purchased from Amphastar. This solution is added to the concentrated solution of human amylin and of excipients so as to obtain the intended final composition. The final pH is adjusted to 7.4 by adding NaOH / HCl.

Example BX2: Preparation of a 0.6 mg / mL human amylin solution and 100 IU / mL human insulin containing co-polyamino acid BB15, m-cresol (29 mM), glycerin (174 mM) and zinc chloride (229 μΜ) at pH 7.4.

[000499] A concentrated solution of co-polyamino acid BB15 and of excipients is prepared by adding concentrated solutions of excipients (m-cresol, glycerin, zinc chloride) to a concentrated solution of co-polyamino acid BB15.

A concentrated human amylin solution at 3 mg / ml and then a human insulin solution at 500 IU / ml are added to the concentrated solution of co-polyamino acid BB15 and of excipients so as to obtain the final composition targeted (table 2). The final pH is adjusted to 7.4 by adding NaOH / HCl.

The BX1, BX6, BX10 and BX11 solutions are prepared according to the above protocol.

Solution BB15 / human amylin ratio Co-polyamino acid BB15 concentration Visual aspect of the solution mol / mol mg / MI mM BX1 * ; <·; Turbid BX6 6 3.6 0.88 Crystal clear BX10 10 6 1.47 Crystal clear BX11 17 10.5 2.57 Crystal clear

Table 2: Compositions and visual appearance of solutions of human amylin and human insulin at pH 7.4 at different concentrations of co-polyamino acid BB15.

In the presence of co-polyamino acid BB15, a clear solution of human amylin (0.6 mg / ml) and human insulin (100 IU / ml) is obtained at pH 7.4.

m

Example BN1: Preparation of a 0.4 mg / mL pramlintide solution and 100 IU / mL human insulin containing phenol (30 mM), glycerin (174 mM), glycylglycine (8 mM) and zinc chloride (229 pM) at pH 7.4.

A concentrated pramlintide solution at 5 mg / ml is added to a concentrated solution of excipients (m-cresol, glycerin, glycylglycine, zinc chloride). A solution of human insulin at 500 IU / ml is added to this concentrated solution of pramlintide and of excipients so as to obtain the final targeted composition. The final pH is adjusted to 7.4 by adding NaOH / HCl.

Example BRI: Preparation of a solution of pramlintide at 0.9 mg / mL and human insulin at 100 IU / mL containing m-cresol (29 mM), glycerin (174 mM) and zinc chloride ( 229 μΜ) at pH 7.4.

By a process similar to that used in Example BN1, a solution of pramlintide at 0.9 mg / mL and human insulin at 100 IU / mL containing m-cresol (29 mM), glycerin (174 mM) and zinc chloride (229 μΜ) at pH 7.4 is obtained.

Example ΒΝ0: Preparation of a pramlintide solution at 0.4 mg / mL and human insulin at 100 IU / mL containing co-polyamino acid BB15, phenol (30 mM), glycerin (174 mM), glycylglycine (8 mM) and zinc chloride (229 μΜ) at pH 7.4.

[000505] A concentrated solution of co-polyamino acid BB15 and of excipients is prepared by adding concentrated solutions of excipients (m-cresol, glycerin, glycylglycine, zinc chloride) to a concentrated solution of co-polyamino acid BB15.

A concentrated pramlintide solution at 5 mg / mL and then a solution of human insulin at 500 IU / mL are added to this concentrated solution of co-polyamino acid BB15 and of excipients so as to obtain the final composition targeted (table 9). The final pH is adjusted to 7.4 by adding NaOH / HCl.

The BN2 and BN3 solutions are prepared according to the above protocol.

112

Solution Co-polyamino acid BB15 concentration Ratio BB15 / pramlintide Visual appearance of the mixture mg / mL mM mol / mol BN1 - : Turbid BN2 2.4 0.59 6 Crystal clear BN3 4 0.98 10 Crystal clear

Table 9: Compositions and visual appearance of the solutions of pramlintide at 0.4 mg / ml and human insulin at 100 IU / ml at pH 7.4 at different concentrations of copolyamino acid BB15.

In the presence of co-polyamino acid BB15, a clear solution of pramlintide (0.4 mg / ml) and human insulin (100 IU / ml) is obtained at pH 7.4.

Example BRO: Preparation of a pramlintide solution at 0.9 mg / mL and human insulin at 100 ZU / mL containing co-polyamino acid BB15, m-cresol (29 mM), glycerin (174 mM) and zinc chloride (229 pM) at pH 7.4.

By a process similar to that used in the BNO example, a solution of pramlintide at 0.9 mg / ml_ and human insulin at 100 IU / ml containing co-polyamino acid BB15, m-cresol (29 mM), glycerin (174 mM) and zinc chloride (229 μΜ) at pH 7.4 is obtained.

The solutions BR2 to BR4 and BU2 to BU8 are prepared according to the above protocol.

Solution Co-polyamino acid BB15 concentration Ratio BB15 / pramlintîde mol / mol Visual aspect of the solution mg / mL mM BIS Turbid BR2 2.7 0.66 3 Crystal clear BR3 3.6 0.88 4 Crystal clear BR4 4.5 1.10 5 Crystal clear BU2 0.9 0.22 1 Crystal clear BU3 1.8 0.44 2 Crystal clear BU7 5.4 1.32 6 Crystal clear BU8 9 -2.20. 10 Crystal clear

Table 11: Compositions and appearance of the solutions of pramlintide at 0.9 mg / ml and human insulin at 100 IU / ml at pH 7.4 at different concentrations of copolyamino acid BB15.

113 [000511] In the presence of co-polyamino acid BB15, a clear solution of pramlintide (0.9 mg / ml) and human insulin (100 IU / ml) at pH 7.4 is obtained.

Example BGO: Preparation of a pramlintide solution at 0.9 mg / mL and human insulin at 100 IU / mL containing various co-polyamino acids of the invention, m-cresol (29 mM), glycerin ( 174 mM) and zinc chloride (229 pM) at pH 7.4.

By a method similar to the BNO example, a solution of pramlintide at 0.9 mg / ml and human insulin at 100 IU / ml containing a co-polyamino acid of the invention, m-cresol ( 29 mM), glycerin (174 mM) and zinc chloride (229 μΜ) at pH 7.4 is obtained.

The BG2 to BG12 solutions are prepared according to the protocol described above.

Solutio n Co-polyaminoaci of Co-polyamino acid concentration Co-polyamino acid / pramlinti ratio of Visual aspect of the solution mg / mL mM mol / mol BG2 BB15 5 10 1.22 2.45 5.4 10.8 Crystal clear Crystal clear BG3 BB14 5 10 0.98 1.96 4.3 8.6 Crystal clear BG4 AB17 5 10 1.11 1.22 4.9 9.8 Crystal clear Crystal clear BG5 AB15 5 10 0.99 1.99 4.4 8.8 Crystal clear BG6 AB14 5 10 1.48 2.96 6.5 13 Crystal clear BG10 BB18 5 10 0.72 1.44 3.2 6.3 Crystal clear Crystal clear BG11 BB9 5 10 1 2.01 4.4 8.8 Crystal clear Crystal clear BG12 BB2 5 10 1.27 2.54 5.6 11.2 Crystal clear Crystal clear

Table 14: Compositions and visual appearance of solutions of pramlintide at 0.9 mg / ml and human insulin at 100 IU / ml at pH 7.4 at different co-polyamino acid concentrations.

114

Example BD1: Preparation of a pramlintide solution at 0.9 mg / mL and insulin lispro at 100 IU / mL containing m-cresol (29 mM), glycerin (174 mM) and zinc chloride ( 300 pM) at pH 7.4.

A concentrated pramlintide solution at 5 mg / mL is added to a concentrated solution of excipients (m-cresol, glycerin, zinc chloride). A solution of insulin lispro at 500 IU / mL is added to this concentrated solution of pramlintide and of excipients so as to obtain the final composition targeted. The final pH is adjusted to

7.4 by addition of NaOH / HCI.

BDO example: Preparation of a pramlintide solution at 0.9 mg / mL and insulin lispro at 100 IU / mL containing co-polyamino acid BB15, m-cresol (29 mM), glycerin (174 mM) and zinc chloride (300 pM) at pH 7.4.

[000515] A concentrated solution of co-polyamino acid BB15 and of excipients is prepared by adding concentrated solutions of excipients (m-cresol, glycerin, zinc chloride) to a concentrated solution of co-polyamino acid BB15.

A concentrated solution of pramlintide at 5 mg / ml and then a solution of insulin lispro at 500 IU / ml are added to the concentrated solution of co-polyamino acid BB15 and of excipients so as to obtain the final targeted composition. The final pH is adjusted to pH 7.4 by adding NaOH / HCl.

BD3 to BD9 solutions are prepared according to the protocol described above.

Solution BB15 / pramlintide ratio Co-polyamino acid BB15 concentration Visual aspect of the solution mol / mol mg / ml mM BD1 - * Turbid BD3 2 1.8 „ 0.44 Crystal clear BD4 3 2.7 0.66 Crystal clear BD5 4 3.6 0.88 Crystal clear BD6 5 4.5 1.10 Crystal clear BD7 6 5.4 1.32 Crystal clear BD8 10 9 2.20 Crystal clear BD9 15 13.5 3.30 Crystal clear

Table 15: Compositions and visual appearance of the solutions of pramlintide at 0.9 mg / ml and insulin lispro at 100 IU / ml at pH 7.4 at different concentrations of co-polyamino acid BB15.

115 [000518] In the presence of co-polyamino acid BB15, a clear solution of pramlintide (0.9 mg / mL) and insulin lispro (100 IU / mL) at pH 7.4 is obtained.

C. Physico-Chemistry

Results of visual observations of the mixture and of fibrillation measurements by ThT

Principle [000519] The poor stability of a peptide can lead to the formation of amyloid fibrils, defined as ordered macromolecular structures. These can possibly result in the formation of gel within the sample.

The test for monitoring the fluorescence of thioflavin T (ThT) is used to analyze the physical stability of the solutions. Thioflavin is a small probe molecule with a characteristic fluorescence signature when it binds to amyloid-like fibrils (Naiki et al. (1989) Anal. BioChem. 177, 244-249; LeVine (1999) Methods. Enzymol. 309, 274-284).

This method makes it possible to follow the formation of fibrils for low concentrations of ThT in undiluted solutions. This monitoring is carried out under accelerated stability conditions: with stirring and at 37 ° C.

Experimental conditions [000522] The samples were prepared just before the start of the measurement. The preparation of each composition is described in the associated example. Thioflavin T was added to the composition from a concentrated stock solution so as to induce a negligible dilution of the composition. The concentration of Thioflavin T in the composition is 1, 2 or 40 μM depending on the type of composition: 40 μM in the case of human amylin compositions at 0.6 mg / ml, 2 μM in the case of pramlintide compositions at 0.9 mg / mL and 1 pM in pramlintide compositions at 0.4 mg / mL. This concentration is recalled in the legend relating to the table of results of the latency times for each type of composition.

[000523] A volume of 150 μL of the composition was introduced into a well of a 96-well plate. Each composition was analyzed in three tests (triplicate) within the same plate. The plate was sealed with transparent film to avoid evaporation of the composition.

[000524] This plate was then placed in the enclosure of a plate reader (EnVision 2104 Multilabel, Perkin Elmer). The temperature is adjusted to 37 ° C., and lateral agitation of 960 rpm with 1 mm of amplitude is imposed.

116 [000525] A reading of the fluorescence intensity in each well is carried out with an excitation wavelength of 442 nm, and an emission wavelength of 482 nm over time.

The fibrillation process is manifested by a sharp increase in fluorescence after a delay called lag time.

For each well, this delay was determined graphically as the intersection between the baseline of the signa! of fluorescence and the slope of the fluorescence curve as a function of time determined during the large initial increase in fluorescence. The latency time value reported corresponds to the average of the latency time measurements made on three wells.

An example of graphical determination is shown in FIG. 1. [000529] In this figure is shown graphically the determination of the latency time (LT) by monitoring the fluorescence of Thioflavin T, on a curve having in ordered the value of the fluorescence (in ua arbitrary units) and on the abscissa the time in minutes.

Example C1: Stability of human amylin solutions at pH 7.4 in the presence of co-polyamino acid BB15 at different concentrations.

Solution BB15 / human amylin ratio Co-polyamino acid concentration BB15 Standby time (hour) mol / mol mg / ml mM BV1 ... <0.02 BV5 5 3 0.73 > 1 BV6 6 3.6 0.88 > 5 BV7 7 4.2 1.03 > 30 BV8 8 4.8 1.17 > 54 BV9 9 5 _r 4 1.32 > 54 BV10 10 6 1.47 > 72

Table 16: Measurement of the latency time by ThT (40 μΜ) of the solutions BV1 to BV10.

The latency time of a solution of human amylin at pH 7.4 (BV1), devoid of co-polyamino acid, is less than 0.02 hours; the BV5 to BV10 solutions according to the invention, containing BB15 / human amylin molar ratios greater than 5 make it possible to obtain latency times greater than one hour, a molar ratio allowing to obtain latency times greater than 72 hours .

117

Example C2: Stability of Human Amylin and Human Insulin Solutions

100 IU at pH 7.4 in the presence of co-polyamino acid BB15 at different concentrations.

Solution BB15 / human amylin ratio Co-polyamino acid BB15 concentration Standby time (hour) mol / mol mg / ml mM BX1 - - * BX6 6 3.6 0.88 > 0.1 BX10 10 6 1.47 > 0.5 BX11 17.5 ... 10.5 2.57 > 5

* Latency time not measured because turbid solution

Table 17: Measurement of the latency time by ThhT (40 μΜ) of the solutions, BX6, BX10 and BX11.

The solution of human amylin and human insulin at pH 7.4 (BX1) is turbid. The co-polyamino acid BB15 makes it possible to obtain a clear solution containing human amylin in the presence of human insulin at pH 7.4 with latency times greater than 0.1 hour from a BB15 / human amylin molar ratio of 6, and greater than 5 hours for a BB15 / human amylin molar ratio of 17.5.

Example C3: Stability of 3.4 mg / ml solutions of pramlintide at pH 7.4 in the presence of co-polyamino acid BB15 at different concentrations.

Solution Co-polyamino acid BB15 concentration Ratio BB15 / pramlintide Latency (hours) mg / ml mM mol / mol BW1 - - 0.7 BW2 ... 2.4 0.59 6 > 40 BW3 4 0.98 10 > 63

Table 19: Measurement of the latency time by ThhT (1 μΜ) of solutions BW1 to BW3.

The pramlintide solution at pH 7.4 (BW1) devoid of copolyamino acid has a short latency time. The co-polyamino acid BB15 makes it possible to obtain a solution containing pramlintide at pH 7.4 with latency times greater than 40 hours from a BB15 / pramlintide molar ratio of 6.

118

Example C4: Stability of solutions at 0.9 mg / ml of pramlintide at pH 7.4 in the presence of co-polyamino acid BB1S at different concentrations.

Solution BB15 / pramlintide ratio Concentration in BB15 Latency (hours) mol / mol mg / ml mM BY1 - - 0.7 BY2 2 1.8 0.44 > 0.8 BY3 3 2.7 0.66 > 4 BY4 4 3.6 0.88 > 30 BY5 5 _4f5 1.10 > 63 BY6 6 . 5.4 1.22 > 63 BY7 10 9 L32 ........... > 63

Table 18: Measurement of the latency time by THh (2 μM) of solutions BY1 to BY7 [000533] The pramlintide solution at pH 7.4 (BY1) devoid of co-polyamino acid exhibits a short latency time; the latency times of the solutions containing a co-polyamino acid are greater than or equal to the latency times of the composition without co-polyamino acid at a molar ratio of co-polyamino acid BB15 / pramlintide of 2: 1.

Example CS: Stability of pramlintide solutions at 0.9 mg / ml at pH 7.4 in the presence of different co-polyamino acids.

Solut ion Co-polya mino acid Concentration in co-polyamino acid Ratio œ- polyamino acid / pra mlintide Standby time (hours) mg / mL mM mol / mol BY1 - - <0.7 BP2 BB15 5 10 1.22 ................... 2.45 5.4 10.8 > 63 > 63 BP3 BB14 10 1.96 8.6 > 15 BP4 AB17 10 1.22 9.8 > 63 BP10 BB18 5 10 0.72 1.44 3.2 6.3 > 63 > 63 BP11 BB9 5 10 1 2.01 4.4 8.8 > 50 > 63 BP12 BB2 5 10 1.27 2.54 5.6 ............................. 11.2 ............... ......, > 10 > 63

Table 23: Measurement of the latency time by ThT (2 μM) of the compositions BY1, BP2 to

BP12.

119 The pramlintide solution at pH 7.4 (BY1) devoid of copolyamino acid has a short latency time. The co-polyamino acids of the invention make it possible to obtain latency times greater than 1.9 hours under the conditions tested.

Example C6: Stability of solutions of pramlintide at 0.4 mg / ml and of human insulin 100 IU / ml at pH 7.4 containing co-polyamino acid BB15.

Solution Co-polyamino acid BB15 concentration BB15 / pramlintide ratio Standby time (hour) mg / mL mM mol / mol BN1 - ; -> * BN2 2.4 0.59 6 > 19 BN3 4 0.98 10 > 19

♦ Latency time not measured because turbid solution.

Table 25: Measurement of the latency time by ThT (1 μΜ) of the compositions BN1 to BN3.

The pramlintide and human insulin solution at pH 7.4 (BN1) devoid of co-polyamino acid is turbid.

The co-polyamino acid BB15 makes it possible to obtain a clear solution of pramlintide at 0.4 mg / ml and of human insulin 100 IU / ml at pH 7.4 with latency times greater than 19 hours for ratios BB15 / pramlintide molars greater than 6.

120

Example C7: Stability of solutions of pramiintide at 0.9 mg / ml and of human insulin 100 IU / ml at pH 7.4 in the presence of co-polyamino acid BB15 at different concentrations.

Solution ΒΒΙΞ / pramlintide ratio Co-polyamino acid BB15 concentration Standby time (hour) mol / mol mg / ml mM BIS ; - * BU3 2 0.44 > 0.5 BR2 3 2.7 0.66 > 2 BR3 4 3.6 0.88 > 6 BR4 5 4.5 1.10 > 9 BU7 6 5.4 1.32 > 9 BU8 10 9 2.20 > 9

* Latency time not measured because turbid solution.

Table 24: Measurement of the latency time by ThT (2 μΜ) of the BRI to BR4 solutions and

BU3 to BU8.

A solution of pramiintide at 0.9 mg / ml and human insulin 100 IU / ml 10 at pH 7.4 (BRI) devoid of co-polyamino acid is turbid. The clear solutions of pramiintide at 0.9 mg / ml and of human insulin 100 IU / ml at pH 7.4 in the presence of co-polyamino acid BB15 have latency times greater than 0.5 hour at molar ratio BB15 / pramlintide of 2, which can be greater than 9 hours for BB15 / pramlintide molar ratios greater than 5.

121

Example C8: Stability of solutions of pramlintide at 0.9 mg / ml and of human insulin 100 IU / ml at pH 7.4 in the presence of different co-polyamino acids.

Solution Co-polya minoa eide Co-polyamino acid concentration Ratio co- polyamino acid / pramli ntide Standby time (hours) mg / m L mM mol / mol BIS • * BG2 BB15 5 10 1.22 2.45 5.4 10.8 > 9 > 7 BG3 BB14 10 1.96 8.6 > 9 BG4 AB17 5 10 1.11 1.22 4.9 9.8 > 2 > 5 BG5 AB15 10 1.99 8.8 > 2 BG10 BB18 5 10 0.72 1.44 3.2 6.3 > 1 > 4 BG11 BB9 5 10 1 2.01 4.4 8.8 > 4 > 3 BG12 BB2 5 10 1.27 2.54 5.6 11.2 > 5 > 6

* Latency time not measured because turbid solution.

Table 29: Measurement of the latency time by ThT (2 μΜ) of solutions BG1 to BG13.

The pramlintide and human insulin solution at pH 7.4 (BRI) is turbid. The co-polyamino acids make it possible to obtain latency times greater than 1 hour under the conditions tested.

122

Example C9: Stability of solutions of pramlintide at 0.9 mg / ml and insulin lispro 100 IU / ml at pH 7.4 in the presence of co-polyamino acid BB15 at different concentrations.

Solution Ratio BB15 / pramlintide Co-polyamino acid BB15 concentration Latency (hours) mol / mol mg / m! mM BD1 * BD3 2 U. 0.44 0.8 BD4 3 2.7 0.66 > 2 BD5 4 3.6 0.88 > 7 BD6 5 4.5 1.10 > 9 BD7 6 5.4 1.22 > 9 BD8 10 9 1.32 > 9

* Latency time not measured because turbid solution.

Table 30: Measurement of the latency time by ThT (2 μΜ) of the BD1 and BD3 to BD8 solutions.

The pramlintide and lispro insuiine solution at pH 7.4 (BD1) is turbid.

The co-polyamino acids make it possible to obtain latency times greater than 0.8 hours under the conditions tested.

123

Claims (12)

1. Composition in the form of an aqueous solution for injection, the pH of which is between 6.0 and 8.0, comprising at least: a) amylin, an amylin receptor agonist or an amylin analog; b) a co-polyamino acid carrying carboxylate 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: • - [gprU-gpaWgpc) ^ra P Formula I in which - GpR is a radical of formulas II or ΙΓ: HH il H * —N — R — N— * π or * —— R — N— * n '; - GpA is a radical of formulas III or III ': 0 HN— * .JL _a ' HN * m or * H A-N— * m '; GpC is a radical of formula IV: - 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 a 1; 124 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; - 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 co-polyamino acid via a covalent bond between a carbonyl of the hydrophobic radical and a nitrogen atom in N-terminal position of the co-polyamino 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 at 1 then the hydrophobic radical of formula I is linked to the co-polyamino 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 an amine function in the N-terminal position carried by the precursor of the co-polyamino 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 II 'of 1 to 11 carbon atoms, said alkyl radical carrying one or several functions -CONH2, and o an ether or unsubstituted polyether radical comprising from 4 to 14 carbon atoms and from 1 to 5 oxygen atoms; 125 - 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 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 ⁺ ; characterized in that the composition does not comprise a basal insulin whose isoelectric point pi is between 5.8 and 8.5. 2. Composition 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 W-GpA \ - GpC ^ra formula V GpR, GpA, GpC, r and a have the definitions given above. 3. Composition 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 — GpCj ^{r 2} Formula VI in which 126 GpR, GpA, GpC, r and a have the definitions given above. 4. Composition according to any one of the preceding claims, characterized in that the co-polyamino acid carrying carboxylate charges and hydrophobic radicals is chosen from the co-polyamino acids of formula VII according to 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 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 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, • R ₂ 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 a radical -NR'R, R 'and R identical or different being chosen from the group consisting of H, linear or branched or cyclic alkyls in C2 to CIO, benzyl 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 metal cations; 127 • η + 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 co-polyamino acid and 5 <n + m <250. 5. Composition according to claim 4, 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, following formula Vlla: 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 C10, 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 C10 alkyl, benzyl and said R' and R alkyl 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. Composition according to claim 4, 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 below: 128 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. 7. Composition according to any one of the preceding claims, characterized in that the molar ratio of co-polyamino acid / amylin, agonist at the amylin receptor or amylin analog is greater than or equal to 1. 8. Composition according to any one of the preceding claims, characterized in that the amylin, amylin receptor agonist or amylin analog is amylin. 9. Composition according to any one of the preceding claims, characterized in that the amylin, amylin receptor agonist or amylin analog is pramlintide. 10. Composition according to any one of the preceding claims, characterized in that it also comprises a mealtime insulin. 11. Composition according to the preceding claim, characterized in that the co-polyamino acid / insulin molar ratio is greater than or equal to 1. 12. Composition according to any one of claims 1 to 9, characterized in that said composition has a stability measured by ThT greater than that of a reference composition comprising amylin, an agonist at the amylin receptor or an amylin analog but not comprising a co-polyamino acid carrying carboxylate charges and hydrophobic Hy radicals.