Composition in form of aqueous solution comprising at least one macromolecular compound

Abstract

The present invention relates to a composition in the form of an aqueous solution filterable on a membrane having a porosity of 0.22 [mu] m, comprising at least one macromolecular compound consisting of a series of identical or different polysaccharides linked together by a divalent group L. L is a divalent group resulting from a reaction between a cross-linking agent and two reactive functional groups, each reactive functional group being carried by two different polysaccharide chains, and the polysaccharide chains not forming a cyclic structure.

Description

[0001] The present invention relates to the field of polysaccharide-based preparations used as biomaterials, more particularly in the medical and aesthetic fields. In these applications, the preparations must have optimized rheological properties, taking into account the characteristics of the medical device, in particular for injection, the characteristics of the treatment area and the desired therapeutic or cosmetic effect.

[0002] Products in gel form based on cross-linked polysaccharides for aesthetic applications have optimized rheological properties for good stability in the injection area. The gels used have in particular robustness to deformation, which results in a loss tangent TanΔ (Tnδ) of less than 1.00, an elastic modulus G' being much greater than the viscosity modulus G" over a wide deformation range. In particular, the loss tangent of most products used in gel form is less than 0.50 (REF). This property of gel-based products results in certain limitations in terms of injectability, which may limit their range of use. In particular, the use of such products in sensitive areas of the body or face may be tricky or even problematic.

[0003] The present invention relates to the development of compositions in the form of aqueous solutions having very good flowability and being filterable on membranes with a porosity of 0.22 μm, these compositions comprising at least one linear or branched macromolecular compound consisting of a series of polysaccharides.

[0004] In patent application WO2020250128 filed by OFFHEALTH SPA, compositions filterable on membranes with a porosity of 0.2 μm are disclosed, comprising nanoparticles of cross-linked hyaluronic acid obtained by mechanical action of the cross-linked hyaluronic acid by a stirring system. These nanoparticles are suspended in the disclosed compositions. Therefore, these compositions do not contain macromolecules comparable to those obtained by the process of the present invention, and the rheological properties of the compositions have not been evaluated, since they are compositions comprising suspended nanoparticles rather than viscoelastic compositions.

[0005] In patent application WO2016030516 filed by the company GALDERMA SA, the possibility of filtering supernatants resulting from thermal, enzymatic or free radical degradation of gels containing hyaluronic acid on membranes with a porosity of 0.2 μm is presented. Although these compositions are filterable on membranes with a porosity of 0.2 μm, the rheological properties of these filtrates are not comparable to those of the compositions disclosed in the present invention and they are not suitable for the applications mentioned in this application.

[0006] Unexpectedly, the composition according to the invention has the significant advantage of being filterable on membranes with a porosity of 0.22 μm, which makes it possible to incorporate into it thermosensitive compounds, such as in particular peptides, proteins, growth factors, antibodies or vitamins, since the composition obtained can then be sterilized by simple filtration. Existing products based on cross-linked polysaccharide gels are not compatible with thermosensitive compounds, since the gels must be autoclaved or heat sterilized to avoid any risk of infection on the area treated by injection.

[0007] The invention also relates to a composition comprising at least one macromolecular compound consisting of a series of hyaluronic acids, which has in particular good durability after injection and resistance to enzymatic degradation.

[0008] The composition according to the invention allows the preparation of solutions with high polysaccharide concentrations, in particular hyaluronic acid, while maintaining ease of injection.

[0009] The present invention relates to a composition in the form of an aqueous solution filterable on a membrane with a porosity of 0.22 μm, comprising at least one macromolecular compound consisting of a series of identical or different polysaccharides linked together by divalent groups L

[0010] L is a divalent group resulting from the reaction between a cross-linking agent and two reactive functional groups, each reactive functional group is carried by two different polysaccharide chains, and the polysaccharide chains do not form a ring structure.

[0011] In one embodiment, the composition according to the invention is characterized in that the macromolecular compound consists of a series of polysaccharides chosen from the group consisting of hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparin precursors and biologically acceptable salts thereof, alone or in mixture.

[0012] In one embodiment, the composition according to the invention is characterized in that the macromolecular compound consists of hyaluronic acid chains.

[0013] In one embodiment, the composition according to the invention is characterized in that the macromolecular compound consists of heparin precursor chains.

[0014] In one embodiment, the composition according to the invention is characterized in that the macromolecular compound consists of a polysaccharide chain chosen from hyaluronic acid, a heparin precursor or their respective salts.

[0015] The present invention also relates to a method for preparing a composition comprising at least one macromolecular compound composed of a series of polysaccharides (such as hyaluronic acid), and more particularly to a method for preparing a composition comprising at least one macromolecular compound composed of polysaccharide chains, which composition has specific properties, such as filterability on a membrane with a porosity of 0.22 μm.

[0016] The method for preparing the composition according to the invention comprises at least the following steps:

[0017] a) providing polysaccharides;

[0018] b) providing a cross-linking agent;

[0019] c) performing one or more cross-linking steps in the presence of said polysaccharide and said cross-linking agent;

[0020] d) obtaining a cross-linked polysaccharide;

[0021] e) performing one or more steps of cleaving glycosidic bonds;

[0022] f) obtaining a macromolecular compound solution;

[0023] g) The macromolecular compound solution was filtered through a membrane with a porosity of 0.22 μm.

[0024] In one embodiment, the method for preparing a composition according to the invention comprises at least the following steps:

[0025] a) providing polysaccharides;

[0026] b) providing a cross-linking agent;

[0027] c) performing one or more cross-linking steps in the presence of said polysaccharide and said cross-linking agent;

[0028] d) obtaining a cross-linked polysaccharide;

[0029] e) performing a step of cleaving glycosidic bonds;

[0030] f) obtaining a macromolecular compound solution;

[0031] g) A sample of the macromolecular compound solution is filtered through a membrane with a porosity of 0.22 μm. This is an in-process control (IPC) test.

[0032] In one embodiment, the method for preparing a composition according to the invention comprises at least the following steps:

[0033] a) providing polysaccharides;

[0034] b) providing a cross-linking agent;

[0035] c) carrying out one or more crosslinking steps from a solution of at least one polysaccharide in the presence of said polysaccharide and said crosslinking agent, the concentration of said solution of at least one polysaccharide being greater than or equal to 10% by mass relative to the total mass of the crosslinking reaction medium, and said one or more crosslinking steps being carried out linearly or in stages for a reaction time of up to 24 hours at a constant temperature or a variable temperature, said constant temperature or variable temperature being less than or equal to 10° C.;

[0036] d) obtaining a cross-linked polysaccharide;

[0037] e) performing a step of cleaving glycosidic bonds;

[0038] f) obtaining a macromolecular compound solution;

[0039] g) The macromolecular compound solution was filtered through a membrane with a porosity of 0.22 μm.

[0040] In one embodiment, the method for preparing a composition according to the invention comprises at least the following steps:

[0041] a) providing polysaccharides;

[0042] b) providing a cross-linking agent;

[0043] c) carrying out one or more crosslinking steps from a solution of at least one polysaccharide in the presence of said polysaccharide and said crosslinking agent, the concentration of said solution of at least one polysaccharide being greater than or equal to 10% by mass relative to the total mass of the crosslinking reaction medium, and said one or more crosslinking steps being carried out linearly or in stages for a reaction time of up to 24 hours at a constant temperature or a variable temperature, said constant temperature or variable temperature being less than or equal to 10° C.;

[0044] d) obtaining a cross-linked polysaccharide;

[0045] e) performing a step of cleaving glycosidic bonds;

[0046] f) obtaining a macromolecular compound solution;

[0047] g) A sample of the macromolecular compound solution is filtered through a membrane with a porosity of 0.22 μm. This is an in-process control (IPC) test.

[0048] During step d) we obtain a network composed of different polysaccharides linked together by divalent groups L originating from the crosslinking agent.

[0049] In one embodiment, the process according to the invention is characterized in that the crosslinking agent is selected from diepoxides, trimetaphosphates, diamines, dialkoxyamines and dihydrazides.

[0050] In one embodiment, the process according to the invention is characterized in that the crosslinking agent is 1,4-butanediol diglycidyl ether (BDDE).

[0051] In one embodiment, the process according to the invention is characterized in that the crosslinking agent is a trimetaphosphate.

[0052] In one embodiment, the process according to the invention is characterized in that the crosslinker is a diamine.

[0053] In one embodiment, the process according to the invention is characterized in that the cross-linking agent is a dihydrazide.

[0054] In one embodiment, step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide results in a major destruction of the cross-linked network formed during step c).

[0055] In one embodiment, step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide results in complete destruction of the cross-linked network formed during step c).

[0056] During this step e), only the glycosidic bonds are broken, with the exception of the bonds between the divalent groups L resulting from the crosslinking agent and the polysaccharide.

[0057] In one embodiment, the process according to the invention is characterized in that step e) of cleaving the glycosidic bonds is performed by a chemical treatment.

[0058] In one embodiment, the process according to the invention is characterized in that step e) of cleaving the glycosidic bonds is performed by heat treatment.

[0059] In one embodiment, the process according to the invention is characterized in that the heat treatment of step e) to cleave the glycosidic bonds is carried out by steam autoclaving.

[0060] In one embodiment, the process according to the invention is characterized in that step e) of cleaving the glycosidic bonds is performed by radiation treatment.

[0061] In one embodiment, the process according to the invention is characterized in that step e) of cleaving the glycosidic bonds is performed by an enzymatic treatment.

[0062] In one embodiment, the process according to the invention is characterized in that step e) of cleaving the glycosidic bonds is performed by high pressure treatment.

[0063] The present invention also relates to macromolecular compounds of the general formula I:

[0064]

[0065] in:

[0066] -Pi , P i' , P i” , P i”' , P j , P j' , P j” , P j”' , P k , P k' , P k” , P k”' , P l , P l' , P l” and P l”' The same or different polysaccharides

[0067] - n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 2000, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2

[0068] -x, x', x", and x'' are integers greater than or equal to 0

[0069] -L is a divalent group generated by the reaction between the crosslinker and two reactive functional groups, each of which is carried by two different polysaccharide chains

[0070] - Polysaccharide chains do not form ring structures

[0071] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 1500, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0072] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 1000, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0073] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 500, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0074] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 250, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0075] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 150, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0076] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 100, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0077] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 50, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0078] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 30, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0079] In one embodiment, n1, n1', n1", n1'", n2, n2', n2", n2'", n3', n3", n3'", n4, n4', n4", n4'", x, x', x", and x'" are equal to 0, and n3 is equal to 2. This embodiment represents a polynucleotide having the formula (L-P k )—(L—P k ) in its simplest form.

[0080] In another embodiment, n1, n1', n1", n1"', n2', n2", n2"', n3', n3", n3"', n4, n4', n4", n4"' are equal to 0, and n2 and n3 are equal to 1. This embodiment also represents a having the following formula (L-P k )—(L—P j ) in a simple form.

[0081] The macromolecular compound according to the invention is therefore a mixed polymer comprising alternating polysaccharides linked together by divalent groups originating from the crosslinking agent. In fact, during step e) of cleaving the glycosidic bonds, the crosslinking agent becomes a "linking group" and it loses its role as a "crosslinking agent".

[0082] The present invention relates to a composition in the form of an aqueous solution filterable on a membrane with a porosity of 0.22 μm, comprising at least one macromolecular compound consisting of a series of identical or different polysaccharides linked together by divalent groups L (linking groups), characterized in that the macromolecular compound has the general formula I:

[0083]

[0084] in:

[0085] -P i , P i' , P i” , P i”' , P j , P j' , P j” , P j”' , P k , P k', P k” , P k”' , P l , P l' , P l” and P l”' The same or different polysaccharides

[0086] - n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 2000, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2

[0087] -x, x', x", and x'' are integers greater than or equal to 0

[0088] -L is a divalent group generated by the reaction between the crosslinker and two reactive functional groups, each of which is carried by two different polysaccharide chains

[0089] -Polysaccharide chains do not form ring structures.

[0090] In one embodiment, n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 1500, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2.

[0091] When the macromolecular compound according to the present invention in dry form is hydrated by means of an aqueous solution, an aqueous solution of the macromolecular compound is obtained.

[0092] In particular, the macromolecular compounds according to the invention have no swelling capacity.

[0093] In the sense of the present invention, swelling capacity means the fact that a compound placed in dry form in the presence of an aqueous solution, for example a saline solution, has the ability to form a gel of significant volume, of which more than 80% are retained by filtration on a membrane with a porosity of 0.22 μm. The formation of a gel is characterized by the presence of two phases in the medium: a gel or "soft solid" and a solution or supernatant.

[0094] In the presence of water, the macromolecular compounds according to the invention do not form gels.

[0095] In particular, the macromolecular compounds according to the invention do not form hydrogels. In the context of the present application, the term "hydrogel" means a gel consisting of a three-dimensional network of at least one compound, capable of absorbing large amounts of water or aqueous solutions and having specific rheological properties, in particular in terms of viscosity and viscoelasticity.

[0096] In one embodiment, the polysaccharide of step a) is selected from the group of glycosaminoglycans (GAGs).

[0097] In one embodiment, the polysaccharide in step a) is selected from the group of chemically modified, oxidized or substituted polysaccharides.

[0098] In one embodiment, the polysaccharide is selected from the group of glycosaminoglycans (GAGs), such as chondroitin, keratan, heparin, heparin precursors or even hyaluronic acid and mixtures thereof.

[0099] In one embodiment, the polysaccharide is selected from hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, oxidized cellulose, alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparin precursors and biologically acceptable salts thereof, alone or in mixture.

[0100] In one embodiment, the polysaccharide is hyaluronic acid, or a biologically acceptable salt thereof, alone or in a mixture.

[0101] In the context of the present application, hyaluronic acid or one of its biologically acceptable salts, alone or in a mixture, is preferred.

[0102] In one embodiment, the polysaccharide is selected from hyaluronic acid, sodium hyaluronate, and mixtures thereof.

[0103] In one embodiment, the polysaccharide is hyaluronic acid.

[0104] In one embodiment, the polysaccharide is selected from sodium hyaluronate and potassium hyaluronate.

[0105] In one embodiment, the polysaccharide is sodium hyaluronate.

[0106] In the context of the present application, sodium hyaluronate is a particularly preferred polysaccharide.

[0107] In one embodiment, the polysaccharide is hyaluronic acid or one of its salts chemically modified by substitution.

[0108] In one embodiment, the polysaccharide is hyaluronic acid or one of its salts substituted with groups providing lipophilic or hydrating properties, for example substituted hyaluronic acid as described in patent application FR 2 983 483 in the name of the Applicant.

[0109] In one embodiment, the polysaccharide is a heparin precursor, alone or in a mixture, or a biologically acceptable salt thereof.

[0110] In the context of the present application, a heparin precursor or one of its biologically acceptable salts, alone or in a mixture, is preferred.

[0111] In one embodiment, the polysaccharide is selected from the group consisting of heparin precursor, heparin precursor sodium, and mixtures thereof.

[0112] In one embodiment, the polysaccharide is a heparin precursor.

[0113] In one embodiment, the polysaccharide is selected from sodium heparin precursor and potassium heparin precursor.

[0114] In one embodiment, the polysaccharide is sodium heparin precursor.

[0115] In the context of the present application, sodium heparin precursor is a particularly preferred polysaccharide.

[0116] In one embodiment, the polysaccharide is a chemically modified heparin precursor or a salt thereof.

[0117] In one embodiment, the polysaccharide is selected from cellulose and cellulose derivatives.

[0118] In one embodiment, the polysaccharide is selected from cellulose derivatives, in particular including hydroxypropyl cellulose, hydroxypropyl methyl cellulose, ethyl methyl cellulose and carboxymethyl cellulose.

[0119] In one embodiment, the polysaccharide is cellulose.

[0120] In one embodiment, the polysaccharide is a cellulose derivative.

[0121] In one embodiment, the polysaccharide used in step a) of the process for preparing a composition according to the invention is a mixture of polysaccharides.

[0122] In the context of the present application, all the polysaccharides listed can be brought together in the form of a mixture during step a), whether they are polysaccharides of the same nature (e.g. a mixture of hyaluronic acid with different molecular weights) or polysaccharides of different nature (e.g. a mixture of hyaluronic acid and chitosan). During the cross-linking step, co-crosslinks can exist between the different polysaccharides.

[0123] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is hyaluronic acid or a mixture of hyaluronates.

[0124] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of two hyaluronic acids or hyaluronates.

[0125] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of three hyaluronic acids or hyaluronates.

[0126] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of 4 hyaluronic acids or hyaluronates.

[0127] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a heparin precursor or a mixture of heparin precursor salts.

[0128] In one embodiment, the process for preparing the composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of 2 heparin precursors or heparin precursor salts.

[0129] In one embodiment, the process for preparing the composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of 3 heparin precursors or heparin precursor salts.

[0130] In one embodiment, the process for preparing the composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of 4 heparin precursors or heparin precursor salts.

[0131] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of hyaluronic acid and a heparin precursor or a salt thereof.

[0132] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of hyaluronic acid and a heparin precursor or a salt thereof.

[0133] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of two hyaluronic acids and a heparin precursor or a salt thereof.

[0134] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of hyaluronic acid and two heparin precursors or salts thereof.

[0135] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of two hyaluronic acids and two heparin precursors or their salts.

[0136] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of hyaluronic acid or a salt thereof and cellulose.

[0137] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of hyaluronic acid or a salt thereof and cellulose.

[0138] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of two hyaluronic acids or salts thereof and cellulose.

[0139] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of hyaluronic acid or a salt thereof and two celluloses.

[0140] In one embodiment, the process for preparing a composition according to the invention is characterized in that the polysaccharide of step a) is a mixture of two hyaluronic acids or salts thereof and two celluloses.

[0141] In the context of this application, Mw or "molecular weight" is referred to as the weight average molecular weight of the polysaccharide measured in Daltons.

[0142] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 0.01 MDa to 10 MDa (0.01 MDa≤Mw≤10 MDa).

[0143] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 0.01 MDa to 8 MDa.

[0144] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 0.01 MDa to 5 MDa.

[0145] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 0.01 MDa to 3.5 MDa.

[0146] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 0.5 MDa to 3.5 MDa.

[0147] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 2.75 MDa to 3.25 MDa.

[0148] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 0.75 MDa to 1.25 MDa.

[0149] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 2 MDa to 5 MDa.

[0150] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 2 MDa to 4 MDa.

[0151] In one embodiment, the polysaccharide or a salt thereof has a molecular weight of 0.5 MDa to 2 MDa.

[0152] In one embodiment, the polysaccharide acid or a salt thereof has a molecular weight of 0.5 MDa to 1.5 MDa.

[0153] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 0.01 MDa to 10 MDa.

[0154] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 0.01 MDa to 5 MDa.

[0155] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 0.01 MDa to 3.5 MDa.

[0156] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 0.5 MDa to 3.5 MDa.

[0157] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 2.75 MDa to 3.25 MDa.

[0158] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 0.75 MDa to 1.25 MDa.

[0159] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 2 MDa to 5 MDa.

[0160] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 2 MDa to 4 MDa.

[0161] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 0.5 MDa to 2 MDa.

[0162] In one embodiment, the hyaluronic acid or a salt thereof has a molecular weight of 0.5 MDa to 1.5 MDa.

[0163] In one embodiment, the heparin precursor or a salt thereof has a molecular weight of 0.01 MDa to 10 MDa.

[0164] In one embodiment, the heparin precursor or a salt thereof has a molecular weight of 0.01 MDa to 5 MDa.

[0165] In one embodiment, the heparin precursor or a salt thereof has a molecular weight of 0.02 MDa to 3 MDa.

[0166] In one embodiment, the heparin precursor or a salt thereof has a molecular weight of 0.02 MDa to 2 MDa.

[0167] To prepare the composition according to the invention, one or more crosslinking steps c) are carried out between the polysaccharide and at least one crosslinking agent.

[0168] In one embodiment, the process for preparing the composition according to the invention is characterized in that bringing together the polysaccharide and at least one crosslinking agent takes place in a solvent.

[0169] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is chosen from diepoxides such as ethylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether (BDDE), 1,2,3,4-diepoxybutane or 1,2,7,8-diepoxyoctane; dialkyl sulfones; divinyl sulfone; formaldehyde; epichlorohydrin or even glutaraldehyde; carbodiimides such as 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC); trimetaphosphates such as sodium trimetaphosphate, calcium trimetaphosphate or even barium trimetaphosphate.

[0170] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is chosen from diepoxides such as ethylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether (BDDE), 1,2,3,4-diepoxybutane or 1,2,7,8-diepoxyoctane; trimetaphosphates such as sodium trimetaphosphate, calcium trimetaphosphate or even barium trimetaphosphate.

[0171] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is chosen from ethylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether (BDDE), 1,2,3,4-diepoxybutane or 1,2,7,8-diepoxyoctane.

[0172] In one embodiment, the process for preparing a composition according to the invention is characterized in that the at least one crosslinking agent of step c) is chosen from bifunctional polyethylene glycols (PEG) carrying an epoxide at each end of the polymer chain.

[0173] In one embodiment, the process for preparing a composition according to the invention is characterized in that the at least one crosslinking agent of step c) is chosen from difunctional polyethylene glycols (PEG) carrying an epoxy group at each end of the polymer chain, chosen from polyethylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether and polybutylene glycol diglycidyl ether.

[0174] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is chosen from trimetaphosphates, such as sodium trimetaphosphate, calcium trimetaphosphate or even barium trimetaphosphate.

[0175] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent in step c) is chosen from epoxides, such as 1,4-butanediol diglycidyl ether (BDDE), epihalohydrins, divinyl sulfone (DVS).

[0176] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent in step c) is divinyl sulfone (DVS).

[0177] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent in step c) is 1,4-butanediol diglycidyl ether (BDDE).

[0178] In the context of the present application, BDDE is particularly preferred.

[0179] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is chosen from compounds carrying two functional groups chosen from amine functions, alkoxyamines and hydrazides.

[0180] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent in step c) is chosen from diamines.

[0181] In one embodiment, the diamine used as crosslinking agent in step c) of the process for preparing a composition according to the invention is selected from diaminotrehalose, diaminosucrose, chitobiose, diaminolactose and diaminoraffinose.

[0182] In one embodiment, the diamine used as cross-linking agent in step c) of the process for preparing a composition according to the invention is diaminotrehalose.

[0183] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent in step c) is chosen from dialkoxyamines.

[0184] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent in step c) is chosen from dihydrazides.

[0185] In one embodiment, the dihydrazide used as cross-linking agent in step c) of the process for preparing the composition according to the invention is adipic acid dihydrazide.

[0186] In one embodiment, the or each cross-linking step in cross-linking step c) requires prior solubilization of the polysaccharide.

[0187] In one embodiment, the polysaccharide is added to the solution in solid form to dissolve the polysaccharide.

[0188] In one embodiment, the solution is added to the polysaccharide in solid form to dissolve the polysaccharide.

[0189] In one embodiment, the or each cross-linking step in step c) does not require solubilization of the polysaccharide.

[0190] The polysaccharide is dissolved by adding water or a saline solution, such as a phosphate buffer solution, such as PBS, or by adding sodium hydroxide or an acid solution to obtain a pH compatible with the performance of the cross-linking step or each of the steps.

[0191] In one embodiment, the polysaccharide is dissolved by adding water or a saline solution, such as a phosphate buffer solution, such as PBS, or by adding a solution of sodium hydroxide or an acid to obtain a pH compatible with the performance of the step or each of the cross-linking steps, wherein the water or saline solution also contains at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0192] In one embodiment, the process for obtaining the composition according to the invention is characterized in that, at the latest during step c), a step of adjusting the pH to the crosslinking pH is carried out.

[0193] The pH adjustment is performed by adding a solution of an inorganic acid, preferably such as hydrochloric acid, or an inorganic base, preferably such as soda or potash, the acid and base being added in such an amount that the target crosslinking pH can be achieved.

[0194] In one embodiment, during one or each of the cross-linking steps c), a step of adjusting the pH to a cross-linking pH suitable for the cross-linking agent is performed.

[0195] In one embodiment, at the latest during the or each cross-linking step in cross-linking step c), a pH adjustment step to a cross-linking pH of greater than 10 is performed.

[0196] In one embodiment, at the latest during the or each cross-linking step in cross-linking step c), a pH adjustment step to a cross-linking pH of less than 3 is performed.

[0197] In one embodiment, the target cross-linking pH during the or each cross-linking step in cross-linking step c) is obtained by adding sodium hydroxide solution having a concentration of at most 0.25 N or 1 mass %.

[0198] In one embodiment, at the latest during the or each cross-linking step in cross-linking step c), a pH adjustment step is performed to a cross-linking pH of greater than 10, the cross-linking agent being BDDE.

[0199] In one embodiment, at the latest during the or each cross-linking step in cross-linking step c), a pH adjustment step is performed to a cross-linking pH of less than 3, the cross-linking agent being BDDE.

[0200] In one embodiment, at the latest during the cross-linking step or each of the cross-linking steps c), a step of adjusting the pH to a cross-linking pH of greater than 10 is performed.

[0201] Cross-linking begins when the following three conditions are met: the presence of a polysaccharide, the presence of a cross-linking agent, and the reaction medium is at an appropriate pH.

[0202] In one embodiment, the or each of the crosslinking steps in crosslinking step c) is characterized in that the initiation of crosslinking is brought about by adding said crosslinking agent.

[0203] In one embodiment the or each of the cross-linking steps in cross-linking step c) is characterized in that initiation of cross-linking is caused by adding said polysaccharide.

[0204] In one embodiment, the or each of the cross-linking steps in cross-linking step c) is characterized in that initiation of cross-linking is caused by applying a cross-linking pH.

[0205] In one embodiment, the process for preparing the composition according to the invention is characterized in that, after step c), a step of adjusting the pH to a pH between 6 and 8 is carried out.

[0206] According to the pH of the reaction medium at the end of the crosslinking reaction of step c), the pH adjustment is carried out by adding preferably a solution of an inorganic acid (for example hydrochloric acid) or preferably an inorganic base (for example soda or potash), the acid and base being added in such an amount that a pH of 6 to 8 can be achieved.

[0207] In one embodiment, the process for preparing the composition according to the invention is characterized in that, after step c), a step of adjusting the pH to a pH between 6 and 8 is carried out by adding at least one acid, said at least one acid being hydrochloric acid (HCl).

[0208] The process for preparing the composition according to the invention is characterized in that the or each of the crosslinking steps c) is carried out linearly or in stages at a constant or variable temperature less than or equal to 50°C.

[0209] The process for preparing the composition according to the invention is characterized in that the or each of the crosslinking steps c) is carried out linearly or in stages at a constant or variable temperature less than or equal to 30°C.

[0210] The process for preparing the composition according to the invention is characterized in that the or each of the crosslinking steps c) is carried out linearly or in stages at a constant or variable temperature less than or equal to 20°C.

[0211] The process for preparing the composition according to the invention is characterized in that the or each of the crosslinking steps c) is carried out linearly or in stages at a constant or variable temperature less than or equal to 15°C.

[0212] The process for preparing the composition according to the invention is characterized in that the or each of the crosslinking steps c) is carried out linearly or in stages at a constant or variable temperature less than or equal to 10°C.

[0213] The process for preparing the composition according to the invention is characterized in that the or each of the crosslinking steps c) is carried out linearly or in stages at a constant or variable temperature less than or equal to 5°C.

[0214] The freezing temperature of the reaction medium means the temperature at which the medium becomes solid. For aqueous media this temperature will be at 0°C or slightly lower, depending on the salt concentration of the medium.

[0215] In one embodiment, the or each of the cross-linking steps c) is carried out linearly or in stages at a constant temperature or a variable temperature between the solidification temperature and 10°C.

[0216] In one embodiment, the or each of the cross-linking steps c) is carried out linearly or in stages at a constant temperature or a variable temperature between the solidification temperature and 5°C.

[0217] In one embodiment, the or each of the cross-linking steps c) is carried out at a constant temperature of 50°C.

[0218] In one embodiment, the or each of the cross-linking steps c) is carried out at a constant temperature of 30°C.

[0219] In one embodiment, the or each of the cross-linking steps c) is carried out at a constant temperature of 20°C.

[0220] In one embodiment, the or each of the cross-linking steps c) is carried out at a constant temperature of 15°C.

[0221] In one embodiment, the or each of the cross-linking steps c) is carried out at a constant temperature of 9°C.

[0222] In one embodiment, the or each of the cross-linking steps c) is carried out at a constant temperature of 5°C.

[0223] In one embodiment, the or each of the cross-linking steps c) is carried out at a constant temperature of 2°C.

[0224] In one embodiment, prior to the or each crosslinking step in crosslinking step c), a cooling step to the crosslinking temperature is performed.

[0225] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 and a temperature less than or equal to 50°C.

[0226] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 and a temperature less than or equal to 30°C.

[0227] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 and a temperature less than or equal to 20°C.

[0228] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 and a temperature less than or equal to 15°C.

[0229] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 and a temperature less than or equal to 10°C.

[0230] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 and a temperature less than or equal to 5°C.

[0231] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 and a temperature less than or equal to 2°C.

[0232] In one embodiment, the process for preparing the composition according to the invention is characterized in that the duration of the crosslinking step c) is between 10 minutes and 26 hours.

[0233] In one embodiment, the process for preparing the composition according to the invention is characterized in that the duration of the crosslinking step c) is between 10 minutes and 18 hours.

[0234] In one embodiment, the process for preparing the composition according to the invention is characterized in that the duration of the crosslinking step c) is from 10 minutes to 12 hours.

[0235] In one embodiment, the process for preparing the composition according to the invention is characterized in that the duration of the crosslinking step c) is between 10 minutes and 5 hours.

[0236] In one embodiment, the process for preparing the composition according to the invention is characterized in that the duration of the crosslinking step c) is between 10 minutes and 3 hours.

[0237] In one embodiment, the process for preparing the composition according to the invention is characterized in that the duration of the crosslinking step c) is between 30 minutes and 3 hours.

[0238] In one embodiment, the process for preparing the composition according to the invention is characterized in that the crosslinking step c) is carried out at 2° C. for a period of 24 hours.

[0239] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 and at a temperature of 2° C. for a period of 24 hours.

[0240] In one embodiment, the process for preparing the composition according to the invention is characterized in that the crosslinking step c) is carried out at 9° C. for a period of 3 hours.

[0241] In one embodiment, the process for preparing the composition according to the invention is characterized in that the at least one crosslinking agent of step c) is 1,4-butanediol diglycidyl ether (BDDE) and that step c) is carried out at a pH greater than 10 at a temperature of 9° C. for a period of 3 hours.

[0242] When several successive crosslinkings are carried out in step c) of the process for preparing the composition according to the invention, the time mentioned is the total time (sum of the times of the successive crosslinkings).

[0243] In one embodiment, during step c), the performance of the cross-linking step in the presence of said polysaccharide and said cross-linking agent takes place in a medium in which said polysaccharide is hydrated and / or swollen by the addition of water or a saline solution, such as a phosphate buffer solution, such as PBS.

[0244] In one embodiment, during step c), the performance of the cross-linking step in the presence of said polysaccharide and said cross-linking agent takes place in a medium in which said polysaccharide is hydrated and / or swollen by the addition of water or a saline solution, such as a phosphate buffered solution, such as PBS, which also comprises at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0245] During step c) of the process for preparing the composition according to the invention, the crosslinking rate (X) can be theoretically calculated using the following formula:

[0246]

[0247] Thus, for example, if the medium contains 100 disaccharide patterns, and the medium also contains 10 cross-linker molecules, the cross-linking ratio (X) will be as follows: X = 10 / 100 = 0.1. This cross-linking ratio is therefore not affected by the degree of polymerization, nor by the molecular weight of the polysaccharide selected, nor by the proportion of cross-linker that actually reacts with at least one functional group of the polysaccharide. This is a theoretical determination that takes into account only the amount of cross-linker and repeating units that come together.

[0248] In one embodiment, the cross-linking ratio X is from 0.001 to 0.20.

[0249] In one embodiment, the cross-linking ratio X is from 0.01 to 0.15.

[0250] In one embodiment, the cross-linking ratio X is from 0.01 to 0.12.

[0251] In one embodiment, the cross-linking ratio X is from 0.03 to 0.10.

[0252] In one embodiment, the cross-linking ratio X is from 0.04 to 0.08.

[0253] The crosslinking can also be assessed ex post facto (after crosslinking) via the degree of modification (Mod). Thus, in contrast to the crosslinking ratio X, Mod takes into account the proportion of crosslinking agent that actually reacted with at least one functional group of the polysaccharide.

[0254] The degree of modification can be expressed as follows:

[0255]

[0256] When the polysaccharide is hyaluronic acid, the repeating units (or monomers) are in a disaccharide pattern.

[0257] The determination of the values ​​of the numerator and the denominator depends on the selected polysaccharide and the selected cross-linking agent and is well known to the person skilled in the art. For example, in the specific case of a formulation based on hyaluronic acid cross-linked with BDDE, the method described in the publication L. Nord, A. Emilson, C. Sturesson, AH Kenne, Degree of Modification of Hyaluronic Acid Dermal Fillers, 18th EADV Congress, Berlin, 2009 can be used.

[0258] In the specific case of formulations based on hyaluronic acid cross-linked with BDDE, the degree of modification can be expressed as follows:

[0259]

[0260] For example, a formulation based on hyaluronic acid cross-linked with BDDE has a Mod of 1% meaning that for every 100 disaccharide patterns it has one BDDE molecule (single or double linked).

[0261] In one embodiment, the degree of modification of the cross-linked polysaccharide is less than 5%.

[0262] In one embodiment, the degree of modification of the cross-linked polysaccharide is less than 4%.

[0263] In one embodiment, the degree of modification of the cross-linked polysaccharide is less than 3%.

[0264] In one embodiment, the degree of modification of the cross-linked polysaccharide is less than 2%.

[0265] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of at least 1% by mass relative to the total mass of the cross-linking reaction medium.

[0266] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of at least 5% by mass relative to the total mass of the cross-linking reaction medium.

[0267] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of at least 10% by mass relative to the total mass of the cross-linking reaction medium.

[0268] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of at least 15% by mass relative to the total mass of the cross-linking reaction medium.

[0269] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of 1% to 60% by mass relative to the total mass of the cross-linking reaction medium.

[0270] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of 5% to 60% by mass relative to the total mass of the cross-linking reaction medium.

[0271] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of 10% to 60% by mass relative to the total mass of the cross-linking reaction medium.

[0272] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, with a polysaccharide concentration ranging from 10% to 40% by mass relative to the total mass of the reaction medium.

[0273] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of 10% to 25% by mass relative to the total mass of the reaction medium.

[0274] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, for example at a polysaccharide concentration of 15% to 25% by mass relative to the total mass of the reaction medium.

[0275] In one embodiment, during step c), the crosslinking step is carried out in a crosslinking reaction medium in the presence of hyaluronic acid or one of its biologically acceptable salts, alone or in a mixture, and said crosslinking agent, the concentration of hyaluronic acid being between 1% and 60% by mass relative to the total mass of the reaction medium.

[0276] In one embodiment, during step c), the crosslinking step is carried out in the presence of hyaluronic acid or one of its biologically acceptable salts and said crosslinking agent, alone or in a mixture, in a crosslinking reaction medium at a concentration ranging from 5% to 60% by weight relative to the total mass of the reaction medium.

[0277] In one embodiment, during step c), the crosslinking step is carried out in the presence of hyaluronic acid or one of its biologically acceptable salts and said crosslinking agent, alone or in a mixture, in a crosslinking reaction medium at a concentration of hyaluronic acid ranging from 10% to 60% by weight relative to the total mass of the reaction medium.

[0278] In one embodiment, during step c), the crosslinking step is carried out in the presence of hyaluronic acid or one of its biologically acceptable salts and said crosslinking agent, alone or in a mixture, at a concentration of hyaluronic acid ranging from 10% to 40% by mass relative to the total mass of the reaction medium.

[0279] In one embodiment, during step c), the crosslinking step is carried out in the presence of hyaluronic acid or one of its biologically acceptable salts and said crosslinking agent, alone or in a mixture, at a concentration of 10 to 25% by mass relative to the total mass of the reaction medium.

[0280] In one embodiment, during step c), the crosslinking step is carried out in the presence of hyaluronic acid or one of its biologically acceptable salts and said crosslinking agent, alone or in a mixture, at a concentration of 15 to 25% by mass relative to the total mass of the reaction medium.

[0281] In one embodiment, during step c), the crosslinking step is carried out in the presence of a heparin precursor or one of its biologically acceptable salts, alone or in a mixture, and said crosslinking agent in a crosslinking reaction medium at a concentration of between 1% and 60% by weight relative to the total mass of the reaction medium.

[0282] In one embodiment, during step c), the crosslinking step is carried out in the presence of a heparin precursor or one of its biologically acceptable salts, alone or in a mixture, and said crosslinking agent in a crosslinking reaction medium at a heparin precursor concentration ranging from 5% to 60% by weight relative to the total mass of the reaction medium.

[0283] In one embodiment, during step c), the crosslinking step is carried out in the presence of a heparin precursor or one of its biologically acceptable salts, alone or in a mixture, and said crosslinking agent in a crosslinking reaction medium at a heparin precursor concentration of 10% to 60% by weight relative to the total mass of the reaction medium.

[0284] In one embodiment, during step c), the crosslinking step is carried out in the presence of a heparin precursor or one of its biologically acceptable salts, alone or in a mixture, and said crosslinking agent, the concentration of the heparin precursor being between 10% and 40% by mass relative to the total mass of the reaction medium.

[0285] In one embodiment, during step c), the crosslinking step is carried out in the presence of a heparin precursor or one of its biologically acceptable salts and said crosslinking agent, alone or in a mixture, at a concentration of 10% to 25% by weight relative to the total mass of the reaction medium.

[0286] In one embodiment, during step c), the crosslinking step is carried out in the presence of a heparin precursor or one of its biologically acceptable salts and said crosslinking agent, alone or in a mixture, at a concentration of 15% to 25% by weight relative to the total mass of the reaction medium.

[0287] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, the cross-linking reaction medium comprising sodium hydroxide (NaOH).

[0288] In one embodiment, during step c), the crosslinking step is carried out in the presence of said polysaccharide and said crosslinking agent, with a soda concentration ranging from 0.5% to 1.5% by mass relative to the total mass of the reaction medium.

[0289] In one embodiment, during step c), the crosslinking step is carried out in the presence of said polysaccharide and said crosslinking agent, with a soda concentration ranging from 0.5% to 1% by mass relative to the total mass of the reaction medium.

[0290] In one embodiment, during step c), the crosslinking step is carried out in the presence of said polysaccharide and said crosslinking agent, with a soda concentration ranging from 0.7% to 0.9% by mass relative to the total mass of the reaction medium.

[0291] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, the cross-linking reaction medium comprising hydrochloric acid (HCl).

[0292] In one embodiment, during step c), the cross-linking step is carried out in the presence of said polysaccharide and said cross-linking agent, the concentration of hydrochloric acid in the reaction medium being between 0.001M and 0.05M.

[0293] In one embodiment, the cross-linking step c) comprises at least one activation step of the chemical functional groups carried by the polysaccharide.

[0294] In one embodiment, the cross-linking step c) comprises at least one step of activating the carboxylic acid functions carried by the polysaccharide.

[0295] In one embodiment, the step of activating the carboxylic acid functions carried by the polysaccharide is carried out using a coupling agent used in peptide chemistry.

[0296] In one embodiment, the step of activating the carboxylic acid functional groups carried by the polysaccharide is carried out using a coupling agent for peptide chemistry selected from the group consisting of triazines, carbodiimides, imidazoles type coupling agents as well as Oxyma and COMU.

[0297] In one embodiment, the triazine-type coupling agent is selected from 4-(4,6-dimethoxy[1.3.5]triazin-2-yl)-4-methylmorpholine chloride (DMTMM) and 2-chloro-4,6-dimethoxy-1,3,5-triazine (CMT).

[0298] In one embodiment, the coupling agent is 4-(4,6-dimethoxy[1.3.5]triazin-2-yl)-4-methylmorpholine Chloride (DMTMM).

[0299] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a step of eliminating the crosslinking agent is carried out.

[0300] In one embodiment, the process for preparing a composition according to the invention is characterized in that, before step d), a purification step of the cross-linked polysaccharide is carried out.

[0301] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a purification step of the cross-linked polysaccharide is carried out by dialysis.

[0302] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a step of purifying the cross-linked polysaccharide is carried out by dialysis using a solution or a dialysis solvent selected from phosphate buffer (eg PBS) and water.

[0303] In one embodiment, the method for preparing a composition according to the invention is characterized in that, prior to step d), a step of purifying the cross-linked polysaccharide is carried out by dialysis using a solution or a dialysis solvent selected from a phosphate buffer (e.g. PBS) and water, the solution or the dialysis solvent also comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0304] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a purification step of the cross-linked polysaccharide is carried out by washing.

[0305] In one embodiment, the step of purifying the cross-linked polysaccharide is performed by washing with a solution or solvent selected from phosphate buffered saline (eg PBS) and water.

[0306] In one embodiment, the step of purifying the cross-linked polysaccharide is performed by washing with a solution or solvent selected from phosphate buffered saline (eg PBS) and water, said solution or solvent also comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0307] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a purification step of the cross-linked polysaccharide is carried out by precipitation in salt form.

[0308] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a purification step of the cross-linked polysaccharide is carried out by precipitation from the solution obtained after adding PBS or NaCl salt.

[0309] In one embodiment, the process for preparing a composition according to the invention is characterized in that, before step d), a purification step of the cross-linked polysaccharide is carried out by precipitation from the solution obtained after the addition of PBS or NaCl salt, said solution also comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0310] In one embodiment, the step of purifying the cross-linked polysaccharide by precipitation is performed by adding at least one organic solvent to the cross-linked polysaccharide.

[0311] In one embodiment, the at least one organic solvent added to the cross-linked polysaccharide in the precipitation purification step is selected from the group of alcohols.

[0312] In one embodiment, the alcohol added to the cross-linked polysaccharide during the precipitation purification step is ethanol.

[0313] In one embodiment, the step of purifying the cross-linked polysaccharide by precipitation occurs in a hydroalcoholic mixture.

[0314] In one embodiment, the precipitated cross-linked polysaccharide is isolated by filtration.

[0315] In one embodiment, the cross-linked polysaccharide that is precipitated and separated by filtration is washed.

[0316] In one embodiment, the cross-linked polysaccharide that has been precipitated and isolated by filtration is dried.

[0317] In one embodiment, the cross-linked polysaccharide precipitated and separated by filtration is dried in the atmosphere at room temperature.

[0318] In one embodiment, the cross-linked polysaccharide precipitated and recovered by filtration is dried under vacuum at room temperature.

[0319] In one embodiment, the cross-linked polysaccharide is lyophilized.

[0320] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out by means of a solution or solvent chosen from phosphate buffer (eg PBS) and water.

[0321] In one embodiment, the process for preparing a composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out by means of a solution or solvent chosen from phosphate buffer (e.g. PBS) and water, and water, said solution or solvent also comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0322] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a concentration of 2 to 200 mg / g of the cross-linked polysaccharide relative to the total mass of the composition.

[0323] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a cross-linked polysaccharide concentration of 2 to 75 mg / g relative to the total mass of the composition.

[0324] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a cross-linked polysaccharide concentration of 5 to 50 mg / g relative to the total mass of the composition.

[0325] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a concentration of 10 to 40 mg / g of cross-linked polysaccharide relative to the total mass of the composition.

[0326] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a cross-linked polysaccharide concentration of about 100 mg / g relative to the total mass of the composition.

[0327] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a cross-linked polysaccharide concentration of about 80 mg / g relative to the total mass of the composition.

[0328] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a cross-linked polysaccharide concentration of about 60 mg / g relative to the total mass of the composition.

[0329] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a cross-linked polysaccharide concentration of about 40 mg / g relative to the total mass of the composition.

[0330] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a cross-linked polysaccharide concentration of about 20 mg / g relative to the total mass of the composition.

[0331] In one embodiment, the process for preparing the composition according to the invention is characterized in that, before step d), a dilution step of the cross-linked polysaccharide is carried out to obtain a cross-linked polysaccharide concentration of about 10 mg / g relative to the total mass of the composition.

[0332] In one embodiment, the polysaccharide obtained in step d) has a tangent Tan Δ (Tn δ) ≥ 0.50 before the bond cleavage step of step e).

[0333] In one embodiment, the polysaccharide obtained in step d) has a tangent Tan Δ (Tn δ) ≥ 0.70 before the bond cleavage step of step e).

[0334] The process for preparing a composition according to the invention comprises at least one step of cleavage of glycosidic bonds, step e), carried out on at least one cross-linked polysaccharide and making it possible to obtain the macromolecular compound according to the invention.

[0335] In one embodiment, step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide is based on a controlled degradation mechanism.

[0336] In one embodiment, step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide is performed by a treatment selected from the group consisting of chemical treatment, treatment by enzymes, treatment by radiation and heat treatment.

[0337] In one embodiment, step e) of cleaving the glycosidic bonds of at least one cross-linked polysaccharide is performed by chemical treatment.

[0338] In one embodiment, the chemical treatment comprises exposing at least one cross-linked polysaccharide to an aqueous solution at an alkaline pH.

[0339] In one embodiment, the aqueous solution at alkaline pH is an aqueous solution of sodium hydroxide (soda).

[0340] In one embodiment, the chemical treatment comprises exposing at least one cross-linked polysaccharide to an aqueous solution having an acidic pH.

[0341] In one embodiment, the aqueous solution at acidic pH comprises an acid selected from the group consisting of acetic acid, hydrochloric acid, sulfuric acid, and phosphoric acid.

[0342] In one embodiment, the aqueous solution having an acidic pH is an aqueous acetic acid solution.

[0343] In one embodiment, the aqueous solution having an acidic pH is an aqueous hydrochloric acid solution.

[0344] In one embodiment, the aqueous solution having an acidic pH is an aqueous sulfuric acid solution.

[0345] In one embodiment, the aqueous solution having an acidic pH is an aqueous phosphoric acid solution.

[0346] In one embodiment, the chemical treatment comprises exposing at least one cross-linked polysaccharide to an oxidizing agent.

[0347] In one embodiment, the oxidizing agent is selected from hydrogen peroxide and sodium hypochlorite.

[0348] In one embodiment, the oxidizing agent is hydrogen peroxide.

[0349] In one embodiment, the oxidizing agent is sodium hypochlorite.

[0350] In one embodiment, the chemical treatment comprises subjecting the cross-linked polysaccharide to a redox depolymerization process.

[0351] In one embodiment, the redox depolymerization process is carried out in the presence of at least one of the compounds Fe2+, Fe3+, ascorbic acid and H2O2.

[0352] In one embodiment, the chemical treatment of the at least one cross-linked polysaccharide is performed at room temperature without heating.

[0353] In one embodiment, the chemical treatment of the at least one cross-linked polysaccharide is performed under heating.

[0354] In one embodiment, the chemical treatment of the at least one cross-linked polysaccharide is performed at low temperature using a cooling system.

[0355] In one embodiment, step e) of cleaving the glycosidic bonds of at least one cross-linked polysaccharide is performed by treatment with at least one enzyme.

[0356] In one embodiment, step e) of cleaving glycosidic bonds of at least one cross-linked polysaccharide is performed by treating with a mixture of enzymes.

[0357] In one embodiment, step e) of cleaving the glycosidic bonds carried out on at least one cross-linked polysaccharide is carried out by treatment with at least one hydrolase type enzyme.

[0358] In one embodiment, step e) of cleaving the glycosidic bonds carried out on at least one cross-linked polysaccharide is carried out by treatment with at least one lyase type enzyme.

[0359] In one embodiment, the enzyme used in step e) for cleaving glycosidic bonds applied to at least one cross-linked polysaccharide is selected from the group of hyaluronidases.

[0360] In one embodiment, the hyaluronidase used in step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide is chosen from the enzymes HYAL1 and HYAL2.

[0361] In one embodiment, the hyaluronidase used in step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide is HYAL1.

[0362] In one embodiment, the hyaluronidase used in step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide is HYAL2.

[0363] In one embodiment, the enzyme used in step e) of cleaving glycosidic bonds applied to at least one cross-linked polysaccharide is an enzyme selected from the group of bacterial enzymes.

[0364] In one embodiment, the enzyme used in step e) for cleaving glycosidic bonds applied to at least one cross-linked polysaccharide is an enzyme selected from the group of bacterial enzymes including N-acetylhexosaminidase.

[0365] In one embodiment, the N-acetylhexosaminidase is selected from the group consisting of chondroitinase ABC (CASE) and chondroitinase AC.

[0366] In one embodiment, the N-acetylhexosaminidase is chondroitinase ABC.

[0367] In one embodiment, the N-acetylhexosaminidase is chondroitinase AC.

[0368] In one embodiment, step e) of cleaving the glycosidic bonds of at least one cross-linked polysaccharide is performed by radiation treatment.

[0369] In one embodiment, the radiation treatment comprises exposing at least one cross-linked polysaccharide to gamma radiation.

[0370] In one embodiment, the radiation treatment comprises exposing at least one cross-linked polysaccharide to beta radiation.

[0371] In one embodiment, the radiation treatment comprises exposing at least one cross-linked polysaccharide to a beam of accelerated electrons (e-beam).

[0372] In one embodiment, step e) of cleaving the glycosidic bonds of at least one cross-linked polysaccharide is performed by heat treatment.

[0373] In one embodiment, the heat treatment is a step of exposing the at least one cross-linked polysaccharide to a temperature of at least 60° C. for at least 2 hours.

[0374] In one embodiment, the heat treatment is a step of exposing the at least one cross-linked polysaccharide to moist heat.

[0375] In one embodiment, the heat treatment comprising exposing the at least one cross-linked polysaccharide to moist heat is performed by steam autoclaving.

[0376] In one embodiment, steam autoclaving is performed at between 120°C and 130°C.

[0377] In one embodiment, steam autoclaving is performed with a final time equivalent (F0) of at least 30 minutes.

[0378] In one embodiment, steam autoclaving is performed with a final time equivalent (F0) of at least 50 minutes.

[0379] In one embodiment, steam autoclaving is performed with a final time equivalent (F0) of at least 90 minutes.

[0380] In one embodiment, the thermal treatment comprises exposing the at least one cross-linked polysaccharide to dry heat.

[0381] In one embodiment, step e) of cleaving glycosidic bonds of at least one cross-linked polysaccharide is performed by high pressure treatment.

[0382] In one embodiment, step e) of cleaving glycosidic bonds is performed on at least one purified cross-linked polysaccharide.

[0383] In one embodiment, step e) of cleaving glycosidic bonds is performed on at least one unpurified cross-linked polysaccharide.

[0384] In one embodiment, the process for preparing the composition according to the invention is characterized in that, after step e), a purification step of the macromolecular compound obtained is carried out.

[0385] In one embodiment, the step of purifying the macromolecular compound is a step of eliminating the cross-linking agent.

[0386] In one embodiment, the process for preparing the composition according to the invention is characterized in that, after step e), a purification step is carried out by dialysis.

[0387] In one embodiment, the method for preparing the composition according to the invention is characterized in that, after step e), a step of purifying the macromolecular compound solution is carried out by dialysis using a solution selected from phosphate buffer (eg PBS) and water or a dialysis solvent.

[0388] In one embodiment, the method for preparing a composition according to the invention is characterized in that, after step e), a step of purifying the solution of the macromolecular compound is carried out by dialysis using a solution or a dialysis solvent selected from a phosphate buffer (e.g. PBS) and water, the solution or the dialysis solvent also comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0389] In one embodiment, the process for preparing the composition according to the invention is characterized in that, after step e), a step of purification of the solution of the macromolecular compound is carried out by washing.

[0390] In one embodiment, the step of purifying the solution of the macromolecular compound is performed by washing with a solution or solvent selected from phosphate buffered saline (eg PBS) and water.

[0391] In one embodiment, the step of purifying the solution of the macromolecular compound is performed by washing with a solution or solvent selected from phosphate buffered saline (eg PBS) and water, said solution or solvent also comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0392] In one embodiment, the process for preparing a composition according to the invention is characterized in that, after step e), a purification step is carried out by tangential filtration.

[0393] In one embodiment, the purification step by tangential filtration comprises at least one ultrafiltration step and at least one diafiltration step.

[0394] In one embodiment, the purification step by tangential filtration comprises at least one ultrafiltration step.

[0395] In one embodiment, the purification step by tangential filtration comprises at least one diafiltration step.

[0396] In one embodiment, the composition of the solution of the macromolecular compound is modified during the purification step by tangential filtration.

[0397] In one embodiment, the composition of the solution of the macromolecular compound is concentrated during the purification step by tangential filtration.

[0398] In one embodiment, the process for preparing the composition according to the invention is characterized in that, after step e), a purification step is carried out by filtration on at least one hollow fiber membrane.

[0399] In one embodiment, the process for preparing the composition according to the invention is characterized in that, after step e), a purification step of the macromolecular compound is carried out by precipitation in salt form.

[0400] In one embodiment, the process for preparing the composition according to the invention is characterized in that, after step e), a purification step of the macromolecular compound is carried out by precipitation from the solution obtained after addition of PBS or NaCl salt.

[0401] In one embodiment, the process for preparing a composition according to the invention is characterized in that, after step e), a purification step of the macromolecular compound is carried out by precipitation from the solution obtained after addition of PBS or NaCl salt, said solution also comprising at least one active ingredient, for example an antioxidant and / or a local anesthetic.

[0402] In one embodiment, the step of purifying the macromolecular compound by precipitation is performed by adding at least one organic solvent to the macromolecular compound solution.

[0403] In one embodiment, the at least one organic solvent added to the macromolecular compound solution in the precipitation purification step is selected from the group of alcohols.

[0404] In one embodiment, during the precipitation purification step, ethanol is added to the macromolecular compound solution.

[0405] In one embodiment, the step of purifying the macromolecular compound by precipitation occurs in a hydroalcoholic mixture.

[0406] In one embodiment, the precipitated macromolecular compound is recovered by filtration.

[0407] In one embodiment, the macromolecular compound precipitated and recovered by filtration is washed.

[0408] In one embodiment, the macromolecular compound precipitated and recovered by filtration is dried.

[0409] In one embodiment, the macromolecular compound precipitated and recovered by filtration is dried under ambient atmosphere.

[0410] In one embodiment, the macromolecular compound precipitated and recovered by filtration is dried under vacuum.

[0411] The method for preparing the composition according to the present invention further comprises at least one step of filtering the aqueous macromolecular compound solution produced in step e) on a membrane having a porosity of 0.22 μm, step g), the filtering step being performed on the aqueous macromolecular compound solution or on a sample of the aqueous macromolecular compound solution.

[0412] In one embodiment, the filtration step performed on a membrane with a porosity of 0.22 μm is a terminal sterilization step.

[0413] In one embodiment, the filtration step on a membrane with a porosity of 0.22 μm is performed once.

[0414] In one embodiment, the filtration step on a membrane with a porosity of 0.22 μm is performed twice.

[0415] In one embodiment, the filtration step on a membrane with a porosity of 0.22 μm is performed three times.

[0416] In one embodiment, the filtration step performed on a membrane with a porosity of 0.22 μm is a means of controlling the process during the process or "in process control" (IPC) and is performed on a sample of the aqueous solution of the macromolecular compound obtained after step e) of the method.

[0417] In one embodiment, after the sample dilution step, the sample of the aqueous solution of the macromolecular compound obtained after step e) of the method is subjected to filtration IPC on a membrane having a porosity of 0.22 μm.

[0418] In one embodiment, if step e) of cleaving the glycosidic bonds is carried out by steam autoclaving, the process for preparing the composition according to the invention does not comprise a filtration step of the macromolecular compound solution on a membrane with a porosity of 0.22 μm.

[0419] In one embodiment, if step e) of cleaving the glycosidic bonds is performed by steam autoclaving, the method for preparing the composition comprises the steps of obtaining a sample of the aqueous solution of the macromolecular compound after step e) and testing the sample for filterability on a membrane with a porosity of 0.22 μm (IPC).

[0420] In one embodiment, the composition according to the invention is characterized by a G' at 1 Hz less than or equal to 300 Pa.

[0421] In one embodiment, the composition according to the invention is characterized by a G' at 1 Hz less than or equal to 150 Pa.

[0422] In one embodiment, the composition according to the invention is characterized by a G' at 1 Hz less than or equal to 100 Pa.

[0423] In one embodiment, the composition according to the invention is characterized by a G' at 1 Hz less than or equal to 85 Pa.

[0424] In one embodiment, the composition according to the invention is characterized by a G' at 1 Hz less than or equal to 70 Pa.

[0425] In one embodiment, the composition according to the invention is characterized by a Tan Δ (Tn δ) greater than or equal to 1.00.

[0426] In one embodiment, the composition according to the invention is characterized by a Tan Δ(Tn δ) of 1.00 to 1.80 (1.00≤Tan Δ(Tn δ)≤1.80).

[0427] In one embodiment, the composition according to the invention is characterized by a Tan Δ (Tn δ) of 1.00 to 1.50 (1.00≤Tan Δ (Tn δ)≤1.50).

[0428] In one embodiment, the composition according to the invention is characterized by a Tan Δ (Tn δ) of 1.00 to 1.30 (1.00≤Tan Δ (Tn δ)≤1.30).

[0429] The invention also relates to a process for the preparation of a formulation comprising a composition according to the invention.

[0430] In one embodiment, the method for preparing a formulation from the composition of the invention further comprises at least one dilution or dissolution step.

[0431] In one embodiment, the diluting or dissolving step is performed by adding water or a saline solution, such as a phosphate buffered solution, such as PBS.

[0432] In one embodiment, the diluting or dissolving step is performed by adding water or a saline solution, such as a phosphate buffered solution, such as PBS, which also comprises at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0433] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a macromolecular compound concentration of 2 to 200 mg / g relative to the total mass of the formulation.

[0434] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a macromolecular compound concentration of 2 to 75 mg / g relative to the total mass of the formulation.

[0435] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a macromolecular compound concentration of 5 to 50 mg / g relative to the total mass of the formulation.

[0436] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a macromolecular compound concentration of 10 to 40 mg / g relative to the total mass of the formulation.

[0437] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a concentration of the macromolecular compound of about 100 mg / g relative to the total mass of the formulation.

[0438] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a concentration of the macromolecular compound of about 80 mg / g relative to the total mass of the formulation.

[0439] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a concentration of the macromolecular compound of about 60 mg / g relative to the total mass of the formulation.

[0440] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a concentration of the macromolecular compound of about 40 mg / g relative to the total mass of the formulation.

[0441] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a concentration of the macromolecular compound of about 20 mg / g relative to the total mass of the formulation.

[0442] In one embodiment, the method for preparing a formulation comprising a composition according to the invention further comprises at least one dilution or dissolution step to obtain a concentration of the macromolecular compound of about 10 mg / g relative to the total mass of the formulation.

[0443] In one embodiment, the process for preparing a composition according to the invention further comprises at least one step of adding at least one active ingredient.

[0444] In one embodiment, at least one active ingredient is added in powder form.

[0445] In one embodiment, at least one active ingredient is added in the form of a solution or a suspension.

[0446] In one embodiment, at least one active ingredient is added in the form of a solution or suspension in a solvent or solution selected from the group consisting of: water, saline solution, such as phosphate buffered solution, such as PBS.

[0447] In one embodiment, at least one active ingredient is added before step e) of the process for preparing a composition according to the invention.

[0448] In one embodiment, at least one active ingredient is added after step e) of the process for preparing a composition according to the invention.

[0449] In one embodiment, at least one active ingredient is added before step g) of the method according to the composition of the invention.

[0450] In one embodiment, at least one active ingredient is added after step g) of the process according to the composition of the invention.

[0451] In one embodiment, the preparation obtained by adding at least one active ingredient after step g) of the process according to the invention is sterilized by filtration through a membrane with a porosity of 0.22 μm.

[0452] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one active ingredient selected from local anesthetics, vitamin C derivatives, anti-inflammatory substances, antioxidants and mixtures thereof.

[0453] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic.

[0454] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic to obtain a local anesthetic concentration ranging from 0.1% to 5% relative to the total mass of the formulation.

[0455] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic to obtain a local anesthetic concentration ranging from 0.1% to 4% relative to the total mass of the formulation.

[0456] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic to obtain a local anesthetic concentration of 0.1% to 2% relative to the total mass of the formulation.

[0457] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic to obtain a local anesthetic concentration ranging from 0.1% to 1% relative to the total mass of the formulation.

[0458] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic to obtain a local anesthetic concentration ranging from 0.1% to 0.5% relative to the total mass of the formulation.

[0459] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic to obtain a local anesthetic concentration of about 0.3% relative to the total mass of the formulation.

[0460] In one embodiment, the local anesthetic is selected from the group of aminoesters.

[0461] In one embodiment, the aminoester is selected from the group comprising procaine, benzocaine, chloroprocaine and tetracaine in base or salt form (eg hydrochloride form).

[0462] In one embodiment, the local anesthetic is selected from the group of aminoamides.

[0463] In one embodiment, the aminoamide is selected from the group comprising lidocaine, mepivacaine, prilocaine, articaine, atocaine, bupivacaine, etidocaine and ropivacaine in base or salt form (eg hydrochloride form).

[0464] In one embodiment, the local anesthetic is selected from the group of amino ethers.

[0465] In one embodiment, the aminoether is selected from the group comprising diaminocaine and pramoxine in base or salt form, such as in hydrochloride or cyclamate form.

[0466] In one embodiment, the amino ether is selected from lidocaine, mepivacaine, and salts thereof, and isolated isomers thereof.

[0467] In one embodiment, the aminoether is lidocaine.

[0468] In one embodiment, the aminoether is lidocaine or a pharmaceutically acceptable salt thereof.

[0469] In one embodiment, the amino ether is lidocaine hydrochloride.

[0470] In one embodiment, the aminoether is mepivacaine.

[0471] In one embodiment, the aminoether is mepivacaine or a pharmaceutically acceptable salt thereof.

[0472] In one embodiment, the aminoether is selected from racemic mepivacaine hydrochloride, racemic mepivacaine hydrochloride, (r)-mepivacaine hydrochloride, (s)-mepivacaine hydrochloride, (r)-mepivacaine and (s)-mepivacaine, or one of their pharmaceutically acceptable salts.

[0473] In one embodiment, the amino ether is mepivacaine hydrochloride.

[0474] In one embodiment, the amino ether is (r)-mepivacaine hydrochloride.

[0475] In one embodiment, the amino ether is (S)-mepivacaine hydrochloride.

[0476] In one embodiment, the amino ether is racemic mepivacaine hydrochloride.

[0477] In one embodiment, the amino ether is (r)-mepivacaine.

[0478] In one embodiment, the amino ether is (s)-mepivacaine.

[0479] In one embodiment, the amino ether is racemic mepivacaine.

[0480] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one aminoether local anesthetic selected from lidocaine, mepivacaine, and mixtures thereof.

[0481] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being lidocaine.

[0482] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being lidocaine, so as to obtain a lidocaine concentration ranging from 0.1% to 5% relative to the total mass of the formulation.

[0483] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being lidocaine, so as to obtain a lidocaine concentration ranging from 0.1% to 4% relative to the total mass of the formulation.

[0484] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being lidocaine, so as to obtain a lidocaine concentration ranging from 0.1% to 2% relative to the total mass of the formulation.

[0485] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being lidocaine, so as to obtain a lidocaine concentration ranging from 0.1% to 1% relative to the total mass of the formulation.

[0486] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being lidocaine, so as to obtain a lidocaine concentration ranging from 0.1% to 0.5% relative to the total mass of the formulation.

[0487] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being lidocaine, to obtain a local anesthetic concentration of about 0.3% relative to the total mass of the formulation.

[0488] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being mepivacaine.

[0489] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being mepivacaine, to obtain a mepivacaine concentration ranging from 0.1% to 5% relative to the total mass of the formulation.

[0490] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being mepivacaine, to obtain a mepivacaine concentration ranging from 0.1% to 4% relative to the total mass of the formulation.

[0491] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being mepivacaine, to obtain a mepivacaine concentration ranging from 0.1% to 2% relative to the total mass of the formulation.

[0492] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being mepivacaine, to obtain a mepivacaine concentration ranging from 0.1% to 1% relative to the total mass of the formulation.

[0493] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being mepivacaine, to obtain a mepivacaine concentration ranging from 0.1% to 0.5% relative to the total mass of the formulation.

[0494] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being mepivacaine, to obtain a local anesthetic concentration of about 0.3% relative to the total mass of the formulation.

[0495] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one local anesthetic, said at least one local anesthetic being dyclonine in base or salt form (eg in hydrochloride form).

[0496] In one embodiment, the method for preparing a formulation comprising at least one composition according to the present invention further comprises at least one step of adding at least one local anesthetic selected from chlorobutanol, guanidine and polidocanol.

[0497] In one embodiment, the local anesthetic is chlorobutanol.

[0498] In one embodiment, the local anesthetic is guanfilcarnitine.

[0499] In one embodiment, the local anesthetic is polidocanol.

[0500] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance.

[0501] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance selected from steroidal anti-inflammatory substances and non-steroidal anti-inflammatory substances.

[0502] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance selected from the group consisting of non-steroidal anti-inflammatory substances.

[0503] In one embodiment, the method for preparing a formulation comprising at least one composition according to the present invention further comprises at least one step of adding at least one anti-inflammatory substance selected from the group consisting of salicylated anti-inflammatory substances, propionic acid derivatives, indole derivatives, pyrazole derivatives, oxicams and coxibs.

[0504] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance selected from the group consisting of diclofenac, nimesulide, niflumic acid, mefenamic acid and nabumetone, alone or in a mixture.

[0505] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one salicylated anti-inflammatory substance selected from the group comprising diflunisal, benorilate and aspirin, alone or in a mixture.

[0506] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance selected from the group of propionic acid derivatives comprising alminoprofen, ketoprofen, ibuprofen, naproxen, flurbiprofen and tiaprofenic acid, alone or in a mixture.

[0507] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance selected from the group of indole derivatives comprising indomethacin, sulindac and etodolac, alone or in a mixture.

[0508] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance selected from the group of pyrazole derivatives including in particular phenylbutazone.

[0509] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance selected from the group of oxicams comprising piroxicam, tenoxicam and meloxicam, alone or in a mixture.

[0510] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance selected from the group of coxibs comprising celecoxib, etoricoxib and rofecoxib, alone or in mixture.

[0511] In one embodiment, the concentration of the non-steroidal anti-inflammatory substance in a preparation comprising at least one composition according to the invention is from 0.01 mg / g to 2000 mg / g.

[0512] In one embodiment, the concentration of the non-steroidal anti-inflammatory substance in a preparation comprising at least one composition according to the invention is from 0.1 mg / g to 1000 mg / g.

[0513] In one embodiment, the concentration of the non-steroidal anti-inflammatory substance in a preparation comprising at least one composition according to the invention is from 0.5 mg / g to 500 mg / g.

[0514] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one steroidal anti-inflammatory substance.

[0515] In one embodiment, the method for preparing a formulation comprising at least one composition according to the present invention further comprises at least one step of adding at least one steroidal anti-inflammatory substance selected from the group consisting of dexamethasone, prednisolone, corticosterone, budesonide, sulfasalazine, mesalazine, cetirizine, diphenhydramine, antipyrine, methyl salicylate, loratadine, thymol, carvacrol, bisabolol, allantoin, eucalyptol, phenazone (antipyrine), propyphenazone, alone or in a mixture.

[0516] In one embodiment, the concentration of the steroidal anti-inflammatory substance in a preparation comprising at least one composition according to the invention is from 0.01 mg / g to 2000 mg / g.

[0517] In one embodiment, the concentration of the steroidal anti-inflammatory substance in a preparation comprising at least one composition according to the invention is from 0.1 mg / g to 1000 mg / g.

[0518] In one embodiment, the concentration of the steroidal anti-inflammatory substance in a preparation comprising at least one composition according to the invention is from 0.5 mg / g to 500 mg / g.

[0519] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance chosen from sucrose octasulfate and its salts.

[0520] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-inflammatory substance chosen from sucrose octasulfate and its sodium and potassium salts.

[0521] In one embodiment, the method for preparing a formulation comprising at least one composition according to the present invention further comprises at least one step of adding at least one water-soluble salt of the anti-inflammatory substance sucrose octasulfate, wherein the water-soluble salt of sucrose octasulfate is selected from alkali metal salts, alkaline earth metal salts, silver salts, ammonium salts, and amino acid salts.

[0522] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one water-soluble salt of the anti-inflammatory substance sucrose octasulfate, wherein the water-soluble salt of sucrose octasulfate is selected from alkali metal salts or alkaline earth metal salts.

[0523] In one embodiment, the method for preparing a formulation comprising at least one composition according to the present invention further comprises at least one step of adding at least one water-soluble salt of the anti-inflammatory substance sucrose octasulfate, wherein the water-soluble salt of sucrose octasulfate is selected from the sodium salt of sucrose octasulfate or the potassium salt of sucrose octasulfate.

[0524] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antimicrobial substance.

[0525] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antimicrobial substance selected from the group consisting of gentamicin, silver sulfadiazine, metronidazole, fucidine, bacitracin, eosin, povidone iodine, copper gluconate, zinc gluconate, manganese gluconate or their salts, alone or in a mixture.

[0526] In one embodiment, the concentration of the antimicrobial substance in a formulation comprising at least one composition according to the invention is from 0.1 mg / g to 200 mg / g.

[0527] In one embodiment, the concentration of the antimicrobial substance in a formulation comprising at least one composition according to the invention is from 0.5 mg / g to 100 mg / g.

[0528] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one glycoside or glycoside derivative.

[0529] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one glycoside or glycoside derivative selected from the group consisting of D-pyranose glucoside, 1,4-glycoside, esculin, hesperidin, diosmin, arbutin, skimmin or aloin, alone or in mixture.

[0530] In one embodiment, the concentration of glycosides in a preparation comprising at least one composition according to the invention is between 0.1 mg / g and 200 mg / g.

[0531] In one embodiment, the concentration of glycosides in a preparation comprising at least one composition according to the invention is between 0.5 mg / g and 100 mg / g.

[0532] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent.

[0533] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent chosen from the macroelements.

[0534] In one embodiment, the method for preparing a formulation comprising at least one composition according to the present invention further comprises at least one step of adding at least one macroelement selected from the group consisting of iron gluconate, calcium, copper, zinc, manganese, magnesium or potassium, alone or in a mixture, trimethylsilanol, trimethylsilanolate, potassium trimethylsilanolate, methylsilanol mannuronate, sodium monoethyltrisilanol o-hydroxybenzoate.

[0535] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent chosen from essential amino acids.

[0536] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one essential amino acid selected from the group consisting of isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine, alone or in mixture.

[0537] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent chosen from semi-essential amino acids.

[0538] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one semi-essential amino acid selected from the group comprising arginine and histidine, alone or in a mixture.

[0539] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent chosen from non-essential amino acids.

[0540] In one embodiment, the method for preparing a formulation comprising at least one composition according to the present invention further comprises at least one step of adding at least one non-essential amino acid selected from the group consisting of alanine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, proline, serine, tyrosine, alone or in mixture.

[0541] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent chosen from vitamins.

[0542] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one vitamin selected from retinol, thiamine, riboflavin, niacinamide, adenine, calcium pantothenate, pyridoxine, inositol, biotin, folic acid, para-aminobenzoic acid, cobalamin, vitamin C, choline chloride, alone or in a mixture.

[0543] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent chosen from nucleic acids.

[0544] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one nucleic acid selected from deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, methylcytosine, alone or in the form of a mixture.

[0545] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent chosen from coenzymes.

[0546] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one coenzyme selected from thiamine pyrophosphate, coenzyme A, FAD, NAD, NADP, UTP, alone or in the form of a mixture.

[0547] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent selected from deoxythymidine, glutathione, sodium pyruvate, lipoic acid and putrescine, alone or in a mixture.

[0548] In one embodiment, the concentration of the moisturizing agent or tissue regenerating agent in the formulation comprising at least one composition according to the invention is from 0.01 mg / g to 500 mg / g.

[0549] In one embodiment, the concentration of the moisturizing agent or tissue regenerating agent in the formulation comprising at least one composition according to the invention is from 0.1 mg / g to 200 mg / g.

[0550] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant.

[0551] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant selected from the group of polyols.

[0552] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol chosen from mannitol, sorbitol, propylene glycol, xylitol, glycerol, maltitol, lactitol and erythritol.

[0553] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol chosen from mannitol, sorbitol, maltitol and glycerol, alone or in mixture.

[0554] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol chosen from mannitol, sorbitol and maltitol, alone or in mixture.

[0555] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol to obtain a polyol concentration ranging from 0.1 mg / ml to 50 mg / ml relative to the total mass of the formulation.

[0556] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol to obtain a polyol concentration of 5 to 40 mg / ml relative to the total mass of the formulation.

[0557] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol to obtain a polyol concentration of 10 to 40 mg / ml relative to the total mass of the formulation.

[0558] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol to obtain a polyol concentration of 20 to 40 mg / ml relative to the total mass of the formulation.

[0559] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol to obtain a polyol concentration of 30 to 40 mg / ml relative to the total mass of the formulation.

[0560] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol, said at least one polyol being mannitol.

[0561] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding mannitol to obtain a mannitol concentration of 5 to 40 mg / ml relative to the total mass of the formulation.

[0562] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding mannitol to obtain a mannitol concentration of 10 to 40 mg / ml relative to the total mass of the formulation.

[0563] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding mannitol to obtain a mannitol concentration of 20 to 40 mg / ml relative to the total mass of the formulation.

[0564] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding mannitol to obtain a mannitol concentration of 30 to 40 mg / ml relative to the total mass of the formulation.

[0565] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol, said at least one polyol being sorbitol.

[0566] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding sorbitol to obtain a sorbitol concentration of 5 to 40 mg / ml relative to the total mass of the formulation.

[0567] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding sorbitol to obtain a sorbitol concentration of 10 to 40 mg / ml relative to the total mass of the formulation.

[0568] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding sorbitol to obtain a sorbitol concentration of 20 to 40 mg / ml relative to the total mass of the formulation.

[0569] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding sorbitol to obtain a sorbitol concentration of 30 to 40 mg / ml relative to the total mass of the formulation.

[0570] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol, said at least one polyol being maltitol.

[0571] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one polyol, said at least one polyol being glycerol.

[0572] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding a mixture of mannitol and sorbitol.

[0573] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant selected from the group of vitamin C derivatives.

[0574] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant selected from the group of vitamin C derivatives comprising magnesium ascorbyl phosphate, sodium ascorbyl phosphate, ascorbic acid-2-glucoside and mixtures thereof.

[0575] In one embodiment, the at least one vitamin C derivative is magnesium ascorbyl phosphate.

[0576] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant chosen from the group of vitamin E derivatives and tocopherol.

[0577] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant chosen from the group of carotenoids and retinoids and derivatives thereof.

[0578] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant selected from the group consisting of carotenoids and retinoids and their derivatives comprising retinol, retinoic acid, retinal, retinol esters and carotenes.

[0579] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant selected from the group of tripeptides.

[0580] In one embodiment, the tripeptide mimetic is glutathione.

[0581] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant selected from the group consisting of different forms of coenzyme Q10, ubiquinone or ubiquinol.

[0582] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one vitamin.

[0583] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one vitamin selected from the group consisting of retinol, thiamine, riboflavin, niacinamide, dexpenthenol, pyridoxine, ascorbic acid, ergocalciferol, tocopherol, biotin and folic acid, alone or in mixture.

[0584] In one embodiment, the vitamin concentration in a preparation comprising at least one composition according to the invention is from 0.01 mg / g to 200 mg / g.

[0585] In one embodiment, the vitamin concentration in a preparation comprising at least one composition according to the invention is from 0.01 mg / g to 100 mg / g.

[0586] In one embodiment, the concentration of vitamins in a preparation comprising at least one composition according to the invention is from 0.5 mg / g to 50 mg / g.

[0587] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one amino acid.

[0588] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one amino acid chosen from essential amino acids, semi-essential amino acids and non-essential amino acids, alone or in mixture.

[0589] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one amino acid selected from the group of essential amino acids.

[0590] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one essential amino acid selected from the group consisting of isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine, alone or in mixture.

[0591] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one amino acid selected from the group of semi-essential amino acids.

[0592] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one semi-essential amino acid selected from the group comprising arginine and histidine, alone or in a mixture.

[0593] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one amino acid selected from the group of non-essential amino acids.

[0594] In one embodiment, the method for preparing a formulation comprising at least one composition according to the present invention further comprises at least one step of adding at least one non-essential amino acid selected from the group consisting of alanine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, proline, serine, tyrosine, alone or in mixture.

[0595] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one amino acid selected from the group comprising hydroxyproline, taurine and ornithine, alone or in mixture.

[0596] In one embodiment, the concentration of amino acids in a formulation comprising at least one composition according to the invention is from 0.01 mg / g to 150 mg / g.

[0597] In one embodiment, the concentration of amino acids in a formulation comprising at least one composition according to the invention is from 0.01 mg / g to 100 mg / g.

[0598] In one embodiment, the concentration of amino acids in a formulation comprising at least one composition according to the invention is from 0.5 mg / g to 50 mg / g.

[0599] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one vasoconstrictor.

[0600] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one vasoconstrictor selected from the group consisting of naphazoline, epinephrine, methoxyamine, methylnorepinephrine, norepinephrine, oxymethazoline, phenylephrine, pseudoephedrine, synephrine, cirazoline and xylometazoline.

[0601] In one embodiment, the concentration of the vasoconstrictor in a formulation comprising at least one composition according to the invention is between 0.01 mg / g and 3 mg / g.

[0602] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one vasodilator.

[0603] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one vasodilator selected from the group comprising adenosine, niacin, minoxidil and diazoxide, alone or in a mixture.

[0604] In one embodiment, the concentration of the vasodilator in a formulation comprising at least one composition according to the invention is between 0.01 mg / g and 10 mg / g.

[0605] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-hemorrhagic or hemostatic agent.

[0606] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one anti-hemorrhagic or hemostatic agent selected from the group comprising aminocaproic acid or tranexamic acid, alone or in a mixture.

[0607] In one embodiment, the concentration of the anti-hemorrhagic or hemostatic agent in a formulation comprising at least one composition according to the invention is from 0.01 mg / g to 5 mg / g.

[0608] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant and at least one local anesthetic.

[0609] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant selected from the group of polyols and at least one local anesthetic selected from the group of aminoamides.

[0610] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant, at least one vitamin and at least one tissue regeneration agent.

[0611] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one amino acid, at least one vitamin and at least one tissue regeneration agent.

[0612] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one antioxidant, at least one amino acid, at least one vitamin and at least one tissue regeneration agent.

[0613] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one active ingredient chosen from heat-sensitive active ingredients.

[0614] In one embodiment, at least one heat-sensitive active ingredient is added after step e) of the process for preparing at least one composition according to the invention.

[0615] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding so-called heat-sensitive active ingredients of natural origin.

[0616] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention also comprises at least one step of adding a so-called thermosensitive active ingredient of synthetic origin.

[0617] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one thermosensitive active ingredient selected from peptides, hormones, proteins, growth factors, antibodies and vitamins, alone or in a mixture.

[0618] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one thermosensitive active ingredient chosen from peptides.

[0619] In one embodiment, the added heat-sensitive active ingredient is a peptide.

[0620] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one thermosensitive active ingredient chosen from hormones.

[0621] In one embodiment, the added heat-sensitive active ingredient is a hormone.

[0622] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one thermosensitive active ingredient chosen from proteins.

[0623] In one embodiment, the added heat-sensitive active ingredient is a protein.

[0624] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one thermosensitive active ingredient chosen from growth factors.

[0625] In one embodiment, the added thermosensitive active ingredient is a growth factor.

[0626] In one embodiment, the method for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one thermosensitive active ingredient selected from antibodies.

[0627] In one embodiment, the added heat-sensitive active ingredient is an antibody.

[0628] In one embodiment, the process for preparing a formulation comprising at least one composition according to the invention further comprises at least one step of adding at least one heat-sensitive active ingredient chosen from vitamins.

[0629] In one embodiment, the vitamins are selected from the group comprising retinol, thiamine, riboflavin, niacinamide, dexpanthenol, pyridoxine, ascorbic acid, ergocalciferol, tocopherol, biotin and folic acid, alone or in admixture.

[0630] The invention also relates to preparations comprising at least one composition according to the invention.

[0631] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is between 2 mg / g and 100 mg / g relative to the total mass of the formulation.

[0632] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is between 2 mg / g and 75 mg / g relative to the total mass of the formulation.

[0633] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is between 5 mg / g and 50 mg / g relative to the total mass of the formulation.

[0634] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is between 10 mg / g and 40 mg / g relative to the total mass of the formulation.

[0635] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is about 100 mg / g relative to the total mass of the formulation.

[0636] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is about 80 mg / g relative to the total mass of the formulation.

[0637] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is about 60 mg / g relative to the total mass of the formulation.

[0638] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is about 40 mg / g relative to the total mass of the formulation.

[0639] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is about 20 mg / g relative to the total mass of the formulation.

[0640] In one embodiment, the formulation is characterized in that the concentration of the macromolecular compound is about 10 mg / g relative to the total mass of the formulation.

[0641] In one embodiment, the formulation is characterized in that it is injectable.

[0642] In one embodiment, the formulation is characterized in that it is sterile.

[0643] In one embodiment, the formulation is characterized in that it is injectable and sterile.

[0644] The preparations obtained on the basis of the compositions according to the invention have numerous applications.

[0645] Among medical applications, we would list, for example, injections to replace defective biological fluids (e.g. injections in joints to replace synovial fluid), postoperative injections to avoid postoperative adhesions, periurethral injections to treat incontinence, and postoperative injections for presbyopia. Among aesthetic applications, we would list, for example, injections for filling wrinkles, fine lines and skin imperfections, or for increasing volume, for example, the volume of lips, cheekbones, etc.

[0646] More particularly, the applications targeted are those commonly used in the case of injectable viscoelastics and polysaccharides used or potentially useful in the following pathologies or treatments:

[0647] - Aesthetic injections of the face: to fill wrinkles, skin imperfections or volume (zygomatic bones, chin, lips);

[0648] - Body enhancement injections: breast and buttocks enlargement, G-spot enlargement, vaginoplasty, labia reconstruction, penis size increase;

[0649] - In joint surgery and dental surgery, for example for filling periodontal pockets;

[0650] - Treatment of osteoarthritis, injections into the joints to replace or supplement defective synovial fluid;

[0651] - periurethral injections for the treatment of urinary incontinence due to sphincter insufficiency;

[0652] - Postoperative injections, particularly to avoid peritoneal adhesions;

[0653] -Injections after presbyopia surgery using laser scleral incisions;

[0654] - Injection into the vitreous cavity;

[0655] - Injections during cataract surgery;

[0656] -Injections for the treatment of vaginal dryness;

[0657] -Injection into interstitial spaces;

[0658] -Injections into the genitals.

[0659] More particularly, in aesthetic surgery, the preparations obtained by the process that is the subject of the invention can be used, thanks to their viscoelastic and lasting properties:

[0660] - For filling fine, medium or deep wrinkles and is injected with a small diameter (e.g., 27 gauge) needle;

[0661] - As a plumping agent, where injection is performed using a larger diameter (eg, 22-26 gauge) and longer (eg, 30-40 mm) needle; in this case, its cohesive nature will ensure that it remains at the injection site.

[0662] These examples of use are by no means limiting, and the preparations obtained according to the method of the subject matter of the invention are more generally intended for use in:

[0663] - filling volume;

[0664] - Generate space within certain tissues, thus promoting their optimal functioning;

[0665] -Replaces defective physiological fluids.

[0666] The macromolecular compound according to the present invention is not used in a preparation as a mixture with a natural polysaccharide. In particular, the macromolecular compound according to the present invention is not used in a preparation as a mixture with hyaluronic acid or a heparin precursor.

[0667] In one embodiment, the macromolecular compounds according to the invention are used in the formulation in a mixture with at least one crosslinking polymer.

[0668] In one embodiment, the macromolecular compound according to the invention is used in a formulation in a mixture with at least one cross-linked polymer selected from the group of cross-linked polysaccharides.

[0669] In one embodiment, the macromolecular compounds according to the invention are used in a mixture with at least one cross-linked polysaccharide selected from hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparin precursors and biologically acceptable salts thereof, alone or in a mixture.

[0670] In one embodiment, the macromolecular compounds according to the invention are used alone or in a mixture with at least one other polymer for the preparation of crosslinked macromolecular compounds.

[0671] In one embodiment, the macromolecular compound according to the invention is used in a mixture with at least one polysaccharide for the preparation of a cross-linked macromolecular compound.

[0672] In one embodiment, the polysaccharides used together with the macromolecular compounds according to the present invention for preparing cross-linked macromolecular compounds are selected from hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparin precursors and biologically acceptable salts thereof, alone or in mixture, and these polysaccharides are cross-linked or not cross-linked.

[0673] In one embodiment, at least one polysaccharide used together with the macromolecular compound according to the invention for preparing a cross-linked macromolecular compound is selected from hyaluronic acid, heparin precursors and biologically acceptable salts thereof, alone or in a mixture, these polysaccharides being cross-linked or not.

[0674] In one embodiment, the polysaccharide used together with the macromolecular compound according to the present invention to prepare the cross-linked macromolecular compound is hyaluronic acid or a salt of hyaluronic acid.

[0675] In one embodiment, the polysaccharide used together with the macromolecular compound according to the present invention for preparing the cross-linked macromolecular compound is a mixture of hyaluronic acid or salts of hyaluronic acid.

[0676] In one embodiment, the polysaccharide used together with the macromolecular compound according to the present invention to prepare the cross-linked macromolecular compound is hyaluronic acid or a salt of cross-linked hyaluronic acid.

[0677] In one embodiment, the polysaccharide used together with the macromolecular compound according to the present invention to prepare the cross-linked macromolecular compound is a heparin precursor or a salt of a heparin precursor.

[0678] In one embodiment, the polysaccharide used together with the macromolecular compound according to the present invention to prepare the cross-linked macromolecular compound is a heparin precursor or a mixture of heparin precursor salts.

[0679] In one embodiment, the polysaccharide used together with the macromolecular compound according to the present invention to prepare the cross-linked macromolecular compound is a heparin precursor or a salt of a cross-linked heparin precursor.

[0680] In one embodiment, the macromolecular compound according to the invention is used to prepare at least one prodrug polymer.

[0681] In one embodiment, the macromolecular compound according to the invention is chemically modified by reaction with at least one active ingredient for the preparation of a prodrug polymer.

[0682] In one embodiment, the prodrug polymers obtained from the macromolecular compounds according to the invention are used in the field of oncology. BRIEF DESCRIPTION OF THE DRAWINGS

[0683] Figure 1 : Graph showing the evolution of the log (intrinsic viscosity - dL / g) curve as a function of log (molecular weight - Da) for each of the compositions studied in the examples.

[0684] Figure 2: Diagrams showing the network formed during the crosslinking step and its degradation during the steps of the claimed method. Diagram A represents the complete network obtained in step d) of the claimed method, the thick lines schematically representing the polysaccharide chains connected together and the thin lines schematically representing the divalent groups L generated by the crosslinking agent connected to the polysaccharide chains. Diagram B represents the cleavage of glycosidic bonds at different points of the previous network according to the claimed method, after the bond cleavage step e) represented by arrow 1. The thick lines schematically represent the broken polysaccharide chains, the dots represent the newly formed hydroxyl functional groups, and the thin lines schematically represent the divalent L groups generated by the crosslinking agent. Finally, diagram C represents the mixed polymer of macromolecular compounds (arrow 2) generated by the bond cleavage step e) according to the invention unfolding in solution; wherein the thick lines schematically represent the broken polysaccharide chains, the dots represent the newly formed hydroxyl functional groups, and the thin lines schematically represent the divalent L groups generated by the crosslinking agent. Example

[0685] As part of the examples, a number of parameters were measured.

[0686] Determination of rheological parameters G′, G″ and Tan Δ: DHR-2 Instrument TA Equipment. Cone geometry with an angle of 2° and a diameter of 40 mm. Frequency sweep oscillation method, 0.8% strain in the frequency range of 0.08 Hz to 5 Hz.

[0687] SEC determination: Viscotek GPCmax II instrument equipped with TDA305 detector (RI, RALS / LALS, viscometer) and Shodex OHpak SB-806HQ + SB-805HQ columns. The measurements were performed at 37°C in the presence of PBS analytical buffer.

[0688] Preparation of the composition according to the invention

[0689] Example 1

[0690] Injectable grade sodium hyaluronate fibers (10.4 g) with a weight average molecular weight of 1 MDa were weighed into a container. An aqueous solution of 1% sodium hydroxide in water was added to hydrate the sodium hyaluronate fibers. The reaction medium was homogenized by alternating manual mechanical stirring and standing for 50 minutes. 0.43 g BDDE was added. The reaction medium containing 10.4 g sodium hyaluronate fibers, 55.7 g sodium hydroxide and 0.43 g BDDE was homogenized again by manual mechanical stirring and then placed in a water bath at 2° C. pre-thermostatically controlled for 24 hours.

[0691] At the end of cross-linking, the reaction medium is neutralized by adding 1N HCl and phosphate buffer under mechanical stirring. The obtained gel is then dialyzed against phosphate buffer until a hyaluronic acid concentration of 27.5 mg / g is obtained. Phosphate buffer is prepared by dissolving 0.23 g NaH2PO4·2H2O, 1.12 g Na2HPO4, 42.5 g NaOH in water for injection (EPPI) added in sufficient amount to obtain 5 L buffer solution. After mechanical homogenization and deaeration, the obtained gel (TanΔ at 1 Hz is 0.54) cannot be filtered on a membrane with a porosity of 0.22 μm; it is dispensed into a 1 mL glass syringe and treated by steam sterilization at 127 ° C with F0 for 50 minutes. After this heat treatment, the syringe contains a solution of macromolecular compounds. A sample is taken from the syringe and a filterability test is performed on a membrane with a porosity of 0.22 μm using the sample. The filterability test was positive: the macromolecular compound solution could be filtered through a membrane with a porosity of 0.22 μm.

[0692] Example 2

[0693] The composition according to the invention is prepared by a method similar to that described in Example 1, but immediately after the addition of BDDE and homogenization, the reaction medium is placed in a water bath previously thermostated at 9°C for 3 hours at this temperature. As described in Example 1, before the heat treatment, the gel (which has a TanΔ of 0.68 at 1 Hz) cannot be filtered on a membrane with a porosity of 0.22 μm; it is dispensed into a syringe. After the heat treatment, the syringe contains 27.5 mg / g of a solution of the macromolecular compound. A sample is taken from the syringe and a filterability test is performed on a membrane with a porosity of 0.22 μm with this sample. The filterability test is positive: the solution of the macromolecular compound can be filtered on a membrane with a porosity of 0.22 μm.

[0694] Reference example 3

[0695] Injectable grade sodium hyaluronate fibers (10.4 g) with a weight average molecular weight of 1 MDa were weighed into a container. An aqueous solution of 1% sodium hydroxide in water was added to hydrate the sodium hyaluronate fibers. The reaction medium was homogenized by alternating manual mechanical stirring and standing for 50 minutes. Phosphate buffer was added to replace BDDE. Phosphate buffer was prepared by dissolving 0.23 g NaH2PO4·2H2O, 1.12 g Na2HPO4, 42.5 g NaOH in water for injection (EPPI) added in sufficient amount to obtain 5 L of buffer solution. The reaction medium containing 2.6 g sodium hyaluronate fibers, 13.9 g sodium hydroxide and 0.11 g buffer and homogenized by manual mechanical stirring was then placed in a pre-thermalized water bath at 9° C. for 3 hours.

[0696] At the end of this 3 hours of stirring, the reaction medium is neutralized by adding 1N HCl and phosphate buffer under mechanical stirring. The obtained gel is then dialyzed against phosphate buffer until a hyaluronic acid concentration of 27.5 mg / g is obtained. After mechanical homogenization and deaeration, the obtained gel (TanΔ at 1 Hz is 1.59) can be filtered on a membrane with a porosity of 0.22 μm; it is dispensed into a 1 mL glass syringe and treated by steam sterilization at 127 ° C with 50 minutes of F0. After this heat treatment, the syringe contains a macromolecular compound solution. A sample is taken from the syringe and a filterability test is performed on a membrane with a porosity of 0.22 μm with the sample. The filterability test is positive: the macromolecular compound solution can be filtered on a membrane with a porosity of 0.22 μm.

[0697] Properties of the composition according to the invention

[0698] The rheological properties determined for the compositions according to the invention are given in Table 1 below.

[0699]

[0700] Table 1

[0701] The composition according to the invention is filterable on a membrane with a porosity of 0.22 μm and has a Tan Δ greater than or equal to 1.00.

[0702] Hydrodynamic behavior of the composition according to the invention compared to the reference composition 3

[0703] The compositions were chromatographed by steric exclusion to determine the coefficients of the Mark-Houwink relationship which is written as:

[0704] [η] = KM a

[0705] [η] is the intrinsic viscosity.

[0706] M is the viscosity average molar mass.

[0707] This linear relationship makes it easy to determine the values ​​of coefficients a and K.

[0708] If we express log([η]) as a function of log(M), we obtain a straight line with slope a and intercept log(K).

[0709] exist Figure 1 For each of the compositions, log(intrinsic viscosity - dL / g) was plotted as a function of log(molecular weight - Da) and the linear correlation was calculated.

[0710] The measured and calculated values ​​are summarized in Table 2 below.

[0711]

[0712] Table 2

[0713] It was observed that the hyaluronic acid of the composition according to the present invention had an intrinsic viscosity and a Mark-Houwink coefficient similar to those of the hyaluronic acid of the composition of Reference Example 3 and was therefore no longer cross-linked.

[0714] Nevertheless, they present very different G' and TanΔ, suitable for the above applications.

[0715] Example 4

[0716] Injectable grade sodium hyaluronate fibers (12.5 g) with a weight average molecular weight of 1 MDa were weighed into a container. An aqueous solution of 1% sodium hydroxide in water was added to hydrate the sodium hyaluronate fibers. The reaction medium was homogenized by alternating manual mechanical stirring and standing for 50 minutes. 0.56 g BDDE was added. The reaction medium containing 12.5 g sodium hyaluronate fibers, 74.4 g sodium hydroxide and 0.56 g BDDE was homogenized again by manual mechanical stirring and then placed in a pre-thermostatically controlled water bath at 8° C. for 3 hours.

[0717] At the end of cross-linking, the reaction medium is neutralized by adding 1N HCl and phosphate buffer under mechanical stirring. The phosphate buffer is prepared by dissolving 5.40 g NaH2PO4·2H2O, 67.60 g Na2HPO4, 240 g NaOH and 4200 g mannitol in water for injection (EPPI) added in sufficient amount to obtain 120 L of buffer solution. The obtained gel is then dialyzed against a phosphate buffer containing 35 g / L of mannitol until a hyaluronic acid concentration of 27.5 mg / g is obtained.

[0718] The gel thus obtained could not be filtered on a membrane with a porosity of 0.22 μm.

[0719] The gel (which had a Tan Delta of 0.70 at 1 Hz) was then divided into aliquots.

[0720] A first aliquot of 10.0 g of the gel was acidified with 350 μL of 1 N HCl to obtain a pH of 3. It was degraded at 50° C. for 32 hours. The gel was then neutralized to pH 7.3 with 1 N NaOH. Its rheological properties were measured.

[0721] A second aliquot of the gel was exposed for 397 hours to radiation emitted by a 370 nm UV lamp at a power of 310 W. After completion of the exposure, the rheological properties of the gel obtained were measured.

[0722] A third aliquot of 10.0 g of gel was thermostated at 37° C. in a syringe. A second syringe containing 100 μL of a hyaluronidase solution at a concentration of 1750 units was thermostated at 37° C. The gel was mixed with hyaluronidase by connecting the two syringes via a double Luer connector and then homogenizing by moving back and forth between the syringes for 30 seconds. The mixture was then placed at 37° C. for 45 minutes. At the end of this enzymatic degradation, the degradation was stopped by immersion in a bath at 100° C. for 4 minutes, then brought back to 25° C. using an ice bath, and its rheological properties were measured.

[0723] The results of all rheological measurements are summarized in Table 3 below.

[0724]

[0725] Table 3

[0726] It was observed that regardless of the degradation method applied, the TanΔ(Tnδ) of the obtained compounds at 1 Hz was strictly greater than 1.00 (TanΔ(Tnδ)>1.00).

[0727] Different aliquots obtained following different degradation methods could all be filtered on a membrane with a porosity of 0.22 μm.

[0728] Example 5

[0729] The composition according to the invention is prepared by a method similar to that described in Example 4 until crosslinking. At the end of crosslinking, the reaction medium is neutralized by adding 1N HCl and phosphate buffer under mechanical stirring. The phosphate buffer is prepared by dissolving 0.23 g NaH2PO4·2H2O, 1.12 g Na2HPO4, 42.5 g NaOH in water for injection (EPPI) added in sufficient amount to obtain 5 L of buffer solution. The gel obtained is then dialyzed against phosphate buffer until a hyaluronic acid concentration of 20.0 mg / g is obtained.

[0730] The gel thus obtained could not be filtered on a membrane with a porosity of 0.22 μm.

[0731] The gel (which had a Tan Delta of 0.79 at 1 Hz) was then divided into aliquots.

[0732] A first aliquot of 10.0 g of the gel was placed in contact with a solution containing 37.5 μL of an aqueous solution of ferric sulfate heptahydrate at a concentration of 15 mg / g and 75 μL of 30% by volume hydrogen peroxide and homogenized by 60 syringe / syringe round trips to achieve free radical degradation after the Fenton reaction. After a contact time of 4 hours and 20 minutes, the gel was purified by dialysis against phosphate buffer. At the end of 2 dialysis baths lasting a total of 41 hours and 30 minutes, its rheological properties were measured.

[0733] A second aliquot of 10.0 g of the gel was acidified with 250 μL of 1 N HCl to obtain a pH of 3. It was degraded at 50° C. for 14 hours and 19 minutes. The gel was then neutralized to pH 7.3 with 1 N NaOH. Its rheological properties were measured.

[0734] A third aliquot of 10.0 g of gel was thermostated at 37° C. in a syringe. A second syringe containing 50 μL of a hyaluronidase solution at a concentration of 1750 units was thermostated at 37° C. The gel was mixed with the hyaluronidase by connecting the two syringes via a double Luer connector and then homogenizing by moving back and forth between the syringes for 30 seconds. The mixture was then placed at 37° C. for 25 minutes. At the end of this enzymatic degradation, the degradation was stopped by immersion in a bath at 100° C. for 4 minutes, then brought back to 25° C. using an ice bath, and its rheological properties were measured.

[0735] All rheological measurement results are summarized in Table 4 below.

[0736]

[0737] Table 4

[0738] It was observed that regardless of the degradation method applied, the TanΔ(Tnδ) of the obtained compounds at 1 Hz was strictly greater than 1.00 (TanΔ(Tnδ)>1.00).

[0739] Different aliquots obtained following different degradation methods could all be filtered on a membrane with a porosity of 0.22 μm.

Claims

1. A composition in the form of an aqueous solution filterable on a membrane with a porosity of 0.22 μm, comprising at least one macromolecular compound consisting of a series of identical or different polysaccharides linked together by divalent groups L.

2. The composition according to claim 1, characterized in that the macromolecular compound has the general formula I: in: -P i , P i' , P i” , P i”' , P j , P j' , P j” , P j”' , P k , P k' , P k” , P k”' , P l , P l' , P l” and P l”' are the same or different polysaccharides, - n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 2000, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2, - x, x', x", and x'" are integers greater than or equal to 0, -L is a divalent group resulting from the reaction between a crosslinker and two reactive functional groups, each of which is carried by two different polysaccharide chains, -Polysaccharide chains do not form ring structures.

3. The composition according to claim 1, characterized in that the macromolecular compound consists of a series of polysaccharides selected from the following: Hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparin precursors and biologically acceptable salts thereof, alone or in mixtures.

4. A macromolecular compound of general formula I: in: -P i , P i' , P i” , P i”' , P j , P j' , P j” , P j”' , P k , P k' , P k” , P k”' , P l , P l' , P l” and P l”' are the same or different polysaccharides, - n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' are integers from 0 to 2000, and at least one of n1, n1', n1", n1"', n2, n2', n2", n2"', n3, n3', n3", n3"', n4, n4', n4" and n4"' is greater than or equal to 2, - x, x', x", and x'" are integers greater than or equal to 0, -L is a divalent group resulting from the reaction between a crosslinker and two reactive functional groups, each of which is carried by two different polysaccharide chains, -Polysaccharide chains do not form ring structures.

5. A method for preparing the composition according to claim 1, comprising at least the following steps: a) providing at least one polysaccharide b) providing at least one cross-linking agent c) performing one or more cross-linking steps in the presence of said polysaccharide and said cross-linking agent; d) Obtaining cross-linked polysaccharides e) performing one or more steps to cleave glycosidic bonds f) Obtaining a macromolecular compound solution g) filtering the macromolecular compound solution through a membrane with a porosity of 0.22 μm.

6. The method according to claim 5, characterized in that the crosslinking agent is selected from diepoxides, trimetaphosphates, diamines, dialkoxyamines and dihydrazides.

7. Process according to any one of claims 5 and 6, characterised in that step e) of cleaving the glycosidic bonds is carried out by chemical treatment.

8. Process according to any one of claims 5 and 6, characterised in that step e) of cleaving the glycosidic bonds is carried out by heat treatment.

9. Process according to any one of claims 5 and 6, characterised in that step e) of cleaving the glycosidic bonds is carried out by radiation treatment.

10. Process according to any one of claims 5 and 6, characterised in that step e) of cleaving the glycosidic bonds is carried out by enzymatic treatment.

11. Process according to any one of claims 5 and 6, characterised in that step e) of cleaving the glycosidic bonds is carried out by high pressure treatment. 12 . The composition according to claim 1 , characterized in that Tan Δ (Tn δ) at 1 Hz is ≥ 1.

00.

13. A preparation comprising at least one composition according to any one of claims 1 to 3. The preparation according to claim 13, characterized in that the preparation further comprises at least one active ingredient.

14. Preparation according to claim 13, characterized in that the active ingredient is selected from local anesthetics, antioxidants, anti-inflammatory agents, vitamins, amino acids, vasoconstrictors, vasodilators, anti-hemorrhagic or hemostatic agents, antimicrobial agents, glycosides and their derivatives, moisturizers or tissue regeneration agents, alone or in a mixture.

Images