A composition in the form of an aqueous solution containing at least one polymer compound

Abstract

The present invention relates to a composition in an aqueous solution, said composition being filterable through a membrane with a porosity of 0.22 μm, comprising at least one polymeric compound consisting of a sequence of identical or different polysaccharides linked together by a divalent radical L, the radical L resulting from a reaction between a crosslinker and two reactive functional groups, the two reactive functional groups being generated by two polysaccharide chains, the sequence of the polysaccharide chains not forming a ring structure.

Description

[Technical field]

[0001] The present invention relates to the field of polysaccharide-based formulations used as biomaterials, in particular in the medical and cosmetic fields, for which the formulations must have optimized properties in terms of flowability, taking into account the characteristics of the medical devices used for injection, the site of treatment and the desired therapeutic or cosmetic effect. [Background technology]

[0002] Crosslinked polysaccharide-based products in gel form used for cosmetic applications have rheological properties optimized to have good stability at the injection site. The gels used are particularly robust to deformations, with the result that the loss tangent Tan Δ (Tnδ) is less than 1.00 and the elastic modulus G' is much greater than the viscous modulus G'' for a wide range of deformations. The majority of products used, particularly in gel form, have a loss tangent less than 0.50 (REF). This characteristic of gel-based products imposes certain limitations with regard to injectability, limiting the scope of use. The use of such products, especially in sensitive areas of the body or face, is delicate or even problematic. Summary of the Invention [Problem to be solved by the invention]

[0003] The present invention relates to the development of a composition in the form of an aqueous solution having very good fluidity and filterable through a membrane with a porosity of 0.22 μm, comprising at least one linear or branched polymeric compound consisting of polysaccharide sequences.

[0004] WO2020250128 filed by OFFHEALTH SPA discloses a composition that contains nanoparticles of crosslinked hyaluronic acid obtained by mechanically acting on crosslinked hyaluronic acid with a stirring system, and that can be filtered through a 0.2 μm porous membrane. These nanoparticles are suspended in the disclosed composition. Therefore, these compositions do not contain a polymer equivalent to the polymer obtained by the method of the present invention, and are compositions containing nanoparticles in a suspension, and are not compositions with viscoelasticity, so the fluidity of the composition cannot be evaluated.

[0005] WO2016030516 filed by GALDERMA SA discloses the possibility of filtration through a 0.2 μm porous membrane, and the supernatant resulting from thermal, enzymatic or radical decomposition of a gel containing hyaluronic acid. Although these compositions can be filtered through a 0.2 μm porous membrane, the fluidity of these filtrates is not equivalent to that of the compositions disclosed in the present invention, and they are not suitable for the uses mentioned in this specification.

[0006] Surprisingly, the compositions of the present invention have the significant advantage of being filterable through a membrane with a porosity of 0.22 μm, and can contain temperature-sensitive compounds, such as peptides, proteins, growth factors, antibodies or vitamins, because the compositions obtained can be sterilized by simple filtration. Existing products based on cross-linked polysaccharide gels are not compatible with compounds that are sensitive to high temperatures, since autoclaving or heat sterilization of the gel is required to avoid the risk of infection at the treated site by injection.

[0007] The present invention also relates to a composition comprising at least one polymeric compound consisting of a sequence of hyaluronic acid family, which has particularly good durability after injection and resistance to enzymatic degradation.

[0008] The compositions of the present invention also allow for the preparation of highly concentrated solutions of polysaccharides, particularly hyaluronic acid, while remaining easily injectable.

[0009] The present invention relates to a composition which, in aqueous solution, is filterable through a membrane of 0.22 μm porosity and which comprises at least one polymeric compound consisting of a sequence of identical or different polysaccharides linked by divalent radicals L.

[0010] L is a divalent radical resulting from the reaction of a crosslinker with two reactive functional groups, each of which is on two separate polysaccharide chains, and the polysaccharide chains do not form a cyclic structure.

[0011] In one embodiment, the composition of the invention is characterized in that the polymeric compound comprises a polysaccharide sequence selected from the group consisting of hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, alginic acid, xanthan, carrageenan, chitosan, chondroitin, heparosan and bioacceptable salts thereof, either alone or in mixtures.

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

[0013] In one embodiment, the composition of the invention is characterized in that the polymeric compound consists of chains of heparosan.

[0014] In one embodiment, the composition of the invention is characterized in that the polymeric compound consists of polysaccharide chains selected from the group consisting of hyaluronic acid, heparosan or reactive salts thereof.

[0015] The invention also relates to a process for the preparation of a composition comprising at least one polymeric compound consisting of sequences of polysaccharides, for example hyaluronic acid, and more particularly a preparation process which makes it possible to obtain a composition comprising at least one polymeric compound consisting of polysaccharide chains and which has particular properties, such as filterability through a membrane of 0.22 μm porosity.

[0016] The process for preparing the composition of the invention comprises at least the following steps: a) providing a polysaccharide; b) providing a cross-linking agent; c) carrying out one or more cross-linking steps in the presence of said polysaccharide and said cross-linking agent; d) obtaining cross-linked polysaccharides; e) carrying out one or more steps of cleaving glycosidic bonds; f) obtaining a polymer solution; g) Filtering the polymer solution through a membrane with a porosity of 0.22 μm.

[0017] In one embodiment, the process for preparing the composition of the invention comprises at least the following steps: a) providing a polysaccharide; b) providing a cross-linking agent; c) carrying out one or more cross-linking steps in the presence of said polysaccharide and said cross-linking agent. d) obtaining cross-linked polysaccharides; e) carrying out a step of cleaving glycosidic bonds; f) obtaining a polymer solution; g) Filtering a sample of the polymer solution through a membrane of 0.22 μm porosity. This is an Inductively Coupled Process (ICP) test.

[0018] In one embodiment, the process for preparing the composition of the invention comprises at least the following steps: a) providing a polysaccharide; b) providing a cross-linking agent; c) carrying out one or more crosslinking steps in the presence of said polysaccharide and said crosslinking agent; the solution is carried out at a concentration of at least one polysaccharide of at least 10% by weight relative to the total weight of the crosslinking reaction medium, at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being less than or equal to 10° C. for a reaction time of up to 24 hours. d) obtaining cross-linked polysaccharides; e) carrying out a step of cleaving glycosidic bonds; f) obtaining a polymer solution; g) Filtering the polymer solution through a membrane with a porosity of 0.22 μm.

[0019] In one embodiment, the process for preparing the composition of the invention comprises at least the following steps: a) providing a polysaccharide; b) providing a cross-linking agent; c) carrying out one or more crosslinking steps in the presence of said polysaccharide and said crosslinking agent; the solution is carried out at a concentration of at least one polysaccharide of at least 10% by weight relative to the total weight of the crosslinking reaction medium, at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being less than or equal to 10° C. for a reaction time of up to 24 hours. d) obtaining cross-linked polysaccharides; e) carrying out a step of cleaving glycosidic bonds; f) obtaining a polymer solution; g) Filter a sample of the polymer solution through a membrane of 0.22 μm porosity. This is an in-process control (ICP) test.

[0020] During step d), a network of different polysaccharides is obtained cross-linked by divalent radicals L coming from the cross-linking agent.

[0021] In one embodiment, the process of the invention is characterized in that the crosslinking agent is selected from the group consisting of bis-epoxides, trimetaphosphates, diamines, dialkoxyamines and dihydrazides.

[0022] In one embodiment, the process of the invention is characterized in that the cross-linking agent is 1,4-butanediol diglycidyl ether (BDDE).

[0023] In one embodiment, the process of the invention is characterized in that the cross-linking agent is a trimetaphosphate.

[0024] In one embodiment, the process of the invention is characterized in that the crosslinker is a diamine.

[0025] In one embodiment, the process of the invention is characterized in that the cross-linking agent is a dihydrazide.

[0026] In one embodiment, step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide results in the destruction of the majority of the cross-linked network formed during step c).

[0027] In one embodiment, step e) of cleaving glycosidic bonds carried out on at least one cross-linked polysaccharide results in the complete destruction of the cross-linked network formed during step c).

[0028] During this step e), only the glycosidic bonds are cleaved, except for the bond between the crosslinker and the divalent radical L resulting from the polysaccharide.

[0029] In one embodiment the inventive process is characterised in that step e) of cleaving the glycosidic bonds is carried out by chemical treatment.

[0030] In one embodiment the inventive method is characterised in that step e) of cleaving the glycosidic bonds is carried out by heat treatment.

[0031] In one embodiment, the inventive process is characterized in that the heat treatment of step e) to cleave the glycosidic bonds is carried out by steam autoclave.

[0032] In one embodiment the method of the invention is characterized in that step e) of cleaving glycosidic bonds is carried out by radiation treatment.

[0033] In one embodiment the method of the invention is characterised in that step e) of cleaving the glycosidic bonds is carried out by enzymatic treatment.

[0034] In one embodiment the inventive method is characterized in that step e) of cleaving glycosidic bonds is carried out by high pressure treatment.

[0035] The invention also relates to polymeric compounds of general formula (I). TIFF2024544753000001.tif72170 ((P in the formula i , Pi' , 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 between 0 and 2000, and at least one of n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'' and n4''' is 2 or greater; x, x', x'', and x'' are integers equal to or greater than 0, L is a divalent radical resulting from the reaction of the crosslinker with two reactive functional groups, each of which is contained on two separate polysaccharide chains; Polysaccharide chains do not form ring structures.)

[0036] In one embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 1500, and at least one of n1, n1', n1'', n1'''', n2, n2', n2'', n2''', n3, n3', n3'', n3'''', n4, n4', n4'', and n4'''' is 2 or greater.

[0037] In one embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 1000, and at least one of n1, n1', n1'', n1'''', n2, n2', n2'', n2'''', n3, n3', n3'', n3'''', n4, n4', n4'', and n4'''' is 2 or greater.

[0038] In one embodiment, n1, n1', n1", n1'", n2, n2', n2", n2'", n3, n3', n3", n3'", n4, n4', n4", and n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 500, and at least one of n1, n1', n1'', n1'''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' is 2 or greater.

[0039] In one embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 250, and at least one of n1, n1', n1'', n1'''', n2, n2', n2'', n2'''', n3, n3', n3'', n3'''', n4, n4', n4'', and n4'''' is 2 or greater.

[0040] In one embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 150, and at least one of n1, n1', n1'', n1'''', n2, n2', n2'', n2'''', n3, n3', n3'', n3'''', n4, n4', n4'', and n4'''' is 2 or greater.

[0041] In one embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 100, and at least one of n1, n1', n1'', n1'''', n2, n2', n2'', n2'''', n3, n3', n3'', n3'''', n4, n4', n4'', and n4'''' is 2 or greater.

[0042] In one embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 50, and at least one of n1, n1', n1'', n1'''', n2, n2', n2'', n2'''', n3, n3', n3'', n3'''', n4, n4', n4'', and n4'''' is 2 or greater.

[0043] In one embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 30, and at least one of n1, n1', n1'', n1'''', n2, n2', n2'', n2'''', n3, n3', n3'', n3'''', n4, n4', n4'', and n4'''' is 2 or greater.

[0044] In one embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''',x, x', x'', and x''' are 0 and n3 is 2. This embodiment represents the simplest form of a polymeric compound and can be represented by the following formula (LP k )-(LP k ).

[0045] In another embodiment, n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are 0 and n2 and n3 are 1. This embodiment also represents a simple form of the polymeric compound and can be represented by the following formula (LP k )-(LP j ).

[0046] The polymeric compound of the invention is therefore a mixed polymer containing alternating polysaccharides linked by divalent radicals originating from the crosslinker. Indeed, during step e) of cleavage of the glycosidic bonds, the crosslinker becomes a "linker" and loses its role as a "crosslinker".

[0047] The invention relates to a composition which, in aqueous solution, is filterable through a membrane of 0.22 μm porosity and which comprises at least one polymeric compound consisting of a sequence of identical or different polysaccharides linked by a divalent radical L (linker), characterized in that the polymeric compound has the general formula (I): TIFF2024544753000002.tif72170 (In the formula, 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 between 0 and 2000, at least one of n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'' and n4''' is 2 or greater, and x, x', x'' and x'' are integers 0 or greater; L is a divalent radical resulting from the reaction of the crosslinker with two reactive functional groups, each of which is contained on two separate polysaccharide chains; Polysaccharide chains do not form ring structures.)

[0048] In one embodiment, n1, n1', n1", n1'", n2, n2', n2", n2'", n3, n3', n3", n3'", n4, n4', n4", and n4'" are integers between 0 and 1500, and at least one of n1, n1', n1", n1'", n2, n2', n2", n2'", n3, n3', n3", n3'", n4, n4', n4", and n4'" is 2 or greater.

[0049] When the polymeric compound of the invention in the dry state is hydrated with an aqueous solution, an aqueous solution of the polymeric compound is obtained.

[0050] In particular, the polymeric compounds of the invention do not have swelling capabilities.

[0051] In the sense of the invention, swelling ability refers to the fact that a compound in dry form placed in the presence of an aqueous solution, for example a saline solution, has the ability to form a gel of a significant volume, more than 80% of the volume of which is restricted by filtration through a membrane of 0.22 μm porosity. Gel formation is characterized by the presence of two phases in the medium: a gel or "soft solid" and a solution or supernatant.

[0052] In the presence of water, the polymeric compounds of the invention do not form gels.

[0053] In particular, the polymeric compounds of the invention do not form hydrogels. In the context of this specification, the term "hydrogel" means a gel consisting of a three-dimensional network of at least one compound, capable of absorbing a large amount of water or an aqueous solution and having specific flow properties, in particular with regard to viscosity and viscoelasticity.

[0054] In one embodiment, said polysaccharide in step a) is selected from the group of glycosaminoglycans (GAGs).

[0055] In one embodiment, said polysaccharide in step a) is selected from the group of chemically modified, oxidized or substituted polysaccharides.

[0056] In one embodiment, the polysaccharide is selected from the group of glycosaminoglycans (GAGs), such as chondroitin, keratan, heparin, heparosan or hyaluronic acid and mixtures thereof.

[0057] In one embodiment, the polysaccharide is selected from the group consisting of hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, oxidized cellulose, alginic acid, xanthan, carrageenan, chitosan, chondroitin, heparosan, and bioacceptable salts thereof, either alone or in mixtures.

[0058] In one embodiment, the polysaccharide is hyaluronic acid or one of its bioacceptable salts, either alone or in mixture.

[0059] In the present context, hyaluronic acid or one of its bioacceptable salts, either alone or in mixture, is preferred.

[0060] In one embodiment, the polysaccharide is selected from the group consisting of hyaluronic acid, sodium hyaluronate, and mixtures thereof.

[0061] In one embodiment, the polysaccharide is hyaluronic acid.

[0062] In one embodiment, the polysaccharide is selected from the group consisting of sodium hyaluronate and potassium hyaluronate.

[0063] In one embodiment, the polysaccharide is sodium hyaluronate.

[0064] In the present context, sodium hyaluronate is a particularly preferred polysaccharide.

[0065] In one embodiment, the polysaccharide is hyaluronic acid or one of its salts chemically modified by substitution.

[0066] In one embodiment, the polysaccharide is hyaluronic acid or one of its salts substituted by groups that confer lipophilicity or hydrophilicity, such as the substituted hyaluronic acids described in patent application FR2 983483 of the applicant.

[0067] In one embodiment, the polysaccharide is heparosan, or one of its bioacceptable salts, alone or in mixture.

[0068] In the context of this specification, heparosan or one of its bioacceptable salts, either alone or in mixture, is preferred.

[0069] In one embodiment, the polysaccharide is selected from the group consisting of heparosan, sodium heparosan, or a mixture thereof.

[0070] In one embodiment, the polysaccharide is heparosan.

[0071] In one embodiment, the polysaccharide is selected from the group consisting of sodium heparosan or potassium heparosan.

[0072] In one embodiment, the polysaccharide is sodium heparosan.

[0073] In the present context, sodium heparosan is a particularly preferred polysaccharide.

[0074] In one embodiment, the polysaccharide is a chemically modified heparosan, or one of its salts.

[0075] In one embodiment, the polysaccharide is selected from the group consisting of cellulose or cellulose derivatives.

[0076] In one embodiment, the polysaccharide is selected from the group consisting of cellulose cellulose derivatives, in particular hydroxypropyl cellulose, hydroxypropyl methyl cellulose, ethyl methyl cellulose and carboxymethyl cellulose.

[0077] In one embodiment, the polysaccharide is cellulose.

[0078] In one embodiment, the polysaccharide is a cellulose derivative.

[0079] In one embodiment, said polysaccharide in step a) of the process for preparing the composition of the invention is a mixture of polysaccharides.

[0080] In the present context, all the polysaccharides mentioned are brought together in a mixture during step a), whether they are of the same nature (for example a mixture of hyaluronic acid of different molecular weights) or of different nature (for example a mixture of hyaluronic acid and chitosan). There may be mutual crosslinks between the different polysaccharides during the crosslinking step.

[0081] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of hyaluronic acids or salts of hyaluronic acids.

[0082] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of two hyaluronic acids or salts of hyaluronic acids.

[0083] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of three hyaluronic acids or salts of hyaluronic acids.

[0084] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of four hyaluronic acids or salts of hyaluronic acids.

[0085] In one embodiment, the process for the preparation of the inventive composition is characterized in that in step a) said polysaccharides are heparosans or a mixture of salts of heparosans.

[0086] In one embodiment, the process for the preparation of the inventive composition is characterized in that in step a) said polysaccharide is a mixture of two heparosans or salts of heparosans.

[0087] In one embodiment, the process for the preparation of the inventive composition is characterized in that in step a) said polysaccharide is a mixture of three heparosans or salts of heparosans.

[0088] In one embodiment, the process for the preparation of the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of four heparosans or salts of heparosans.

[0089] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharides are a mixture of hyaluronic acids and heparosans or salts thereof.

[0090] In one embodiment, the process for the preparation of the inventive composition is characterized in that in step a) said polysaccharide is a mixture of one hyaluronic acid and one heparosan or a salt thereof.

[0091] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of two hyaluronic acids and one heparosan or a salt thereof.

[0092] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of one hyaluronic acid and two heparosans or salts thereof.

[0093] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of two hyaluronic acids and two heparosans or salts thereof.

[0094] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharides are a mixture of hyaluronic acids or salts thereof and celluloses.

[0095] In one embodiment, the process for preparing the inventive composition is characterized in that in step a) said polysaccharide is a mixture of one hyaluronic acid or a salt thereof and one cellulose.

[0096] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of two hyaluronic acids or salts thereof and one cellulose.

[0097] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of one type of hyaluronic acid or a salt thereof and two types of celluloses.

[0098] In one embodiment, the process for preparing the composition of the invention is characterized in that in step a) said polysaccharide is a mixture of two hyaluronic acids or salts thereof and two celluloses.

[0099] In the context of this specification, Mw or "molecular weight" refers to the weight average molecular weight of a polysaccharide, measured in Daltons.

[0100] In one embodiment, the polysaccharide or one of its salts has a molecular weight of 0.01 MDa to 10 MDa (0.01 MDa≦Mw≦10 MDa).

[0101] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 0.01 MDa and 8 MDa.

[0102] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 0.01 MDa and 5 MDa.

[0103] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 0.01 MDa and 3.5 MDa.

[0104] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 0.5 MDa and 3.5 MDa.

[0105] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 2.75 MDa and 3.25 MDa.

[0106] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 0.75 MDa and 1.25 MDa.

[0107] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 2 MDa and 5 MDa.

[0108] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 2 MDa and 4 MDa.

[0109] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 0.5 MDa and 2 MDa.

[0110] In one embodiment, the polysaccharide or one of its salts has a molecular weight between 0.5 MDa and 1.5 MDa.

[0111] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 0.01 MDa and 10 MDa.

[0112] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 0.01 MDa and 5 MDa.

[0113] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 0.01 MDa and 3.5 MDa.

[0114] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 0.5 MDa and 3.5 MDa.

[0115] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 2.75 MDa and 3.25 MDa.

[0116] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 0.75 MDa and 1.25 MDa.

[0117] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 2 MDa and 5 MDa.

[0118] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 2 MDa and 4 MDa.

[0119] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 0.5 MDa and 2 MDa.

[0120] In one embodiment, the hyaluronic acid or one of its salts has a molecular weight between 0.5 MDa and 1.5 MDa.

[0121] In one embodiment, the heparosan or one of its salts has a molecular weight between 0.01 MDa and 10 MDa.

[0122] In one embodiment, the heparosan or one of its salts has a molecular weight between 0.01 MDa and 5 MDa.

[0123] In one embodiment, the heparosan or one of its salts has a molecular weight between 0.02 MDa and 3 MDa.

[0124] In one embodiment, the heparosan or one of its salts has a molecular weight between 0.02 MDa and 2 MDa.

[0125] For the preparation of the inventive composition, one or more cross-linking steps c) are carried out between the polysaccharide and at least one cross-linking agent.

[0126] In one embodiment, the process for the preparation of the inventive composition is characterized in that the mixing of said polysaccharide and at least one crosslinking agent is carried out in a solvent.

[0127] In one embodiment, the process for the preparation of the inventive composition is characterized in that the at least one crosslinker in step c) is selected from the group consisting of ethylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether (BDDE), bis-epoxy such as 1,2,3,4-diepoxybutane or 1,2,7,8-diepoxyoctane, dialkylsulfones, divinylsulfones, formaldehyde, epichlorohydrin or glutaraldehyde, carbodiimides such as 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC), trimetaphosphates such as sodium trimetaphosphate, calcium trimetaphosphate or barium trimetaphosphate.

[0128] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinker in step c) is selected from the group consisting of ethylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether (BDDE), bis-epoxy such as 1,2,3,4-diepoxybutane or 1,2,7,8-diepoxyoctane, trimetaphosphates such as sodium trimetaphosphate, calcium trimetaphosphate or barium trimetaphosphate.

[0129] In one embodiment, the process for preparing the inventive composition is characterized in that said at least one crosslinker in step c) is selected from the group consisting of ethylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether (BDDE), 1,2,3,4-diepoxybutane or 1,2,7,8-diepoxyoctane.

[0130] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinker in step c) is selected from the group consisting of bifunctional polyethylene glycols (PEGs) containing one epoxide at each end of the polymer chain.

[0131] In one embodiment, the process for preparing the inventive composition is characterized in that said at least one crosslinker in step c) is selected from the group consisting of difunctional polyethylene glycols (PEGs) comprising one epoxy group at each end of the polymer chain and selected from the group consisting of polyethylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether and polytetramethylene glycol diglycidyl ether.

[0132] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinking agent in step c) is selected from the group consisting of trimetaphosphates, such as sodium trimetaphosphate, calcium trimetaphosphate or barium trimetaphosphate.

[0133] In one embodiment, the process for preparing the inventive composition is characterized in that said at least one crosslinker in step c) is selected from the group consisting of epoxides, such as 1,4-butanediol diglycidyl ether (BDDE), epihalohydrins, divinyl sulfone (DVS).

[0134] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinker in step c) is divinylsulfone (DVS).

[0135] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinking agent in step c) is 1,4-butanediol diglycidyl ether (BDDE).

[0136] In the present context, BDDE is particularly preferred.

[0137] In one embodiment, the process for preparing the inventive composition is characterized in that said at least one crosslinker in step c) is selected from compounds containing two functional groups selected from the group consisting of an amine function, alkoxyamines and hydrazides.

[0138] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinking agent in step c) is selected from diamines.

[0139] In one embodiment, the diamine used as one cross-linking agent in step c) of the process for preparing the inventive composition is selected from the group consisting of diaminotrehalose, diaminosucrose, chitobiose, diaminolactose and diaminoraffinose.

[0140] In one embodiment, the diamine used as cross-linking agent in step c) of the process for preparing the inventive composition is diaminotrehalose.

[0141] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinking agent in step c) is selected from dialkoxyamines.

[0142] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinking agent in step c) is selected from dihydrazides.

[0143] In one embodiment, the dihydrazide used as crosslinker in step c) of the process for preparing the inventive composition is adipic dihydrazide.

[0144] In one embodiment, the cross-linking step or each step of cross-linking step c) requires that said polysaccharide is pre-dissolved.

[0145] In one embodiment, the polysaccharide is added in solid form to the solution to dissolve said polysaccharide.

[0146] In one embodiment, a solution is added to the polysaccharide in a solid state to dissolve said polysaccharide.

[0147] In one embodiment the cross-linking step or each step of cross-linking step c) does not require that the polysaccharide is in solution.

[0148] The polysaccharide is dissolved by adding water or an aqueous saline solution, such as a phosphate buffer solution, e.g. PBS, or by adding a sodium hydroxide solution or an acidic solution to obtain a pH appropriate for carrying out the crosslinking step or steps.

[0149] In one embodiment, the polysaccharide is dissolved by adding water or an aqueous saline solution, such as a phosphate buffer, e.g. PBS, further comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic, or by adding a sodium hydroxide solution or an acidic solution to obtain a pH appropriate for carrying out the crosslinking step or steps.

[0150] In one embodiment, the process for the preparation of the inventive composition is characterized in that at the latest during step c) a step of adjusting the pH to a crosslinking pH is carried out.

[0151] The adjustment of the pH is preferably carried out by adding an inorganic acid solution, such as hydrochloric acid, or preferably an inorganic base, such as sodium oxide or potassium oxide, said acids and bases being added in predetermined amounts making it possible to obtain the target crosslinking pH.

[0152] In one embodiment, a step of adjusting the pH during the or each crosslinking step c) is carried out to a crosslinking pH compatible with said crosslinking agent.

[0153] In one embodiment, a step of adjusting the pH is carried out at the latest during the crosslinking step or during each step of crosslinking step c) to obtain a pH of the crosslinking above 10.

[0154] In one embodiment, a step of adjusting the pH is carried out at the latest during the crosslinking step or during each step of crosslinking step c) to obtain a pH of the crosslinking of less than 3.

[0155] In one embodiment, during the crosslinking step or each step of crosslinking step c), aqueous sodium hydroxide solution with a concentration of up to 0.25 N or 1% by weight is added to obtain the targeted crosslinking pH.

[0156] In one embodiment, the step of adjusting the pH at the latest during the crosslinking step or during each step of crosslinking step c) is carried out at a crosslinking pH above 10, said crosslinker being BDDE.

[0157] In one embodiment the step of adjusting the pH at the latest during the crosslinking step or during each step of crosslinking step c) is carried out at a crosslinking pH of less than 3, said crosslinker being BDDE.

[0158] In one embodiment, a step of adjusting the pH to a crosslinking pH is carried out at the latest during the crosslinking step or during each step of crosslinking step c), said crosslinking pH being greater than 10.

[0159] Crosslinking begins when three conditions are met: the presence of polysaccharide, the presence of a crosslinker, and a reaction medium with the appropriate pH.

[0160] In one embodiment the crosslinking step or each step of crosslinking step c) is characterized in that initiation of crosslinking occurs by addition of said crosslinking agent.

[0161] In one embodiment the cross-linking step or each step of cross-linking step c) is characterized in that initiation of cross-linking occurs by addition of said polysaccharide.

[0162] In one embodiment the crosslinking step or each step of crosslinking step c) is characterized in that the initiation of crosslinking occurs by application of a crosslinking pH.

[0163] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step c) a step of adjusting the pH to a pH between 6 and 8 is carried out.

[0164] Depending on the pH of the reaction medium at the end of the crosslinking reaction of step c), the adjustment of the pH is preferably carried out by adding a solution of an inorganic acid, such as hydrochloric acid, or preferably an inorganic base, such as sodium oxide or potassium oxide, the said acids and bases being added in amounts making it possible to obtain a pH between 6 and 8.

[0165] In one embodiment, the process for the preparation of the inventive composition is characterized in that the step of adjusting the pH to a pH between 6 and 8 after step c) is carried out by adding at least one acid, which is hydrochloric acid (HCl).

[0166] The process for preparing the composition of the invention is characterized in that the crosslinking step or each step of the crosslinking step c) is carried out at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being below 50°C.

[0167] The process for preparing the composition of the invention is characterized in that the crosslinking step or each step of the crosslinking step c) is carried out at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being below 30°C.

[0168] The process for preparing the composition of the invention is characterized in that the crosslinking step or each step of the crosslinking step c) is carried out at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being below 20°C.

[0169] The process for preparing the composition of the invention is characterized in that the crosslinking step or each step of the crosslinking step c) is carried out at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being below 15°C.

[0170] The process for preparing the composition of the invention is characterized in that the crosslinking step or each step of the crosslinking step c) is carried out at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being below 10°C.

[0171] The process for preparing the composition of the invention is characterized in that the crosslinking step or each step of the crosslinking step c) is carried out at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being below 5°C.

[0172] The solidification temperature of a reaction medium means the temperature at which the medium becomes solid. For aqueous media, this temperature is 0° C. or slightly lower depending on the salt concentration of said medium.

[0173] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being between the solidification temperature and 10°C.

[0174] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature or at a temperature which varies linearly or stepwise, said constant or variable temperature being between the solidification temperature and 5°C.

[0175] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature of 50°C.

[0176] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature of 30°C.

[0177] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature of 20°C.

[0178] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature of 15°C.

[0179] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature of 9°C.

[0180] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature of 5°C.

[0181] In one embodiment, the crosslinking step or each step of crosslinking step c) is carried out at a constant temperature of 2°C.

[0182] In one embodiment, a cooling step is carried out at the crosslinking temperature before the or each crosslinking step c).

[0183] In one embodiment, the process for preparing the inventive composition is characterized in that the at least one crosslinking agent in 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 less than or equal to 50° C.

[0184] In one embodiment, the process for preparing the inventive composition is characterized in that the at least one crosslinking agent in 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 less than or equal to 30° C.

[0185] In one embodiment, the process for preparing the inventive composition is characterized in that the at least one crosslinking agent in 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 less than or equal to 20° C.

[0186] In one embodiment, the process for preparing the inventive composition is characterized in that the at least one crosslinking agent in 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 less than or equal to 15° C.

[0187] In one embodiment, the process for preparing the inventive composition is characterized in that the at least one crosslinking agent in 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 less than or equal to 10° C.

[0188] In one embodiment, the process for preparing the inventive composition is characterized in that the at least one crosslinking agent in 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 less than or equal to 5° C.

[0189] In one embodiment, the process for preparing the inventive composition is characterized in that the at least one crosslinking agent in 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 less than or equal to 2° C.

[0190] In one embodiment, the process for the preparation of the inventive composition is characterized in that the crosslinking step c) has a duration between 10 minutes and 26 hours.

[0191] In one embodiment, the process for the preparation of the inventive composition is characterized in that the crosslinking step c) has a duration between 10 minutes and 18 hours.

[0192] In one embodiment, the process for the preparation of the inventive composition is characterized in that the crosslinking step c) has a duration between 10 minutes and 12 hours.

[0193] In one embodiment, the process for the preparation of the inventive composition is characterized in that the crosslinking step c) has a duration between 10 minutes and 5 hours.

[0194] In one embodiment, the process for the preparation of the inventive composition is characterized in that the crosslinking step c) has a duration between 10 minutes and 3 hours.

[0195] In one embodiment, the process for the preparation of the inventive composition is characterized in that the crosslinking step c) has a duration between 30 minutes and 3 hours.

[0196] In one embodiment, the process for the preparation of the inventive composition is characterized in that the crosslinking step c) is carried out at 2° C. for 24 hours.

[0197] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinking agent in step c) is 1,4-butanediol diglycidyl ether (BDDE) and said step c) is carried out at a pH above 10 and at 2° C. for 24 hours.

[0198] In one embodiment, the process for the preparation of the inventive composition is characterized in that the crosslinking step c) is carried out at 9° C. for 3 hours.

[0199] In one embodiment, the process for the preparation of the inventive composition is characterized in that said at least one crosslinking agent in step c) is 1,4-butanediol diglycidyl ether (BDDE) and said step c) is carried out at a pH above 10 and at 9° C. for 3 hours.

[0200] If several successive crosslinkings are carried out in step c) of the process for the preparation of the inventive composition, the times stated are the total times (sum of the times of the successive crosslinkings).

[0201] In one embodiment, during step c), carrying out the cross-linking step in the presence of said polysaccharide and said cross-linking agent occurs in a medium in which said polysaccharide is hydrated or swells by addition of water or an aqueous saline solution, such as a phosphate buffer, e.g. PBS.

[0202] In one embodiment, during step c), carrying out 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 or swells by addition of water or an aqueous saline solution, such as a phosphate buffer, e.g. PBS, further comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0203] In step c) of the process for preparing the inventive composition, the crosslinking rate (X) can be theoretically calculated using the following formula: X = moles of cross-linker introduced into the reaction medium / moles of repeating units (disaccharide pattern) introduced into the reaction medium

[0204] Thus, for example, if a medium contains 100 disaccharide patterns and said medium also contains 10 molecules of cross-linker, the cross-linking percentage (X) is: X=10 / 100=0.1. This percentage of cross-linking is therefore independent of the degree of polymerization, the molecular weight of the polysaccharide selected, and the proportion of cross-linker that actually reacts with at least one functional group of the polysaccharide. It is theoretically determined by considering only the amount of cross-linker and repeating units mixed.

[0205] In one embodiment, the crosslinking percentage X is between 0.001 and 0.20.

[0206] In one embodiment, the crosslinking percentage X is between 0.01 and 0.15.

[0207] In one embodiment, the crosslinking percentage X is between 0.01 and 0.12.

[0208] In one embodiment, the crosslinking percentage X is between 0.03 and 0.10.

[0209] In one embodiment, the crosslinking percentage X is between 0.04 and 0.08.

[0210] Crosslinking can also be evaluated a posteriori (after crosslinking) by the degree of modification (Mod), which thus takes into account the proportion of crosslinker that actually reacts with at least one functional group of the polysaccharide as opposed to the crosslinking rate X.

[0211] The degree of modification can be expressed as follows: Mod(%)=number of moles of crosslinker linked to at least one disaccharide pattern by at least one covalent bond / number of moles of repeating units in the reaction medium*100

[0212] The repeating units (monomers) are a disaccharide pattern when the polysaccharide is hyaluronic acid.

[0213] The determination of the numerator and denominator values ​​is known to the skilled artisan depending on the selected polysaccharide and the selected crosslinker. For example, in the specific case of preparations based on hyaluronic acid crosslinked with BDDE, the method described in the publication of L. Nord, A. Emilson, C. Sturesson and AH Kenne, Degree of Modification of Hyaluronic Acid Dermal Fillers, Congress EADV 18th Congress, Berlin, 2009, can be used.

[0214] In the specific case of a formulation based on hyaluronic acid crosslinked with BDDE, the degree of modification can be expressed as follows: Mod(%) = moles of BDDE linked to at least one disaccharide pattern of hyaluronic acid by at least one covalent bond / moles of repeating units (disaccharide pattern of hyaluronic acid) in the reaction medium * 100

[0215] For example, a formulation based on hyaluronic acid cross-linked with BDDE and having a Mod of 1% means that the formulation has one BDDE molecule (singly or doubly linked) per 100 disaccharide patterns.

[0216] In one embodiment, the degree of modification of said cross-linked polysaccharides is less than 5%.

[0217] In one embodiment, the degree of modification of the cross-linked polysaccharides is less than 4%.

[0218] In one embodiment, the degree of modification of the cross-linked polysaccharides is less than 3%.

[0219] In one embodiment, the degree of modification of the cross-linked polysaccharide is less than 2%.

[0220] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of polysaccharide is, for example, at least 1% by weight, relative to the total weight of the cross-linking reaction medium.

[0221] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of polysaccharide is, for example, at least 5% by weight, relative to the total weight of the cross-linking reaction medium.

[0222] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of polysaccharide is, for example, at least 10% by weight, relative to the total weight of the cross-linking reaction medium.

[0223] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of polysaccharide is, for example, at least 15% by weight, relative to the total weight of the cross-linking reaction medium.

[0224] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of the polysaccharide is, for example, between 1% and 60% by weight, relative to the total weight of the cross-linking reaction medium.

[0225] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of the polysaccharide is, for example, between 5% and 60% by weight relative to the total weight of the cross-linking reaction medium.

[0226] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of the polysaccharide is, for example, between 10% and 60% by weight relative to the total weight of the cross-linking reaction medium.

[0227] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of polysaccharide is for example between 10% and 40% by weight relative to the total weight of the reaction medium.

[0228] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of the polysaccharide is, for example, between 10% and 25% by weight relative to the total weight of the reaction medium.

[0229] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the concentration of polysaccharide is for example between 15% and 25% by weight relative to the total weight of the reaction medium.

[0230] In one embodiment, hyaluronic acid or one of its bioacceptable salts is present in the crosslinking reaction medium, alone or in a mixture, and during step c), the crosslinking step in the presence of said crosslinker, the concentration of hyaluronic acid is between 1% and 60% by weight, relative to the total weight of the reaction medium.

[0231] In one embodiment, hyaluronic acid or one of its bioacceptable salts is present in the crosslinking reaction medium, alone or in a mixture, and during step c), the crosslinking step in the presence of said crosslinker, the concentration of hyaluronic acid is between 5% and 60% by weight relative to the total mass of the reaction medium.

[0232] In one embodiment, hyaluronic acid or one of its bioacceptable salts is present in the crosslinking reaction medium, alone or in a mixture, and during step c), the crosslinking step in the presence of said crosslinker, the concentration of hyaluronic acid is between 10% and 60% by weight relative to the total mass of the reaction medium.

[0233] In one embodiment, hyaluronic acid or one of its bioacceptable salts is present, alone or in a mixture, and during step c), the crosslinking step, in the presence of said crosslinker, the concentration of hyaluronic acid is between 10% and 40% by weight relative to the total mass of the reaction medium.

[0234] In one embodiment, hyaluronic acid or one of its bioacceptable salts is present, alone or in a mixture, and during step c), the crosslinking step, in the presence of said crosslinker, the concentration of hyaluronic acid is between 10% and 25% by weight relative to the total mass of the reaction medium.

[0235] In one embodiment, hyaluronic acid or one of its bioacceptable salts is present, alone or in a mixture, and during step c), the crosslinking step, in the presence of said crosslinker, the concentration of hyaluronic acid is between 15% and 25% by weight relative to the total mass of the reaction medium.

[0236] In one embodiment, heparosan or one of its bioacceptable salts is present in the crosslinking reaction medium, alone or in a mixture, and during step c), the crosslinking step in the presence of said crosslinker, the concentration of heparosan is between 1% and 60% by weight, relative to the total mass of the reaction medium.

[0237] In one embodiment, heparosan or one of its bioacceptable salts is present in the crosslinking reaction medium, alone or in a mixture, and during step c), the crosslinking step in the presence of said crosslinker, the concentration of heparosan is between 5% and 60% by weight, relative to the total mass of the reaction medium.

[0238] In one embodiment, heparosan or one of its bioacceptable salts is present in the crosslinking reaction medium, alone or in a mixture, and during step c), the crosslinking step in the presence of said crosslinker, the concentration of heparosan is between 10% and 60% by weight, relative to the total mass of the reaction medium.

[0239] In one embodiment, heparosan or one of its bioacceptable salts is present, alone or in a mixture, and in the presence of said crosslinker, during step c), the crosslinking step, the concentration of heparosan is between 10% and 40% by weight, relative to the total weight of the reaction medium.

[0240] In one embodiment, heparosan or one of its bioacceptable salts is present, alone or in a mixture, and during step c), the crosslinking step, in the presence of said crosslinker, the concentration of heparosan is between 10% and 25% by weight, relative to the total mass of the reaction medium.

[0241] In one embodiment, heparosan or one of its bioacceptable salts is present, alone or in a mixture, and during step c), the crosslinking step, in the presence of said crosslinker, the concentration of heparosan is between 15% and 25% by weight relative to the total mass of the reaction medium.

[0242] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the cross-linking reaction medium comprises sodium hydroxide (NaOH).

[0243] In one embodiment, during step c), crosslinking in the presence of said polysaccharide and said crosslinking agent, the concentration of sodium oxide is between 0.5% and 1.5% by weight relative to the total weight of the reaction medium.

[0244] In one embodiment, during step c), crosslinking in the presence of said polysaccharide and said crosslinking agent, the concentration of sodium oxide is between 0.5% and 1% by weight relative to the total weight of the reaction medium.

[0245] In one embodiment, during step c), crosslinking in the presence of said polysaccharide and said crosslinking agent, the concentration of sodium oxide is between 0.7% and 0.9% by weight relative to the total weight of the reaction medium.

[0246] In one embodiment, during step c), the cross-linking step in the presence of said polysaccharide and said cross-linking agent, the cross-linking reaction medium comprises hydrochloric acid (HCl).

[0247] In one embodiment, during step c), cross-linking in the presence of said polysaccharide and said cross-linking agent, the concentration of hydrochloric acid is between 0.001 M and 0.05 M in the reaction medium.

[0248] In one embodiment, the cross-linking step c) comprises at least one step of activating chemical functional groups that the polysaccharide comprises.

[0249] In one embodiment, the cross-linking step c) comprises at least one step of activating the carboxylic acid groups that the polysaccharide comprises.

[0250] In one embodiment, the step of activating the carboxylic acid groups contained in the polysaccharide is carried out using a coupling agent used in peptide chemistry.

[0251] In one embodiment, the step of activating the carboxylic acid groups contained in the polysaccharide is carried out using a coupling agent selected from the group of triazine, carbodiimide, imidazolium type coupling agents as well as Oxyma and COMU in peptide chemistry.

[0252] In one embodiment, the triazine-type coupling agent is selected from the group consisting of 4-(4,6-dimethoxy[1,3,5]triazin-2-yl)-4-methylmorpholinium chloride (DMTMM) and 2-chloro-4,6-dimethoxy-1,3,5-triazine (CMT).

[0253] In one embodiment, the coupling agent is 4-(4,6-dimethoxy[1,3,5]triazin-2-yl)-4-methylmorpholinium chloride (DMTMM).

[0254] In one embodiment, the process for the preparation of the inventive composition is characterized in that before step d), a step of removing said crosslinking agent is carried out.

[0255] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a step of purifying the cross-linked polysaccharide is carried out.

[0256] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a step of purifying the cross-linked polysaccharide by dialysis is carried out.

[0257] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a step of purifying the crosslinked polysaccharide is carried out by dialysis with a solution or dialysis solvent selected from the group consisting of phosphate buffer, e.g. PBS and water.

[0258] In one embodiment, the process for the preparation of the inventive composition is characterized in that prior to step d), a step of purifying the crosslinked polysaccharide is carried out by dialysis with a solution or dialysis solvent selected from the group consisting of phosphate buffer, e.g. PBS and water, further comprising at least one active ingredient, e.g. an antioxidant and / or a local anesthetic.

[0259] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a step of purifying the crosslinked polysaccharide by washing is carried out.

[0260] In one embodiment, the step of purification of the cross-linked polysaccharide is carried out by washing with a solution or solvent selected from the group consisting of a phosphate buffer, e.g., PBS, and water.

[0261] In one embodiment, the step of purification of the crosslinked polysaccharide is further carried out by washing with a solution or solvent selected from the group consisting of phosphate buffer, e.g., PBS, and water, which further contains at least one active ingredient, e.g., an antioxidant and / or a local anesthetic.

[0262] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a purification step of the crosslinked polysaccharide is carried out by precipitation in salified conditions.

[0263] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a purification step of the crosslinked polysaccharide is carried out by precipitation from a solution obtained after addition of PBS or NaCl salt.

[0264] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a purification step of the crosslinked polysaccharide is carried out by precipitation from a solution obtained after addition of PBS or NaCl salt and further comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0265] In one embodiment, the step of purification of the cross-linked polysaccharide by precipitation is carried out by adding at least one organic solvent to the cross-linked polysaccharide.

[0266] In one embodiment, the at least one organic solvent added to the crosslinked polysaccharide in the precipitation purification step is selected from the group of alcohols.

[0267] In one embodiment, the alcohol added to the cross-linked polysaccharide in the precipitation purification step is ethanol.

[0268] In one embodiment, the purification step of the cross-linked polysaccharide by precipitation occurs in an aqueous alcohol mixture.

[0269] In one embodiment, the precipitated cross-linked polysaccharide is isolated by filtration.

[0270] In one embodiment, the cross-linked polysaccharide that has been precipitated and isolated by filtration is washed.

[0271] In one embodiment, the cross-linked polysaccharide that has been precipitated and isolated by filtration is dried.

[0272] In one embodiment, the cross-linked polysaccharide that has been precipitated and isolated by filtration is dried under air and at room temperature.

[0273] In one embodiment, the cross-linked polysaccharide that has been precipitated and isolated by filtration is dried under vacuum and at room temperature.

[0274] In one embodiment, the cross-linked polysaccharide is lyophilized.

[0275] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a step of diluting the crosslinked polysaccharide is carried out with a solution or solvent selected from the group consisting of phosphate buffers, e.g. PBS, and water.

[0276] In one embodiment, the process for the preparation of the inventive composition is characterized in that, prior to step d), a step of diluting the crosslinked polysaccharide is carried out with a solution or solvent selected from the group consisting of phosphate buffer, e.g. PBS and water, and water, further comprising at least one active ingredient, e.g. an antioxidant and / or a local anesthetic.

[0277] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out such that the concentration of the cross-linked polysaccharide is between 2 mg / g and 200 mg / g relative to the total weight of the composition.

[0278] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out so as to obtain a concentration of the cross-linked polysaccharide between 2 mg / g and 75 mg / g relative to the total weight of the composition.

[0279] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out so as to obtain a concentration of the cross-linked polysaccharide between 5 mg / g and 50 mg / g relative to the total weight of the composition.

[0280] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out so as to obtain a concentration of the cross-linked polysaccharide ranging from 10 mg / g to 40 mg / g relative to the total weight of the composition.

[0281] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out such that the concentration of the cross-linked polysaccharide is about 100 mg / g relative to the total weight of the composition.

[0282] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out such that the concentration of the cross-linked polysaccharide is about 80 mg / g relative to the total weight of the composition.

[0283] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out such that the concentration of the cross-linked polysaccharide is about 60 mg / g relative to the total weight of the composition.

[0284] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out such that the concentration of the cross-linked polysaccharide is about 40 mg / g relative to the total weight of the composition.

[0285] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out such that the concentration of the cross-linked polysaccharide is about 20 mg / g relative to the total weight of the composition.

[0286] In one embodiment, the process for preparing the inventive composition is characterized in that, prior to step d), a step of diluting the cross-linked polysaccharide is carried out such that the concentration of the cross-linked polysaccharide is about 10 mg / g relative to the total weight of the composition.

[0287] In one embodiment, the polysaccharide obtained in step d) has a tangent Tan Δ(Tnδ)≧0.50 before the bond cleavage step in step e).

[0288] In one embodiment, the polysaccharide obtained in step d) has a tangent Tan Δ(Tnδ)≧0.70 before the bond cleavage step in step e).

[0289] The process for the preparation of the inventive composition may comprise at least one glycosidic bond cleavage step, step e), carried out on at least one crosslinked polysaccharide to obtain the inventive polymeric compound.

[0290] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is based on a controlled degradation mechanism.

[0291] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by a treatment selected from the group of chemical treatment, enzymatic treatment, radiation treatment and heat treatment.

[0292] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by chemical treatment.

[0293] In one embodiment, the chemical treatment comprises exposing the at least one cross-linked polysaccharide to an aqueous solution at a basic pH.

[0294] In one embodiment, the aqueous solution at basic pH is an aqueous solution of sodium hydroxide (sodium oxide).

[0295] In one embodiment, the chemical treatment comprises exposing the at least one cross-linked polysaccharide to an aqueous solution having an acidic pH.

[0296] In one embodiment, the aqueous solution at an acidic pH comprises an acid selected from the group consisting of acetic acid, hydrochloric acid, sulfuric acid, and phosphoric acid.

[0297] In one embodiment, the aqueous solution at an acidic pH is an aqueous solution of acetic acid.

[0298] In one embodiment, the aqueous solution with an acidic pH is an aqueous hydrochloric acid solution.

[0299] In one embodiment, the aqueous solution at an acidic pH is an aqueous solution of sulfuric acid.

[0300] In one embodiment, the aqueous solution at an acidic pH is an aqueous phosphoric acid solution.

[0301] In one embodiment, the chemical treatment comprises exposing the at least one cross-linked polysaccharide to an oxidizing agent.

[0302] In one embodiment, the oxidizing agent is selected from the group consisting of hydrogen peroxide and sodium hypochlorite.

[0303] In one embodiment, the oxidizing agent is hydrogen peroxide.

[0304] In one embodiment, the oxidizing agent is sodium hypochlorite.

[0305] In one embodiment, the chemical treatment comprises subjecting the cross-linked polysaccharide to an oxidative-reductive depolymerization process.

[0306] In one embodiment, the oxidative-reductive depolymerization method is carried out in the presence of at least one of the compounds Fe2+, Fe3+, ascorbic acid, and H2O2.

[0307] In one embodiment, the chemical treatment of the at least one cross-linked polysaccharide is carried out at room temperature, without heating.

[0308] In one embodiment, the chemical treatment of the at least one cross-linked polysaccharide is carried out by heating.

[0309] In one embodiment, the chemical treatment of the at least one cross-linked polysaccharide is carried out at low temperature and using a cooling system.

[0310] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by treatment with at least one enzyme.

[0311] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by treatment with a mixture of enzymes.

[0312] In one embodiment the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by treatment with at least one hydrolase type enzyme.

[0313] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by treatment with at least one lyase-type enzyme.

[0314] In one embodiment, the enzyme used in glycosidic bond cleavage step e) on at least one cross-linked polysaccharide is selected from the group of hyaluronan degrading enzymes.

[0315] In one embodiment, the hyaluronan degrading enzyme used in the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is selected from the group consisting of the enzymes HYAL1 and HYAL2.

[0316] In one embodiment, the hyaluronan degrading enzyme used in the glycosidic bond cleavage step e) carried out on at least one cross-linked polysaccharide is HYAL1.

[0317] In one embodiment, the hyaluronan degrading enzyme used in the glycosidic bond cleavage step e) carried out on at least one cross-linked polysaccharide is HYAL2.

[0318] In one embodiment, the enzyme used in glycosidic bond cleavage step e) on the at least one cross-linked polysaccharide is an enzyme selected from the group of bacterial enzymes.

[0319] In one embodiment, the enzyme used in the glycosidic bond cleavage step e) on the at least one cross-linked polysaccharide is an enzyme selected from the group of bacterial enzymes consisting of N-acetylhexosaminidases.

[0320] In one embodiment, the N-acetylhexosaminidase enzyme is selected from the group consisting of chondroitinase ABC (CASE) and chondroitinase AC.

[0321] In one embodiment, the N-acetylhexosaminidase is chondroitinase ABC (CASE).

[0322] In one embodiment, the N-acetylhexosaminidase is chondroitinase AC.

[0323] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by radiation treatment.

[0324] In one embodiment, the radiation treatment comprises exposing the at least one cross-linked polysaccharide to gamma radiation.

[0325] In one embodiment, the radiation treatment comprises exposing the at least one cross-linked polysaccharide to beta radiation.

[0326] In one embodiment, the radiation treatment comprises exposing the at least one cross-linked polysaccharide to an accelerated electron beam (e-beam).

[0327] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by heat treatment.

[0328] In one embodiment, the heat treatment comprises exposing the at least one crosslinked polysaccharide to a temperature of at least 60° C. for at least 2 hours.

[0329] In one embodiment, the heat treatment is a step of exposing the at least one crosslinked polysaccharide to moist heat.

[0330] In one embodiment, this heat treatment, which comprises exposing the at least one crosslinked polysaccharide to moist heat, is carried out in a steam autoclave.

[0331] In one embodiment, the steam autoclave is run at between 120°C and 130°C.

[0332] In one embodiment, the steam autoclaving is carried out for a final time equivalent (F0) of at least 30 minutes.

[0333] In one embodiment, the steam autoclaving is carried out for a final time equivalent (F0) of at least 50 minutes.

[0334] In one embodiment, the steam autoclave is run for a final time equivalent (F0) of at least 90 minutes.

[0335] In one embodiment, the heat treatment comprises exposing the at least one cross-linked polysaccharide to dry heat.

[0336] In one embodiment, the glycosidic bond cleavage step e) carried out on the at least one cross-linked polysaccharide is carried out by high pressure treatment.

[0337] In one embodiment, the glycosidic bond cleavage step e) is carried out on at least one purified cross-linked polysaccharide.

[0338] In one embodiment, the glycosidic bond cleavage step e) is carried out on at least one unpurified, cross-linked polysaccharide.

[0339] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e) a step of purifying the obtained polymeric compound is carried out.

[0340] In one embodiment, the step of purifying the polymeric compound comprises removing said crosslinker.

[0341] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e) a purification step is carried out by dialysis.

[0342] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e), a step of purifying the macromolecular compound solution is carried out by dialysis using a solution or a dialysis solvent selected from the group consisting of phosphate buffers, e.g. PBS and water.

[0343] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e), a step of purifying the macromolecular compound solution is carried out by dialysis using a solution or a dialysis solvent selected from the group consisting of phosphate buffers, e.g. PBS and water, further comprising at least one active ingredient, e.g. an antioxidant and / or a local anesthetic.

[0344] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e) a step of purifying the polymer solution is carried out by washing.

[0345] In one embodiment, the step of purifying the macromolecular compound solution is carried out by washing with a solution or solvent selected from the group consisting of phosphate buffer, e.g., PBS, and water.

[0346] In one embodiment, the step of purifying the polymer solution is carried out by washing with a solution or solvent selected from the group consisting of phosphate buffer, e.g., PBS, and water, further comprising at least one active ingredient, e.g., an antioxidant and / or a local anesthetic.

[0347] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e) a purification step is carried out by tangential filtration.

[0348] In one embodiment, the purification step by tangential filtration comprises at least one ultrafiltration step and at least one diafiltration step.

[0349] In one embodiment, the purification step by tangential filtration comprises at least one ultrafiltration step.

[0350] In one embodiment, the purification step by tangential filtration comprises at least one diafiltration step.

[0351] In one embodiment, the composition of the macromolecular compound solution is modified during a purification step by tangential filtration.

[0352] In one embodiment, the composition of the macromolecular compound solution is concentrated during a purification step by tangential filtration.

[0353] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e) a purification step is carried out by filtration through at least one hollow fiber membrane.

[0354] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e) a purification step of the polymeric compound is carried out by precipitation in salified conditions.

[0355] In one embodiment, the process for the preparation of the inventive composition 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.

[0356] In one embodiment, the process for the preparation of the inventive composition is characterized in that after step e), a purification step of the polymeric compound is carried out by precipitation from a solution obtained after addition of PBS or NaCl salt and further comprising at least one active ingredient, such as an antioxidant and / or a local anesthetic.

[0357] In one embodiment, the step of purifying the polymer by precipitation is carried out by adding at least one organic solvent to the polymer solution.

[0358] In one embodiment, at least one organic solvent added to the polymer solution in the precipitation purification is selected from the group of alcohols.

[0359] In one embodiment, ethanol is added to the polymer solution in the precipitation purification step.

[0360] In one embodiment, the purification step of the polymeric compound by precipitation occurs in an aqueous alcohol mixture.

[0361] In one embodiment, the precipitated polymeric compound is recovered by filtration.

[0362] In one embodiment, the polymeric compound that is precipitated and recovered by filtration is washed.

[0363] In one embodiment, the polymeric compound that is precipitated and recovered by filtration is dried.

[0364] In one embodiment, the polymeric compound precipitated and recovered by filtration is dried under ambient air.

[0365] In one embodiment, the polymeric compound precipitated and recovered by filtration is dried under vacuum.

[0366] The process for preparing the composition of the invention further comprises at least one step of filtering the aqueous polymer solution resulting from step e) through a membrane of 0.22 μm porosity, said filtering step being performed on the aqueous polymer solution or on a sample of the aqueous polymer solution.

[0367] In one embodiment, the step of filtering through a membrane of 0.22 μm porosity is the terminal sterilization step.

[0368] In one embodiment, the step of filtering through a membrane of 0.22 μm porosity is performed once.

[0369] In one embodiment, the step of filtering through a membrane of 0.22 μm porosity is carried out twice.

[0370] In one embodiment, the step of filtering through a membrane of 0.22 μm porosity is performed three times.

[0371] In one embodiment, the filtration step through a membrane of 0.22 μm porosity is an "in-process control" (IPC) measure for controlling the process during the process and is performed on a sample of the aqueous polymer solution obtained after process step e).

[0372] In one embodiment, a filtration IPC on a membrane of 0.22 μm porosity is carried out on a sample of the aqueous macromolecular solution obtained after process step e) after the sample dilution step.

[0373] In one embodiment, when the glycosidic bond cleavage step e) is carried out in a steam autoclave, the process for preparing the inventive composition does not include a step of filtering the aqueous polymer solution through a membrane of 0.22 μm porosity.

[0374] In one embodiment, when the glycosidic bond cleavage step e) is carried out in a steam autoclave, the process for the preparation of the inventive composition comprises the steps of taking a sample of the aqueous polymer solution after step e) and filtration test (IPC) of the sample through a membrane of 0.22 μm porosity.

[0375] In one embodiment, the composition of the invention is characterized by a G' of 300 Pa or less at 1 Hz.

[0376] In one embodiment, the composition of the invention is characterized by a G' of 150 Pa or less at 1 Hz.

[0377] In one embodiment, the composition of the invention is characterized by a G' of 100 Pa or less at 1 Hz.

[0378] In one embodiment, the composition of the invention is characterized by a G' of 85 Pa or less at 1 Hz.

[0379] In one embodiment, the composition of the invention is characterized by a G' of 70 Pa or less at 1 Hz.

[0380] In one embodiment, the compositions of the invention are characterized by having a Tan Δ (Tn δ) of 1.00 or greater.

[0381] In one embodiment, the compositions of the invention are characterized by a Tan Δ(Tnδ) between 1.00 and 1.80 (1.00≦Tan Δ(Tnδ)≦1.80).

[0382] In one embodiment, the composition of the invention is characterized by a Tan Δ(Tnδ) between 1.00 and 1.50 (1.00≦Tan Δ(Tnδ)≦1.50).

[0383] In one embodiment, the compositions of the invention are characterized by a Tan Δ(Tnδ) between 1.00 and 1.30 (1.00≦Tan Δ(Tnδ)≦1.30).

[0384] The invention also relates to processes for the preparation of formulations containing the compositions of the invention.

[0385] In one embodiment, the process for preparing a formulation from the composition of the invention further comprises at least one dilution or dissolution step.

[0386] In one embodiment, the dilution or dissolution step is carried out by adding water or an aqueous saline solution, such as a phosphate buffer, for example PBS.

[0387] In one embodiment, the dilution or dissolution step is carried out by adding water or an aqueous saline solution, such as a phosphate buffer, e.g., PBS, further comprising at least one active ingredient, e.g., an antioxidant and / or a local anesthetic.

[0388] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a macromolecular compound concentration between 2 mg / g and 200 mg / g, based on the total mass of the formulation.

[0389] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a macromolecular compound concentration between 2 mg / g and 75 mg / g, based on the total mass of the formulation.

[0390] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a macromolecular compound concentration of between 5 mg / g and 50 mg / g based on the total mass of the formulation.

[0391] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a macromolecular compound concentration of between 10 mg / g and 40 mg / g, based on the total mass of the formulation.

[0392] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a polymer concentration of about 100 mg / g based on the total mass of the formulation.

[0393] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a polymer concentration of about 80 mg / g based on the total mass of the formulation.

[0394] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a polymer concentration of about 60 mg / g based on the total mass of the formulation.

[0395] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a polymer concentration of about 40 mg / g based on the total mass of the formulation.

[0396] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a polymer concentration of about 20 mg / g based on the total mass of the formulation.

[0397] In one embodiment, the process for preparing a formulation comprising the composition of the invention further comprises at least one dilution or dissolution step to obtain a polymer concentration of about 10 mg / g based on the total mass of the formulation.

[0398] In one embodiment, the process for preparing the inventive composition further comprises at least one step of adding at least one active ingredient.

[0399] In one embodiment, at least one active ingredient is added in powder form.

[0400] In one embodiment, the at least one active ingredient is added in the form of a solution or suspension.

[0401] In one embodiment, at least one active ingredient is provided in a solution or suspension in a solvent or solution selected from the group consisting of water or saline solutions, for example phosphate buffers such as PBS.

[0402] In one embodiment, at least one active ingredient is added prior to step e) of the process for preparing the inventive composition.

[0403] In one embodiment, the at least one active ingredient is added after step e) of the process for preparing the inventive composition.

[0404] In one embodiment, at least one active ingredient is added prior to step g) of the process for preparing the inventive composition.

[0405] In one embodiment, at least one active ingredient is added after step g) of the process for preparing the inventive composition.

[0406] In one embodiment, at least one active ingredient is added after step g) of the process for preparing the inventive composition and the resulting formulation is sterilized by filtration through a membrane of 0.22 μm porosity.

[0407] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one active ingredient selected from the group consisting of local anesthetics, derivatives of vitamin C, anti-inflammatory agents, antioxidants, and mixtures thereof.

[0408] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic agent.

[0409] In one embodiment, the process for preparing a formulation comprising at least one composition of the invention further comprises at least one step of adding at least one local anesthetic to a concentration of between 0.1% and 5% of the local anesthetic relative to the total mass of the formulation.

[0410] In one embodiment, the process for preparing a formulation comprising at least one composition of the invention further comprises at least one step of adding at least one local anesthetic to a concentration of between 0.1% and 4% of the local anesthetic relative to the total mass of the formulation.

[0411] In one embodiment, the process for preparing a formulation comprising at least one composition of the invention further comprises at least one step of adding at least one local anesthetic to a concentration of between 0.1% and 2% of the local anesthetic relative to the total mass of the formulation.

[0412] In one embodiment, the process for preparing a formulation comprising at least one composition of the invention further comprises at least one step of adding at least one local anesthetic to a concentration of between 0.1% and 1% of the local anesthetic relative to the total mass of the formulation.

[0413] In one embodiment, the process for preparing a formulation comprising at least one composition of the invention further comprises at least one step of adding at least one local anesthetic to a concentration of between 0.1% and 0.5% of the local anesthetic relative to the total mass of the formulation.

[0414] In one embodiment, the method for preparing a formulation comprising at least one composition of the invention further comprises at least one step of adding at least one local anesthetic to the formulation to provide a concentration of about 0.3% of the local anesthetic based on the total mass of the formulation.

[0415] In one embodiment, the local anesthetic is selected from the group of amino esters.

[0416] In one embodiment, the amino ester is selected from the group comprising procaine, benzocaine, chloroprocaine and tetracaine in their base or salt form, for example in their hydrochloride form.

[0417] In one embodiment, the local anesthetic is selected from the group of aminoamides.

[0418] In one embodiment, the aminoamide is selected from the group comprising lidocaine, mepivacaine, prilocaine, articaine, aptocaine, bupivacaine, etidocaine and ropivacaine in their base or salt form, for example in their hydrochloride form.

[0419] In one embodiment, the local anesthetic is selected from the group of amino ethers.

[0420] In one embodiment, the aminoether is selected from the group comprising diamocine and bromocaine in their base or salt form, for example as hydrochlorides or cyclamates.

[0421] In one embodiment, the aminoether is selected from the group consisting of lidocaine, mepivacaine and its salts and isolated isomers.

[0422] In one embodiment, the aminoether is lidocaine.

[0423] In one embodiment, the aminoether is lidocaine or a pharma- ceutically acceptable salt thereof.

[0424] In one embodiment, the aminoether is lidocaine hydrochloride.

[0425] In one embodiment, the aminoether is mepivacaine.

[0426] In one embodiment, the aminoether is mepivacaine or a pharma- ceutically acceptable salt thereof.

[0427] In one embodiment, the aminoether is selected from the group consisting of racemic mepivacaine hydrochloride, racemic mepivacaine hydrochloride, (r)-mepivacaine hydrochloride, (s)-mepivacaine hydrochloride, r)-mepivacaine, and (s)-mepivacaine, or one of its pharma- ceutically acceptable salts.

[0428] In one embodiment, the aminoether is mepivacaine hydrochloride.

[0429] In one embodiment, the aminoether is (r)-mepivacaine hydrochloride.

[0430] In one embodiment, the aminoether is (s)-mepivacaine hydrochloride.

[0431] In one embodiment, the aminoether is racemic mepivacaine hydrochloride.

[0432] In one embodiment, the aminoether is (r)-mepivacaine.

[0433] In one embodiment, the aminoether is (s)-mepivacaine.

[0434] In one embodiment, the aminoether is racemic mepivacaine.

[0435] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one aminoether local anesthetic selected from the group consisting of lidocaine, mepivacaine, and mixtures thereof.

[0436] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine.

[0437] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of lidocaine is between 0.1% and 5% relative to the total mass of the formulation.

[0438] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of lidocaine is between 0.1% and 4% relative to the total mass of the formulation.

[0439] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of lidocaine is between 0.1% and 2% relative to the total mass of the formulation.

[0440] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of lidocaine is between 0.1% and 1% by weight of the total formulation.

[0441] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of lidocaine is between 0.1% and 0.5% relative to the total mass of the formulation.

[0442] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of lidocaine is about 0.3% based on the total weight of the formulation.

[0443] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is mepivacaine.

[0444] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of mepivacaine is between 0.1% and 5% relative to the total mass of the formulation.

[0445] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of mepivacaine is between 0.1% and 4% relative to the total mass of the formulation.

[0446] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of mepivacaine is between 0.1% and 2% relative to the total mass of the formulation.

[0447] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of mepivacaine is between 0.1% and 1% relative to the total mass of the formulation.

[0448] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of mepivacaine is between 0.1% and 0.5% relative to the total mass of the formulation.

[0449] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic, which is lidocaine, such that the concentration of mepivacaine is about 0.3% relative to the total mass of the formulation.

[0450] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic agent, which is dyclonine in its base or salt form, e.g., its hydrochloride salt form.

[0451] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one local anesthetic selected from the group consisting of chlorobutanol, guafecainol, and polidocanol.

[0452] In one embodiment, the local anesthetic is chlorobutanol.

[0453] In one embodiment, the local anesthetic is guafecainol.

[0454] In one embodiment, the local anesthetic is polidocanol.

[0455] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent.

[0456] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group consisting of steroidal and non-steroidal anti-inflammatory agents.

[0457] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group consisting of non-steroidal anti-inflammatory agents.

[0458] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group comprising salicylic acid anti-inflammatory agents, propionic acid derivatives, indole derivatives, pyrazole derivatives, oximes, and coxibs.

[0459] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group comprising diclofenac, nimesulide, niflumic acid, mefenamic acid and nabumetone, either alone or in mixture.

[0460] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one salicylated anti-inflammatory agent selected from the group comprising diflunisal, benorylate and aspirin, either alone or in admixture.

[0461] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention comprises at least one step of adding at least one anti-inflammatory agent selected from the group of propionic acid derivatives including aluminoprofen, ketoprofen, ibuprofen, naproxen, flurbiprofen and tiaprofenic acid, either alone or in mixtures.

[0462] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group of indole derivatives including indomethacin, sulindac, and etodolac, either alone or in mixture.

[0463] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory substance, in particular selected from the group of pyrazole derivatives including phenylbutazone.

[0464] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group of oxicams, including piroxicam, tenoxicam, and meloxicam, either alone or in mixture.

[0465] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group of coxibs, including celecoxib, etoricoxib, and rofecoxib, either alone or in mixtures.

[0466] In one embodiment, the concentration of the nonsteroidal anti-inflammatory agent in a formulation comprising at least one composition of the invention is between 0.01 mg / g and 2000 mg / g.

[0467] In one embodiment, the concentration of the nonsteroidal anti-inflammatory agent in a formulation comprising at least one composition of the invention is between 0.1 mg / g and 1000 mg / g.

[0468] In one embodiment, the concentration of the nonsteroidal anti-inflammatory agent in a formulation comprising at least one composition of the invention is between 0.5 mg / g and 500 mg / g.

[0469] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one steroidal anti-inflammatory agent.

[0470] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one steroidal anti-inflammatory agent selected from the group consisting of dexamethasone, prednisolone, corticosterone, budesonide, sulfasalazine, mesalamine, cetirizine, diphenhydramine, antipyrine, methyl salicylate, loratadine, thymol, carvacrol, bisabolol, allantoin, eucalyptol, phenazone (antipyrine) propyphenazone, alone or in mixtures.

[0471] In one embodiment, the concentration of the steroidal anti-inflammatory agent in a formulation comprising at least one composition of the invention is between 0.01 mg / g and 2000 mg / g.

[0472] In one embodiment, the concentration of the steroidal anti-inflammatory agent in a formulation comprising at least one composition of the invention is between 0.1 mg / g and 1000 mg / g.

[0473] In one embodiment, the concentration of the steroidal anti-inflammatory agent in a formulation comprising at least one composition of the invention is between 0.5 mg / g and 500 mg / g.

[0474] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group consisting of sucrose octasulfate and salts thereof.

[0475] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent selected from the group consisting of sucrose octasulfate and its sodium salt and its potassium salt.

[0476] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent, a water-soluble salt of sucrose octasulfate, selected from the group consisting of alkali metal salts, alkaline earth metal salts, silver salts, ammonium salts, and amino acid salts.

[0477] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent, a water-soluble salt of sucrose octasulfate, selected from the group consisting of alkali metal salts or alkaline earth metal salts.

[0478] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-inflammatory agent, a water-soluble salt of sucrose octasulfate, selected from the group consisting of the sodium salt of sucrose octasulfate or the potassium salt of sucrose octasulfate.

[0479] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antimicrobial agent.

[0480] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antimicrobial agent selected from the group comprising gentamicin, silver sulfadiazine, metronidazole, fucidin, eosin, povidone-iodine, copper gluconate, zinc gluconate, manganese gluconate, or salts thereof, either alone or in mixtures.

[0481] In one embodiment, the concentration of the antimicrobial agent in a formulation comprising at least one composition of the invention is between 0.1 mg / g and 200 mg / g.

[0482] In one embodiment, the concentration of the antimicrobial agent in a formulation comprising at least one composition of the invention is between 0.5 mg / g and 100 mg / g.

[0483] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one glucoside or glucoside derivative.

[0484] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one glucoside or glucoside derivative selected from the group comprising D-glucopyranose, 1,4 glucoside, esculin, hesperidin, diosmin, arbutin, skinmin or aloin, either alone or in mixture.

[0485] In one embodiment, the concentration of glucosides in a formulation comprising at least one composition of the invention is between 0.1 mg / g and 200 mg / g.

[0486] In one embodiment, the concentration of glucosides in a formulation comprising at least one composition of the invention is between 0.5 mg / g and 100 mg / g.

[0487] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing agent or tissue regenerating agent.

[0488] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of macroelements.

[0489] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of iron, calcium, copper, zinc, manganese, magnesium or potassium gluconate, trimethylsilanol, trimethylsilanolate salts, potassium trimethylsilanol, methylsilanol mannuronate, sodium monoethylsilanol orthohydroxybenzoate, alone or in mixture.

[0490] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of essential amino acids.

[0491] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one essential amino acid selected from the group comprising isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine, either alone or in mixtures.

[0492] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of semi-essential amino acids.

[0493] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of 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, either alone or in mixture.

[0494] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of non-essential amino acids.

[0495] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one non-essential amino acid selected from the group including alanine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, proline, serine, tyrosine, either alone or in mixtures.

[0496] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of vitamins.

[0497] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one vitamin selected from the group consisting of retinol, thiamine, riboflavin, nicotinamide, adenine, calcium pantothenate, pyridoxine, inositol, biotin, folic acid, para-aminobenzoic acid, cobalamin, vitamin C, and choline chloride, either alone or in admixture.

[0498] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of nucleic acids.

[0499] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one nucleic acid selected from the group consisting of deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, methylcytosine, either alone or in mixtures.

[0500] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of coenzymes.

[0501] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one coenzyme selected from the group consisting of thiamine pyrophosphate, coenzyme A, FAD, NAD, NADP, UTP, either alone or in mixtures.

[0502] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one moisturizing or tissue regenerating agent selected from the group consisting of deoxythymidine, glutathione, sodium pyruvate, lipoic acid, and putrescine, either alone or in mixtures.

[0503] In one embodiment, the concentration of the moisturizing or tissue regenerating agent in a formulation comprising at least one composition of the invention is between 0.01 mg / g and 500 mg / g.

[0504] In one embodiment, the concentration of the moisturizing or tissue regenerating agent in a formulation comprising at least one composition of the invention is between 0.1 mg / g and 200 mg / g.

[0505] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant.

[0506] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant selected from the group of polyols.

[0507] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol selected from the group consisting of mannitol, sorbitol, propylene glycol, xylitol, glycerol, maltitol, lactitol, and erythritol.

[0508] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol selected from the group consisting of mannitol, sorbitol, maltitol and glycerol, either alone or in mixture.

[0509] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol selected from the group consisting of mannitol, sorbitol, and maltitol, either alone or in mixture.

[0510] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol such that the concentration of the polyol is between 0.1 mg / ml and 50 mg / ml relative to the total mass of the formulation.

[0511] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol such that the concentration of the polyol is between 5 mg / ml and 40 mg / ml, relative to the total mass of the formulation.

[0512] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol such that the concentration of the polyol is between 10 mg / ml and 40 mg / ml, relative to the total mass of the formulation.

[0513] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol such that the concentration of the polyol is between 20 mg / ml and 40 mg / ml, relative to the total mass of the formulation.

[0514] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol such that the concentration of the polyol is between 30 mg / ml and 40 mg / ml, relative to the total mass of the formulation.

[0515] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol which is mannitol.

[0516] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding mannitol to a concentration between 5 mg / ml and 40 mg / ml, relative to the total mass of the formulation.

[0517] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding mannitol to a concentration between 10 mg / ml and 40 mg / ml, relative to the total mass of the formulation.

[0518] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding mannitol to a concentration between 20 mg / ml and 40 mg / ml, relative to the total mass of the formulation.

[0519] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding mannitol to a concentration of between 30 mg / ml and 40 mg / ml, relative to the total mass of the formulation.

[0520] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol which is sorbitol.

[0521] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding sorbitol to a concentration between 5 mg / ml and 40 mg / ml of sorbitol relative to the total mass of the formulation.

[0522] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding sorbitol to a concentration between 10 mg / ml and 40 mg / ml of sorbitol relative to the total mass of the formulation.

[0523] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding sorbitol to a concentration between 20 mg / ml and 40 mg / ml of sorbitol relative to the total mass of the formulation.

[0524] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding sorbitol to a concentration between 30 mg / ml and 40 mg / ml, relative to the total weight of the formulation.

[0525] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol which is maltitol.

[0526] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one polyol which is glycerol.

[0527] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding a mixture of mannitol and sorbitol.

[0528] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant selected from the group of vitamin C derivatives.

[0529] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant selected from the group of vitamin C derivatives including magnesium ascorbyl phosphate, sodium ascorbyl phosphate, ascorbic acid-2-glucoside, and mixtures thereof.

[0530] In one embodiment, the at least one vitamin C derivative is magnesium ascorbyl phosphate.

[0531] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant selected from the group of vitamin E derivatives and tocopherols.

[0532] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant selected from the group of carotenoids and retinoids and their derivatives.

[0533] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant selected from the group of carotenoids and retinoids and their derivatives, including retinol, retinoic acid, retinal, retinol esters, and carotene.

[0534] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant selected from the group of pseudotripeptides.

[0535] In one embodiment, the tripeptide mimetic is glutathione.

[0536] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant selected from the group comprising different forms of coenzyme Q10, ubiquitinone and ubiquinol.

[0537] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one vitamin.

[0538] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one vitamin selected from the group comprising retinol, thiamine, riboflavin, nicotinamide, dexpanthenol, pyridoxine, ascorbic acid, ergocalciferol, tocopherol, biotin, and folic acid, either alone or in mixtures.

[0539] In one embodiment, the concentration of vitamins in a formulation comprising at least one composition of the invention is between 0.01 mg / g and 200 mg / g.

[0540] In one embodiment, the concentration of vitamins in a formulation containing at least one of the compositions of the invention is between 0.01 mg / g and 100 mg / g.

[0541] In one embodiment, the concentration of vitamins in a formulation containing at least one composition of the invention is between 0.5 mg / g and 50 mg / g.

[0542] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one amino acid.

[0543] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one amino acid selected from the group consisting of essential amino acids, semi-essential amino acids and non-essential amino acids, either alone or in mixtures.

[0544] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one amino acid selected from the group of essential amino acids.

[0545] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one essential amino acid selected from the group comprising isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine, either alone or in mixtures.

[0546] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one amino acid selected from the group of semi-essential amino acids.

[0547] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of 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, either alone or in mixture.

[0548] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one amino acid selected from the group of non-essential amino acids.

[0549] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one non-essential amino acid selected from the group comprising alanine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, proline, serine, tyrosine, either alone or in mixtures.

[0550] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one amino acid selected from the group comprising hydroxyproline, taurine and ornithine, either alone or in mixture.

[0551] In one embodiment, the concentration of amino acids in a formulation comprising at least one composition of the invention is between 0.01 mg / g and 150 mg / g.

[0552] In one embodiment, the concentration of amino acids in a formulation comprising at least one composition of the invention is between 0.01 mg / g and 100 mg / g.

[0553] In one embodiment, the concentration of amino acids in a formulation comprising at least one of the compositions of the invention is between 0.5 mg / g and 50 mg / g.

[0554] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one vasoconstrictor.

[0555] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one vasoconstrictor selected from the group including naphazoline, epinephrine, methoxamine, methylnorepinephrine, norepinephrine, oxymetazoline, phenylephrine, pseudoephedrine, synephrine, cirazoline, and xylometazoline.

[0556] In one embodiment, the concentration of the vasoconstrictor in a formulation containing at least one composition of the invention is between 0.01 mg / g and 3 mg / g.

[0557] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one vasodilator.

[0558] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one vasodilator selected from the group comprising adenosine, nicotinic acid, minoxidil, and diazoxide, either alone or in mixture.

[0559] In one embodiment, the concentration of the vasodilator in a formulation comprising at least one composition of the invention is between 0.01 mg / g and 10 mg / g.

[0560] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one anti-hemorrhagic or hemostatic agent.

[0561] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antihemorrhagic or hemostatic agent selected from the group comprising aminocaproic acid or tranexamic acid, either alone or in admixture.

[0562] In one embodiment, the concentration of the antihemorrhagic or hemostatic agent in a formulation comprising at least one composition of the invention is between 0.01 mg / g and 5 mg / g.

[0563] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant and at least one local anesthetic.

[0564] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of 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.

[0565] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one antioxidant, at least one vitamin, and at least one tissue regenerative agent.

[0566] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one amino acid, at least one vitamin, and at least one tissue regenerative agent.

[0567] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of 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 regenerative agent.

[0568] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one active ingredient selected from the group consisting of high temperature sensitive active ingredients.

[0569] In one embodiment, the at least one high temperature sensitive active ingredient is added after step e) of the process for preparing at least one composition of the invention.

[0570] In one embodiment, the process for the preparation of a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding a so-called temperature-sensitive active ingredient of natural origin.

[0571] In one embodiment, the process for the preparation of a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding a so-called temperature-sensitive active ingredient of synthetic origin.

[0572] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one temperature-sensitive active ingredient selected from the group consisting of peptides, hormones, proteins, growth factors, antibodies and vitamins, either alone or in mixtures.

[0573] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one temperature sensitive active ingredient selected from the group consisting of peptides.

[0574] In one embodiment, the added temperature sensitive active ingredient is a peptide.

[0575] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one high temperature sensitive active ingredient selected from the group consisting of hormones.

[0576] In one embodiment, the added temperature sensitive active ingredient is a hormone.

[0577] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one temperature sensitive active ingredient selected from the group consisting of proteins.

[0578] In one embodiment, the added temperature sensitive active ingredient is a protein.

[0579] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one temperature sensitive active ingredient selected from the group consisting of growth factors.

[0580] In one embodiment, the added temperature sensitive active ingredient is a growth factor.

[0581] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one temperature sensitive active ingredient selected from the group consisting of antibodies.

[0582] In one embodiment, the temperature sensitive active ingredient added is an antibody.

[0583] In one embodiment, the process for preparing a formulation comprising at least one of the compositions of the invention further comprises at least one step of adding at least one high temperature sensitive active ingredient selected from the group consisting of vitamins.

[0584] In one embodiment, the vitamins are selected from the group comprising retinol, thiamine, riboflavin, nicotinamide, dexpanthenol, pyridoxine, ascorbic acid, ergocalciferol, tocopherol, biotin, and folic acid, either alone or in mixtures.

[0585] The invention also relates to formulations comprising at least one of the compositions of the invention.

[0586] In one embodiment, the formulation is characterized in that the concentration of polymeric compound is between 2 mg / g and 100 mg / g, relative to the total mass of said formulation.

[0587] In one embodiment, the formulation is characterized in that the concentration of polymeric compound is between 2 mg / g and 75 mg / g, relative to the total mass of said formulation.

[0588] In one embodiment, the formulation is characterized in that the concentration of polymeric compound is between 5 mg / g and 50 mg / g, relative to the total mass of said formulation.

[0589] In one embodiment, the formulation is characterized in that the concentration of polymeric compound is between 10 mg / g and 40 mg / g, relative to the total mass of said formulation.

[0590] In one embodiment, the formulation is characterized by a concentration of polymeric compound of about 100 mg / g, relative to the total mass of said formulation.

[0591] In one embodiment, the formulation is characterized by a concentration of polymeric compound of about 80 mg / g, relative to the total mass of said formulation.

[0592] In one embodiment, the formulation is characterized by a concentration of polymeric compound of about 60 mg / g, relative to the total mass of said formulation.

[0593] In one embodiment, the formulation is characterized by a concentration of polymeric compound of about 40 mg / g, relative to the total mass of said formulation.

[0594] In one embodiment, the formulation is characterized by a concentration of polymeric compound of about 20 mg / g, relative to the total mass of said formulation.

[0595] In one embodiment, the formulation is characterized by a concentration of polymeric compound of about 10 mg / g, relative to the total mass of said formulation.

[0596] In one embodiment, the formulation is characterized as being injectable.

[0597] In one embodiment, the formulation is characterized as being sterile.

[0598] In one embodiment, the formulation is injectable and characterized as being sterile.

[0599] The formulations obtained based on the compositions of the invention have many uses.

[0600] Medical applications include, for example, injections into joints to replace defective fluids, for example, to replace joint fluid, post-operative injections, for example, to avoid post-operative adhesions, periurethral injections for the treatment of incontinence, and post-op surgery injections. Cosmetic applications include, for example, injections to fill wrinkles, fine lines and skin defects, or to increase the volume of, for example, lips, cheekbones, etc.

[0601] The applications of interest are more particularly those commonly used in the context of injectable viscoelastics and polysaccharides that are used, or potentially can be used, in the treatment of the following conditions or conditions: Cosmetic injections to the face: filling wrinkles, skin defects or volumes (cheekbones, chin, lips) Volume injections for various body stages: breast and buttock augmentation, G-spot augmentation, vaginoplasty, labia reconstruction, penis size increase Joint surgery or dental surgery, e.g. for filling periodontal pockets Treating osteoarthritis, injections into joints to replace or replenish defective joint fluid Periurethral injections for the treatment of urinary incontinence due to sphincter dysfunction Postoperative injections, especially to avoid peritoneal adhesions Injection after presbyopia surgery using laser scleral ablation Injection into the vitreous cavity Injection during cataract surgery Injections for the treatment of cases of vaginal dryness Injection into tissue spaces Genital injection

[0602] More particularly, the preparations obtained with the process object of the invention may be used in cosmetic surgery, due to their viscoelastic and persistent properties. It is injected with a fine diameter needle (e.g., 27 gauge) to fill in fine, medium, or deep wrinkles. As a volumizer for injections using larger diameter, eg, 22 to 26 gauge, and longer (eg, 30 to 40 mm) needles, where cohesiveness ensures retention at the injection site.

[0603] These examples of use are in no way limiting, and the preparations obtained according to the process of the invention are intended more broadly. To fill to increase volume. Creating space within a particular tissue to facilitate optimal functioning. To replace defective physiological fluids.

[0604] The polymeric compounds of the invention are not used in combination with native polysaccharides in the formulation, and in particular, the polymeric compounds of the invention are not used in combination with hyaluronic acid or heparosan in the formulation.

[0605] In one embodiment, the polymeric compounds of the invention are used in combination with at least one cross-linking polymer in the formulation.

[0606] In one embodiment, the polymeric compounds of the invention are used in combination with at least one cross-linked polymer selected from the group of cross-linked polysaccharides in the formulation.

[0607] In one embodiment, the polymeric compound of the invention is used in admixture with at least one cross-linked polysaccharide selected from the group consisting of hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, alginic acid, xanthan, carrageenan, chitosan, chondroitin, heparosan, and bioacceptable salts thereof, either alone or in mixtures.

[0608] In one embodiment, the polymeric compounds of the invention are used to prepare crosslinked polymeric compounds, either alone or in admixture with at least one other polymer.

[0609] In one embodiment, the polymeric compounds of the invention are mixed with at least one polysaccharide to prepare a crosslinked polymeric compound.

[0610] In one embodiment, the polysaccharides used in conjunction with the polymeric compounds of the invention to prepare crosslinked polymeric compounds are selected from the group consisting of hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, alginic acid, xanthan, carrageenan, chitosan, chondroitin, heparosan, and bioacceptable salts thereof, either alone or in mixtures, and these polysaccharides may be crosslinked or uncrosslinked.

[0611] In one embodiment, the at least one polysaccharide used together with the inventive polymeric compounds to prepare crosslinked polymeric compounds is selected from the group consisting of hyaluronic acid, heparosan and their bioacceptable salts, either alone or in mixtures, and these polysaccharides may be crosslinked or uncrosslinked.

[0612] In one embodiment, the polysaccharide used with the polymeric compounds of the invention in preparing crosslinked polymeric compounds is hyaluronic acid or a salt of hyaluronic acid.

[0613] In one embodiment, the polysaccharide used with the inventive polymeric compounds in preparing the crosslinked polymeric compounds is hyaluronic acid or a mixture of salts of hyaluronic acid.

[0614] In one embodiment, the polysaccharide used with the polymeric compounds of the invention in preparing the crosslinked polymeric compounds is a salt of hyaluronic acid or crosslinked hyaluronic acid.

[0615] In one embodiment, the polysaccharide used with the inventive polymeric compounds in preparing the crosslinked polymeric compounds is heparosan or a salt of heparosan.

[0616] In one embodiment, the polysaccharide used with the inventive polymeric compounds in preparing the crosslinked polymeric compounds is heparosan or a mixture of salts of heparosan.

[0617] In one embodiment, the polysaccharide used with the inventive polymeric compounds in the preparation of crosslinked polymeric compounds is heparosan or a salt of crosslinked heparosan.

[0618] In one embodiment, the polymeric compounds of the invention are used in the preparation of at least one prodrug polymer.

[0619] In one embodiment, the polymeric compounds of the invention are chemically modified by reaction with at least one active ingredient to prepare a prodrug polymer.

[0620] In one embodiment, the prodrug polymers derived from the polymeric compounds of the invention are used in the field of oncology. [Brief description of the drawings]

[0621] FIG. 1 is a graph showing the development of the curve of log(specific viscosity dL / g) as a function of log(molecular weight Da) for each composition investigated in the examples.

[0622] FIG. 2 shows the network formed during the cross-linking step and its degradation during the steps of the claimed process. Diagram A represents the complete network obtained in step d) of the claimed process, where the thick lines represent the polysaccharide chains connected to each other and the thin lines represent the divalent radicals L derived from the cross-linker connected to the polysaccharide chains. Diagram B represents the cleavage of the glycosidic bonds at different points of the previous network after the bond cleavage step e), indicated by arrow 1, according to the claimed process. The thick lines represent the cleaved polysaccharide chains, the dots represent the newly formed hydroxyl groups and the thin lines represent the divalent radicals L derived from the cross-linker. Finally, diagram C represents the mixed polymers spread in solution, arrow 2, of the polymeric compound resulting from the bond cleavage step e) of the invention. The thick lines represent the cleaved polysaccharide chains, the dots represent the newly formed hydroxyl groups and the thin lines represent the divalent radicals L derived from the cross-linker. Working Example

[0623] As part of the examples several parameters were measured.

[0624] Measurement of flow parameters G', G" and Tan Δ Equipment: DHR-2, manufactured by TA Instruments Cone geometry with angle 2° and diameter 40mm. Frequency sweep oscillation method, 0.8% distortion over the frequency range of 0.08 to 5Hz

[0625] SEC measurements: Viscotek GPCmax II instrument equipped with a TDA305 detector (RI, RALS / LALS, viscometer) and Shodex OHpak SB-806HQ+SB-805HQ columns. Measurements were performed at 37°C in the presence of PBS assay buffer. [Preparation of the composition of the invention] Example 1

[0626] Sodium hyaluronate fibers (10.4 g) of injectable grade with a weight average molecular weight of 1 MDa are weighed into a container. A 1% aqueous solution of sodium hydroxide is added to hydrate the sodium hyaluronate fibers. The reaction medium is homogenized by alternating current manual mechanical stirring and left for 50 minutes. 0.43 g of BDDE is added. The reaction medium containing 10.4 g of sodium hyaluronate fibers, 55.7 g of sodium hydroxide and 0.43 g of BDDE is again homogenized by manual mechanical stirring and left for 24 hours in a water bath previously thermostated at 2°C.

[0627] At the end of crosslinking, the reaction medium is neutralized by adding 1N HCl and phosphate buffer with mechanical stirring. The gel obtained is dialyzed against phosphate buffer until the concentration of hyaluronic acid is 27.5 mg / g. The phosphate buffer is prepared by dissolving 0.23 g NaH2PO4·2H2O, 1.12 g Na2HPO4, 42.5 g NaOH in a sufficient amount of water for injection (EPPI) to obtain 5 L of buffer. After mechanical homogenization and bubble removal, the gel obtained, with a TanΔ of 0.54 at 1 Hz, cannot be filtered through a membrane with a porosity of 0.22 μm. It is dispensed into a 1 ml glass syringe and steam sterilized at 127 °C and FO value of 50 min. After heat treatment, the syringe contains a solution of the polymeric compound. A sample is taken from the syringe and a filterability test is performed on this sample through a membrane with a porosity of 0.22 μm. The filterability test is positive. A solution of a polymer compound can be filtered through a membrane with a porosity of 0.22 μm. Example 2

[0628] The composition of the invention is prepared in a similar manner to that described in Example 1, except that after the addition of BDDE and homogenization, the reaction medium is immediately placed in a water bath previously thermostated at 9° C. for 3 hours at this temperature. As described in Example 1, before heat treatment, the gel with a Tan Δ of 0.68 at 1 Hz is not filterable through a membrane with a porosity of 0.22 μm. It is dispensed into a syringe. After heat treatment, the syringe contains 27.5 mg / g of polymer solution. A sample is taken from the syringe and a filterability test is carried out on this sample through a membrane with a porosity of 0.22 μm. The filterability test is positive. The polymer solution is filterable through a membrane with a porosity of 0.22 μm. Reference example 3

[0629] Sodium hyaluronate fibers (10.4 g) of injectable grade with a weight average molecular weight of 1 MDa are weighed into a container. The sodium hyaluronate fibers are hydrated by adding 1% aqueous sodium hydroxide solution. The reaction medium is homogenized by alternating current manual mechanical stirring and left for 50 minutes. Phosphate buffer is added instead of BDDE. The phosphate buffer is prepared by dissolving 0.23 g NaH2PO4·2H2O, 1.12 g Na2HPO4, 42.5 g NaOH in a sufficient amount of water for injection (EPPI) to obtain 5 L buffer. The reaction medium containing 2.6 g sodium hyaluronate fibers, 13.9 g sodium hydroxide and 0.11 g buffer, homogenized again by manual mechanical stirring, is left for 3 hours in a water bath previously thermostated at 9 °C.

[0630] After 3 hours of stirring, the reaction medium is neutralized by adding 1N HCl and phosphate buffer with mechanical stirring. The gel obtained is dialyzed against phosphate buffer until the concentration of hyaluronic acid is 27.5 mg / g. After mechanical homogenization and bubble removal, the gel obtained, with a Tan Δ of 1.59 at 1 Hz, can be filtered through a membrane with a porosity of 0.22 μm. It is dispensed into a 1 ml glass syringe and steam sterilized at 127 °C and FO value of 50 min. After heat treatment, the syringe contains the solution of the polymer. A sample is taken from the syringe and a filterability test with a membrane with a porosity of 0.22 μm is performed on this sample. The filterability test is positive. The solution of the polymer is filterable through a membrane with a porosity of 0.22 μm. Nature of the composition of the invention

[0631] The measured flow properties of the compositions of the invention are shown in Table 1 below. TIFF2024544753000003.tif54170

[0632] The compositions of the invention are filterable through a membrane of 0.22 μm porosity and have a Tan Δ of 1.00 or greater. Hydraulic behavior of the inventive composition compared to reference example 3

[0633] The compositions are subjected to steric exclusion chromatography analysis to determine the coefficients of the Mark-Hwink relationship below. [η] = KM a [η] is the intrinsic viscosity M is the viscosity average molecular weight The linear relationship makes it easy to determine the values ​​of the coefficients and K. If we express log([η]) as a function of log(M), we obtain a line of slope and intercept log(K).

[0634] In FIG. 1, a curve (intrinsic viscosity-dL / g) is plotted as a function of log(molecular weight-Da) for each of the compositions and the linear correlation is calculated.

[0635] The measured and calculated values ​​are shown in Table 2 below. TIFF2024544753000004.tif49170

[0636] It can be seen that the hyaluronic acids of the inventive composition have an intrinsic viscosity and a Mark-Hwink coefficient similar to that of the hyaluronic acid of the composition of Reference Example 3 and are therefore no longer crosslinked. Nevertheless, they exhibit very different G' and Tan Δ and are suitable for the above mentioned applications. Example 4

[0637] Sodium hyaluronate fibers (12.5 g) of injectable grade with a weight average molecular weight of 1 MDa are weighed into a container. A 1% aqueous solution of sodium hydroxide is added to hydrate the sodium hyaluronate fibers. The reaction medium is homogenized by alternating current manual mechanical stirring and left for 50 minutes. 0.56 g of BDDE is added. The reaction medium containing 12.5 g of sodium hyaluronate fibers, 74.4 g of sodium hydroxide and 0.56 g of BDDE is again homogenized by manual mechanical stirring and left for 3 hours in a water bath previously thermostated at 8°C.

[0638] At the end of the crosslinking, the reaction medium is neutralized by adding 1N HCl and phosphate buffer with 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 a sufficient amount of water for injection (EPPI) to obtain 120 L of buffer. The resulting gel is dialyzed against phosphate buffer containing 35 g / L mannitol until the concentration of hyaluronic acid is 27.5 mg / g.

[0639] The resulting gel cannot be filtered through a membrane of 0.22 μm porosity.

[0640] The gel with a Tan Δ of 0.70 at 1 Hz is divided into aliquots.

[0641] A first aliquot of 10.0 g of gel is acidified with 350 μl of 1N HCl to pH 3. The first aliquot is digested for 32 hours at 50° C. The gel is neutralized with 1N NaOH to pH 7.3. The flow properties are measured.

[0642] A second aliquot of the gel is exposed for 397 hours to radiation emitted by a 370 nm UV lamp at a power of 370 W. After the exposure is over, the fluidity of the resulting gel is measured.

[0643] A third aliquot of 10.0 g of gel was placed in a syringe and thermostated at 37 °C. The temperature is adjusted to 37°C. A second syringe containing 100 μl of hyaluronan degrading enzyme solution with a strength of 1750 units is thermostatically controlled at 37°C. The two syringes are connected by a double luer connector and the hyaluronan degrading enzyme and gel are mixed and homogenized by moving back and forth from syringe to syringe for 30 seconds. The mixture is placed at 37°C for 45 minutes. At the end of the enzymatic digestion, the digestion is stopped by immersion in a 100°C water bath for 4 minutes and returned to 25°C using an ice bath, and the flowability is measured.

[0644] The results of all the flow measurements are summarized in Table 3 below. TIFF2024544753000005.tif38170

[0645] Regardless of the decomposition method applied, the Tan Δ(Tn δ) at 1 Hz of the obtained compounds is strictly greater than 1.00 (Tan Δ(Tn δ)>1.00).

[0646] The different aliquots obtained according to the different digestion methods are all filterable through a membrane with a porosity of 0.22 μm. Example 5

[0647] The composition of the invention is prepared in a similar manner to that described in Example 4 up to the crosslinking stage. At the end of the crosslinking stage, the reaction medium is neutralized by adding 1N HCl and phosphate buffer with mechanical stirring. The phosphate buffer is prepared by dissolving 0.23 g of NaH2PO4·2H2O, 1.12 g of Na2HPO4, 42.5 g of NaOH in a sufficient amount of water for injection (EPPI) to obtain 5 L of buffer. The gel obtained is dialyzed against the phosphate buffer until the concentration of hyaluronic acid is 20.0 mg / g.

[0648] The resulting gel cannot be filtered through a membrane of 0.22 μm porosity.

[0649] The gel with a Tan Δ of 0.79 at 1 Hz is divided into aliquots.

[0650] A first aliquot of 10.0 g of the gel is contacted with a solution containing 37.5 μl of aqueous iron phosphate heptahydrate at a concentration of 15 mg / g and 75 μl of 30% by volume hydrogen peroxide, and homogenized by 60 syringe-to-syringe strokes to induce radical decomposition after the Fenton reaction. After 4 hours and 20 minutes of contact, the gel is purified by dialysis against a phosphate buffer solution. The fluidity is measured at the end of two dialysis baths lasting a total of 41 hours and 30 minutes.

[0651] A second aliquot of 10.0 g of gel is acidified with 250 μl of 1N HCl to pH 3. The second aliquot is digested for 14 h 19 min at 50° C. Neutralize with 1N NaOH to pH 7.3. The fluidity is measured.

[0652] A third aliquot of 10.0 g of gel was placed in a syringe and thermostated at 37 °C. The second syringe containing 50 μl of hyaluronan degrading enzyme solution with a strength of 1750 units is thermostatically controlled at 37°C. The two syringes are connected by a double luer connector to mix the hyaluronan degrading enzyme and the gel, homogenizing them by moving back and forth from syringe to syringe for 30 seconds. The mixture is placed at 37°C for 25 minutes. At the end of the enzymatic digestion, the digestion is stopped by immersion in a water bath at 100°C for 4 minutes, and the temperature is returned to 25°C using an ice bath, and the flowability is measured.

[0653] The results of all the flow measurements are summarized in Table 4 below. TIFF2024544753000006.tif38170

[0654] Regardless of the decomposition method applied, the Tan Δ(Tn δ) at 1 Hz of the obtained compounds is strictly greater than 1.00 (Tan Δ(Tn δ)>1.00).

[0655] The different aliquots obtained according to the different digestion methods are all filterable through a membrane with a porosity of 0.22 μm.

Claims

1. A formulation comprising a composition in aqueous solution, the composition is filterable through a membrane with a porosity of 0.22 μm and comprises at least one polymeric compound consisting of a sequence of identical or different polysaccharides linked by divalent radicals L; A formulation characterized in that the concentration of the polymeric compound is between 2 mg / g and 200 mg / g, relative to the total mass of the formulation.

2. 2. The formulation according to claim 1, characterized in that the polymeric compound is selected from compounds of general formula (I): (P in the formula 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, and n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 2000, and at least one of n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' is 2 or greater; x, x', x'' and x''' are integers of 0 or greater; L is a divalent radical resulting from the reaction of the crosslinker with two reactive functional groups, each reactive functional group being contained in two separate polysaccharide chains; The polysaccharide chains do not form ring structures.

3. The polymer compound is characterized in that it is composed of a polysaccharide sequence selected from the group consisting of hyaluronic acid, keratan, heparin, cellulose, cellulose derivatives, alginic acid, xanthan, carrageenan, chitosan, chondroitin, heparosan, and bioacceptable salts thereof, either singly or in mixtures. The formulation of claim 1.

4. A polymeric compound of general formula (I): (P in the formula 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, and n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' are integers between 0 and 2000, and at least one of n1, n1', n1'', n1''', n2, n2', n2'', n2''', n3, n3', n3'', n3''', n4, n4', n4'', and n4''' is 2 or greater; x, x', x'' and x''' are integers of 0 or greater; L is a divalent radical resulting from the reaction of the crosslinker with two reactive functional groups, each reactive functional group being contained in two separate polysaccharide chains; Polysaccharide chains do not form ring structures.

5. A method for preparing the composition of claim 1 in the form of an aqueous solution, comprising: the composition is filterable through a membrane with a porosity of 0.22 μm and comprises at least one polymeric compound consisting of a sequence of identical or different polysaccharides linked by divalent radicals L; A manufacturing method comprising at least the following steps: a) providing at least one polysaccharide; b) providing at least one cross-linking agent; c) carrying out one or more cross-linking steps in the presence of said polysaccharide and said cross-linking agent, and carrying out a step of diluting said cross-linked polysaccharide so that the concentration of said cross-linked polysaccharide is between 2 mg / g and 200 mg / g relative to the total weight of said composition; d) obtaining cross-linked polysaccharides; e) carrying out one or more steps of cleaving glycosidic bonds; f) obtaining a solution of the polymer compound; g) filtering the polymer solution through a membrane with a porosity of 0.22 μm;

6. The method of claim 5, wherein the crosslinking agent is selected from the group consisting of bis-epoxides, trimetaphosphates, diamines, dialkoxyamines, and dihydrazides.

7. 7. The method according to claim 5 or 6, characterized in that step e) of glycosidic bond cleavage is carried out by chemical treatment.

8. 7. The method according to claim 5 or 6, characterized in that step e) of glycosidic bond cleavage is carried out by heat treatment.

9. 7. The method according to claim 5 or 6, characterized in that step e) of glycosidic bond cleavage is carried out by radiation treatment.

10. 7. The method according to claim 5 or 6, characterized in that step e) of glycosidic bond cleavage is carried out by enzymatic treatment.

11. 7. The method according to claim 5 or 6, characterized in that step e) of glycosidic bond cleavage is carried out by high pressure treatment.

12. 12. A composition obtained by the process according to any one of claims 5 to 11, characterized in that Tan Δ (Tnδ) ≥ 1.00 at 1 Hz.

13. 4. A formulation according to any one of claims 1 to 3, characterized in that it further comprises at least one active ingredient.

14. 14. The formulation according to claim 13, characterized in that the active ingredient is selected from the group consisting of local anesthetics, antioxidants, anti-inflammatory agents, vitamins, amino acids, vasoconstrictors, vasodilators, antihemorrhagic or hemostatic agents, antibacterial agents, glycosides and derivatives thereof, moisturizers, or tissue regenerating agents, alone or in mixture.

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