FR3162003A1 - BIOMATERIAL COMPOSITION COMPRISING POLYHYDROXYBUTYRATE IN PARTICULAR FORM - Google Patents

Abstract

BIOMATERIAL COMPOSITION COMPRISING A POLYHYDROXYBUTYRATE IN PARTICLE FORM The present invention relates to an injectable biomaterial, and more particularly to a filler or implant consisting of a composition comprising at least one biocompatible polymer and PHA particles. The present invention therefore relates to a composition comprising a biocompatible polymer and PHA particles. It also relates to PHA particles and their preparation process. It also relates to the process for preparing said injectable composition comprising a biocompatible polymer and PHA particles. It also relates to methods for implanting said injectable composition comprising PHA particles.

Description

BIOMATERIAL COMPOSITION COMPRISING POLYHYDROXYBUTYRATE IN PARTICULAR FORM

The invention relates to the field of biomaterial formulations, particularly in the medical and aesthetic fields. In these applications, the formulations must exhibit optimized properties in terms of rheology, taking into account the characteristics of the medical device used for injection, the characteristics of the treated area, and the desired therapeutic or cosmetic effect, as well as in terms of persistence to achieve the most lasting effect possible.

Human skin tissue is composed of fibrous proteins such as collagen and an extracellular matrix (ECM) containing fibronectin, laminin, and glycosaminoglycans (GAGs). When skin tissue is damaged—that is, when it has defects, for example, due to aging—a common method is to restore function and form by injecting a filler material made of synthetic polymers or biological tissues into the affected area. This technique can restore tissue, fill defects, improve wrinkles, or correct contours.

The raw materials for fillers used in facial plastic surgery include autologous dermal implants, collagen, or hyaluronic acid (HA).

To improve the performance of dermal fillers, attempts have been made to manufacture fillers containing microspheres made of biopolymers with a slow decomposition rate. Among these, fillers containing polycaprolactone (PCL) are absorbed by the body. Their very slow decomposition rate is why they are used as semi-permanent fillers. However, these products do have drawbacks, such as the generation of byproducts that can cause inflammatory reactions.

It is therefore necessary to develop filling compositions based on a new material that not only has high biocompatibility, immediate and lasting effects, but also has the advantage of generating a high-quality and properly organized neo-tissue.

Polyhydroxyalkanoates (PHAs) are diverse biopolyesters with a similar structure and different side-chain groups, synthesized by a variety of microorganisms. Due to their excellent biodegradability and biocompatibility, PHAs have been used in numerous applications, including medical implants, bioengineering, and regenerative medicine.

The preparation of PHA-based particles that can be incorporated into filler materials, which are by design aqueous formulations, presents numerous technical difficulties, ranging from the difficulty of preparing particles with a size distribution compatible with the preparation of injectable formulations under conditions that meet the precision requirement necessary for wrinkle filling.

The present invention relates to an injectable biomaterial and more particularly to a filler or implant consisting of a composition comprising at least one biocompatible polymer and PHA particles.

The present invention therefore relates to a composition comprising a biocompatible polymer and PHA particles.

It also concerns PHA particles and their production process.

It also relates to the preparation process of said injectable composition comprising a biocompatible polymer and PHA particles.

It also relates to the methods of implanting said injectable composition comprising PHA particles.

In one embodiment, the composition according to the invention is in solvent-free form, in particular water-free, for example in lyophilized form.

In one embodiment, the composition is in the form of an injectable aqueous composition.

When the composition is in aqueous form, the biocompatible polymer is in the form of a gel or hydrogel, and the PHA particles are suspended in the gel.

In one embodiment, the composition is in the form of a bipartite composition, one part of which comprises an aqueous polymeric phase and the other part comprises particles suspended in an organic phase miscible in the aqueous polymeric phase.

In one embodiment, the composition is in the form of a bipartite composition, one part of which comprises an aqueous polymeric phase and the other part comprises the particles in dry form.

In the context of this application, a “hydrogel” is defined as a polymeric gel consisting of a three-dimensional network made up of at least one polymer, capable of absorbing a large quantity of water or aqueous solution and which exhibits particular rheological properties, particularly in terms of viscosity and viscoelasticity.

The said network can be formed by grafting and/or chemical crosslinking by creating bonds between polymer chains, these bonds being covalent bonds.

This network can also be obtained through transient physical interactions, for example ionic, hydrophobic or hydrogen bonds.

When the composition is in aqueous form, the aqueous phase consists of water or an aqueous saline solution, for example a phosphate buffer solution, for example PBS or a biological type buffer used in cell culture of the HEPES type ((hydroxyethyl)piperazinyl]ethanesulfonic acid)

Dissolving the biocompatible polymer or the composition according to the invention or in solvent-free form is carried out by adding water or an aqueous saline solution, for example a phosphate buffer solution, for example PBS.

In one embodiment, the biocompatible polymer is dissolved by adding water or an aqueous saline solution, for example a phosphate buffer solution, for example PBS, further comprising at least one active ingredient, for example an antioxidant and/or a local anesthetic.

In this application, the term "filler" or "filler product" refers to a product intended to be injected into the dermal or subcutaneous layer of the skin in order to visually improve the wrinkled area of the face, and to serve to maintain volume by itself without pharmacological action.

On the other hand, implants are biomaterials used to replace and repair damaged organs in the human body.

Therefore, although the filler and the implant have the same composition (biocompatible polymer, PHA particles), their application objectives and effects are different.

In one embodiment, said biocompatible polymer is a polysaccharide and/or a protein and/or a polypeptide alone or in mixture.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 2 mg/g and 200 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 2 mg/g and 75 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 5 mg/g and 50 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 2 mg/g and 50 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 4 mg/g and 40 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 5 mg/g and 30 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 10 mg/g and 30 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 10 mg/g and 40 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 2 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 4 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 5 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 6 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 8 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 10 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 14 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 20 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 40 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 60 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 80 mg/g.

In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 100 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 200 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 75 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is between 5 mg/g and 50 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 50 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is between 4 mg/g and 40 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is between 5 mg/g and 30 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is between 10 mg/g and 30 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is between 10 mg/g and 40 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 2 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 4 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 5 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 6 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 8 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 10 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 14 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 20 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 40 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 60 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 80 mg/g.

In one embodiment, the PHA particle content in the injectable aqueous composition is 100 mg/g.

In one embodiment, the polysaccharide is selected from the group consisting of hyaluronic acid, keratin, heparin, cellulose, cellulose derivatives (including methylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, ethylmethylcellulose, carboxymethylcellulose), alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparosan, and their biologically acceptable salts, alone or in mixture.

In one embodiment, said polysaccharide is hyaluronic acid.

In one embodiment, said polysaccharide is hyaluronic acid or one of its salts, alone or in mixture.

In one embodiment, said polysaccharide is hyaluronic acid in the form of sodium or potassium salt.

In one embodiment, said polysaccharide is hyaluronic acid in the form of sodium salt.

In one embodiment, said polysaccharide is non-crosslinked hyaluronic acid or one of its salts, alone or in mixture

In one embodiment, said polysaccharide is cross-linked hyaluronic acid or one of its salts, alone or in mixture.

In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of at least one crosslinking agent.

In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of at least one bi- or polyfunctional crosslinking agent.

In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of at least one bifunctional crosslinking agent being butanedioldiglycidyl ether (BDDE) or 1,2,7,8-diepoxyoctane or divinylsulfone.

In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of BDDE.

In one embodiment, said polysaccharide is cross-linked hyaluronic acid or one of its salts, alone or in mixture, and in that said cross-linking is carried out by means of sodium or potassium trimetaphosphate.

When the polysaccharide, and more specifically hyaluronic acid, is cross-linked using a cross-linking agent, the degree of cross-linking (x) is theoretically calculated using the following formula:

With regard to hyaluronic acid, the repeating unit is a disaccharidic motif.

In one embodiment, the cross-linked hyaluronic acid has a cross-linking ratio x between 0.001 and 0.5.

In one embodiment, the cross-linked hyaluronic acid has a cross-linking ratio x between 0.01 and 0.4.

In one embodiment, the cross-linked hyaluronic acid has a cross-linking ratio x between 0.1 and 0.3.

In one embodiment, the cross-linked hyaluronic acid has a cross-linking ratio x of 0.06.

In one embodiment, the cross-linked hyaluronic acid has a cross-linking ratio x of 0.07.

In one embodiment, the cross-linked hyaluronic acid has a cross-linking ratio x of 0.12.

In one embodiment, said polysaccharide is co-crosslinked hyaluronic acid or one of its salts, alone or in mixture.

In one embodiment, said polysaccharide is chemically modified hyaluronic acid by substitution, crosslinked or non-crosslinked, or one of its salts, alone or in mixture.

In one embodiment, said polysaccharide is doubly cross-linked hyaluronic acid as described in patent application WO2000/046253 in the name of Fermentech medical limited.

In one embodiment, said polysaccharide is a mixture of hyaluronic acids, or one of their salts, cross-linked and non-cross-linked.

In one embodiment, said polysaccharide is a cross-linked mixture of hyaluronic acids, or one of their salts.

In one embodiment, said polysaccharide is a cross-linked mixture of hyaluronic acids, or one of their salts, such as that described in patent application WO2009/071697 in the name of the applicant.

In one embodiment, said polysaccharide is a mixture of hyaluronic acids, obtained by mixing several hyaluronic acids, or one of their salts, of different molecular masses prior to their cross-linking, as described in patent application WO2004092222 in the name of Cornéal industrie.

In one embodiment, the polysaccharide is hyaluronic acid or one of its salts, substituted by a group providing lipophilic or hydrating properties, such as substituted hyaluronic acids as described in patent application FR2983483 filed by the applicant. This application describes a process for the simultaneous substitution and crosslinking of a polysaccharide via its hydroxyl groups in an aqueous phase, characterized in that it comprises the following steps: (i) a polysaccharide is placed in an aqueous medium, (ii) it is in contact with at least one precursor of a substituent, (iii) it is in contact with a crosslinking agent, and (iv) the substituted and crosslinked polysaccharide is obtained and isolated. These process steps can be readily implemented within the scope of the present invention.

In one embodiment, said polysaccharide is hyaluronic acid or one of its salts, grafted with glycerol, for example as described in application WO2017162676 on behalf of MERZ.

In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is within a range of 0.01 MDa to 5 MDa (0.01 ≤Mw ≤5 MDa).

In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is within a range of 0.1 MDa to 3.5 MDa (0.1 ≤Mw ≤3.5 MDa).

In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is within a range of 1 MDa to 3 MDa (1 ≤Mw ≤3 MDa).

In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 1 MDa.

In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 3 MDa.

In one embodiment, the concentration of hyaluronic acid [HA] is between 2 mg/g and 50 mg/g of total weight of said injectable aqueous composition.

In one embodiment, the concentration of hyaluronic acid [HA] is between 4 mg/g and 40 mg/g of total weight of said injectable aqueous composition.

In one embodiment, the concentration of hyaluronic acid [HA] is between 5 mg/g and 30 mg/g of total weight of said injectable aqueous composition.

In one embodiment, the concentration of hyaluronic acid [HA] is between 10 mg/g and 30 mg/g of total weight of said injectable aqueous composition.

In one embodiment, the concentration of hyaluronic acid [HA] is 20 mg/g of total weight of said injectable aqueous composition.

In one embodiment, the concentration of hyaluronic acid [HA] is 14 mg/g of total weight of said injectable aqueous composition.

In one embodiment, the concentration of hyaluronic acid [HA] is 10 mg/g of total weight of said injectable aqueous composition.

Furthermore, according to one embodiment of the invention, the composition comprises at least one protein and/or polypeptide alone or in mixture with polysaccharides.

According to one embodiment of the invention, the protein(s) and/or polypeptide(s) are selected from collagen, extracellular matrix proteins, structural proteins, blood-derived proteins, glycoproteins, lipoproteins, natural proteins, synthetic proteins, intracellular proteins, extracellular proteins, membrane proteins, and all combinations thereof.

According to one embodiment of the invention, collagen is selected from native collagen, collagen types I, II, III, V, XI, XXIV, collagen types VIII and X, collagen type IV, collagen type VI, collagen type VII, collagen types XIII, XVII, XXIII and XXV, collagen types XV and XVIII, fibrillar collagen, collagen associated with fibrils types IX, XII, XIV, XVI, XIX, XX, XXI, XXII and XXVI and all their combinations.

Other forms of collagen that can be used include fibrillar atelopeptide collagen, telopeptide-containing collagen, freeze-dried collagen, mammalian collagen, recombinant collagen, reconstituted collagen, atelopeptide collagen obtained from a vertebrate species, recombinant collagen, collagen produced by genetically modified eukaryotic or prokaryotic cells or by genetically modified organisms, purified collagen and reconstituted purified collagen, collagen derived from genetically modified plants, proto-collagen and all their combinations.

PHAs are chosen from first generation PHAs such as poly(3-hydroxybutyric acid) (PHB) and poly(4-hydroxybutyric acid) (P4HB), second generation poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), third generation poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx or PHBHx), fourth generation poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB or P3HB4HB) or fifth generation poly(3-hydroxybutyricacid-co-3-hydroxyvalericacid-co-3-hydroxyhexanoic acid) (PHBVHHx or PHBVHx), alone or in mixture.

In one embodiment, PHA is P4HB.

In one embodiment, the PHA particles have a size strictly less than 100 µm.

In one embodiment, the PHA particles have a size strictly less than 80 µm.

In one embodiment, the PHA particles have a size strictly less than 60 µm.

In one embodiment, the PHA particles have a size between 5 and 100 µm.

In one embodiment, the PHA particles have a size between 5 and 80 µm.

In one embodiment, the PHA particles have a size between 5 and 60 µm.

In one embodiment, the PHA particles have a size between 20 and 100 µm.

In one embodiment, the PHA particles have a size between 40 and 100 µm.

In one embodiment, the PHA particles have a size between 20 and 90 µm.

In one embodiment, the PHA particles have a size between 40 and 90 µm.

In one embodiment, the PHA particles have a size between 20 and 80 µm.

In one embodiment, the PHA particles have a size between 20 and 60 µm.

Controlling the particle size distribution during the particle formation process is fundamental because the PHA particles are incorporated into a biomaterial which is an injectable composition.

PHA particles are preferably prepared directly by a solvent evaporation technique, a double emulsion technique, or by microfluidization, using methods known from the literature. (Koosha, doctoral thesis, 1989, Univ. Nottingham, UK, Diss. Abstr. Int. B 51:1206 (1990); Bruhn & Müeller, Proceed Intern. Symp. Control. Rel. Bioact. Mater. 18:668-69 (1991); Conti, et al, J Microencapsulation,9:153-66 (1992); Ogawa, et al, Chem. Bull., 36:1095-103 (1988);

In one embodiment, the processes for preparing PHA particles are chosen from solvent evaporation techniques of a dispersed phase of an emulsion, said emulsion being obtained classically by mixing, by ultrasonic emulsification or by membrane emulsification, but also from conventional spray-drying techniques or possibly electrostatic spraying and/or microfluidization.

In one embodiment, the particles are prepared by microfluidization using the droplet generation device described in patent application WO2019/007965 in the name of the UNIVERSITE LIBRE DE BRUXELLES, the contents of which are incorporated by reference.

In one embodiment, PHA is solubilized in an organic solvent that is immiscible and/or soluble in water, such as dichloromethane, for example at a concentration of between 1 and 10% of PHA in dichloromethane.

Water is added to the emulsion generation system to form droplets which are then collected, rinsed to remove solvents and then dried to obtain PHA particles.

In one embodiment, a surfactant or film-forming agent such as polyvinyl alcohol is added to the aqueous phase.

The dried particles, possibly calibrated by sieving, are then incorporated into the biocompatible polymer composition using various processes.

The PHA particles are suspended in water or an aqueous saline solution, for example a phosphate buffer solution, for example PBS, and then this suspension is incorporated into a composition comprising a biocompatible polymer in solution or in the form of a gel or hydrogel in water or in an aqueous saline solution, for example a phosphate buffer solution, for example PBS, to obtain a composition according to the invention.

To preserve this composition, it can be lyophilized and then rehydrated with water or an aqueous saline solution, for example, a phosphate buffer solution (PBS), before injection. The invention also relates to the composition comprising the biocompatible polymer and PHA in lyophilized form. This composition must be rehydrated before use.

In one embodiment, the PHA particles are suspended in an organic liquid such as glycerol, preferably polyethylene glycol having a molecular mass less than 1000 g/mol, or dimethyl sulfoxide.

In one embodiment, the PHA particles are incorporated without having been suspended directly into an aqueous composition of biocompatible polymer in the form of a gel or hydrogel to obtain, by means of a mixing device, an injectable composition according to the invention.

In one embodiment the mixing device consists of two syringes connected at the time of incorporation and the mixing is carried out by making several back and forth movements until a homogeneous suspension is obtained.

In one embodiment, the mixing device is a conventional mixer and the incorporation of particles is carried out by mixing until a homogeneous composition is obtained.

The product can be incorporated by the practitioner just before use. Alternatively, it can be incorporated at the end of the manufacturing process, and the resulting composition will then be subjected to a freeze-drying process to rehydrate it before injection.

In one embodiment, the composition according to the invention further comprises an active ingredient selected from the group consisting of local anesthetics, vitamin C derivatives, anti-inflammatories, antioxidants, and mixtures thereof.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic.

In one embodiment, the composition according to the invention further comprises a local anesthetic at a local anesthetic concentration of between 0.1 and 5%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises a local anesthetic at a local anesthetic concentration of between 0.1 and 4%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises a local anesthetic at a local anesthetic concentration of between 0.1 and 2%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises a local anesthetic to obtain a local anesthetic concentration of between 0.1 and 1%, relative to the total mass of said formulation.

In one embodiment, the composition according to the invention further comprises a local anesthetic to obtain a local anesthetic concentration of between 0.1 and 0.5%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises local anesthetic at a local anesthetic concentration of approximately 0.3%, relative to the total mass of said composition.

In one embodiment, the local anesthetic is chosen from the amino-ester group.

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

In one embodiment, the local anesthetic is chosen from the amino-amide group.

In one embodiment, the amino-amide is selected from the group comprising lidocaine, mepivacaine, prilocaine, articaine, aptocaine, bupivacaine, etidocaine and ropivacaine in base or salt form, for example in hydrochloride form.

In one embodiment, the local anesthetic is chosen from the amino-ether group.

In one embodiment, the amino-ether is chosen from the group comprising diamocaine and pramocaine in base or salt form, for example in hydrochloride or cyclamate form.

In one embodiment, the amino-ether is chosen from the group consisting of lidocaine, mepivacaine, and their isolated salts and isomers.

In one embodiment, the amino-ether is lidocaine.

In one embodiment, the amino-ether is lidocaine or one of its pharmaceutically acceptable salts.

In one embodiment, the amino-ether is lidocaine hydrochloride.

In one embodiment, the amino-ether is mepivacaine.

In one embodiment, the amino-ether is mepivacaine or one of its pharmaceutically acceptable salts.

In one embodiment, the amino ether 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 their pharmaceutically acceptable salts.

In one embodiment, the amino-ether is mepivacaine hydrochloride.

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

In one embodiment, the amino-ether is (s)-mepivacaine hydrochloride.

In one embodiment, the amino-ether is racemic mepivacaine hydrochloride.

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

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

In one embodiment, the amino-ether is racemic mepivacaine.

In one embodiment, the composition according to the invention further comprises at least one amino-ether local anesthetic selected from the group consisting of lidocaine, mepivacaine, and mixtures thereof.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 5%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 4%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 2%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 1%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 0.5%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of approximately 0.3%, %, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely mepivacaine.

In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 5%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 4%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 2%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 1%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 0.5%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a mepivacaine concentration of approximately 0.3%, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic being diclonine in base or salt form, for example in hydrochloride form.

In one embodiment, the composition according to the invention further comprises at least one local anesthetic selected from the group consisting of chlorobutanol, guafecainol and polidocanol.

In one embodiment, the local anesthetic is chlorobutanol.

In one embodiment, the local anesthetic is guafecainol.

In one embodiment, the local anesthetic is polidocanol.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group consisting of steroidal and non-steroidal anti-inflammatory drugs.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory drug selected from the group consisting of non-steroidal anti-inflammatory drugs.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group comprising salicylate anti-inflammatories, propionic derivatives, indolic derivatives, pyrazole derivatives, oxicams and coxibs.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group comprising diclofenac, nimesulfide, niflumic acid, mefenamic acid and nabumetone, alone or in mixture.

In one embodiment, the composition according to the invention further comprises at least one salicylate anti-inflammatory selected from the group comprising diflunisal, benorilate and aspirin, alone or in mixture.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of propionic derivatives including alminoprofen, ketoprofen, ibuprofen, naproxen, flurbiprofen and tiaprofenic acid, alone or in mixture.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of indole derivatives including indomethacin, sulindac and etodolac, alone or in mixture.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of pyrazole derivatives including in particular phenylbutazone.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of oxicams including piroxicam, tenoxicam and meloxicam, alone or in mixture.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of coxibs comprising celecoxib, etoricoxib and rofecoxib, alone or in mixture.

In one embodiment, the concentration of non-steroidal anti-inflammatory drugs in the composition according to the invention is between 0.01 and 2000 mg/g.

In one embodiment, the concentration of non-steroidal anti-inflammatory drugs in the composition according to the invention is between 0.1 and 1000 mg/g.

In one embodiment, the concentration of non-steroidal anti-inflammatory drugs in the composition according to the invention is between 0.5 and 500 mg/g.

In one embodiment, the composition according to the invention further comprises at least one steroidal anti-inflammatory.

In one embodiment, the composition according to the invention further comprises at least one steroidal anti-inflammatory selected from the group including dexamethasone, prednisolone, corticosterone, budesonide, sulfasalazine, mesalamine, cetirizine, diphenhydramine, antipyrine, methyl salicylate, loratadine, thymol, carvacrol, bisabolol, allantoin, eucalyptol, phenazone (antipyrine), propyphenazone, alone or in mixture.

In one embodiment, the concentration of steroidal anti-inflammatory drugs in the composition according to the invention is between 0.01 and 2000 mg/g.

In one embodiment, the concentration of steroidal anti-inflammatory drugs in the composition according to the invention is between 0.1 and 1000 mg/g.

In one embodiment, the concentration of steroidal anti-inflammatory drugs in the composition according to the invention is between 0.5 and 500 mg/g.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group consisting of sucrose octasulfate and its salts.

In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group consisting of sucrose octasulfate and its sodium and potassium salts.

In one embodiment, the composition according to the invention further comprises at least one water-soluble anti-inflammatory salt of sucrose octasulfate selected from the group consisting of alkali metal salts, alkaline earth metal salts, silver salts, ammonium salts, amino acid salts.

In one embodiment, the composition according to the invention further comprises at least one water-soluble anti-inflammatory salt of sucrose octasulfate selected from the group consisting of alkali metal salts or alkaline earth metal salts.

In one embodiment, the composition according to the invention further comprises at least one water-soluble anti-inflammatory salt of sucrose octasulfate selected from the group consisting of sodium salt of sucrose octasulfate or potassium salt of sucrose octasulfate.

In one embodiment, the composition according to the invention further comprises at least one antimicrobial.

In one embodiment, the composition according to the invention further comprises at least one antimicrobial selected from the group including gentamicin, silver sulfadiazine, metronidazole, fucidin, bacitracin, eosin, povidone-iodine, copper gluconate, zinc gluconate, manganese gluconate or their salts, alone or in mixture.

In one embodiment, the concentration of antimicrobials in the composition according to the invention is between 0.1 and 200 mg/g.

In one embodiment, the concentration of antimicrobials in the composition according to the invention is between 0.5 and 100 mg/g.

In one embodiment, the composition according to the invention further comprises at least one glycoside or a glycoside derivative.

In one embodiment, the composition according to the invention further comprises at least one glycoside or a glycoside derivative selected from the group comprising D-glucopyranose, 1,4-glycoside, esculin, hesperidin, diosmin, arbutin, skimmine or aloin, alone or in mixtures.

In one embodiment, the concentration of glycosides in the composition according to the invention is between 0.1 and 200 mg/g.

In one embodiment, the concentration of glycosides in the composition according to the invention is between 0.5 and 100 mg/g.

In one embodiment, the composition according to the invention further comprises at least antioxidant.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of polyols.

In one embodiment, the composition according to the invention further comprises at least one polyol selected from the group consisting of mannitol, sorbitol, propylene glycol, xylitol, glycerol, maltitol, lactitol and erythritol.

In one embodiment, the composition according to the invention further comprises at least one polyol selected from the group consisting of mannitol, sorbitol, maltitol and glycerol, alone or in mixture.

In one embodiment, the composition according to the invention further comprises at least one polyol selected from the group consisting of mannitol, sorbitol and maltitol, alone or in mixture.

In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 0.1 mg/ml and 50 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 5 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 10 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 20 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 30 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol at a concentration between 5 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol at a concentration between 10 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol at a concentration between 20 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol at a concentration of between 30 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol at a concentration between 5 mg/ml and 40 mg/ml, relative to the total mass of said composition.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol at a concentration of between 10 mg/ml and 40 mg/ml, relative to the total mass of said composition

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol at a concentration of between 20 mg/ml and 40 mg/ml, relative to the total mass of said composition

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol at a concentration of between 30 mg/ml and 40 mg/ml, relative to the total mass of said composition

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being maltitol.

In one embodiment, the biomaterial according to the invention further comprises at least one polyol, said polyol being glycerol.

In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being a mixture of mannitol and sorbitol.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of vitamin C derivatives.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of vitamin C derivatives including magnesium ascorbyl phosphate, sodium ascorbyl phosphate, ascorbyl-2-glucoside, and mixtures thereof.

In one embodiment, said at least one vitamin C derivative is magnesium ascorbyl phosphate.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of vitamin E derivatives and tocopherols.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of carotenoids and retinoids and their derivatives.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of carotenoids and retinoids and their derivatives including retinol, retinoic acid, retinal, retinol esters and carotene.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of pseudo-tripeptides.

In one embodiment, the pseudo-tripeptide is glutathione.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group comprising the various forms of coenzyme Q10, ubiquinone or ubiquinol.

In one embodiment, the composition according to the invention further comprises at least one vitamin.

In one embodiment, the composition according to the invention further comprises at least one vitamin selected from the group comprising retinol, thiamine, riboflavin, nicotinamide, dexpenthenol, piridoxine, ascorbic acid, ergocalciferol, tocopherol, biotin and folic acid alone or in mixture.

In one embodiment, the concentration of vitamins in the composition according to the invention is between 0.01 and 200 mg/g.

In one embodiment, the composition according to the invention further comprises at least one vasoconstrictor.

In one embodiment, the composition according to the invention further comprises at least one vasoconstrictor selected from the group comprising naphazoline, epinephrine, methoxamine, methylnorepinephrine, norepinephrine, oxymethazoline, phenylephrine, pseudoephedrine, synephrine, cirazolin and xylomethazoline.

In one embodiment, the concentration of vasoconstrictors in the composition according to the invention is between 0.01 and 3 mg/g.

In one embodiment, the composition according to the invention further comprises at least one vasodilator.

In one embodiment, the composition according to the invention further comprises at least one vasodilator selected from the group comprising adenosine, nicotinic acid, minoxidil and diazoxide, alone or in mixture.

In one embodiment, the concentration of vasodilators in the composition according to the invention is between 0.01 and 10 mg/g.

In one embodiment, the composition according to the invention further comprises at least one anti-hemorrhagic or hemostatic agent.

In one embodiment, the composition according to the invention further comprises at least one antihemorrhagic or hemostatic agent selected from the group comprising aminocaproic acid or tranexamic acid, alone or in mixture.

In one embodiment, the concentration of antihemorrhagic or hemostatic agents in the composition according to the invention is between 0.01 and 5 mg/g.

In one embodiment, the composition according to the invention further comprises at least one antioxidant and at least one local anesthetic.

In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of polyols and at least one local anesthetic selected from the group of amino-amides.

In one embodiment, the composition according to the invention is characterized in that it is injectable.

In one embodiment, the composition according to the invention is characterized in that it is sterile.

In one embodiment, the composition according to the invention is characterized in that it is injectable and sterile.

The composition according to the invention has numerous applications.

Medical applications include injections to replace deficient biological fluids, such as synovial fluid in joints, injections following surgery to prevent post-surgical adhesions, periurethral injections to treat incontinence, and injections following presbyopia surgery. Aesthetic applications include injections to fill wrinkles, fine lines, and skin imperfections, or to augment volume, such as that of the lips, cheekbones, etc.

• injections esthétiques au niveau du visage : de comblement de rides, défauts cutanés ou volumatrices (pommettes, menton, lèvres) ;
• injections volumatrices au niveau du corps : augmentation des seins et des fesses, augmentation du point G, vaginoplastie, reconstruction des lèvres vaginales, augmentation de la taille du pénis ;
• en chirurgie articulaire et en chirurgie dentaire pour le comblement des poches parodontales par exemple.
• traitement de l’arthrose, injection dans l’articulation en remplacement ou en complément du liquide synovial déficient ;
• injection péri-urétrale pour le traitement de l'incontinence urinaire par insuffisance sphinctérienne ;
• injection post-chirurgicale pour éviter les adhésions péritonéales notamment ;
• injection suite à une chirurgie de la presbytie par incisions sclérales au laser ;
• injection dans la cavité vitréenne ;
• injection au cours de la chirurgie de la cataracte ;
• injection pour le traitement des cas de sécheresses vaginales ;
• injection dans les espaces tissulaires ;
• injection dans les parties génitales.

The applications targeted are more specifically those commonly used with injectable viscoelastics and polysaccharides or collagen used or potentially usable in the following pathologies or treatments:

• cosmetic injections on the face: for filling wrinkles, skin defects or for volumizing (cheekbones, chin, lips);
• volumizing injections for the body: breast and buttock augmentation, G-spot augmentation, vaginoplasty, vaginal lip reconstruction, penis enlargement;
• in joint surgery and in dental surgery for filling periodontal pockets for example.
• treatment of osteoarthritis, injection into the joint as a replacement or supplement for deficient synovial fluid;
• periurethral injection for the treatment of urinary incontinence due to sphincter insufficiency;
• post-surgical injection to prevent peritoneal adhesions in particular;
• injection following presbyopia surgery via scleral incisions with laser;
• injection into the vitreous cavity;
• injection during cataract surgery;
• injection for the treatment of cases of vaginal dryness;
• injection into tissue spaces;
• injection into the genitals.

• pour le comblement des rides fines, moyennes ou profondes, et être injectées avec des aiguilles de diamètre fin (27 Gauge par exemple) ;
• comme volumateur avec une injection par des aiguilles de diamètre plus important, de 22 à 26 Gauge par exemple, et plus longues (30 à 40 mm par exemple); dans ce cas, son caractère cohésif permettra de garantir son maintien à l'emplacement de l'injection.

More specifically, in cosmetic surgery, depending on its viscoelastic and residual properties, the composition that is the subject of the invention may be used:

• for filling fine, medium or deep wrinkles, and to be injected with fine diameter needles (27 Gauge for example);
• as a volumizer with injection using larger diameter needles, 22 to 26 Gauge for example, and longer (30 to 40 mm for example); in this case, its cohesive nature will ensure that it remains in place at the injection site.

• combler des volumes ;
• générer des espaces au sein de certains tissus, favorisant ainsi leur fonctionnement optimal ;
• remplacer des liquides physiologiques déficients.

These examples of use are by no means limiting, and the composition that is the subject of the invention can be used for:

• filling volumes;
• to generate spaces within certain tissues, thereby promoting their optimal functioning;
• to replace deficient physiological fluids.

Examples

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

Particle preparation Example A: Particle preparation

A solution of P4HB in dichloromethane with 5% P4HB is prepared and introduced into the device described in patent application WO2019/007965 in the name of the FREE UNIVERSITY OF BRUSSELS.

Water containing 1% polyvinyl alcohol is introduced as a continuous phase in the above-mentioned device.

The flow rate is set at 50 µL/min and particles are obtained which are then washed with water to extract the solvent until it is eliminated and then dried under an airflow.

There is no coalescence and the beads are strong enough to be handled after 30 minutes.

Drying is continued until completion and the particle size is measured; the diameter is 58 µm.

Example 1 : Preparation of Phosphate/NaCl buffer solution

The buffer solution is prepared by introducing 0.225 g of NaH ₂ PO ₄ ∙2H ₂ O, 1.115 g of Na ₂ HPO ₄ , and 42.5 g of NaCl into a 5 L volumetric flask which is filled to the mark with water for injection (WFI).

Example 2 : Composition of non-crosslinked hyaluronic acid

The hyaluronic acid (HA) fibers used in the following examples were purchased from the company HTL. They have an average molecular mass of 2.5 MDa and a moisture content of 13.7%.

0.704 g of wet HA fibers (13.7%) are introduced into 20.30 g of buffer solution, to obtain a [ _{non-crosslinked} HA] concentration of 30 mg.g ^-1 . The composition is hydrated overnight at room temperature.

The following day, the mixture is blended for 4 x 3 minutes with a 1.5-minute rest period between each blending step. The mixture is then hydrated for 48 hours in a cold environment.

The mixture is remixed for 4 x 3 minutes with a rest period of 1.5 minutes between each mixing step. This mixture is then used as is in the following compositions.

Example 3 : Preparation of a composition including CMC and P4HB microbeads

A composition containing carboxymethylcellulose (CMC) is prepared by solubilizing 454 mg of CMC (AQUALON product type 7MF PH BET or 7LF PH BET degree of substitution 0.7 with a viscosity at 2% between 20 and 700 mPa.sec) in 20 mL of buffer solution prepared according to the protocol of example 1.

The composition obtained has a concentration of [CMC] of 22.5 mg.g ^-1 .

162.2 mg of P4HB microbeads with a diameter of 58 µm, obtained in Example A, are introduced into 4.09 mL of the previously obtained CMC composition to form a CMC composition comprising P4HB microbeads

Example 4: Preparation of a composition including RH Collagen III and P4HB microbeads

A composition containing RH Collagen III is prepared by solubilizing 700 mg of RH Collagen III (Product DEMULCENT Purity≥95%, collagen content 90-110%) in 10.3 mL of buffer solution prepared according to the protocol of example 1.

The composition obtained has a concentration of [RH Collagen III] of 68 mg.g-1.

173.2 mg of P4HB microbeads, with a diameter of 58 µm, obtained in Example A, are introduced into 4.33 mL of the buffer solution containing RH Collagen III to obtain an RH Collagen III composition comprising P4HB microbeads.

Example 5 : Preparation of a composition comprising non-crosslinked hyaluronic acid and P4HB microbeads

A composition containing P4HB is prepared by suspending 79.5 mg of P4HB microbeads, 58 µm in diameter, obtained in Example A in 2 mL of buffer solution prepared according to the protocol of Example 1.

The composition obtained has a concentration of [P4HB] of 39.75 mg.g ^-1 .

1 mL of this composition is mixed with 1.048 g of the non-crosslinked hyaluronic acid composition obtained in Example 2. To carry out the mixing, the two compositions are introduced into two 2.5 mL syringes which are connected together, and subsequently, back-and-forth movements are carried out until a homogeneous composition of non-crosslinked hyaluronic acid containing P4HB microbeads is obtained.

Example 6 : Freeze-drying of compositions

Two grams of each of the compositions in examples 3, 4, and 5 are taken and placed in the freeze dryer. Freeze drying is carried out for 12 hours at a temperature of -55 °C, under a pressure of 100 mTorr.

After freeze-drying, the freeze-dried compositions are kept cold.

Example 7: Preparation of injectable compositions

The following non-limiting examples present distinct embodiments for preparing injectable compositions comprising different polysaccharides mixed with P4HB microbeads.

One embodiment consists of carrying out rehydration just before the injection of the mixtures obtained in example 6, using water for injection (WFI).

A second embodiment consists of mixing just before injection an effective quantity of P4HB microbeads obtained in example A with an injectable composition comprising at least one polysaccharide.

A third embodiment consists of preparing a mixture just before injection of a suspension in injectable water comprising P4HB microbeads obtained in example A with an injectable composition comprising at least one polysaccharide.

Example 7a

Composition 7a, comprising RH Collagen III and P4HB microbeads, 58 µm in diameter, obtained in Example A, is prepared by hydrating 2 g of the lyophilized composition obtained in Example 6 from the composition obtained in Example 4 in 2 mL of water for injection.

This composition is divided into 4 portions. These are introduced into 4 1 mL syringes.

Example 7b

Composition 7b, comprising non-crosslinked hyaluronic acid (HA) and P4HB microbeads, 58 µm in diameter, obtained in Example A, is prepared by hydrating 2 g of the lyophilized composition obtained in Example 6 from the composition obtained in Example 5 in 2 mL of water for injection.

This composition is divided into 3 portions. These are introduced into 3 1 mL syringes.

Example 7c

Composition 7c, comprising carboxymethylcellulose (CMC) and P4HB microbeads with a diameter of 58 µm, obtained in Example A, is prepared by hydrating 2 g of the lyophilized composition obtained in Example 6 from the composition obtained in Example 3 in 2 mL of water for injection.

This composition is divided into 2 portions. These are introduced into 2 1 mL syringes.

Example 7d

Composition 7d comprising STYLAGE XXL, marketed by VIVACY, prepared according to the process described in patent application WO2009/071697, carboxymethylcellulose (CMC) and P4HB microbeads, 58 µm in diameter, obtained in Example A, is prepared by mixing the lyophilized composition obtained in Example 6 from the composition obtained in Example 3 and STYLAGE XXL in a mass ratio of 1:1.

This composition is introduced into a 1 mL syringe.

Example 7th

Compositions 7e containing cross-linked hyaluronic acid and P4HB were made by mixing commercial products from the STYLAGE range, marketed by the company VIVACY, and P4HB in the form of solid microbeads with a diameter of 58 µm obtained in example A.

Each of the compositions 7e.1 to 7e.4 was made by mixing 1 mL of STYLAGE XXL product prepared according to the process described in patent application WO2009/071697, contained in a syringe, and a defined mass of P4HB microbeads having a diameter of 58 µm obtained in example A, contained in a second syringe, to obtain compositions having varying concentrations of [P4HB].

The two components are mixed by connecting the two syringes end to end and making several back-and-forth movements until a homogeneous suspension is obtained.

Each of the compositions 7e.5 to 7e.7 was made by mixing 0.8 mL of STYLAGE S product prepared according to the process described in patent application WO2009/071697, contained in a syringe, and a defined mass of P4HB microbeads having a diameter of 58 µm obtained in example A, contained in a second syringe, to obtain compositions having varying concentrations of [P4HB].

The two components are mixed by connecting the two syringes end to end and making several back-and-forth movements until a homogeneous suspension is obtained.

The compositions and various technical characteristics of compositions 7e.1 to 7e.7 are summarized in the table below.

compositions commercial products volume (mL) [HA] (mg.g ^-1 ) P4HB mass (mg) [P4HB] (mg.g ^-1 ) 7th.1 XXL STYLE 1 21 59.7 59.7 7th.2 XXL STYLE 1 21 61.2 61.2 7th.3 XXL STYLE 1 21 99.7 99.7 7th.4 XXL STYLE 1 21 160.3 160.3 7th.5 STYLING S 0.8 16 20.2 25.3 7th.6 STYLING S 0.8 16 39.0 47.5 7th.7 STYLING S 0.8 16 61.0 76.3

Example 7f

A third embodiment consists of mixing, just before injection, a solution containing P4HB microbeads obtained in example A with an injectable composition containing at least one polysaccharide.

Compositions 7f containing crosslinked hyaluronic acid and P4HB in the presence of a biocompatible solvent were made by mixing commercial products from the STYLAGE range, marketed by the company VIVACY, and an organic solvent containing P4HB microbeads with a diameter of 58 µm obtained in example A.

In a first step, a defined mass of P4HB microbeads is weighed and introduced into a defined volume of organic solvent and the resulting suspension is introduced into a syringe 1.

Each of the compositions 7f.1 and 7f.2 were made by mixing 0.8 mL of STYLAGE S product prepared according to the process described in patent application WO2009/071697, contained in a syringe 2, and the suspension obtained previously, contained in syringe 1, by connecting the two syringes end to end and making several back-and-forth movements until a homogeneous suspension was obtained.

The compositions and various technical characteristics of compositions 7f.1 and 7f.2 are summarized in the table below.

compositions commercial products organic solvents [P4HB] (mg.g ^-1 ) volume (mL) [HA] (mg.g ^-1 ) volume (mL) P4HB mass (mg) 7f.1 STYLING S PEG 400 100.0 0.8 16 0.21 101.2 7f.2 STYLING S glycerol 101.5 0.8 16 0.18 101.2

Example 8: Injectability Tests

Example 8a

Composition 7a, containing RH Collagen III and P4HB microbeads with a diameter of 58 µm, was divided and introduced into four 1 mL syringes and tested for injectability. The results of these tests are presented in the table below. Composition 7a: RH Collagen III + P4HB (58µm microbead) needles injectable composition Remarks diameter length 23 G 30 mm Yes injectable without any problem 25 G 25 mm Yes presence of P4HB agglomerates in the syringe before the needle 26 G 13 mm Yes presence of P4HB agglomerates increases injection force 27 G 13 mm Yes presence of P4HB agglomerates

Tests have shown that composition 7a is injectable regardless of the injection device used. It remains injectable even when the diameter of the needle used is significantly reduced, and even in the presence of P4HB microbead agglomerates.

Example 8b

Composition 7b, containing non-crosslinked HA and P4HB microbeads with a diameter of 58 µm, was divided and introduced into three 1 mL syringes and tested for injectability. The results of these tests are presented in the table below.

Composition 7b: Non-crosslinked HA + P4HB (58µm microbead) needles injectable composition Remarks diameter length 23 G 30 mm Yes injectable without any notable problems 25 G 16 mm Yes injectable without any notable problems 26 G 13 mm Yes injectable without any notable problems

Tests have shown that composition 7b is injectable regardless of the injection device used. It remains injectable even when the diameter of the needle used is significantly reduced. Unlike previous cases, no agglomeration of P4HB microbeads was observed.

Example 8c

Composition 7c, containing CMC and P4HB microbeads with a diameter of 58 µm, was divided and introduced into two 1 mL syringes and tested for injectability. The results of these tests are presented in the table below.

composition 7c: CMC + P4HB (58µm microbead) needles injectable composition Remarks diameter length 23 G 30 mm Yes injectable without any notable problems 25 G 16 mm Yes presence of P4HB agglomerates in the syringe before the needle

Tests have shown that composition 7c is injectable regardless of the injection device used. It remains injectable even when the diameter of the needle used is significantly reduced, and even in the presence of P4HB microbead agglomerates.

Example 8d

Composition 7d, comprising STYLAGE XXL Lido prepared according to the process described in patent application WO2009/071697 and marketed by VIVACY, carboxymethylcellulose (CMC), and P4HB microbeads with a diameter of 58 µm, divided and introduced into two 1 mL syringes, was tested for injectability. The results of these tests are presented in the table below.

composition 7d: STYLAGE LIDO XXL + CMC + P4HB (58µm microbead) needles injectable composition Remarks diameter length 27 G 13 mm Yes injectable without any notable problems

The tests carried out show that the 7d composition is injectable through a needle with a diameter of 27 G.

Example 8

The injectability of compositions 7e.1 to 7e.3, comprising the STYLAGE XXL product prepared according to the process described in patent application WO2009/071697, marketed by the company VIVACY, and varying concentrations of P4HB microbeads, having a diameter of 58 µm, was evaluated and the results of these tests are presented in the table below.

7th grade compositions: STYLAGE XXL + P4HB (58µm microbead) concentrations (mg.g ^-1 ) needles injectable composition Remarks [HA] [P4HB] diameter length composition 7e.1 25 G 13 mm Yes injectable without any notable problems 21 59.7 composition 7e.2 27 G 13 mm Yes injectable without any notable problems 21 61.2 composition 7e.3 27 G 13 mm Yes injectable without any notable problems 21 99.7

The tests carried out show that compositions 7e.1 to 7e.3 containing cross-linked hyaluronic acid and varying concentrations of P4HB microbeads are injectable through needles with a diameter of 27 G, even when the concentration of microbeads increases sharply up to 99.7 mg.g ^-1 .

Example 8f

The injectability of compositions 7f.1 and 7f.2, comprising the STYLAGE S product prepared according to the process described in patent application WO2009/071697, marketed by the company VIVACY, P4HB microbeads, having a diameter of 58 µm, and a biocompatible solvent, was evaluated and the results of these tests are presented in the table below.

Composition 7f: STYLAGE S + P4HB (58µm microbead) + solvent compositions needles injectable composition Remarks [HA] (mg.g ^-1 ) [P4HB] (mg.g ^-1 ) solvent diameter long.
(mm) composition 7f.1 27 G 13 Yes injectable without any notable problems 12.8 100.0 PEG 400 composition 7f.2 27 G 13 Yes injectable without any notable problems 12.8 101.5 glycerol The tests carried out show that compositions 7f.1 and 7f.2, containing cross-linked hyaluronic acid and P4HB microbeads, at a concentration of approximately 100 mg/g^-1, and an organic solvent are injectable through needles having a diameter of 27 G.

Claims (19)

Injectable biomaterial consisting of a composition comprising at least one biocompatible polymer and PHA particles. Biomaterial according to claim 1, characterized in that the composition is in solvent-free form, in particular water-free, for example in lyophilized form. Biomaterial according to claim 1, characterized in that the composition is in aqueous form, the biocompatible polymer is in gel or hydrogel form, and the PHA particles are suspended in the gel. Biomaterial according to any one of the preceding claims, characterized in that said biocompatible polymer is a polysaccharide and/or a protein and/or a polypeptide alone or in mixture Biomaterial according to any one of claims 1, 3 or 4, characterized in that the concentration of biocompatible polymer in the composition is between 2 mg/g and 75 mg/g. Biomaterial according to any one of claims 1, 3, 4 or 5, characterized in that the PHA particle content in the composition is between 2 mg/g and 200 mg/g Biomaterial according to any one of the preceding claims, characterized in that the polysaccharide is selected from the group consisting of hyaluronic acid, keratin, heparin, cellulose, cellulose derivatives (in particular hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylmethylcellulose, carboxymethylcellulose), alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparosan, and their biologically acceptable salts, alone or in mixture. Biomaterial according to any one of claims 4 to 7, characterized in that said polysaccharide is hyaluronic acid or one of its salts, alone or in mixture. Biomaterial according to any one of claims 4 to 8, characterized in that said polysaccharide is non-crosslinked hyaluronic acid or one of its salts, alone or in mixture Biomaterial according to any one of claims 4 to 8, characterized in that said polysaccharide is cross-linked hyaluronic acid or one of its salts, alone or in mixture. Biomaterial according to any one of claims 8 to 10, characterized in that said polysaccharide has the average molecular mass Mw of at least one hyaluronic acid within a range of 0.01 MDa to 5 MDa (0.01 ≤Mw ≤5 MDa). Biomaterial according to any one of claims 8 to 11, characterized in that the concentration of hyaluronic acid [HA] is between 2 mg/g and 50 mg/g of total weight of said composition according to any one of claims 1, 3, 4 or 5. Biomaterial according to any one of the preceding claims, characterized in that the composition comprises at least one protein and/or polypeptide alone or in mixture with polysaccharides. Biomaterial according to claim 13, characterized in that the protein(s) and/or polypeptide(s) are selected from collagen, extracellular matrix proteins, structural proteins, blood-derived proteins, glycoproteins, lipoproteins, natural proteins, synthetic proteins, intracellular proteins, extracellular proteins, membrane proteins, and all combinations thereof. Biomaterial according to any one of the preceding claims, characterized in that the PHAs are selected from first-generation PHAs such as poly(3-hydroxybutyric acid) (PHB) and poly(4-hydroxybutyric acid) (P4HB), second-generation poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), third-generation poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx or PHBHx), fourth-generation poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB or P3HB4HB) or fifth-generation poly(3-hydroxybutyricacid-co-3-hydroxyvalericacid-co-3-hydroxyhexanoic acid) (PHBVHHx or PHBVHx), alone or in mixture. Biomaterial according to any one of the preceding claims, characterized in that the PHA is the P4HB. Biomaterial according to any one of the preceding claims, characterized in that the PHA particles have a size strictly less than 100 µm. Biomaterial according to any one of the preceding claims, characterized in that the PHA particles are prepared by microfluidization Biomaterial according to any one of the preceding claims, characterized in that the composition further comprises an active ingredient selected from the group consisting of local anesthetics, vitamin C derivatives, anti-inflammatories, antioxidants, and mixtures thereof.