WO2019002371A1 - Gel de glycosaminoglycane avec tampon bis-tris - Google Patents

Gel de glycosaminoglycane avec tampon bis-tris Download PDF

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Publication number
WO2019002371A1
WO2019002371A1 PCT/EP2018/067256 EP2018067256W WO2019002371A1 WO 2019002371 A1 WO2019002371 A1 WO 2019002371A1 EP 2018067256 W EP2018067256 W EP 2018067256W WO 2019002371 A1 WO2019002371 A1 WO 2019002371A1
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bis
composition
glycosaminoglycan
gel
concentration
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PCT/EP2018/067256
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English (en)
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Craig Steven Harris
Laura JING JING
Katarina Edsman
Anders Karlsson
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Nestlé Skin Health Sa
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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
    • A61K31/167—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide having the nitrogen of a carboxamide group directly attached to the aromatic ring, e.g. lidocaine, paracetamol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/738—Cross-linked polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0069—Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof

Definitions

  • the present invention relates to the field of hydrogels containing cross- linked polysaccharides and the use of such hydrogels in medical and/or cosmetic applications. More specifically, it relates to injectable hydrogels containing cross-linked hyaluronic acid.
  • hyaluronic acid is a naturally occurring polysaccharide belonging to the group of glycosaminoglycans (GAGs).
  • GAGs glycosaminoglycans
  • Hyaluronic acid and the other GAGs are negatively charged heteropolysaccharide chains which have a capacity to absorb large amounts of water.
  • Hyaluronic acid and products derived from hyaluronic acid are widely used in the biomedical and cosmetic fields, for instance during viscosurgery and as a dermal filler.
  • Water-absorbing gels are widely used in the biomedical field. They are generally prepared by chemical cross-linking of polymers to infinite networks. While native hyaluronic acid and certain cross-linked hyaluronic acid products absorb water until they are completely dissolved, cross-linked hyaluronic acid gels typically absorb a certain amount of water until they are saturated, i.e. they have a finite liquid retention capacity, or swelling degree.
  • hyaluronic acid Since hyaluronic acid is present with identical chemical structure except for its molecular mass in most living organisms, it gives a minimum of reactions and allows for advanced medical uses. Cross-linking and/or other modifications of the hyaluronic acid molecule is necessary to improve its duration in vivo. Furthermore, such modifications affect the liquid retention capacity of the hyaluronic acid molecule. As a consequence thereof, hyaluronic acid has been the subject of many modification attempts. Summary of the invention
  • An object of the present invention is to provide improved injectable hydrogel compositions, preferably hyaluronic acid based hydrogel
  • compositions for use as dermal fillers are provided.
  • An object of the present invention is to provide improved injectable hydrogel compositions, preferably hyaluronic acid based hydrogel
  • compositions which exhibit decreased degradation of the composition during autoclaving.
  • An object of the present invention is to provide improved injectable hydrogel compositions, preferably hyaluronic acid based hydrogel
  • compositions which exhibit increased stability after autoclaving.
  • Hydrogel compositions such as hyaluronic acid based hydrogel compositions, for use in injection need to be sterilized before use. Sterilization is generally performed by heat treatment, such as autoclaving. The heat treatment generally leads to a reduction of the rigidity or viscosity of the composition.
  • a method of preparing a sterilized injectable hydrogel composition comprising the steps: a) providing a covalently crosslinked glycosaminoglycan,
  • injectable means that the composition is provided in a form which is suitable for parenteral injection, e.g. into soft tissue, such as skin, of a subject or patient.
  • An injectable composition should be sterile and free from components that may cause adverse reactions when introduced into soft tissue, such as the skin, of a subject or patient. This implies that no, or only very mild, immune response occurs in the treated individual. That is, no or only very mild undesirable local or systemic effects occur in the treated individual.
  • the glycosaminoglycan is selected from the group consisting of hyaluronic acid, heparosan, chondroitin and chondroitin sulfate, and mixtures thereof. According to some embodiments, the glycosaminoglycan is selected from the group consisting of hyaluronic acid, chondroitin and chondroitin sulfate, and mixtures thereof. In a preferred embodiment, the glycosaminoglycan is hyaluronic acid.
  • the present inventors have identified that addition of Bis-Tris buffer to a hydrogel composition comprising crosslinked glycosaminoglycan leads to a significant reduction in degradation during autoclaving and storage compared to identical compostions in water or other buffer solutions.
  • the composition exhibits increased stability compared to an identical composition without the Bis-Tris buffer.
  • the composition exhibits increased stability compared to an identical composition in water instead of the Bis-Tris buffer.
  • the Bis-Tris buffer the
  • composition exhibits increased stability compared to an identical composition in phosphate buffer instead of the Bis-Tris buffer.
  • stability is used to denote the ability of the sterilized injectable hydrogel composition to resist degradation during storage and handling prior to use. It is known that the addition of constituents to a glycosaminoglycan, such as hyaluronic acid or hyaluronic acid gel, may affect the stability of said glycosaminoglycan. Stability of a hydrogel composition comprising a glycosaminoglycan can be determined by a range of different methods. Methods for determining stability include, but are not limited to, assessing homogeneity, color, clarity, pH, gel content and rheological properties of the composition.
  • Stability of a hydrogel composition comprising a glycosaminoglycan is often determined by observing or measuring one or more of said parameters over time. Stability may for example be determined by measuring the viscosity and/or elastic modulus G' of the composition over time. Viscosity may for example be measured as the "Zero shear viscosity, rjo" by rotational viscometry using an Ares G2 rheometer (Measuring system cone plate or parallel plates, Gap 1 .00 mm). Other methods of measuring viscosity may also be applicable.
  • the elastic modulus G' may for example be measured using a Ares G2 Reometer (Measure system parallel plates, Gap 1 .00 mm) by performing a strain sweep to find the linear viscoelastic region (LVR) and then measuring the viscoelastic properties within the LVR. Other methods of measuring elastic modulus G' may also be applicable.
  • Ares G2 Reometer Measure system parallel plates, Gap 1 .00 mm
  • (2-[Bis(2-hydroxyethyl)amino]-2-(hydroxymethyl) propane-1 ,3-diol) is an amine buffer with a useful pH range of 5.8 - 7.2.
  • the concentration of Bis-Tris in the solution of step b) may generally be in the range of 1 to 200 mM, but it is rarely necessary to have a
  • the concentration of Bis-Tris in the solution of step b) is in the range of 1 to 100 mM, preferably in the range of 1 to 50 mM, and more preferably in the range of 1 to 20 mM. In some embodiments, the concentration of Bis-Tris in the solution of step b) is in the range of 5 to 20 mM.
  • the Bis-Tris solution further comprises NaCI at physiological concentration.
  • the NaCI concentration in the solution is in the range of 130-140 mM.
  • Bis-Tris buffer also has advantages when a divalent cation is present in the hydrogel composition. It is sometimes desired to add divalent cations, such as Zn 2+ to injectable hydrogel compositions. Other buffers, such as for example phosphate buffer, have been found to cause precipitation of the Zn 2+ in the hydrogel. The use of Bis-Tris buffer has been found to prevent this problem, allowing the addition of divalent cations to the hydrogel composition without precipitation.
  • the solution of step b) further comprises a divalent cation.
  • the divalent cation is preferably selected from the group consisting of Ca 2+ , Cu 2+ , Mg 2+ and Zn 2+ .
  • the divalent cation is Zn 2+ .
  • the solution of step b) comprises Zn 2+ at a concentration in the range of 0.05 to 4 mM, preferably in the range of 0.05 to 2 mM.
  • the glycosaminoglycan of the composition is preferably a hyaluronic acid.
  • hyaluronic acid encompasses all variants and combinations of variants of hyaluronic acid, hyaluronate or hyaluronan, of various chain lengths and charge states, as well as with various chemical modifications, including crosslinking. That is, the term also encompasses the various hyaluronate salts of hyaluronic acid with various counter ions, such as sodium hyaluronate.
  • Various modifications of the hyaluronic acid are also encompassed by the term, such as oxidation, e.g.
  • oxidation of -CH2OH groups to -CHO and/or -COOH periodate oxidation of vicinal hydroxyl groups, optionally followed by reduction, e.g. reduction of -CHO to -CH2OH or coupling with amines to form imines followed by reduction to secondary amines; sulphation; deamidation, optionally followed by deamination or amide formation with new acids; esterification; crosslinking; substitutions with various compounds, e.g. using a crosslinking agent or a carbodiimide assisted coupling; including coupling of different molecules, such as proteins, peptides and active drug components, to hyaluronic acid; and deacetylation.
  • modifications are isourea, hydrazide, bromocyan, monoepoxide and monosulfone couplings.
  • the hyaluronic acid can be obtained from various sources of animal and non-animal origin.
  • Sources of non-animal origin include yeast and preferably bacteria.
  • the molecular weight of a single hyaluronic acid molecule is typically in the range of 0.1 -10 MDa, but other molecular weights are possible.
  • the concentration of the glycosaminoglycan is in the range of 1 to 100 mg/ml. In some embodiments the concentration of the glycosaminoglycan is in the range of 2 to 50 mg/ml. In specific embodiments the concentration of the glycosaminoglycan is in the range of 5 to 30 mg/ml or in the range of 10 to 30 mg/ml.
  • the glycosaminoglycan is covalently crosslinked.
  • the covalently crosslinked glycosaminoglycan may be obtained by covalently crosslinking a glycosaminoglycan using a bi- or polyfunctional crosslinking agent, or it may be obtained by so called linker free crosslinking where a coupling agent is used to form covalent bonds directly between functional groups already present in the glycosaminoglycan, but where the coupling agent does not form part of the crosslink.
  • Crosslinking of the glycosaminoglycan may be achieved by
  • the crosslinking agent may for example selected from the group consisting of divinyl sulfone, multiepoxides and diepoxides.
  • the crosslinking agent is selected from the group consisting of 1 ,4-butanediol diglycidyl ether (BDDE), 1 ,2-ethanediol diglycidyl ether (EDDE) and diepoxyoctane.
  • the crosslinking agent is 1 ,4-butanediol diglycidyl ether (BDDE).
  • crosslinking of the glycosaminoglycan is achieved by amide coupling of glycosaminoglycan molecules.
  • Amide coupling using a using a di- or multiamine functional crosslinker together with a coupling agent is an attractive route to preparing crosslinked
  • Crosslinking can be achieved using a non-carbohydrate based di- or multinucleophilic crosslinker, for example hexamethylenediamine (HMDA), or a carbohydrate based di- or multinucleophilic crosslinker, for example diaminotrehalose (DATH) together with a glycosaminoglycan.
  • Crosslinking can also be achieved using an at least partially deacetylated glycosaminoglycan, either alone or in combination with a second glycosaminoglycan, whereby the deacetylated glycosaminoglycan itself acts as the di- or multinucleophilic crosslinker.
  • Diaminotrehalose (DATH) can be synthesized as described in "Synthetic Carbohydrate Polymers Containing Trehalose Residues in the Main Chain: Preparation and Characteristic Properties”; Keisuke Kurita, * Naoko Masuda, Sadafumi Aibe, Kaori Murakami, Shigeru Ishii, and Shin- Ichiro Nishimurat; Macromolecules 1994, 27, 7544-7549.
  • Crosslinking of the glycosaminoglycan may for example be achieved in aqueous media using a crosslinker comprising at least two nucleophilic functional groups, for example amine groups, capable of forming covalent bonds directly with carboxylic acid groups of GAG molecules by a reaction involving the use of a coupling agent.
  • a crosslinker comprising at least two nucleophilic functional groups, for example amine groups, capable of forming covalent bonds directly with carboxylic acid groups of GAG molecules by a reaction involving the use of a coupling agent.
  • the crosslinker comprising at least two nucleophilic functional groups may for example be a non-carbohydrate based di- or multinucleophilic crosslinker or a carbohydrate based di- or multinucleophilic crosslinker.
  • Carbohydrate based di- or multinucleophilic crosslinkers are preferred, since they provide a hydrogel product based entirely on carbohydrate type structures or derivatives thereof, which minimizes the disturbance of the crosslinking on the native properties of the glycosaminoglycans.
  • the crosslinker itself can also contribute to maintained or increased properties of the hydrogel, for example when crosslinking with a structure that correlates to hyaluronic acid or when crosslinking with a structure with high water retention properties.
  • the carbohydrate based di- or multinucleophilic crosslinker may for example be selected from the group consisting of di- or multinucleophilic functional di-, tri-, tetra-, oligosaccharides, and polysaccharides.
  • the di- or multinucleophilic crosslinker is an at least partially deacetylated glycosaminoglycan, i.e. an acetylated glycosaminoglycan which has been at least partially deacetylated to provide a glycosaminoglycan having free amine groups.
  • deacetylated glycosaminoglycan can be crosslinked either alone or in combination with a second glycosaminoglycan, whereby the deacetylated glycosaminoglycan itself acts as the di- or multinucleophilic crosslinker.
  • the coupling agent may for example be selected from the group consisting of triazine-based coupling agents, carbodiimide coupling agents, imidazolium-derived coupling reagents, Oxyma and COMU.
  • a preferred coupling agent is a triazine-based coupling agent, including 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 (CDMT), preferably DMTMM.
  • Another preferred coupling agent is a carbodiimide coupling agent, preferably N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) combined with N-hydroxysuccinimide (NHS).
  • EDC N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
  • NHS N-hydroxysuccinimide
  • the crosslinked GAG is obtained by:
  • the sterilized injectable composition formed using the inventive method is a hydrogel. That is, it can be regarded as a water-insoluble, but substantially dilute crosslinked system of glycosaminoglycan molecules when subjected to a liquid, typically an aqueous liquid.
  • the sterilized injectable hydrogel composition contains mostly liquid by weight and can e.g. contain 90-99.9% water, but it behaves like a solid due to a three-dimensional crosslinked hyaluronic acid network within the liquid. Due to its significant liquid content, the gel is structurally flexible and similar to natural tissue, which makes it very useful as a scaffold in tissue engineering and for tissue augmentation.
  • the hydrogel composition is preferably biocompatible. This implies that no, or only very mild, immune response occurs in the treated individual. That is, no or only very mild undesirable local or systemic effects occur in the treated individual.
  • crosslinking of a glycosaminoglycan such as hyaluronic acid, to form the crosslinked glycosaminoglycan may for example be achieved by modification with a crosslinking agent, for example BDDE (1 ,4- butandiol diglycidylether).
  • BDDE 1,4- butandiol diglycidylether
  • the glycosaminoglycan concentration and the extent of crosslinking affects the mechanical properties, e.g. the elastic modulus G', and stability properties of the hydrogel.
  • glycosaminoglycan gels are often characterized in terms of "degree of modification".
  • the degree of modification of hyaluronic acid gels generally range between 0.1 and 15 mole%.
  • the hyaluronic acid gel has a degree of modification of 12 mole% or less, such as 12 mole% or less, such as 10 mole% or less, for example in the range of 0.1 to 12 mole%, such as in the range of 0.2 to 10 mole%, such as in the range of 0.3 to 8 mole%.
  • the hyaluronic acid gel has a degree of modification of 12 mole% or less, such as 12 mole% or less, such as 10 mole% or less, for example in the range of 2 to 12 mole%, such as in the range of 3 to 10 mole%, such as in the range of 4 to 8 mole%.
  • the hyaluronic acid gel has a degree of modification of 2 mole% or less, such as 1 .5 mole% or less, such as 1 .25 mole% or less, for example in the range of 0.1 to 2 mole%, such as in the range of 0.2 to 1 .5 mole%, such as in the range of 0.3 to 1 .25 mole%.
  • the degree of modification (mole%) describes the amount of crosslinking agent(s) that is bound to glycosaminoglycan, i.e. molar amount of bound crosslinking agent(s) relative to the total molar amount of repeating glycosaminoglycan disaccharide units.
  • the degree of modification reflects to what degree the glycosaminoglycan has been chemically modified by the crosslinking agent.
  • Reaction conditions for crosslinking and suitable analytical techniques for determining the degree of modification are all well known to the person skilled in the art, who easily can adjust these and other relevant factors and thereby provide suitable conditions to obtain a degree of modification in the range of 0.1 -15% and verify the resulting product characteristics with respect to the degree of modification.
  • a BDDE (1 ,4- butandiol diglycidylether) crosslinked hyaluronic acid gel may for example be prepared according to the method described in Examples 1 and 2 of published international patent application WO 9704012.
  • the crosslinked glycosaminoglycan of the composition is present in the form of a crosslinked hyaluronic acid crosslinked by a crosslinking agent, wherein the concentration of said hyaluronic acid is in the range of 2 to 50 mg/ml and the degree of modification with said
  • crosslinking agent is in the range of 0.1 to 2 mole%.
  • the injectable hydrogel composition may further comprise a
  • a local anesthetic is a drug that causes reversible local anesthesia and a loss of nociception. When it is used on specific nerve pathways (nerve block), effects such as analgesia (loss of pain sensation) and paralysis (loss of muscle power) can be achieved.
  • the local anesthetic may be added to the composition to reduce pain or discomfort experienced by the patient due to the injection procedure.
  • the local anesthetic is selected from the group consisting of amide and ester type local anesthetics, for example bupivacaine, butanilicaine, carticaine, cinchocaine (dibucaine), clibucaine, ethyl parapiperidinoacetylaminobenzoate, etidocaine, lignocaine (lidocaine), mepivacaine, oxethazaine, prilocaine, ropivacaine, tolycaine, trimecaine, vadocaine, articaine, levobupivacaine, amylocaine, cocaine, propanocaine, clormecaine, cyclomethycaine, proxymetacaine, amethocaine (tetracaine), benzocaine, butacaine, butoxycaine, butyl aminobenzoate, chloroprocaine, dimethocaine (larocaine), oxybuprocaine, piperocaine, parethoxycaine
  • the local anesthetic is lidocaine.
  • the local anesthetic is lidocaine.
  • Lidocaine is a well-known substance, which has been used extensively as a local anesthetic in injectable formulations, such as hyaluronic acid
  • the concentration of the amide or ester local anesthetic may be selected by the skilled person within the therapeutically relevant concentration ranges of each specific local anesthetic or a combination thereof. In some embodiments the concentration of said local anesthetic is in the range of 0.1 to 30 mg/ml. In certain embodiments the concentration of said local anesthetic is in the range of 0.5 to 10 mg/ml.
  • the lidocaine When lidocaine is used as the local anesthetic, the lidocaine may preferably be present in a concentration in the range of 1 to 5 mg/ml, more preferably in the range of 2 to 4 mg/ml, such as in a concentration of about 3 mg/ml.
  • the method described herein involves sterilization of the composition by autoclaving, i.e sterilization using saturated steam.
  • the autoclaving may be performed at an Fo-value > 4.
  • the autoclaving may preferably be performed at an Fo-value in the range of 10 to 50, such as in the range of 20- 30.
  • the Fo value of a saturated steam sterilisation process is the lethality expressed in terms of the equivalent time in minutes at a temperature of 121 °C delivered by the process to the product in its final container with reference to microorganisms posessing a Z-value of 10.
  • the composition comprises hyaluronic acid at a concentration in the range of 2-50 mg/ml, and Bis-Tris buffer at a concentration in the range of 1 -50 mM, and sterilization is performed by autoclaving at an Fo-value > 4.
  • the sterilized injectable hydrogel compositions formed according to the inventive method exhibit increased stability compared to identical compositions without the Bis- Tris buffer.
  • the composition exhibits increased stability compared to an identical composition in water instead of the Bis-Tris buffer.
  • the composition exhibits increased stability compared to an identical composition in phosphate buffer instead of the Bis-Tris buffer.
  • a sterilized injectable hydrogel composition obtainable by the method described above. According to other aspects illustrated herein, there is provided a sterilized injectable hydrogel composition comprising:
  • the present inventors have identified that addition of Bis-Tris buffer to a hydrogel composition comprising crosslinked glycosaminoglycan leads to a significant reduction in degradation during autoclaving and storage compared to identical compostions in water or other buffer solutions.
  • the composition exhibits increased stability compared to an identical composition without the Bis-Tris buffer.
  • the composition exhibits increased stability compared to an identical composition in water instead of the Bis-Tris buffer.
  • the composition exhibits increased stability compared to an identical composition in phosphate buffer instead of the Bis-Tris buffer.
  • the concentration of Bis-Tris in the composition is in the range of 1 to 100 mM, preferably in the range of 1 to 50 mM, more preferably in the range of 1 to 20 mM. In some embodiments, the
  • the concentration of Bis-Tris in the composition is in the range of 5 to 20 mM, In some embodiments, the Bis-Tris solution further comprises NaCI at physiological concentration. Typically, the NaCI concentration in the solution is in the range of 130-140 mM.
  • the composition further comprises a divalent cation.
  • the divalent cation is preferably selected from the group consisting of Ca 2+ , Cu 2+ , Mg 2+ and Zn 2+ .
  • the divalent cation is Zn 2+ .
  • the composition comprises Zn 2+ at a
  • the glycosaminoglycan is selected from the group consisting of hyaluronic acid, heparosan, chondroitin and chondroitin sulfate, and mixtures thereof. According to some embodiments, the glycosaminoglycan is selected from the group consisting of hyaluronic acid, chondroitin and chondroitin sulfate, and mixtures thereof. In a preferred embodiment, the glycosaminoglycan is hyaluronic acid.
  • the glycosaminoglycan is covalently crosslinked.
  • the covalently crosslinked glycosaminoglycan may be obtained by covalently crosslinking a glycosaminoglycan using a bi- or polyfunctional crosslinking agent, or it may be obtained by so called linker free crosslinking where a coupling agent is used to form covalent bonds directly between functional groups already present in the glycosaminoglycan, but where the coupling agent does not form part of the crosslink.
  • Crosslinking of the glycosaminoglycan may be achieved by
  • the crosslinking agent may for example selected from the group consisting of divinyl sulfone, multiepoxides and diepoxides.
  • the crosslinking agent is selected from the group consisting of 1 ,4-butanediol diglycidyl ether (BDDE), 1 ,2-ethanediol diglycidyl ether (EDDE) and diepoxyoctane.
  • the crosslinking agent is 1 ,4-butanediol diglycidyl ether (BDDE).
  • crosslinking of the glycosaminoglycan is achieved by amide coupling of glycosaminoglycan molecules.
  • Amide coupling using a using a di- or multiamine functional crosslinker together with a coupling agent is an attractive route to preparing crosslinked
  • Crosslinking can be achieved using a non-carbohydrate based di- or multinucleophilic crosslinker, for example hexamethylenediamine (HMDA), or a carbohydrate based di- or multinucleophilic crosslinker, for example diaminotrehalose (DATH) together with a glycosaminoglycan.
  • Crosslinking can also be achieved using an at least partially deacetylated glycosaminoglycan, either alone or in combination with a second glycosaminoglycan, whereby the deacetylated glycosaminoglycan itself acts as the di- or multinucleophilic crosslinker.
  • Crosslinking of the glycosaminoglycan may for example be achieved in aqueous media using a crosslinker comprising at least two nucleophilic functional groups, for example amine groups, capable of forming covalent bonds directly with carboxylic acid groups of GAG molecules by a reaction involving the use of a coupling agent.
  • a crosslinker comprising at least two nucleophilic functional groups, for example amine groups, capable of forming covalent bonds directly with carboxylic acid groups of GAG molecules by a reaction involving the use of a coupling agent.
  • the covalently crosslinked glycosaminoglycan has a degree of modification of 2 mole% or less, such as 1 .5 mole% or less, such as 1 .25 mole% or less.
  • the covalently crosslinked glycosaminoglycan has a degree of modification in the range of 0.1 to 2 mole%, such as in the range of 0.2 to 1 .5 mole%, such as in the range of 0.3 to 1 .25 mole%.
  • the concentration of the glycosaminoglycan in the composition is in the range of 1 to 100 mg/ml. In some embodiments, the concentration of the glycosaminoglycan in the composition is in the range of 2 to 50 mg/ml. In some embodiments, the concentration of the
  • glycosaminoglycan in the composition is in the range of 10 to 30 mg/ml.
  • the composition further comprises a
  • the local anesthetic is selected from the group consisting of amide and ester type local anesthetics, for example bupivacaine, butanilicaine, carticaine, cinchocaine (dibucaine), clibucaine, ethyl parapiperidinoacetylaminobenzoate, etidocaine, lignocaine (lidocaine), mepivacaine, oxethazaine, prilocaine, ropivacaine, tolycaine, trimecaine, vadocaine, articaine, levobupivacaine, amylocaine, cocaine, propanocaine, clormecaine, cyclomethycaine, proxymetacaine, amethocaine (tetracaine), benzocaine, butacaine, butoxycaine, butyl aminobenzoate, chloroprocaine, dimethocaine (larocaine), oxybuprocaine, piperocaine, parethoxycaine
  • compositions described herein have preferably been subjected to sterilization by autoclaving, i.e sterilization using saturated steam.
  • the autoclaving may be performed at an Fo-value > 4.
  • the Fo value of a saturated steam sterilisation process is the lethality expressed in terms of the equivalent time in minutes at a temperature of 121 °C delivered by the process to the product in its final container with reference to microorganisms posessing a Z-value of 10.
  • the inventive composition preferably exhibits increased stability compared to an identical composition without the Bis-Tris buffer.
  • composition may be further defined as described above with reference to the method of preparing the sterilized injectable hydrogel composition.
  • the sterilized injectable hydrogel compositions according to the invention may be provided in the form of a pre-filled syringe, i.e. a syringe that is pre-filled with the injectable hydrogel composition and autoclaved.
  • the sterilized injectable hydrogel compositions as described herein may advantageously be used for the transport or administration and slow or controlled release of various parmaceutical or cosmetic substances.
  • the sterilized injectable hydrogel compositions described herein may be employed in medical as well as non-medical, e.g. purely cosmetic, procedures by injection of the composition into soft tissues of a patient or subject.
  • the compositions have been found useful in, e.g., soft tissue augmentation, for example filling of wrinkles, by hyaluronic acid gel injection.
  • the compositions have also been found useful in a cosmetic treatment, referred to herein as skin revitalization, whereby small quantities of the hyaluronic acid composition are injected into the dermis at a number of injection sites distributed over an area of the skin to be treated, resulting in improved skin tone and skin elasticity.
  • Skin revitalization is a simple
  • composition is useful, for example in the treatment of various dermatological conditions.
  • an injectable hyaluronic acid composition as described above for use in a dermatological treatment selected from the group consisting of wound healing, treatment of dry skin conditions or sun-damaged skin, treatment of hyper pigmentation disorders, treatment and prevention of hair loss, and treatment of conditions that have inflammation as a component of the disease process, such as psoriasis and asteototic eczema.
  • an injectable hyaluronic acid composition as described above for use in the manufacture of a medicament for use in a dermatological treatment selected from the group consisting of wound healing, treatment of dry skin conditions or sun-damaged skin, treatment of hyper pigmentation disorders, treatment and prevention of hair loss, and treatment of conditions that have
  • inflammation as a component of the disease process, such as psoriasis and asteototic eczema.
  • an injectable hyaluronic acid composition as described above for cosmetic, non-medical, treatment of a subject by injection of the composition into the skin of the subject may be for improving the appearance of the skin, preventing and/or treating hair loss, filling wrinkles or contouring the face or body of a subject.
  • the cosmetic, nonmedical, use does not involve treatment of any form of disease or medical condition. Examples of improving the appearance of the skin include, but are not limited to, treatment of sun-damaged or aged skin, skin revitalization, skin whitening and treatment of hyper pigmentation disorders such as senile freckles, melasma and ephelides.
  • Figure 1 a is a diagram showing the elastic modulus G' as a function of Bis- Tris concentration in Example 1 .
  • Figure 1 b is a diagram showing the Gel Content as a function of Bis-Tris concentration in Example 1 .
  • Figure 2a is a diagram showing the elastic modulus G' as a function of Bis- Tris concentration in Example 2.
  • Figure 2b is a diagram showing the Gel Content as a function of Bis-Tris concentration in Example 2.
  • Figure 3 is a diagram showing the elastic modulus G' as a function of Bis-Tris concentration in Example 3.
  • Figure 4 is a diagram showing degradation of gels in different buffers and buffer concentrations at 60 °C.
  • Figure 5 is a diagram showing corrected swelling degree (SwCC) of gels crosslinked with DATH in different buffers.
  • Figure 6 is a diagram showing the gel part (GelP) of gels crosslinked with DATH in different buffers.
  • Figure 7 is a diagram showing how much gel part of gels crosslinked with DATH in different buffers that is left after day 14 at 60 °C compared to at day 0.
  • SwC Swelling capacity in saline, total liquid uptake per gram PS (mL/g). SwCC - Corrected swelling degree, total liquid uptake of one gram PS, corrected for GelP (mL/g).
  • GelP - Gel part is a description of the percentage of polysaccharide that is a part of the gel network. A number of 90% means that only 10% of
  • polysaccharide is not a part of the network.
  • polysaccharide in the gel was measured with LC-SEC-UV.
  • DoA Degree of Acetylation.
  • the degree of acetylation is the molar ratio of acetyl groups compared to hyaluronic acid disaccharides.
  • DoA can be calculated from NMR spectra by comparing the integral of the acetyl signal of the hyaluronan disaccharide residues to the integral of the C2-H signal of the deacetylated glucosamine residues according to the equation.
  • a CrR of 1 .0 means that all of the crosslinker has crosslinked.
  • Diaminotrehalose (DATH) was synthesized as described in "Synthetic Carbohydrate Polymers Containing Trehalose Residues in the Main Chain: Preparation and Characteristic Properties”; Keisuke Kurita * Naoko Masuda, Sadafumi Aibe, Kaori Murakami, Shigeru Ishii, and Shin-Ichiro Nishimurat; Macromolecules 1994, 27, 7544-7549.
  • the crosslinked material was swelled in 0.25 M NaOH (1 g material : 9 g 0.25 M NaOH resulting in pH 13) for at least 1 h at room temperature.
  • the gel was neutralized with 1 .2 M HCI to pH 7 and then precipitated with ethanol.
  • the resulting precipitate was washed with 100 mM NaCI in ethanol (70% w/w) to remove excess reagents and then with ethanol (70% w/w) to remove salts and finally with ethanol to remove water.
  • the crosslinked material was swelled in 0.7% NaCI 8 mM phosphate buffer pH 7.4 at room temperature. The pH was adjusted to 7.2-7.5 if needed. The gel was left at 70 °C for 20-24 h and then precipitated with ethanol. The resulting precipitate was washed with 100 mM NaCI in ethanol (70% w/w) to remove excess reagents and then with ethanol (70% w/w) to remove salts and finally with ethanol to remove water. Determination of HA concentration
  • the method for determination of HA content is adopted from the assay test for sodium hyaluronate described in Ph. Eur. 1472.
  • the principle for the method is that a condensation reaction of the furfural derivatives formed by heating in sulphuric acid occurs with the carbazole reagent, forming a purple colored product.
  • the reaction is specific for the D-glucuronic acid part of HA.
  • the absorbance is measured at 530 nm and glucuronic acid is used for standardization.
  • the product formed from the content of D-glucuronic acid (GlcA) in the sample is determined by reaction with carbazole.
  • the stabilized gel of HA is degraded with sulphuric acid at 70°C and diluted with 0.9% NaCI-solution.
  • the solutions are mixed with sulphuric acid at 95°C and thereafter with carbazole reagent.
  • the reactions result in pink colored solutions.
  • the intensity of the colour is measured with a colorimeter at 530 nm, and the absorbance of each sample is directly proportional to the GlcA-content.
  • the HA content is calculated from the GlcAcontent in each sample.
  • GeIC describes in % the proportion of the total HA that is bound in gel form.
  • Gel content is defined as the amount of HA in a sample that does not pass through a 0.22 ⁇ filter.
  • GeIC is calculated from the amount of HA that is collected in the filtrate, here denoted extractable HA.
  • the gel content and the extractable HA content are given in percent of the total amount of HA in the gel sample.
  • the gel content is determined by mixing a certain amount of gel sample with 0.9% NaCI in a test tube. The gel is allowed to swell where after the NaCI-phase is separated from the gel-phase by filtration through a 0.22 ⁇ filter.
  • the concentration of HA in the filtrate is determined according to the procedure for determination of HA concentration.
  • Rheometry in the oscillating mode is used to determine the viscoelastic properties of the swelled gel product.
  • the elastic modulus (G') describes the gel strength in terms of the gels physical resistance to elastic deformation.
  • the viscous modulus (G") describes the gel strength in terms of the gel's physical resistance to viscous deformation.
  • the measurement is performed using an oscillating rheometer. Rheometry measurements are performed as follows. Frequency sweeps are made with a resting time of at least 15 minutes between sample loading and measurement within the linear viscoelastic range.
  • a parallel plate geometry with a diameter of 25 mm is used with a gap of 1 or 2 mm. Amplitude sweeps are made at 1 Hz to determine the linear
  • formulations were made from the same HA gel crosslinked with BDDE.
  • the formulations contained 20 mg/ml HA-gel, 0.9% NaCI and Bis- Tris buffer. The only difference between the three formulations were the Bis- Tris buffer concentration.
  • Formulation 1 contained 1 mM Bis-Tris
  • formulation 2 contained 5 mM Bis-Tris
  • formulation 3 contained 10 mM Bis-Tris.
  • the formulations were filled into 1 ml_ glass syringes and were autoclaved.
  • the elastic modulus G' is shown in figure 1 a as a function of Bis-Tris
  • the elastic modulus increases as a function of Bis-Tris concentration indicate that the Bis-Tris protects the HA gel from degradation during autoclavation. The protection is larger the larger the Bis-Tris
  • the formulations contained 20 mg/ml HA-gel, 0.9% NaCI, 1 mM ZnC and Bis Tris buffer. The only difference between the three formulations were the Bis-Ths buffer concentration.
  • Formulation 1 contained 1 mM Bis- Ths
  • formulation 2 contained 5 mM Bis-Ths
  • formulation 3 contained 10 mM Bis-Ths.
  • the formulations were filled into 1 ml_ glass syringes and were autoclaved. Analysis of the rheological properties and the gel content was made.
  • the elastic modulus G' is shown in figure 2a as a function of Bis-Tris concentration.
  • the elastic modulus increases as a function of Bis-Tris concentration indicate that the Bis-Tris protects the HA gel from degradation during autoclavation. The protection is larger the larger the Bis-Tris concentration is.
  • Gel Content confirms these results.
  • the coupling agent DMTMM was dissolved in Na-phosphate buffer (pH 7.4). if needed pH was adjusted on the DMTMM mixture and the solution was subsequently added to deacetylated HA (DeAcHA).
  • the reaction mixture was homogenized by shaking for 3.5 minutes, by mixing with a spatula or by pressing the mixture though a filter.
  • the reaction mixture was incubated at 23 or 35 °C. After 24 hours, the reaction was stopped by cutting the gel in to small pieces with a spatula or pressing the gel through a filter, the resulting material was subjected to alkaline treatment and subsequent dried in vacuum over night.
  • Table 1 A summary of the conditions for crosslinking is provided in Table 1 .
  • the dried gels were swelled in Bis-Tris buffer (1 mM) in 0.7% NaCI for at least two hours to a concentration of approx. 20 mg/mL.
  • the pH was controlled and adjusted if necessary to 7.4.
  • the gel particles were reduced in size with fine filter.
  • the gel was filled in syringes and the syringes were sterilized by autoclavation (Fo 23).
  • a summary of the gel properties is provided in Table 3.
  • the dried gels were swelled in Bis-Tris buffer (1 mM) in 0.7% NaCI with 1 mM ZnC for at least two hours to a concentration of approx 20 mg/mL.
  • the pH was controlled and adjusted if necessary to 7.4.
  • the gel particles were reduced in size with fine filter.
  • the gel was filled in syringes and the syringes were sterilized by autoclavation (Fo 23).
  • a summary of the gel properties is provided in Table 4.
  • Hyaluronic acid was weighed in a reaction vessel.
  • a stock solution of the crosslinker (DATH) was prepared by dissolving it in buffer pH 7.
  • DMTMM was weighed in a PTFE-container and the crosslinker-solution was added to the DMTMM to dissolve it.
  • the pH of the DMTMM-crosslinker solution was adjusted to 6-7 with 1 .2 M HCI and then added to the HA.
  • the contents was thoroughly homogenized and then incubated at 23 °C for 24 h.
  • the resulting material was pressed through a 1 mm steel mesh two times, swelled in 0.9% NaCI and the pH adjusted to 7.3-7.5 with diluted acid/base.
  • the gel was subjected to heat (70 °C, 24 h) in order to hydrolyze any potentially formed ester bonds.
  • the gel was particle size reduced through a 125 ⁇ mesh followed by precipitation with ethanol and the precipitate was washed with 100 mM NaCI in ethanol (70% w/w) to remove excess reagents and then with ethanol (70% w/w) to remove salts and finally with ethanol to remove water.
  • the precipitate was then dried in vacuum over night.
  • Example 6 The gels produced in Example 6 (6-1 , 6-2 and 6-3) by cross-linking of hyaluronan with diaminotrehalose (DATH) were swelled in various buffers, and the properties of the resulting gels were analyzed.
  • DATH diaminotrehalose
  • the dried gels were swelled in phosphate buffer (1 mM) in 0.9% NaCI with 3 mg/mL lidocaine for at least two hours to a concentration of 20-35 mg/mL.
  • the pH was controlled and adjusted if necessary to 7.4.
  • the gel particles were reduced in size with a fine filter.
  • the gel was filled in syringes and the syringes were sterilized by autoclavation (Fo 23).
  • Table 6 A summary of the gel properties is provided in Table 6. Table 6.
  • the dried gels were swelled in Bis-Tris buffer (1 mM) in 0.9% NaCI with 3 mg/mL lidocaine for at least two hours to a concentration of 20-35 mg/mL.
  • the pH was controlled and adjusted if necessary to 7.4.
  • the gel particles were reduced in size with fine filter.
  • the gel was filled in syringes and the syringes were sterilized by autoclavation (Fo 23).
  • a summary of the gel properties is provided in Table 7.
  • the dried gels were swelled in Bis-Tris buffer (1 mM) in 0.9% NaCI with 1 mM ZnC and 3 mg/mL lidocaine for at least two hours to a concentration of 20- 35 mg/mL.
  • the pH was controlled and adjusted if necessary to 7.4.
  • the gel particles were reduced in size with fine filter.
  • the gel was filled in syringes and the syringes were sterilized by autoclavation (Fo 23).
  • the corrected swelling degree (SwCC) of gels crosslinked with DATH in different buffers is presented in Fig. 5.
  • the gel part (GelP) of gels crosslinked with DATH in different buffers is presented in Fig. 6.
  • the graphs show that the hydrogels are stable after the process, also in the presence of Zn.
  • Gel examples 7-1 to 7-9 manufactured in Example 7 (A)-(C) were further subjected to an accelerated stability study at 60 °C for 7 and 14 days.
  • the gel part was analyzed for each of the batches and compared to the gel part at the beginning of the study (day 0).
  • the equation used to compare how much of the gel part that is left after day 14 compared to day 0 is shown below.
  • DMTMM (10.5 mol% DMTMM/heparosan) and the crosslinker (DATH), 1 .5 mol% DATH/Heparosan) were weighed in Falcon tubes and subsequently dissolved in phosphate buffer ( 1 mM, pH 7.4). The pH of the solutions was adjusted to pH 7-7.5 with 1 .2 M HCI. The DMTMM and DATH solutions were successively added to heparosan (Mw 140 kDa) weighed in a reaction vessel. The suspension was homogenized by shaking for 3.5 minutes and mixing with a spatula. The reaction mixture was placed in an incubator at 23 °C. After 24 hours the reaction was stopped by removal from incubator and the resulting material was pressed through a 1 mm steel mesh two times.
  • the bulk material was split in two factions.
  • the first fraction was swelled in a buffer containing 0.9% NaCI 10 mM bis-tris (pH 7.3) at room temperature to a concentration of 50 mg/nnL and pH was adjusted to 7.2-7.5.
  • the material was left at 70 °C for 24 hours and then particle-size reduced through a fine filter mesh three times.
  • the gel was filled on syringes and sterilized by autoclavation.
  • the second fraction was swelled in buffer containing 1 mM ZnC , 10 mM bis- tris and 0.9% NaCI (pH 7.3) at room temperature to a concentration of 50 mg/nnL and pH was adjusted to 7.2-7.5.
  • the material was left at 70 °C for 24 hours and then particle-size reduced through a fine filter mesh three times.
  • the gel was filled on syringes and sterilized by autoclavation. A summary of the gel properties is provided in Table 9.
  • DMTMM and the crosslinker DATH were weighed in separate Falcon tubes and dissolved in phosphate buffer (1 mM, pH 7.4) and the pH of the solutions were adjusted to pH 7-7.5 with 1 .2 M HCI.
  • DMTMM and DATH solutions were successively added to the chondroitin sulfate (Mw 30 kDA) weighed in a reaction vessel. The suspension was homogenized by shaking for 3.5 minutes and mixing with a spatula. The reaction mixture was incubated at 23 °C. After 24 hours the reaction was stopped by removal from incubator and the resulting material was pressed through a 1 mm steel mesh two times.
  • the bulk material was split in two factions.
  • the first fraction was swelled in a buffer containing 0.9% NaCI 10 mM bis-tris (pH 7.3) at room temperature to a concentration of 50 mg/mL and pH was adjusted to 7.2-7.5.
  • the material was left at 70 °C for 24 hours and then particle-size reduced through a fine filter mesh three times.
  • the gel was filled on syringes and sterilized by
  • the second fraction was swelled in buffer containing 1 mM ZnC , 10 mM bis-tris and 0.9% NaCI (pH 7.3) at room temperature to a concentration of 50 mg/mL and pH was adjusted to 7.2-7.5.
  • the material was left at 70 °C for 24 hours and then particle-size reduced through a fine filter mesh three times.
  • the gel was filled on syringes and sterilized by autoclavation. A summary of the gel properties is provided in Table 10.

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Abstract

L'invention concerne un procédé de préparation d'une composition d'hydrogel injectable stérilisée, comprenant les étapes consistant à : a) utiliser un glycosaminoglycane réticulé de manière covalente, b) faire gonfler le glycosaminoglycane réticulé de manière covalente dans une solution comprenant du tampon Bis-Tris, et c) stériliser la composition d'hydrogel par autoclavage pour former une composition d'hydrogel injectable stérilisée. L'invention concerne également une composition d'hydrogel injectable stérilisée comprenant : i) un glycosaminoglycane réticulé de manière covalente, et ii) un tampon Bis-Tris.
PCT/EP2018/067256 2017-06-28 2018-06-27 Gel de glycosaminoglycane avec tampon bis-tris WO2019002371A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997004012A1 (fr) 1995-07-17 1997-02-06 Q Med Ab Composition de gel a base de polysaccharide
US20120208890A1 (en) * 2010-01-13 2012-08-16 Allergan, Inc. Stable hydrogel compositions including additives
WO2015071433A1 (fr) * 2013-11-18 2015-05-21 Capsum Composition comprenant des capsules gélifiées stabilisées par un tampon
US20150209265A1 (en) * 2014-01-27 2015-07-30 Allergan Holdings France S.A.S. Spherical forms of cross-linked hyaluronic acid and methods of use thereof
EP3156044A1 (fr) * 2015-10-16 2017-04-19 Merz Pharma GmbH & Co. KGaA Compositions de polysaccharide réticulable in situ et ses utilisations

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997004012A1 (fr) 1995-07-17 1997-02-06 Q Med Ab Composition de gel a base de polysaccharide
US20120208890A1 (en) * 2010-01-13 2012-08-16 Allergan, Inc. Stable hydrogel compositions including additives
WO2015071433A1 (fr) * 2013-11-18 2015-05-21 Capsum Composition comprenant des capsules gélifiées stabilisées par un tampon
US20150209265A1 (en) * 2014-01-27 2015-07-30 Allergan Holdings France S.A.S. Spherical forms of cross-linked hyaluronic acid and methods of use thereof
EP3156044A1 (fr) * 2015-10-16 2017-04-19 Merz Pharma GmbH & Co. KGaA Compositions de polysaccharide réticulable in situ et ses utilisations

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
K. DE BOUILLE ET. AL.: "A Review of the Metabolism of 1,4-Butanediol Diglycidyl Ether-Crosslinked Hyaluronic Acid Dermal Fillers.", DERMATOLOOGIC SURGERY, vol. 39, no. 12, 1 December 2013 (2013-12-01), pages 1758 - 1766, XP005203470, DOI: 10.1111/dsu.12301 *
KEISUKE KURITA; NAOKO MASUDA; SADAFUMI AIBE; KAORI MURAKAMI; SHIGERU ISHII; SHIN-ICHIRO NISHIMURAT: "Synthetic Carbohydrate Polymers Containing Trehalose Residues in the Main Chain: Preparation and Characteristic Properties", MACROMOLECULES, vol. 27, 1994, pages 7544 - 7549, XP000485396, DOI: doi:10.1021/ma00104a007

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