KR20140096028A - Dermal filler compositions for fine line treatment - Google Patents

Abstract

The present invention provides a highly injectable, long-lasting hyaluronic acid hydrogel dermal filler composition that is particularly advantageous for the correction of facial wrinkles.

Description

{DERMAL FILLER COMPOSITIONS FOR FINE LINE TREATMENT}

Related application

This application claims priority and benefit of U.S. Provisional Patent Application No. 61 / 534,780, filed September 14, 2011, which is a continuation-in-part of U.S. Patent Application No. 13 / 486,754, filed June 1, 2012, This application is a continuation-in-part of U.S. Patent Application No. 13 / 593,313, filed on August 23, 2012, which claims priority and benefit of U.S. Provisional Patent Application No. 61 / 493,309, filed June 3, The disclosure of which is incorporated herein by reference in its entirety.

The technical field of the present invention

FIELD OF THE INVENTION The present invention relates generally to dermal filler compositions, and more particularly to injectable dermal filler compositions effective for the treatment of fine wrinkles in the skin.

Skin aging is a progressive phenomenon that occurs over time and can be affected by lifestyle factors such as alcohol consumption, tobacco and sun exposure. Aging of facial skin can be characterized by atrophy, sagging and thickening. Atrophy corresponds to a huge reduction in skin tissue thickness. Sagging of the subcutaneous tissue causes excessive skin and ptosis, resulting in ball and eyelid drooping. Thickening refers to excessive weight gain due to swelling of the lower face and neck. These changes are typically associated with drying, loss of elasticity and rough texture.

Hyaluronic acid (HA), also known as hyaluronan, is a non-sulfur oxidized glycosaminoglycan that is widely distributed throughout the human body in joints, epithelium, and nerve tissue. The hyaluronic acid ensures, for example, good hydration, assists in the organization of the extracellular matrix and acts as a filler material; It is common in the skin of different layers when they have various actions such as participating in the tissue damage mechanism. However, depending on the age, the amount of hyaluronic acid, collagen, elastin and other substrate polymers present in the skin is reduced. Repeated exposure to ultraviolet light, such as sunlight, for example, causes dermal cells not only to reduce the production of hyaluronan but also to increase its rate of degradation. This loss of material leads to various skin conditions such as, for example, wrinkles, hollowness, moisture loss and other unwanted conditions that contribute to the appearance of aging.

The injectable epidermal filler has been successfully used to treat aging skin. To treat such a skin condition, the filler can replace the lost endogenous substrate polymer or enhance / promote the function of the existing substrate polymer. The hyaluronic acid-based dermis filler has become increasingly popular because hyaluronic acid is a substance found naturally throughout the human body. These fillers are generally well tolerated, non-permanent, and extremely low risk for a wide variety of skin conditions.

The Tyndall effect is a side effect that occurs in some patients treated with a hyaluronic acid (HA) dermal filler. The Thinle effect is characterized by the appearance of a blue discoloration of the dermal filler in the injected skin area, which represents visible hyaluronic acid visible through the translucent skin. Clinical reports suggest that filler dosing techniques and skin characteristics may affect the signs of this side effect. Fillers with high rigidity and elasticity are successfully used in the correct areas on the face, such as wrinkles, balls, and jaws, without fear of facial discoloration because the material is injected into the middle and deep dermal areas. However, when these filler materials are used to repair wrinkles, for example tear creases, fine lines, laughs or wrinkles that are not deep in the forehead, or are applied too unduly in the upper region of the dermis, Bluish discoloration of the skin is often observed. This phenomenon, which is thought to be the result of the tinned effect, leaves a semi-permanent discoloration of the applied area and sometimes only disappears after the administration of hyaluronidase to break down the filler material. As a result, the Thinle effect is more common in patients treated for undersized wrinkles. Long-term signs of a tin effect that typically lasts for as long as seven months on the skin are the cause of the patient's main concern.

HA-based dermal filler gels have been specially formulated to treat "fine-wrinkled" wrinkles found around the tear grooves, forehead, eye wrinkles, The commercially available HA "fine wrinkle" gels include Juvederm Refine (G '~ 67 Pa; G' / G '~ 0.59, HA concentration 18 mg / ml), Belotero Soft (G' G '/ G' ~ 1.1, HA concentration 20 mg / ml), Emervel Touch (G '~ 56 Pa; G' / G '~ 0.64, HA concentration 20 mg / G "/ G '~ 0.20, HA concentration 16 mg / ml), Teosyal First Lines (G' 59 Pa; G" ~ 0.53, HA concentration 20 mg / ml) and Restylane Touch (G '~ 489 Pa; G "/ G' ~ 0.24, HA concentration 18 mg / ml). These gels are formulated to have a low elastic modulus, for example by lightly crosslinking a small amount of crosslinking agent with a linear HA chain and / or by reducing the final HA concentration of these gels, but most commercially available " Not particularly deep, for example, when injected at a depth of less than about 1 mm, still exhibits a tin effect in some patients.

Collagen gel can be used to treat undersized wrinkles and does not appear to cause tin effects. Collagen gels are not highly preferred because they have a relatively poor duration in the skin and require prior testing on the individual. Radiesse (registered trademark) (calcium hydroxyapatite) is a subcutaneously injectable implant, the principle ingredient of which is synthetic calcium hydroxyapatite, not hyaluronic acid. Unlike the hyaluronic acid dermis filler, calcium hydroxyapatite is not transparent and thus avoids the problem of the tinned effect. However, if placed too deeply, this filler can be seen as a white substance just below the skin. Further, compared to a hyaluronic acid filler, Radiesse (R) requires a larger needle for injection, and is not typically recommended for use in the eye area.

It is preferable to provide a main-use hyaluronic acid dermis filler which does not exhibit a bluish discoloration due to a tin-depletion effect even when it is injected deeply.

The present invention describes a composition and a method of making a HA-based dermis filler that can be administered to the upper dermis without producing any bluish discoloration of the skin or at least noticeable or noticeable bluish discoloration. In addition, many of the presently described filler gels of the present invention have been found to be significantly longer lasting in vivo than currently commercially available gels. In some embodiments of the present invention, an optically clear dermis filler is provided which is useful for improving the appearance of the skin, which adds volume and boldness and reduces the appearance of certain fine wrinkles without "tinling ". The present compositions can be introduced into the wrinkles in the skin, even in the thin skin area, rather than deeply, without causing negative negative blue discoloration associated with many conventional optically clear dermal fillers.

More specifically, in one aspect of the present invention there is provided a long-lasting therapeutic dermal filler composition comprising a polymer generally suitable for the living body, for example, an additive in combination with a crosslinked hyaluronic acid component and a hyaluronic acid component.

In one embodiment, the polymer is a polysaccharide, for example, a hyaluronic acid. The hyaluronic acid comprises a crosslinking component and may further comprise a non-crosslinking component. The additive may be a stabilized form of vitamin, for example vitamin C, e.g. vitamin C, or a vitamin C derivative such as L-ascorbic acid 2-glucoside (AA2G), ascorbyl 3-aminopropyl phosphate Vitagen) or sodium ascorbyl phosphate (AA2P).

In one aspect of the invention, the additive is a vitamin derivative covalently conjugated to the polymer by a suitable reaction process, such as ethanation, amidation or esterification.

In a broad aspect of the present invention, a dermal filler composition is provided, wherein the composition comprises a cross-linking component, a cross-linked hyaluronic acid component, and an additive other than a cross-linking component. The hyaluronic acid component can be chemically conjugated to the additive. Additionally, the composition exhibits a reduced tinned effect when administered to the dermis area of the patient, for substantially the same composition, but without the additive. The composition may further comprise an additive, for example an anesthetic such as lidocaine. In one embodiment, the additive is a vitamin C derivative, e. G., AA2G. In another embodiment, the additive is Vitagen.

In one embodiment, the hyaluronic acid component is chemically conjugated to an additive having a degree of conjugation of from about 3 mol% to about 40 mol, for example, from about 3 mol% to about 10 mol%.

The composition may be substantially optically clear. The composition generally has a G 'value of from about 40 Pa to about 100 Pa, e.g., about 100 Pa or less, and, for example, about 40 Pa or more.

In another aspect of the invention, a method of treating fine wrinkles in a patient's skin is provided. In one embodiment, the method comprises introducing into the patient's skin a composition comprising a hyaluronic acid component, a cross-linking component that bridges the hyaluronic acid, and an additive other than a cross-linking component, wherein the composition is substantially optically clear, Exhibit a reduced tinned effect when administered to the dermis area of the patient, for substantially the same composition, but without the additive.

In another aspect of the invention, there is provided a method of improving the appearance of a face, the method generally comprising administering a substantially optically clear dermal filler that does not exhibit, or does not significantly, . The composition comprises providing a hyaluronic acid, reacting the cross-linking agent with a vitamin C derivative, adding a reacted cross-linking agent and a vitamin C derivative to the hyaluronic acid to form a cross-linked hyaluronic acid composition comprising covalently conjugated vitamin C; And a step of homogenizing and neutralizing the crosslinked hyaluronic acid composition to obtain a used dermal filler composition. In some embodiments, the vitamin C derivative is AA2G. In another embodiment, the vitamin C derivative is vitagen.

In another aspect of the present invention, there is provided a method of reducing the appearance of fine wrinkles in a thin skin area of a patient, the method generally comprising providing a dermis filler composition, i.e., a substantially optically transparent composition comprising a vitamin C or vitamin C derivative Comprising the step of administering to a patient a hyaluronic acid dermal filler composition at a depth of about 1 mm or less. In some embodiments, the composition is injected at a depth of about 0.8 mm or less, about 0.6 mm or less, or about 0.4 mm or less.

In another embodiment of the invention, a dermo-peel composition is provided that comprises a substantially optically clear, generally cross-linked, hyaluronic acid component and a vitamin C derivative covalently conjugated to a hyaluronic acid component. In an exemplary embodiment, the composition has a G 'value of from about 40 Pa to about 100 Pa. In addition, the composition may have a hyaluronic acid concentration of about 18 mg / g to about 30 mg / g. These compositions can be particularly useful and effective in the treatment of fine wrinkles or deep creases even in the skin, for example, very thin skin, for example, skin having a thickness of less than about 1 mm. In some embodiments, the composition of the present invention lasts for at least 3 months, at least 6 months, or 1 year after being introduced into the skin.

These and other aspects of the invention will be more readily understood and appreciated with reference to the following drawings and detailed description.

1 shows the structure of L-ascorbic acid 2-glucoside (AA2G);
Figure 2 shows the structure of ascorbyl 3-aminopropyl phosphate (bitagene);
Figure 3 shows the structure of sodium ascorbyl phosphate (AA2P);
4 shows the structure of 1,4-butanediol diglycidyl ether (BDDE);
Figure 5 shows the structure of pentaerythritol glycidyl ether (star-PEG epoxide);
Figure 6 shows the structure of pentaerythritol (3-aminopropyl) ether (star-PEG amine);
Figure 7 shows a table showing the degree of conjugation and G 'values for various dermal filler compositions according to the present invention;
Figure 8 is a table showing the degree of conjugation, HA concentration and G 'value for the HA-AA2G (BDDE) dermis filler composition according to the present invention;
Figure 9 shows a graphical representation of the observed release percentage of AsA from a solution of AA2G in PBS for time (minutes) for four different a-glucosidase concentrations;
Figure 10 shows a representation of the release profile (sustained release) of free AsA from the conjugated dermal filler according to the invention (mol% versus AA2G conversion at reaction time);
FIGS. 11A and 11B show additional emission data for various dermal fillers in accordance with the present invention; FIGS.
Figure 12 shows an image of the skin after deep injection of the HA-based dermal filler gel of the present invention and some commercially available gels for fine wrinkle application;
Figure 13 shows the visual tin scores of the HA-based dermal filler gels of the present invention and certain commercially available gels for fine wrinkle application;
Figure 14 shows percentages of blue light emitted from the skin of the HA-based dermatophyll gel of the present invention and some commercially available gel for fine wrinkle application;
Figure 15 shows the overall percentage of gels remaining after one week of transplantation of the HA-based dermal filler gel of the present invention and some commercially available gels for fine line application;
Figure 16 shows the overall percentage of gels remaining on the 0th, 12th, 24th, and 40th weeks of the implanted HA-based dermal filler gel of the present invention for fine wrinkle application and some commercially available gels drawing.

In one aspect of the invention, a dermal filler composition is provided, the composition generally comprising a biocompatible polymer, such as a polysaccharide such as a crosslinked hyaluronic acid, and a vitamin C derivative covalently conjugated to the polymer. The compositions provide for sustained release of vitamin C for skin as well as neocollagenesis as well as other therapeutic or cosmetic benefits. When introduced into the skin, e. G. Into the dermis, the composition reacts with endogenous enzymes in the body and, over time, bioactive vitamin C is produced in vivo through enzymatic cleavage. Since vitamin C is released from the composition over a period of weeks or months, its attendant advantages are made available to the body.

The polymer may be selected from the group of proteins, peptides and polymers consisting of polypeptides, polylysine, collagen, pro-collagen, elastin and laminin.

The polymer may be a synthetic polymer having hydroxyl, amine and carboxyl functional groups: poly (vinyl alcohol), polyethylene glycol, polyvinylamine, polyallylamine, deacetylated polyacrylamide, polyacrylic acid and polymethacrylic acid ≪ / RTI > The polymer may be selected from the group of polymers consisting of a dendritic or branched polymer comprising a dendritic polyol and a dendritic polyamine. The polymer may be selected from the group of polymers consisting of solid surfaces having hydroxyl, amine and carboxyl functionalities.

Polymers include, for example, starches and derivatives thereof; Dextran and its derivatives, cellulose and its derivatives; Chitin and chitosan and alginate and derivatives thereof.

In an exemplary embodiment of the present invention, the polymer is glycosaminoglycan. The hydrogel compositions disclosed herein may additionally comprise two or more different glycosaminoglycan polymers. The term "glycosaminoglycans" as used herein is synonymous with "GAG" and "mucopolysaccharides" and refers to long unbranched polysaccharides consisting of repeating disaccharide units. The repeating unit consists of hexosaccharide (hexose) or hexuronic acid and pharmaceutically acceptable salts thereof, linked to hexosamine (a nitrogen containing six-membered sugar). Members of the GAG family differ in the type of hexosamine, hexose or hexuronic acid units they contain, such as glucuronic acid, iduronic acid, galactose, galactosamine, glucosamine, and also the geometry of the glycoside bond . Any glycosaminoglycan polymer is useful in the hydrogel compositions disclosed herein, provided that the glycosaminoglycan polymer improves the skin condition. Non-limiting examples of glycosaminoglycans include chondroitin sulfate, dermatan sulfate, keratan sulfate, hyaluronan. Non-limiting examples of acceptable salts of glycosaminoglycans include sodium salts, potassium salts, magnesium salts, calcium salts, and combinations thereof. Glycosaminoglycans and their obtained polymers useful in the hydrogel compositions and methods disclosed herein are described, for example, in US Patent Publication No. 2003/0148995 to Piron and Tholin (entitled Polysaccharide Crosslinking, Hydrogel Preparation, Resulting Polysaccharides (s) and Hydrogel (s), uses Thereof); Lebreton (name of the invention: Cross-Linking of Low and High Molecular Weight Polysaccharides Preparation of Injectable Monophase Hydrogels); U.S. Patent Application Publication 2008/0089918 to Lebreton, entitled Viscoelastic Solutions Containing Sodium Hyaluronate and Hydroxypropyl Methyl Cellulose, Preparation and Uses; U.S. Patent Publication No. 2010/0028438 to Lebreton et al. Entitled Hyaluronic Acid-Based Gels Including Lidocaine; And United States Patent Publication No. 2006/0194758 (entitled Polysaccharides and Hydrogels thus Obtained); And International Patent Publication WO 2004/073759 to Di Napoli, entitled " Composition and Method for Intradermal Soft Tissue Augmentation ", each of which is incorporated herein by reference in its entirety. GAGs useful in the hydrogel compositions and methods disclosed herein include, for example, hyaluronic acid dermis peeler JUVEDERM, JUVEDERM 30, JUVEDERM, JUVEDERM® Ultra®, JUVEDERM® Ultra XC and JUVEDERM® ULTRA PLUS XC available from Alabin, Calif., Irvine, Calif. 0.0 > Allergan < / RTI > Inc). Table 1 lists representative GAGs.

An aspect of the present invention provides a hydrogel composition comprising, in part, a chondroitin sulfate polymer. The term "chondroitin sulfate polymer " as used herein refers to disaccharides of two alternative monosaccharides of D-glucuronic acid (GlcA) and N- acetyl-D-galactosamine (GalNAc) and their pharmaceutically acceptable salts ≪ RTI ID = 0.0 > sulfated < / RTI > polymers. The chondroitin sulfate polymer may comprise a D-glucuronic acid residue epimerized with L-iduronic acid (IdoA), in which case the resulting disaccharide is referred to as further malonic acid. Chondroitin sulfate polymers may have more than 100 individual sugar chains, each of which may be sulfated in variable amounts and amounts. Chondroitin sulfate polymer is a structural component of critical cartilage and provides its great resistance to compression. Any chondroitin sulfate polymer is useful in the compositions disclosed herein, provided that the chondroitin sulfate polymer improves the skin condition. Non-limiting examples of pharmaceutically acceptable salts of chondroitin sulfate include sodium chondroitin sulfate, potassium chondroitin sulfate, magnesium chondroitin sulfate, calcium chondroitin sulfate, and combinations thereof.

An aspect of the present disclosure provides a hydrogel composition that includes, in part, a keratan sulfate polymer. As used herein, the term "keratan sulfate polymer" refers to a variable length polymer comprising disaccharide units, which themselves are? -D-galactose and N- acetyl-D-galactosamine (GalNAc) and And pharmaceutically acceptable salts thereof. Within the repeat region of keratan sulfate, the disaccharide can be fucosylated and the N-acetylneuraminic acid capts the ends of the chain. Any keratan sulfate polymer is useful in the compositions disclosed herein, provided that the keratan sulfate polymer improves the skin condition. Non-limiting examples of pharmaceutically acceptable salts of keratan sulfate include sodium keratan sulfate, potassium keratan sulfate, magnesium keratan sulfate, calcium keratan sulfate, and combinations thereof.

An aspect of the present disclosure provides a hydrogel composition comprising a partially hyaluronan polymer. The term " hyaluronic acid polymer "as used herein is synonymous with" HA polymer "," hyaluronic acid polymer ", and" hyaluronate polymer " Refers to a polymer that itself comprises D-glucuronic acid and DN-acetylglucosamine monomer and pharmaceutically acceptable salts thereof linked together via alternative? -1,4 and? -1,3 glycoside linkages do. The hyaluronan polymer may be purified from animal and non-animal sources. The polymer of hyaluronan can range in size from about 5,000 Da to about 20,000,000 Da. Any hyaluronan polymer is useful in the compositions disclosed herein, provided that the hyaluronan polymer improves the skin condition. Non-limiting examples of pharmaceutically acceptable salts of hyaluronan include hyaluronan sodium, hyaluronan potassium, hyaluronan magnesium, hyaluronan calcium, and combinations thereof.

An embodiment of the present disclosure provides a hydrogel composition comprising a partially crosslinked glycosaminoglycan polymer. The term "crosslinked" as used herein refers to an intermolecular bond in which an individual polymer molecule or monomer chain is bonded to a more stable structure such as a gel. Thus, the crosslinked glycosaminoglycan polymer has intermolecular bonds in which at least one individual polymer molecule is bonded to each other. Bridging of the glycosaminoglycan polymers typically results in the formation of hydrogels. Such hydrogels have high viscosity and require considerable force to be extruded through fine needles. The glycosaminoglycan polymers disclosed herein can be used in combination with polyfunctional PEG-based crosslinkers, such as, but not limited to, di-aldehydes, including, but not limited to, polyfunctional PEG-based crosslinkers, divinyl sulfone, diglycidyl ether and bis- Can be crosslinked using a disulfide crosslinking agent. Non-limiting examples of hyaluronan cross-linking agents include polyfunctional PEG crosslinker-like pentaerythritol tetraglycidyl ether (PETGE), divinyl sulfone (DVS), 1,4-butanediol diglycidyl ether (BDDE) (EGDGE), 1,2,7,8-diepoxyoctane (DEO), (phenylenebis- (ethyl) -carbodiimide and 1, Hexamethylenebis (ethylcarbodiimide), adipdihydrazide (ADH), bis (sulfosuccinimidyl) suberate (BS), hexamethylenediamine (HMDA), 1- Propyl) -2, 3-epoxycyclohexane, lysine, lysine methyl ester, or combinations thereof. Other useful cross-linking agents are described in Stroumpoulis and Tezel, U.S. Patent Application No. 12 / 910,466, filed October 22, , Entitled " Tunably Crosslinked Polysaccharide Compositions ", which is incorporated herein by reference in its entirety. Lycan polymers are useful in the hydrogel compositions disclosed herein, provided that the glycosaminoglycan polymer improves the skin condition. Non-limiting examples of glycosaminoglycans include chondroitin sulfate, dermatan sulfate, Sulfuric acid, and hyaluronan. Non-limiting examples of acceptable salts of glycosaminoglycans include sodium, potassium, magnesium, calcium salts, and combinations thereof. The hydrogel compositions disclosed herein And the resulting polymers thereof are described in, for example, U.S. Patent Publication No. 2003/0148995 to Piron and Tholin, entitled Polysaccharide Crosslinking, Hydrogel Preparation, Resulting Polysaccharides (s) and Hydrogel (s), uses Thereof); Lebreton (the name of the invention: Cross-Linking of Low and High Molecular Weight Polysaccharides Preparation of Injectable Monophase Hydrogels); U.S. Patent Application Publication 2008/0089918 to Lebreton, entitled Viscoelastic Solutions Containing Sodium Hyaluronate and Hydroxypropyl Methyl Cellulose, Preparation and Uses; U.S. Patent Publication No. 2010/0028438 to Lebreton et al. Entitled Hyaluronic Acid-Based Gels Including Lidocaine; And United States Patent Publication No. 2006/0194758 (entitled Polysaccharides and Hydrogels thus Obtained); And International Patent Publication WO 2004/073759 to Di Napoli, entitled " Composition and Method for Intradermal Soft Tissue Augmentation ", each of which is incorporated herein by reference in its entirety.

An embodiment of the present disclosure provides a hydrogel composition comprising a glycosaminoglycan polymer having a degree of crosslinking in part. The term "degree of crosslinking" as used herein refers to the percentage of glycosaminoglycan polymer monomer units such as, for example, disialic monomer units of hyaluronan bound to a crosslinking agent. The degree of crosslinking is expressed as a weight percentage ratio of crosslinking agent to glycosaminoglycan. In certain advantageous embodiments of the invention, the degree of crosslinking is from about 3% to about 12%, such as from about 5% to about 10%.

In an embodiment, the hydrogel composition comprises a crosslinked glycosaminoglycan polymer, for example, a crosslinked hyaluronic acid, wherein the crosslinked glycosaminoglycan polymer is, for example, from about 18 mg / Is present in the composition at a concentration of about 30 mg / g. In some embodiments, the composition has a total hyaluronic acid concentration of about 24 mg / g or about 25 mg / g.

An embodiment of the present disclosure provides a hydrogel composition comprising a partially low molecular weight hyaluronan polymer, a high molecular weight hyaluronan polymer, or both a low molecular weight and a high molecular weight hyaluronan polymer. As used herein, the term "high molecular weight" when referring to "hyaluronan" refers to a hyaluronan polymer having an average molecular weight of at least 1,000,000 Da. Non-limiting examples of high molecular weight hyaluronan polymers include hyaluronan polymers of about 1,500,000 Da, about 2,000,000 Da, about 2,500,000 Da, about 3,000,000 Da, about 3,500,000 Da, about 4,000,000 Da, about 4,500,000 Da to about 5,000,000 Da, . The term "low molecular weight" as used herein refers to a hyaluronan polymer having an average molecular weight of less than 1,000,000 Da when referring to "hyaluronan. &Quot; Non-limiting examples of low molecular weight hyaluronan polymers include hyaluronan polymers of about 200,000 Da, about 300,000 Da, about 400,000 Da, about 500,000 Da, about 600,000 Da, about 700,000 Da, about 800,000 Da, .

In an embodiment, the composition comprises a low molecular weight crosslinked hyaluronan polymer. In an embodiment of this embodiment, the composition comprises, for example, about 100,000 Da, about 200,000 Da, about 300,000 Da, about 400,000 Da, about 500,000 Da, about 600,000 Da, about 700,000 Da, about 800,000 Da, Of the average molecular weight of the crosslinked hyaluronan polymer. In another embodiment of this embodiment, the composition comprises, for example, at most 100,000 Da, at most 200,000 Da, at most 300,000 Da, at most 400,000 Da, at most 500,000 Da, at most 600,000 Da, at most 700,000 Da, at most 800,000 Da, at most 900,000 Da Da or a crosslinked hyaluronan polymer having an average molecular weight of up to 950,000 Da. In another embodiment of this embodiment, the composition comprises, for example, from about 100,000 Da to about 500,000 Da, from about 200,000 Da to about 500,000 Da, from about 300,000 Da to about 500,000 Da, from about 400,000 Da to about 500,000 Da, Da to about 950,000 Da, about 600,000 Da to about 950,000 Da, about 700,000 Da to about 950,000 Da, about 800,000 Da to about 950,000 Da, about 300,000 Da to about 600,000 Da, about 300,000 Da to about 700,000 Da, About 800,000 Da, or a crosslinked hyaluronan polymer having an average molecular weight of about 400,000 Da to about 700,000 Da.

In another embodiment, the composition comprises a high molecular weight crosslinked hyaluronan polymer. In an embodiment of this embodiment, the composition comprises, for example, about 1,000,000 Da, about 1,500,000 Da, about 2,000,000 Da, about 2,500,000 Da, about 3,000,000 Da, about 3,500,000 Da, about 4,000,000 Da, about 4,500,000 Da, And a crosslinked hyaluronan polymer having an average molecular weight. At least 1,000,000 Da, at least 1,500,000 Da, at least 2,000,000 Da, at least 2,500,000 Da, at least 3,000,000 Da, at least 3,500,000 Da, at least 4,000,000 Da, at least 4,500,000 Da, or at least 5,000,000 Da And a cross-linked hyaluronan polymer having an average molecular weight of Da. In another embodiment of this embodiment, the composition comprises, for example, from about 1,000,000 Da to about 5,000,000 Da, from about 1,500,000 Da to about 5,000,000 Da, from about 2,000,000 Da to about 5,000,000 Da, from about 2,500,000 Da to about 5,000,000 Da, To about 3,000,000 Da, and an average molecular weight of about 2,500,000 Da to about 3,000,000 Da.

In another embodiment, the composition comprises a crosslinked hyaluronan polymer wherein the crosslinked hyaluronan polymer comprises a combination of a high molecular weight hyaluronan polymer and a low molecular weight hyaluronan polymer in various ratios do. In another embodiment, the composition comprises a crosslinked hyaluronan polymer wherein the crosslinked hyaluronan polymer has a molecular weight of about 20: 1, about 15: 1, about 10: 1, about 5: 1, about 1: : 1, about 1: 5, about 1:10, about 1: 15, or about 1: 20, and a low molecular weight hyaluronan polymer.

An embodiment of the present disclosure provides a hydrogel composition comprising a partially uncrosslinked glycosaminoglycan polymer. The term "uncrosslinked ", as used herein, refers to the lack of intermolecular linkages that bond individual glycosaminoglycan polymer molecules or monomer chains. As such, the uncrosslinked glycosaminoglycan polymer is not linked to any other glycosaminoglycan polymer by intermolecular bonding. In an embodiment of this embodiment, the composition is selected from the group consisting of an uncondensed chondroitin sulfate polymer, an uncoupled dermatan sulfate polymer, an uncoupled keratan sulfate polymer, an uncured heparan polymer, an uncoupled heparan sulfate polymer, Lt; / RTI > polymers. Uncrosslinked glycosaminoglycan polymers are water soluble and generally remain fluid in nature. As such, uncrosslinked glycosaminoglycan polymers are often mixed with a glycosaminoglycan polymeric hydrogel composition as a lubricant to facilitate the process of extruding the composition through a fine needle.

In an embodiment, the composition comprises an uncrosslinked glycosaminoglycan polymer, wherein the uncrosslinked glycosaminoglycan polymer is, for example, about 2 mg / g, about 3 mg / g, about 4 About 8 mg / g, about 10 mg / g, about 11 mg / g, about 12 mg / g, about 5 mg / about 13 mg / g, about 14 mg / g, about 15 mg / g, about 16 mg / g, about 17 mg / g, about 18 mg / About 20 mg / g, about 40 mg / g, or about 60 mg / g. In another aspect of this embodiment, the composition provides uncrosslinked glycosaminoglycans, wherein the uncrosslinked glycosaminoglycan has, for example, at least 1 mg / g, at least 2 mg / g G, at least 5 mg / g, at least 10 mg / g, at least 15 mg / g, at least 20 mg / g, at least 25 mg / g, at least 35 mg / g, or at least 40 mg / Lt; / RTI > mg / g. In another embodiment of this embodiment, the composition comprises uncrosslinked glycosaminoglycans, wherein the uncrosslinked glycosaminoglycan has, for example, a maximum of 1 mg / g, a maximum of 2 mg / g, at most 3 mg / g, at most 4 mg / g, at most 5 mg / g, at most 10 mg / g, at most 15 mg / g, at most 20 mg / g or at most 25 mg / g. In another embodiment of this embodiment, the composition comprises uncrosslinked glycosaminoglycan, wherein the uncrosslinked glycosaminoglycan is, for example, from about 1 mg / g to about 60 mg / g, about 10 mg / g to about 40 mg / g, about 7.5 to about 19.5 mg / g, about 8.5 to about 18.5 mg / g, about 9.5 to about 17.5 mg / From about 10.5 mg / g to about 16.5 mg / g, from about 11.5 mg / g to about 15.5 mg / g, or from about 12.5 mg / g to about 14.5 mg / g.

Embodiments herein provide a hydrogel composition essentially free of partially crosslinked glycosaminoglycan polymers. The term "essentially absent" (or "consisting essentially of") as used herein refers to a composition wherein only trace amounts of crosslinked substrate polymer can be detected. In an embodiment of this embodiment, the composition comprises chondroitin sulfate essentially free of bridged chondroitin sulfate polymer, dermatan sulfate essentially free of bridged maltitanium sulfate polymer, keratan sulfate essentially free of bridged keratan sulfate polymer, Heparan which is essentially free of heparan polymer, heparan sulfuric acid essentially free of crosslinked heparan sulfate polymer, or hyaluronan sulfate which is essentially free of crosslinked hyaluronan polymer.

Embodiments herein provide a hydrogel composition that is completely free of partially crosslinked glycosaminoglycan polymers. The term "completely absent" as used herein refers to a composition within the detection range of the instrument or process used, wherein the crosslinked glycosaminoglycan polymer can not be detected or its presence can not be confirmed . In an embodiment of this embodiment, the composition comprises chondroitin sulfate completely free of bridged chondroitin sulfate polymer, dermatan sulfate completely free of bridged maltitanium sulfate polymer, keratan sulfate completely free of bridged keratan sulfate polymer, Heparan completely lacking the polymer, heparan sulfate completely lacking the crosslinked heparan sulfate polymer, or hyaluronan sulfate completely lacking the crosslinked hyaluronan polymer.

An aspect of the present disclosure provides a hydrogel composition comprising a ratio of a partially crosslinked glycosaminoglycan polymer to an uncrosslinked glycosaminoglycan polymer. This ratio of cross-linked and uncrosslinked glycosaminoglycan polymers is known as the gel: fluid ratio. Any gel: fluid ratio is useful in the preparation of the compositions disclosed herein, provided that the ratio disclosed herein improves the skin conditions disclosed herein. Non-limiting examples of gel to fluid ratios in the compositions of the present invention are 100: 0, 98: 2, 90:10, 75:25, 70:30, 60:40, 50:50, 40:60, 30:70, 25:75, 10:90; 2:98, and 0: 100.

In an embodiment of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio is, for example, about 0: 100, about 1:99, about 2:98, about 3:97, about 4:96, about 5:95, about 6:94, about 7:93, about 8:92, about 9:91, or about 10:90 . In another aspect of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio is, for example, up to 1:99, Up to 2:98, Up to 3:97, Up to 4:96, Up to 5:95, Up to 6:94, Up to 7:93, Up to 8:92, Up to 9:91 or Up to 10:90. In an embodiment of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio is, for example, from about 0: 100 to about 3: 97, from about 0: 100 to about 5: 95, or from about 0: 100 to about 10: 90.

In another aspect of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio is, for example, about 15:85, about 20:80, about 25:75, about 30:70, about 35:65, about 40:60, about 45:55, about 50:50, about 55:45, about 60:40, about 65:35, about 70:30, about 75:25, about 80:20, about 85:15, about 90:10, about 95: 5, about 98: 2, or about 100: In another embodiment of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio is, for example, up to 15:85, Maximum 20:80, maximum 25:75, maximum 30:70, maximum 35:65, maximum 40:60, maximum 45:55, maximum 50:50, maximum 55:45, maximum 60:40, maximum 65:35, Maximum 70:30, maximum 75:25, maximum 80:20, maximum 85:15, maximum 90:10, maximum 95: 5, maximum 98: 2 or maximum 100: In another embodiment of this embodiment, the composition comprises a crosslinked glycosaminoglycan polymer and an uncrosslinked glycosaminoglycan polymer, wherein the gel: fluid ratio is, for example, from about 10: About 70:30, about 15:85 to about 70:30, about 10:90 to about 55:45, about 80:20 to about 95: 5, about 90:10 to about 100: 0, about 75:25 About 100: 0, or about 60: 40 to about 100: 0.

The hydrogel compositions disclosed herein may additionally comprise other agents or combinations of agents that provide a beneficial effect when the composition is administered to a subject. Such advantageous agonists include antioxidants, anti-itch agents, anti-cellulite agents, anti-scarring agents, antiinflammatory agents, anesthetics, irritation inhibitors, vasoconstrictors, vasodilators, antihypertensive agents, antifibrinolytic agents, exfoliants, , Antioxidants, or moisturizers.

For purposes of this specification, unless otherwise stated, "%" in the formulation is defined as a weight / weight (ie, w / w) percentage.

Embodiments of the present disclosure provide hydrogel compositions disclosed herein that may optionally include anesthetic in part. Anesthetics are preferably local anesthetics, i.e. anesthetics which cause loss of reversible local anesthesia and infiltration, for example, aminamide local anesthetics and amino ester topical anesthetics. The amount of anesthetic agent included in the compositions disclosed herein is an amount effective to alleviate the pain experienced by an individual upon administration of the composition. As such, the amount of anesthetic included in the compositions disclosed herein is from about 0.1% to about 5% by weight of the total composition. Non-limiting examples of anesthetic agents include lidocaine, ambalocaine, amolanone, amilocaine, benoxinate, benzocaine, betoxycaine, biphenamine, bupivacaine, butacaine, butamben, butanylicaine, But are not limited to, but are not limited to, butoxycycline, carthocine, chloropropane, coco ethylene, cocaine, cyclomethicaine, dibucaine, dimethisoquine, dimethocaine, difetron, dicyclo- Aminobenzoate, ricinocaine mesylate, lepidocaine, lidocaine, mefibacaine, mephylcaine, methabutoxycaine, methyl chloride, myrtecaine, napine , Octacaine, orthocaine, oxetazine, parethoxycaine, phenacaine, phenol, piperocaine, pyridocaine, polydocanol, pramoxine, prilocaine, procaine, propanocaine, Propifocaine, propoxycaine, shu Cocaine, fatigue Cain, and the ropi tanks, salicyl alcohol, tetracaine, tolyl Rica of, tri Mecca, Zola diamine, a combination thereof and salts thereof. Non-limiting examples of amino ester topical anesthetics include, but are not limited to, procaine, chloroprocaine, cocaine, cyclomethicaine, shimetokine (laurocaine), propoxycaine, procaine (novocaine), proparacaine, tetracaine Tokine). Non-limiting examples of aminoamide local anesthetics include but are not limited to articaine, bupivacaine, cinchocaine (dibucaine), ethidocaine, levobupivacaine, lidocaine (lignocaine), mephibacaine, piperocaine, Rocaine, lopivacaine, and tramecane. The compositions disclosed herein may comprise a single anesthetic or multiple anesthetics. A non-limiting example of a local anesthetic is lidocaine / prilocaine (EMLA).

Thus, in an embodiment, the compositions disclosed herein include an anesthetic and salts thereof. In an embodiment of this embodiment, the compositions disclosed herein comprise an aminoamide local anesthetics and salts thereof or amino ester topical anesthetics and salts thereof. In another aspect of this embodiment, the composition disclosed herein is selected from the group consisting of procaine, chloroprocaine, cocaine, cyclomethicaine, simethotaine, propoxycaine, procaine, proparacaine, tetracaine, And any combination of these. In another aspect of this embodiment, the composition disclosed herein is selected from the group consisting of ativa, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mephivacaine, piperocaine, Or a salt thereof, or any combination thereof. In another aspect of this embodiment, the compositions disclosed herein comprise a lidocaine / prilocaine combination.

In another aspect of this embodiment, the compositions disclosed herein comprise, for example, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5% About 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1.0 wt%, about 2.0 wt%, about 3.0 wt%, about 4.0 wt%, about 5.0 wt%, about 6.0 wt%, about 7.0 wt% %, About 8.0 wt.%, About 9.0 wt.%, Or about 10 wt.% Of the anesthetic agent. In another embodiment, the compositions disclosed herein comprise, for example, at least 0.1%, at least 0.2%, at least 0.3%, at least 0.4%, at least 0.5%, at least 0.6%, at least 0.7% At least 0.9 wt%, at least 1.0 wt%, at least 2.0 wt%, at least 3.0 wt%, at least 4.0 wt%, at least 5.0 wt%, at least 6.0 wt%, at least 7.0 wt%, at least 8.0 wt% %, At least 9.0 wt%, or at least 10 wt%. In another embodiment, the compositions disclosed herein comprise, for example, up to 0.1 wt%, up to 0.2 wt%, up to 0.3 wt%, up to 0.4 wt%, up to 0.5 wt%, up to 0.6 wt% Up to 0.9 wt%, up to 1.0 wt%, up to 2.0 wt% up to 3.0 wt% up to 4.0 wt% up to 5.0 wt% up to 6.0 wt% up to 7.0 wt% up to up to 8.0 wt% %, Up to 9.0 wt%, or up to 10 wt%. In a further embodiment, the compositions disclosed herein comprise, for example, from about 0.1% to about 0.5%, from about 0.1% to about 1.0%, from about 0.1% to about 2.0%, from about 0.1% About 0.2 wt% to about 0.9 wt%, about 0.2 wt% to about 1.0 wt%, about 0.2 wt% to about 3.0 wt%, about 0.1 wt% to about 4.0 wt%, about 0.1 wt% From about 0.5 wt% to about 2.0 wt%, from about 0.5 wt% to about 1.0 wt%, or from about 0.5 wt% to about 2.0 wt%.

In another embodiment, the compositions disclosed herein do not include an anesthetic.

In one aspect of the present invention, a polymer, such as a glycosaminoglycan polymer, such as a hyaluronic acid polymer, such as a hyaluronic acid at least partially crosslinked, and an additive or beneficial agent in combination with a polymer, An injectable dermis filler is provided.

Advantageous agonists in combination with polymers may include vitamins, such as vitamin C, for example. Non-limiting examples of suitable forms of vitamin C include sodium, potassium and calcium salts of ascorbic acid and ascorbic acid, fat-soluble esters of ascorbic acid and long chain fatty acids (ascorbyl palmitate or ascorbyl stearate), magnesium ascorbyl phosphate MAP), sodium ascorbyl phosphate (SAP), and ascorbic acid 2-glucoside (AA2G ™), sodium ascorbyl phosphate (AA2P), disodium 2-sodium ascorbyl sulfate, and ascorbyl 3-aminopropyl phosphate ).

In a particularly advantageous embodiment, the beneficial agent is covalently conjugated to the polymer. For example, an advantageous agonist may be vitamin C, or a vitamin C derivative, which is covalently conjugated to the polymer and is present in an amount ranging from about 0.04% to about 5.0% by weight of the total composition, for example, about 0.1% % To about 4.0 wt%, e.g., from about 0.2 wt% to about 2.0 wt% of the total composition. In one embodiment, the amount of vitamin C contained in the compositions disclosed herein is from about 0.3% to about 1.2% by weight of the total composition.

Preferably, the vitamin C that is covalently conjugated to the polymer is selected from the group consisting of ascorbic acid, L-ascorbic acid, L-ascorbic acid 2-sulfate (AA-2S) and L-ascorbic acid 2-phosphate AA-2G), 6-O-acyl-2-O-alpha-D-glucopyranosyl-L-ascorbic acid (2-O- (Ascorbyl 3-aminopropyl phosphate, ascorbyl palmitate), derivatives, and derivatives thereof. The compositions disclosed herein may comprise a single vitamin C agonist or multiple vitamin C agonists.

In another embodiment of the present invention, a dermal filler is provided, wherein the hyaluronic acid is crosslinked with BDDE. In this embodiment, the degree of conjugation may be from about 3 mol% to about 10 mol, and from about 15 mol% to about 40 mol%.

In some embodiments, the dermis filler has sustained bioavailability. For example, when introduced into human skin, dermal fillers effective to release ascorbic acid or other vitamins to humans for at least about 1 month to about 20 months or more are provided.

Aspects of the present invention provide hydrogel compositions disclosed herein that partially exhibit complex modulus, elastic modulus, viscous modulus and / or tan delta. The compositions as disclosed herein are viscoelastic, that is, the composition is composed of an elastomeric component (such as a solid-like, cross-linked glycosaminoglycan polymer) and a viscous component (such as a liquid, Such as an uncrosslinked glycosaminoglycan polymer or carrier phase. The rheological property describing this property is the complex modulus (G *), which defines the overall resistance of the composition to deformation. The complex modulus of elasticity is a complex number with real and imaginary parts: G * = G '+ iG ". The absolute value of G * is Abs (G *) = Sqrt (G' ² + G" ² ). The complex elastic modulus can be defined as the sum of the modulus of elasticity (G ') and the viscosity (G "). [Falcone, et al., Temporary Polysaccharide Dermal Fillers : A Model for Persistence Based on Physical Properties , Dermatol Surg. 35 (8): 1238-1243 (2009); Tezel, cited above, 2008; Kablik, cited above, 2009; Beasley, cited above, 2009]; Each of which is incorporated herein by reference in its entirety.

The modulus of elasticity or modulus refers to the ability of a hydrogel material to resist the tendency of an object to deform or otherwise non-permanently deform when a force is applied to the object. The modulus of elasticity is characterized by the hardness of the composition and is also known as the storage modulus because it accounts for the storage of energy from the movement of the composition. The modulus of elasticity describes the interaction between elasticity and strength (G '= stress / strain) and thus provides a quantitative measure of the hardness or softness of the composition. The elastic modulus of an object is defined as the slope of its stress-strain curve in the region of elastic strain: lambda = stress / strain, where lambda is the modulus of elasticity (Pascal); Stress is the force that causes deformation, divided by the area to which the force is applied; Deformation is the ratio of change to the original state of an object caused by stress. Despite the dependence on the rate at which the force is applied, the stiffer composition will have a higher modulus of elasticity and will take greater force to deform the material to a given distance, for example with an injection. Specifying the way in which stress is measured, including the direction, allows multiple types of elasticity to be defined. The three main elastic moduli are tensile, shear, and bulk modulus.

The viscosity rate is also known as the loss rate, since it accounts for the energy lost as a viscous extinction. Tan δ is the ratio of the viscosity and the modulus, tan δ = G "/ G '. [Falcone, cited above, 2009] For the tan δ values disclosed herein, tan δ is the dynamic modulus at a frequency of 1 Hz The lower tan delta corresponds to a more rigid, harder or more elastic composition.

In another embodiment, the compositions disclosed herein do not exhibit a modulus of elasticity. In an embodiment of this embodiment, the hydrogel composition has a viscosity of about 250 Pa, about 250 Pa, about 100 Pa, about 125 Pa, about 150 Pa, about 175 Pa, about 200 Pa, about 250 Pa, about 300 Pa, about 350 Pa, about 400 Pa, about 450 Pa, about 500 Pa, about 550 Pa, about 600 Pa, about 650 Pa, about 700 Pa, about 750 Pa, about 800 Pa, About 1000 Pa, about 1,200 Pa, about 1,300 Pa, about 1,400 Pa, about 1,500 Pa, about 1,600 Pa, about 1700 Pa, about 1800 Pa, about 1900 Pa, about 2,000 Pa, about 2,100 Pa, about 2,200 Pa, about 2,300 Pa, about 2,400 Pa, or about 2,500 Pa. In another aspect of this embodiment, the hydrogel composition comprises, for example, at least 25Pa, at least 50Pa, at least 75Pa, at least 100Pa, at least 125Pa, at least 150Pa, at least 175Pa, at least 200Pa, at least 250 At least 300 Pa, at least 350 Pa, at least 400 Pa, at least 450 Pa, at least 500 Pa, at least 550 Pa, at least 600 Pa, at least 650 Pa, at least 700 Pa, at least 750 Pa, at least 800 Pa, at least 850 Pa, At least 1,000 Pa, at least 1,200 Pa, at least 1,300 Pa, at least 1,400 Pa, at least 1,500 Pa, at least 1,600 Pa, at least 1700 Pa, at least 1800 Pa, at least 1900 Pa, at least 2,000 Pa, at least 2,100 Pa Pa, at least 2,200 Pa, at least 2,300 Pa, at least 2,400 Pa, or at least 2,500 Pa. In yet another aspect of this embodiment, the hydrogel composition may have a viscosity of, for example, at most 25 Pa, at most 50 Pa, at most 75 Pa, at most 100 Pa, at most 125 Pa, at most 150 Pa, Up to 250Pa, up to 300Pa, up to 350Pa, up to 400Pa, up to 450Pa, up to 500Pa, up to 550Pa, up to 600Pa, up to 650Pa, up to 700Pa, up to 750Pa, up to 800Pa, up to 850 Pa, up to 900 Pa, up to 950 Pa, up to 1,000 Pa, up to 1,200 Pa, up to 1,300 Pa, up to 1,400 Pa, up to 1,500 Pa, or up to 1,600 Pa. In another embodiment of this embodiment, the hydrogel composition has a viscosity of from about 25 Pa to about 150 Pa, from about 25 Pa to about 300 Pa, from about 25 Pa to about 500 Pa, from about 25 Pa to about 800 Pa, From about 500 Pa to about 1,600 Pa, from about 600 Pa to about 1,600 Pa, from about 700 Pa to about 1,600 Pa, from about 800 to about 800 Pa, from about 125 Pa to about 300 Pa, from about 125 Pa to about 500 Pa, From about 1 Pa to about 1,600 Pa, from about 900 Pa to about 1,600 Pa, from about 1,000 Pa to about 1,600 Pa, from about 1,100 Pa to about 1,600 Pa, from about 1,200 Pa to about 1,600 Pa, About 1500 Pa to about 1,800 Pa, about 1,500 Pa to about 1,800 Pa, about 1,500 Pa to about 2,500 Pa, about 2,000 Pa to about 2,500 Pa, about 1,300 Pa to about 1,600 Pa, Pa, from about 1,700Pa to about 2,000Pa, from about 1,800Pa to about 2,100Pa, from about 1,900Pa to about 2,200Pa, from about 2,000Pa to about 2,300Pa, from about 2,100Pa to about 2,400Pa, Represents the modulus of elasticity of about 2,200㎩ to about 2,500㎩.

In another embodiment, the compositions disclosed herein do not exhibit a viscosity ratio. In an embodiment of this embodiment, the hydrogel composition has a viscosity of about 10 Pa, about 20 Pa, about 30 Pa, about 40 Pa, about 50 Pa, about 60 Pa, about 70 Pa, about 80 Pa, about 90 Pa, about 100 Pa, about 150 Pa, about 200 Pa, about 250 Pa, about 300 Pa, about 350 Pa, about 400 Pa, about 450 Pa, about 500 Pa, Or a viscosity of about 700 Pa. In another aspect of this embodiment, the hydrogel composition has a maximum of 10 Pa, a maximum of 20Pa, a maximum of 30Pa, a maximum of 40Pa, a maximum of 50Pa, a maximum of 60Pa, a maximum of 70Pa, Up to 300 Pa, up to 400 Pa, up to 450 Pa, up to 500 Pa, up to 600 Pa, up to 650 Pa, up to 650 Pa Or a viscosity of up to 700 Pa. In another embodiment of this embodiment, the hydrogel composition has a viscosity of from about 10 Pa to about 30 Pa, from about 10 Pa to about 50 Pa, from about 10 Pa to about 100 Pa, from about 10 Pa to about 150 Pa, From about 150 Pa to about 450 Pa, from about 250 Pa to about 550 Pa, from about 350 Pa to about 700 Pa, from about 50 Pa to about 150 Pa, from about 100 Pa to about 100 Pa, from about 50 Pa to about 350 Pa, From about 250 Pa to about 350 Pa, from about 300 Pa to about 400 Pa, from about 350 Pa to about 450 Pa, from about 400 Pa to about 200 Pa, from about 200 Pa to about 200 Pa, from about 150 Pa to about 250 Pa, From about 500 Pa to about 450 Pa to about 550 Pa, from about 500 Pa to about 600 Pa, from about 550 Pa to about 650 Pa, or from about 600 Pa to about 700 Pa.

In another embodiment, the hydrogel compositions disclosed herein exhibit tan delta. In an embodiment of this embodiment, the hydrogel composition has a pH of about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1.0, about 1.1, 2, about 2.3, about 2.4, about 2.5, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4 or about 2.5. In another aspect of this embodiment, the hydrogel composition can have, for example, at most 0.1, at most 0.2, at most 0.3, at most 0.4, at most 0.5, at most 0.6, at most 0.7, at most 0.8, at most 0.9, at most 1.0, A maximum of 1.2, a maximum of 1.3, a maximum of 1.4, a maximum of 1.5, a maximum of 1.6, a maximum of 1.7, a maximum of 1.8, a maximum of 1.9, a maximum of 2.0, a maximum of 2.1, a maximum of 2.2, a maximum of 2.3, a maximum of 2.4 or a maximum of 2.5. In another embodiment of this embodiment, the hydrogel composition has, for example, from about 0.1 to about 0.3, from about 0.3 to about 0.5, from about 0.5 to about 0.8, from about 1.1 to about 1.4, from about 1.4 to about 1.7, from about 0.3 From about 0.1 to about 0.6, from about 0.1 to about 0.5, from about 0.5 to about 0.9, from about 0.1 to about 0.6, from about 0.1 to about 1.0, from about 0.5 to about 1.5, from about 1.0 to about 2.0, or from about 1.5 to about 2.5 .

Embodiments of the present disclosure provide hydrogel compositions disclosed herein having partial transparency and / or translucency. Optical transparency is a physical property that allows visible light to pass through a material, while translucency (also called translucency or translucency) only allows light to pass through it. The opposite characteristic is opaque. Transparent materials are clear, while white translucent materials can not be seen clearly. The hydrogels disclosed herein are preferably optically transparent or at least translucent.

In an embodiment, the hydrogel compositions disclosed herein are optically translucent. In an embodiment of this embodiment, the hydrogel composition may contain, for example, about 75% light, about 80% light, about 85% light, about 90% light, about 95% Of the light. In another aspect of this embodiment, the hydrogel composition is dispersed, for example, to at least 75% of light, at least 80% of light, at least 85% of light, at least 90% of light, or at least 95% . In another embodiment of this embodiment, the hydrogel composition comprises, for example, from about 75% to about 100% light, from about 80% to about 100% light, from about 85% to about 100% % To about 100% of light, or about 95% to about 100% of light. In an embodiment, the hydrogel compositions disclosed herein are optically clear and deliver 100% visible light.

The hydrogel compositions disclosed herein may be further processed by pulverizing the hydrogel into particles and optionally mixed with a carrier such as, for example, water or saline to provide a solution for injection or topical material, oil, lotion, Gels, ointments, creams, slurries, plasters or pastes. As such, the disclosed hydrogel compositions can be single-phase or multi-phase compositions. The hydrogel may have a diameter ranging from about 10 microns to about 1000 microns, such as from about 15 microns to about 30 microns, from about 50 microns to about 75 microns, from about 100 microns to about 150 microns, from about 200 microns to about 300 microns, About 550 占 퐉, about 600 占 퐉 to about 700 占 퐉, about 750 占 퐉 to about 850 占 퐉, or about 900 占 퐉 to about 1,000 占 퐉.

Embodiments herein provide that, in part, the compositions disclosed herein are primary use. The term " injected ", as used herein, refers to a material having the properties necessary to administer the composition to a skin area of an individual using a syringe having fine needles. The term "fine needle" as used herein refers to a needle of 27 gauge or less. The scanning ability of the compositions disclosed herein can be performed by scaling the hydrogel particles as discussed above.

In an embodiment of this embodiment, the hydrogel compositions disclosed herein can be injected through a fine needle. In another aspect of this embodiment, the hydrogel compositions disclosed herein are injectable, for example, through a needle of about 27 gauge, about 30 gauge, or about 32 gauge. In another aspect of this embodiment, the hydrogel compositions disclosed herein are injectable, for example, through needles of no more than 22 gauge, no more than 27 gauge, no more than 30 gauge, or no more than 32 gauge. In another aspect of this embodiment, the hydrogel compositions disclosed herein can have from about 22 gauge to about 35 gauge, from 22 gauge to about 34 gauge, from 22 gauge to about 33 gauge, from 22 gauge to about 32 gauge, From about 22 gauge to about 27 gauge, or from about 27 gauge to about 32 gauge needle.

In embodiments of this embodiment, the hydrogel compositions disclosed herein have a viscosity of about 60 N, about 55 N, about 50 N, about 45 N, about 40 N, about 35 N, about 30 N, about 25 N, about 20 N, Or at an output power of about 15 N. In another aspect of this embodiment, the hydrogel compositions disclosed herein have a viscosity of about 60 N or less, about 55 N or less, about 50 N or less, about 45 N or less, about 40 N or less, about 35 N or less, about 30 N or less, Less than about 15 N, about 10 N or less, or about 5 N or less. In another aspect of this embodiment, the hydrogel compositions disclosed herein have a viscosity of about 60 N or less, about 55 N or less, about 50 N or less, about 45 N or less, about 40 N or less, about 35 N or less, about 30 N or less, Can be injected through a 30 gauge needle having an output power of 20 N or less, about 15 N or less, about 10 N or less, or about 5 N or less. In another aspect of this embodiment, the hydrogel compositions disclosed herein have a viscosity of about 60 N or less, about 55 N or less, about 50 N or less, about 45 N or less, about 40 N or less, about 35 N or less, about 30 N or less, Can be injected through a 32 gauge needle having an output power of 20 N or less, about 15 N or less, about 10 N or less, or about 5 N or less.

Embodiments herein provide a hydrogel composition as described herein that exhibits partial adhesion. Adhesion, also referred to as cohesive force, cohesive force or compressive force, is a physical property of the material and is caused by intermolecular attraction between similar molecules in the material that serves to incorporate the molecules. Adhesion is expressed relative to the gram-force (gmf). The cohesive strength is particularly favorable when the molecular weight ratio of the initial glass glycosaminoglycan polymer, the degree of crosslinking of the glycosaminoglycan polymer, the degree of crosslinking of the glycosaminoglycan polymer, the amount of the residual glass glycosaminoglycan polymer after crosslinking, And the pH of the hydrogel composition. The composition should be sufficiently cohesive as to remain localized at the site of administration. Additionally, in certain applications, sufficient cohesion is important to retain its shape and functionality to the composition in the event of a mechanical loading cycle. Thus, in one embodiment, the compositions disclosed herein exhibit cohesive forces equivalent to water. In another embodiment, the hydrogel compositions disclosed herein exhibit sufficient adhesiveness that remains localized at the site of administration. In another embodiment, the hydrogel compositions disclosed herein exhibit sufficient adhesion to retain their shape. In a further embodiment, the hydrogel compositions disclosed herein exhibit sufficient adhesion to retain its shape and functionality.

Embodiments herein provide a hydrogel composition as disclosed herein that exhibits a physiologically acceptable osmolarity in part. The term "osmolality" as referred to herein refers to the concentration of a solute that is osmotically active in solution. The term "physiologically acceptable osmolality" as referred to herein refers to a characteristic of the normal function of a living organism or the osmolality thereof. As such, the administration of a hydrogel composition as disclosed herein exhibits an osmolar concentration that, when administered to a mammal, has a permanent deleterious or substantially non-prolonged effect. Osmol concentration is expressed relative to the osmolality of the osmotically active solute per liter of solvent (Osmol / l or Osm / l). The osmolality is distinct from the molar concentration because it measures the moles of solute particles that are more osmotically active than the moles of solute. Some compounds can be dissociated in solution, while others are not. The osmolality of the solution can be calculated from the following expression: Osmol / ℓ = Σ φ _i η _i C _i where φ is the osmotic coefficient for calculating the ratio anomaly of the solution; eta is the number of particles (e.g., ions) from which the molecule is dissociated; C is the molar concentration of solute; i is an index representing the identity of a particular solute. The osmolarity of the hydrogel compositions disclosed herein can be measured using conventional methods of treating solutions.

In an embodiment, the hydrogel compositions disclosed herein exhibit physiologically acceptable osmolarity. The term "osmolality" as referred to herein refers to the concentration of osmotically active solute per kilo of solvent in the body. The term "physiologically acceptable osmotic concentration" as used herein refers to the osmotic concentration depending on the characteristics of a living organism or normal function. As such, administration of the hydrogel compositions disclosed herein exhibits an osmotic concentration that has a permanent deleterious or substantially non-long term effect when administered to a mammal. The osmolality is expressed relative to the osmolality of osmotically active solutes per kilogram of solvent (osol / kg or Osm / kg) and is equal to the sum of the molar concentrations of all solutes present in the solution. The osmolality of the solution can be measured using an osmometer. The most commonly used instrument in modern laboratories is the freezing point osmolarity meter. This device measures changes in water vapor pressure (water vapor pressure osmolarity meter) resulting from a change in the freezing point (freezing point osmolarity meter) or an increase in osmotic concentration caused by increasing osmotic concentration.

In an embodiment of this embodiment, the hydrogel composition comprises, for example, about 100 mOsm / l, about 150 mOsm / l, about 200 mOsm / l, about 250 mOsm / l, about 300 mOsm / l, about 400 mOsm / l, about 450 mOsm / l, or about 500 mOsm / l. In another aspect of this embodiment, the hydrogel composition comprises at least 100 mOsm / l, at least 150 mOsm / l, at least 200 mOsm / l, at least 250 mOsm / l, at least 300 mOsm / l, at least 350 mOsm / l, at least 400 mOsm / l, at least 450 mOsm / l or at least 500 mOsm / l. In another embodiment of this embodiment, the hydrogel composition may have a composition of, for example, at most 100 mOsm / l, at most 150 mOsm / l, at most 200 mOsm / l, at most 250 mOsm / l, at most 300 mOsm / / l, a maximum of 400 mOsm / l, a maximum of 450 mOsm / l or a maximum concentration of 500 mOsm / l. In another embodiment of this embodiment, the hydrogel composition has a viscosity of from about 100 mOsm / l to about 500 mOsm / l, from about 200 mOsm / l to about 500 mOsm / l, from about 200 mOsm / / L, from about 300 mOsm / L to about 400 mOsm / L, from about 270 mOsm / L to about 390 mOsm / L, from about 225 mOsm / L to about 350 mOsm / L, from about 250 mOsm / L to about 325 mOsm / L , About 275 mOsm / l to about 300 mOsm / l, or about 285 mOsm / l to about 290 mOsm / l.

Embodiments of the present disclosure provide the hydrogel compositions disclosed herein exhibiting, in part, substantial stability. When referring to the hydrogel compositions disclosed herein, the term "stability" or "stable ", as referred to herein, ≪ / RTI > The term "substantial thermal stability "," substantial thermal stability ", "autoclave stability ", or" steam sterilization stability ", as used herein, Quot; hydrogel "

The stability of the hydrogel compositions disclosed herein can be determined, for example, by subjecting the hydrogel composition to thermal treatment, such as steam sterilization, at normal pressure or under reduced pressure (e.g., autoclaving). Preferably, the heat treatment is carried out at a temperature of at least about 100 DEG C for about 1 minute to about 10 minutes. The substantial stability of the hydrogel compositions disclosed herein is determined by 1) determining the change in the pressure output (DELTA F) of the hydrogel compositions disclosed herein after sterilization, wherein the change at the pressure output of less than 2N (the hydrogel having the specified additive The pressure output of the gel composition) - (the pressure output of the hydrogel composition without the added additives); And / or 2) determining the change in rheological properties of the hydrogel compositions disclosed herein after sterilization, wherein the change in tan δ 1 Hz (tan δ 1 Hz of the gel formulation by the additive) - Gt; 1 < / RTI > Hz) of a gel formulation free of water (not shown). As such, the substantially stable hydrogel compositions disclosed herein retain one or more of the following characteristics after sterilization: homogeneous, pressure power, cohesive strength, hyaluronan concentration, agonist (s) concentration, osmolality, pH , Or other rheological properties desired by the hydrogel prior to heat treatment.

In an embodiment, a hydrogel composition comprising a glycosaminoglycan polymer and at least one of the agents disclosed herein is treated using heat treatment to maintain the desired hydrogel properties disclosed herein. In embodiments of this embodiment, the hydrogel compositions comprising the glycosaminoglycan polymer and the at least one agonist disclosed herein can be used at temperatures of, for example, about 100 캜, about 105 캜, about 110 캜, about 115 캜 , About 120 캜, about 125 캜, or about 130 캜. In another aspect of this embodiment, a hydrogel composition comprising a glycosaminoglycan polymer and at least one agonist disclosed herein is at least 100 캜, at least 105 캜, at least 110 캜, at least 115 Deg.] C, at least 120 [deg.] C, at least 125 [deg.] C or at least 130 [deg.] C. In another embodiment of this embodiment, a hydrogel composition comprising a glycosaminoglycan polymer and at least one of the agents disclosed herein is heated to a temperature of from about 100 < 0 > C to about 120 & About 110 DEG C to about 135 DEG C, about 100 DEG C to about 130 DEG C, about 100 DEG C to about 135 DEG C, about 110 DEG C to about 120 DEG C, about 110 DEG C to about 125 DEG C, From about 120 캜 to about 125 캜, from about 120 캜 to about 130 캜, from about 120 캜 to about 135 캜, from about 125 캜 to about 130 캜, or from about 125 캜 to about 135 캜.

The stability of the hydrogel compositions disclosed herein can be determined, for example, by subjecting the hydrogel composition to thermal treatment, such as steam sterilization, at normal pressure or under reduced pressure (e.g., autoclaving). The long-term stability of the hydrogel compositions disclosed herein can be determined, for example, by subjecting the hydrogel compositions to a heat treatment such as storage at about 45 캜 for about 60 days. The long-term stability of the hydrogel compositions disclosed herein can be determined by 1) evaluating the clarity and color of the hydrogel composition after 45 ° C heat treatment (a clear, colorless hydrogel composition represents a substantially stable hydrogel composition); 2) By determining the change in the pressure output (? F) of the hydrogel compositions disclosed herein after 45 ° C heat treatment (the change in the pressure output less than 2N (the pressure output of the hydrogel composition having the specified additive before the 45 ° C heat treatment) - < / RTI > (the pressure output of the hydrogel composition with the specified additives after 45 < 0 > C heat treatment); And / or 3) determining the change in rheological properties of the hydrogel compositions disclosed herein after sterilization, wherein the change at tan δ 1 Hz of less than 0.1 (tan δ of the gel formulation having the specified additive before 45 ° C heat treatment 1 Hz) - (representing a substantially stable hydrogel composition as measured by tan δ 1 Hz of a gel formulation with the specified additives after 45 ° C heat treatment). As such, the long term stability of the hydrogel compositions disclosed herein is evaluated by retaining one or more of the following characteristics after 45 ° C heat treatment: clarity (transparency and translucency), homogeneity and cohesive force.

In embodiments of this embodiment, the hydrogel composition may be administered at a dosage of, for example, about 3 months, about 6 months, about 9 months, about 12 months, about 15 months, about 18 months, about 21 months, about 24 months, , About 30 months, about 33 months, or about 36 months. In another aspect of this embodiment, the hydrogel composition is at least 3 months, at least 6 months, at least 9 months, at least 12 months, at least 15 months, at least 18 months, at least 21 months, at least 24 months, at least 27 months, at least 30 months, at least 33 months, or at least 36 months at room temperature. In another aspect of this embodiment, the hydrogel composition may be formulated to contain, for example, from about 3 months to about 12 months, from about 3 months to about 18 months, from about 3 months to about 24 months, from about 3 months to about 30 months, From about 6 months to about 30 months, from about 6 months to about 36 months, from about 6 months to about 12 months, from about 6 months to about 18 months, from about 6 months to about 24 months, from about 6 months to about 30 months, from about 6 months to about 36 months, From about 9 months to about 30 months, from about 9 months to about 36 months, from about 12 months to about 18 months, from about 12 months to about 12 months, from about 9 months to about 18 months, from about 9 months to about 24 months, 24 months, from about 12 months to about 30 months, from about 12 months to about 36 months, from about 18 months to about 24 months, from about 18 months to about 30 months, or from about 18 months to about 36 months .

The composition may optionally include other pharmaceutically acceptable ingredients, including, but not limited to, buffers, preservatives, isotonic agents, salts, antioxidants, osmotic agents, emulsifiers, wetting agents and the like, It does not.

Pharmaceutically acceptable buffering agents are buffers that can be used to prepare the hydrogel compositions disclosed herein, provided that the resulting formulation is pharmaceutically acceptable. Non-limiting examples of pharmaceutically acceptable buffers include acetate buffers, borate buffers, citrate buffers, neutral buffered saline, phosphate buffers and phosphate buffered saline. Any concentration of the pharmaceutically acceptable buffer may be useful in formulating the pharmaceutical compositions disclosed herein, provided that a therapeutically effective amount of the active ingredient is recovered using such effective concentrations of buffering agent. Non-limiting examples of physiologically acceptable buffer concentrations range from about 0.1 mM to about 900 mM. The pH of the pharmaceutically acceptable buffer may be adjusted, provided that the resulting formulation is pharmaceutically acceptable. It is understood that if desired, the acid or base can be used to adjust the pH of the pharmaceutical composition. Any buffered pH level may be useful in formulating pharmaceutical compositions, provided that a therapeutically effective amount of the substrate polymer active ingredient is recovered using such effective pH levels. Non-limiting examples of physiologically acceptable pHs range from about pH 5.0 to about pH 8.5. For example, the pH of the hydrogel compositions disclosed herein may be from about 5.0 to about 8.0, or from about 6.5 to about 7.5, from about 7.0 to about 7.4, or from about 7.1 to about 7.3.

Pharmaceutically acceptable preservatives include sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole, and butylated hydroxytoluene. Pharmaceutically acceptable preservatives include, but are not limited to, benzalkonium chloride, chlorobutanol, thimerosal, phenyl mercury acetate, phenyl mercury nitrate, stabilized oxychloro compositions such as PURITE (registered trademark) Allergen, Inc.) and chelating agents such as DTPA or DTPA-bisamide, calcium DTPA, and CaNaDTPA-bisamide.

Pharmaceutically acceptable isotonicity adjusting agents useful in the hydrogel compositions disclosed herein include, for example, sodium chloride and potassium chloride; And salts such as glycerine. The composition may be provided as a salt and may be formed by a number of acids including, but not limited to, hydrochloric acid, sulfuric acid, acetic acid, lactic acid, tartaric acid, malic acid, succinic acid and the like. Salts tend to be more soluble in aqueous or other protonic solvents than the corresponding free base forms. These and other materials known in the pharmaceutical arts are understood to be included in the pharmaceutical compositions disclosed herein. Other non-limiting examples of pharmacologically acceptable ingredients are described, for example, in Ansel, supra, (1999); Gennaro, supra, (2000); Hardman, supra, (2001); And Rowe, supra, (2003), each of which is incorporated herein by reference in its entirety.

Aspects of the present disclosure provide a method of treating the soft tissue condition of an individual by administering, in part, the hydrogel compositions disclosed herein. The term "treating ", as used herein, refers to reducing or eliminating the cosmetic or clinical symptoms of a soft tissue condition that is characterized by soft tissue imperfections, defects, diseases and / or disorders; Or delaying or preventing the onset of a cosmetic or clinical condition of a condition characterized by a soft tissue imperfection, defect, disease and / or disorder. For example, the term "treating" may include, for example, at least 20 percent, at least 30 percent, at least 40 percent, at least 50 percent, at least 60 percent, at least 70 percent, at least 80 percent, at least 90 percent, ≪ / RTI > may be meant to reduce the symptoms of conditions characterized by soft tissue defects, diseases and / or disorders. The efficacy of the hydrogel compositions disclosed herein for treating conditions characterized by soft tissue imperfections, defects, diseases and / or disorders can be determined by observing physiological indicators associated with one or more cosmetic, clinical symptoms and / or conditions have. Improvements in soft tissue imperfections, defects, diseases and / or disorders can also be indicated by reduced need for concurrent therapy. Those skilled in the art will know appropriate symptoms or indicators associated with a particular soft tissue defect, disease and / or disorder, and will know how to determine whether an individual is a candidate for treatment with the compounds or compositions disclosed herein.

The hydrogel composition according to the present invention is administered to an individual. An individual is typically a person of any age, gender, or race. Any entity that is typically a candidate for routine procedures for treating soft tissue disorders is a candidate for the methods disclosed herein. Although the subject experiencing signs of aging skin is an adult, subjects who have experienced premature aging or other skin conditions (e.g., scarring) suitable for treatment may be treated with the hydrogel compositions disclosed herein. In addition, the presently disclosed hydrogel compositions and methods can be applied to individuals seeking small / medium enlargement, shape changes or contour changes of body parts or regions, which are not technically possible by conventional soft tissue implant techniques, or It may not be aesthetically acceptable. Pre-operative evaluation typically involves routine history and physiological testing in addition to prior consent to initiate all relevant risks and benefits of the procedure.

The hydrogel compositions and methods disclosed herein are useful for treating soft tissue conditions. The soft tissue condition includes but is not limited to soft tissue imperfections, defects, diseases and / or disorders. Non-limiting examples of soft tissue conditions include but are not limited to breast imperfections, defects, diseases and / or disorders such as breast imperfections, defects, diseases and / or disorders such as breast enlargement, breast reconstruction, Defects due to graft complications such as hypertension, polish syndrome, capsule contraction and / or rupture; Facial imperfections, defects, diseases and / or disorders such as facial enlargement, facial reconstruction, mesotherapy, Parry-Romberg syndrome, cardiac hyperplastic lupus, dermal divot, scarring, sunken check, Facial folds, wrinkles and / or wrinkles such as thin lips, nasal imperfections or bonds, oropharyngeal imperfections or defects, intricate wrinkles, wrinkles and / or wrinkles, dorsal wrinkles, and / or lingual wrinkles and / or other contour deformities or facial Incomplete; Neck imperfections, defects, diseases and / or disorders; Skin imperfections, defects, diseases and / or disorders; Including lower legs, soles, including other soft tissue imperfections, defects, diseases and / or disorders such as upper arms, lower arms, hands, shoulders, back, torso, Enlargement or reconstruction of foot, eye, genital or other bodily parts, areas or areas, or diseases or disorders affecting these bodily parts, areas or areas; Incontinence, incontinence, other types of incontinence; And gastroesophageal reflux disease (GERD). The term " mesotherapy ", as used herein, is intended to encompass skin, non-surgical cosmetic, and / or subcutaneous administration of subcutaneous, intradermal, subcutaneous and / or subcutaneous injection of an agent to be administered as a multiplicity of small drops into the dermis Refers to an anti-healing technique.

The amount of hydrogel composition used by any of the methods disclosed herein will typically be determined by one skilled in the art to be within the scope of the present invention, as is typically desired and / or improved, the reduction and / or elimination of desired soft tissue disease symptoms, / RTI > and / or the cosmetic effect and the body part or area being treated. The effectiveness of the composition administration can be demonstrated by one or more of the following clinical and / or cosmetic measurements: altered and / or improved soft tissue shape, altered and / or improved soft tissue size, altered and / , Altered and / or improved tissue function, tissue internal growth and / or new collagen deposition, sustained engraftment of the composition, improved patient satisfaction and / or quality of life, and reduced use of transplantable foreign material.

The effectiveness of the compositions and methods for treating facial soft tissues can be demonstrated by one or more of the following clinical and / or cosmetic measurements: facial features such as increased size, shape and / or contour of the lips, An increased size, shape and / or contour of the article; Shape, and / or contour of facial features such as altered size, shape, and / or contour of a lip, ball, or eye region shape; Reduction or elimination of wrinkles, creases or wrinkles in the skin; Resistance to wrinkles, folds or wrinkles in the skin; Rehydration of skin; Increased elasticity to the skin; Reduction or elimination of skin roughness; Increased and / or improved skin tension; Reducing or eliminating stretch marks or marks; Increased and / or improved skin tone, sparkle, brightness and / or brightness; Increased and / or improved skin color, reduction or elimination of pale skin; The duration of the composition; Reduced side effects; Improved patient satisfaction and / or quality of life.

As another example, for incontinence procedures, the effectiveness of the compositions and methods for sphincter support may be demonstrated by one or more of the following clinical measures: reduced incontinence cycle, persistent engraftment, improved patient satisfaction, and / or Quality of life and reduced use of transplantable outpatient fillers.

In an embodiment of this embodiment, the amount of hydrogel composition administered is, for example, about 0.01 g, about 0.05 g, about 0.1 g, about 0.5 g, about 1 g, about 5 g, about 10 g, about 20 g, About 40 g, about 50 g, about 60 g, about 70 g, about 80 g, about 90 g, about 100 g, about 150 g, or about 200 g. In another aspect of this embodiment, the amount of hydrogel composition administered is, for example, from about 0.01 g to about 0.1 g, from about 0.1 g to about 1 g, from about 1 g to about 10 g, from about 10 g to about 100 g, It is about 200g. In another aspect of this embodiment, the amount of hydrogel composition administered is, for example, about 0.01 ml, about 0.05 ml, about 0.1 ml, about 0.5 ml, about 1 ml, about 5 ml, about 10 ml, about 20 About 30 ml, about 40 ml, about 50 ml, about 60 ml, about 70 g, about 80 ml, about 90 ml, about 100 ml, about 150 ml, or about 200 ml. In another aspect of this embodiment, the amount of hydrogel composition administered is from about 0.01 ml to about 0.1 ml, from about 0.1 ml to about 1 ml, from about 1 ml to about 10 ml, from about 10 ml to about 100 ml , Or from about 50 ml to about 200 ml.

The duration of the treatment will typically be determined based on the cosmetic and / or clinical effect desired by the individual and / or physician and the body part or area being treated. In embodiments of this embodiment, the administration of the hydrogel compositions disclosed herein may take place, for example, at about 6 months, at about 7 months, at about 8 months, at about 9 months, at about 10 months, at about 11 months, at about 12 months, About 13 months, about 14 months, about 15 months, about 18 months, or about 24 months. In another aspect of this embodiment, the administration of the hydrogel compositions disclosed herein is for example at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, At least 13 months, at least 14 months, at least 15 months, at least 18 months, or at least 24 months. In another aspect of this embodiment, administration of the hydrogel compositions disclosed herein may be, for example, from about 6 months to about 12 months, from about 6 months to about 15 months, from about 6 months to about 18 months, From about 9 months to about 21 months, from about 6 months to about 24 months, from about 9 months to about 12 months, from about 9 months to about 15 months, from about 9 months to about 18 months, from about 9 months to about 21 months, From about 12 months to about 18 months, from about 12 months to about 18 months, from about 12 months to about 21 months, from about 12 months to about 21 months, from about 12 months to about 18 months, from about 12 months to about 21 months, , From about 15 months to about 24 months, from about 18 months to about 21 months, from about 18 months to about 24 months, or from about 21 months to about 24 months.

An aspect of the present disclosure provides a step of administering, in part, the hydrogel compositions disclosed herein. The term "administering " as used herein refers to any delivery mechanism that provides the compositions disclosed herein for individuals that potentially result clinically, therapeutically or experimentally. The actual delivery mechanism used to administer the composition to an individual depends on the type of skin condition, the location of the skin condition, the cause of the skin condition, the severity of the skin condition, the degree of relief desired, the particular composition employed, , The pharmacokinetics of the particular composition employed, the nature of the other compounds involved in the particular composition employed, the particular route of administration, the particular characteristics, the history of the individual and the risk factors, such as age, weight, general health status, But not limited to, any combination thereof, including, but not limited to, combinations thereof. In an embodiment of this embodiment, the compositions disclosed herein are administered to the skin area of an individual by injection.

The route of administration of the hydrogel composition to an individual patient will typically be determined based on the cosmetic and / or clinical effect desired by the individual and / or physician and the body part or area being treated. The compositions disclosed herein may be administered by any means known to those of ordinary skill in the art, including, but not limited to, needle implanted by local or direct surgical implantation, needle (e.g., hand-compressed pistol) ≪ / RTI > The hydrogel compositions disclosed herein may be administered into a skin area, such as, for example, a dermal area or subcutaneous area. For example, the hydrogel compositions disclosed herein can be injected using a needle having a diameter of about 0.26 mm to about 0.4 mm and a length in the range of about 4 mm to about 14 mm. Alternatively, the needle may be from 21G to 32G and may have a length of from about 4 mm to about 70 mm. Preferably, the needle is a disposable needle. The needle may be combined with a syringe, a catheter, and / or a pistol.

In addition, the compositions disclosed herein may be administered in single or multiple doses. Ultimately, the time used will follow quality control standards. For example, the hydrogel compositions disclosed herein may be administered at least one or more working hours with a working time of a few days or weeks. For example, an individual may be administered the hydrogel compositions disclosed herein every 1, 2, 3, 4, 5, 6 or 7 days or every 1, 2, 3 or 4 weeks. Administration of the hydrogel compositions disclosed herein to an individual may be administered on a monthly or two-month basis, or every 3, 6, 9, or 12 months.

An aspect of the present disclosure provides, in part, a dermal region. The term "dermal region" as referred to herein refers to a region of the skin that includes the epidermis-dermal junction and the deep dermis (papillary region) and deep dermis (reticular region). The skin consists of three main layers: the epidermis, which provides waterproofing and acts as a barrier to infection; A dermis acting as a place for the outer skin of the skin; And hippie (subcutaneous fat layer). The epidermis contains no blood vessels and is nourished by diffusion from the dermis. The main types of cells constituting the epidermis are keratinocytes, melanocytes, Langerhans cells and Merkel cells.

The dermis consists of connective tissue and is a layer of skin under the epidermis that alleviates the impact of the body from stress and deformation. The dermis is firmly bound to the epidermis by the lowermost membrane. It also conceals multiple mechanical receptors / nerve endings that provide a sense of contact and heat. It contains pores, sweat glands, sebaceous glands, apocrine glands, lymph vessels and blood vessels. The blood vessels in the dermis provide nutrients and waste removal from the basal layer of the epidermis as well as from its own cells. The dermis is structurally divided into two regions: a supercritical region adjacent to the epidermis, called the nipple region, and a deeper and thicker region known as the reticular region.

The nipple area consists of loose areolar tissue connective tissue. It is called a finger-shaped protrusion called teat which extends to the epidermis. The nipple has a "rugged" surface that interlocks with the epidermis and provides a dermis that enhances the bond between the two layers of skin. The reticular area lies deeper than the nipple area, and is usually much thicker. It is made up of dense irregular connective tissue and gets its name from the dense concentration of collagenous, elastic and reticular fibers that are woven throughout it. These protein fibers provide their dermis with properties of strength, elongation and elasticity. In addition, hair follicles, sebaceous glands, sweat glands, receptors, nails, and blood vessels are located within the reticular region. The tattoo ink is retained in the dermis. Also, stretch marks from the pregnancy are located in the dermis.

Harpy is placed under the dermis. Its purpose is not only to attach the dermis areas of the base bone and muscles but also to supply blood vessels and nerves to it. It consists of loose connective tissue and elastin. The main cell types are fibroblasts, macrophages and adipocytes (hippies contain 50% of body fat). The fat acts as a pad and an insulator to the body.

In an embodiment of this embodiment, the hydrogel compositions disclosed herein are administered to the skin region of an individual by injection into the dermal region or subcutaneous tissue region. In an embodiment of this embodiment, the hydrogel compositions disclosed herein are administered to the dermal areas of the body by injection, for example, into the epidermal-dermal junctional region, the papillary region, the reticular region, or any combination thereof.

Advantageously, a portion of the composition is particularly useful and effective, for example, in reducing fine lines in a thin skin area of a patient. For example, there is provided a method for the treatment of fine wrinkles comprising the step of administering to a patient a dermal filler composition as described elsewhere herein, at a depth of about 1 mm or less.

Another aspect of the disclosure includes administering a hydrogel composition as described herein to an individual suffering from a partial skin disease, wherein the administration of the composition improves the skin disorder and thereby treats the skin disorder. In an embodiment of this embodiment, the skin disorder comprises administering the hydrogel composition disclosed herein to a subject suffering from skin dehydration, wherein administration of the composition rehydrates the skin, thereby treating skin dehydration. In another aspect of this embodiment, a method of treating a lack of skin elasticity comprises administering to a subject suffering from a skin elastic lack of hydrogel composition disclosed herein, wherein administration of the composition increases skin elasticity, Thereby treating the lack of skin elasticity. In another aspect of this embodiment, a method of treating skin roughness comprises administering the hydrogel composition disclosed herein to an individual suffering from skin roughness, wherein administration of the composition reduces skin roughness, Treat skin roughness. In another aspect of this embodiment, a method of treating a lack of skin roughness comprises administering a hydrogel composition as disclosed herein to a subject suffering from a lack of skin tension, wherein administration of the composition creates skin tension , Thereby treating the lack of skin tension.

In a further aspect of this embodiment, a method of treating skin stretch marks or marks comprises the step of administering the hydrogel compositions disclosed herein to a subject suffering from skin liner or marks, wherein the administration of the composition comprises applying a skin- Thereby eliminating or eliminating skin flaking or marks. In another aspect of this embodiment, a method of treating pale skin comprises administering the hydrogel composition disclosed herein to a subject suffering from pale skin, wherein administration of the composition increases skin tone or light, Thereby treating the pale skin. In another aspect of this embodiment, a method of treating skin wrinkles comprises the step of administering the hydrogel compositions disclosed herein to a subject suffering from skin wrinkles, wherein administration of the composition reduces or eliminates skin wrinkles , Thereby treating the skin wrinkles. In another aspect of this embodiment, a method of treating skin wrinkles comprises the step of administering to a subject a hydrogel composition as disclosed herein, wherein administration of the composition creates skin resistance to the skin wrinkles, Lt; / RTI >

In some embodiments, the dermis filler has sustained bioavailability. For example, when introduced into the human skin (e.g., into the dermis or subcutaneously to the human to repair soft tissue defects in the facial space), ascorbic acid (or other Vitamins) are provided.

For example, an assessment of the degree of conjugation is made to predict sustained vitamin C efficacy with a filler lasting. This evaluation was based on the formulation of AA2G conjugation to HA via ethers. The formulation is stable under physiological conditions, but begins to release ascorbic acid (AsA) by the a-glucosidase attached to the cell membrane. Due to the fact that alpha -glucosidase attaches to the cell membrane, AsA release occurs. Additional release of AsA from HA-AA2G will involve HA degradation to make AA2G available to fibroblasts. Thus, the release of AsA is dependent on the degree of AA2G conjugation and the duration of HA. A gel having a degree of conjugation of approximately 5 mol% was able to release active vitamin C for a period of at least 1 month, for example 3 to 5 months; A gel with 10 mol% conjugation could release active vitamin C for up to 6 to 8 months; A gel with 15 mol% conjugation can release active vitamin C for periods of up to 10 months; 30 mol% can be released by one and a half years.

In an embodiment of the present invention, a vitamin C derivative comprising a star-PEG epoxide and a cross-linked hyaluronic acid and conjugated to hyaluronic acid at a degree of conjugation of about 3 mol% to about 40 mol% (for example, AA2G (3-aminopropyl-L-ascorbyl phosphate), and SAP (sodium ascorbyl phosphate).

This dermal filler is prepared by the reaction temperature and the reaction time ratio suitable for achieving the composition containing AA2G by the 4-arm epoxide (AA2G-4 cancer epoxide), unreacted 4-arm epoxide and free AA2G (Star-PEG epoxide) with ascorbic acid 2-glucoside (AA2G). 4 Arm Epoxide The capped AA2G (AA2G-4 arm epoxide) is conjugated to the hyaluronic acid via an epoxidic group. The unreacted 4-arm epoxide acts as a cross-linking agent for crosslinking the hyaluronic acid and additionally as a conjugation agent for conjugating AA2G.

In another embodiment of the present invention, a vitamin C derivative comprising BDDE and a crosslinked hyaluronic acid and conjugated to hyaluronic acid at a degree of conjugation of about 3 mol% to about 10 mol% (for example, AA2G (ascorbic acid 2-glucoside ), Bitagent (3-aminopropyl-L-ascorbyl phosphate) and SAP (sodium ascorbyl phosphate).

The method of preparing the dermis filler is such that the ratio of BDDE (AA2G-BDDE), unreacted BDDE, and AA2G capped by AA2G to the reaction temperature and reaction time suitable for achieving the composition, (AA2G). BDDE capping AA2G (AA2G-BDDE) is conjugated to the hyaluronic acid via an epoxyl group. Unreacted BDDE serves as a cross-linking agent for cross-linking the hyaluronic acid and as a conjugation agent to further conjugate AA2G.

FIG. 9 is a table showing the effect of the concentration of? -Glucosidase on the AsA release from the AA2G-PBS solution. The conversion of AA2G to AsA depends on the concentration of the alpha -glycosidase. When the [alpha] -glycosidase concentration is 6.3 units per gram of gel, AA2G is almost completely converted to AsA after 15 minutes. When the [alpha] -glycosidase concentration is 4.7 units per gram of gel, AA2G takes 30 minutes to complete conversion to AsA. Further reduced α-glycosidase concentrations led to a slow conversion of AA2G to AsA.

Figure 10 shows a representation of the release profile (sustained release) of free AsA from the conjugated dermal filler according to the invention (mol% versus AA2G conversion at reaction time); AA2G is fully converted to AsA from AA2G / HA mix at 40 minutes. The AA2H / HA conjugate showed the time dependence of AA2G conversion to AsA.

Figures 11A and 11B show additional emission data for various dermal fillers in accordance with the present invention. More specifically, the conversion of AA2G to AsA in the HA-AA2G gel is dependent on the [alpha] -glycosidase concentration. High α-glycosidase concentrations led to rapid conversion of AA2G to AsA. For a given a-glycosidase concentration, the different formulations exhibited different profiles of AA2G to AsA.

In one aspect of the invention, the appearance of the fine wrinkles, for example, wrinkles that are not relatively deep in the skin, such as, but not limited to, eye wrinkles, tear grooves, forehead, A dermis filler is provided which is particularly effective for removing, and removing.

The dermis filler is an important side effect experienced by some dermatologist patients, which is the appearance of a blue discoloration on the injected skin area (the Tinlle effect). The Tindle effect is more common in patients treated for undersized wrinkles. An embodiment of the present invention has been developed that provides a long lasting translucent filler that can be injected deeply to treat fine lines and wrinkles even in the area of relatively thin skin without causing any blue discoloration from the Thinle effect. Fine wrinkles or deep wrinkles are often seen in the skin (forehead, lateral lobe, red sea border / upper lip wrinkles) typically found in the face area where the skin is thinnest, that is, the skin has a dermis thickness of less than 1 mm. It is understood to be a line. On the forehead, the average dermal thickness is about 0.95 mm for normal skin and about 0.81 mm for wrinkled skin. The dermis around the lateral lobe is even thinner (e.g., about 0.61 mm for normal skin, about 0.41 mm for wrinkled skin). The average outer diameter of 30 or 32 gauge needles (needles typically used for fine wrinkle gel application) is from about 0.30 to about 0.24 mm.

The present invention provides dermal filler compositions as described elsewhere herein that do not result in tin effects. For example, the composition of the present invention includes a crosslinking component, a crosslinked hyaluronic acid component, and an additive other than the crosslinking component; Wherein the composition exhibits a reduced tinned effect when administered into the dermal area of a patient for substantially the same composition but without the additive. The composition may be substantially optically clear.

In one embodiment, the additive is a vitamin C derivative, for example AA2G, which can be chemically conjugated to a hyaluronic acid as described elsewhere herein.

In some embodiments, the cross-linking component is BDDE and the degree of conjugation is up to about 3 mol% to about 10 mol%, or up to 15 mol%. In some embodiments, the composition further comprises an anesthetic, e.g., lidocaine, in an amount suitable to provide comfort to the patient during injection.

A method of treating fine wrinkles in a patient ' s skin is also provided. The methods generally involve introducing a composition as described herein to the skin of a patient. For example, the composition comprises a mixture of a hyaluronic acid component, a cross-linking component bridging the hyaluronic acid, and an additive other than a cross-linking component, wherein the composition is substantially optically clear; The dermal filler composition exhibits a reduced tinned effect for substantially the same composition, except that the additive is absent.

In some embodiments of the present invention, the composition comprises a hyaluronic acid component crosslinked with a die or a farad amine crosslinker using EDC chemistry. For example, the crosslinking agent may be HMDA.

In certain embodiments of the present invention, the cross-linking agent is HMDA, wherein the composition has a G 'of up to about 70Pa, a G "/ G' of about 0.65 to about 0.75, an approximate output of about 24N or less, Gt; mg / g. ≪ / RTI >

In certain embodiments of the present invention, the additive is an HA-AA2G conjugate or an HA-bitter conjugate wherein the degree of conjugation is from about 3 mol% to about 10 mol%, or up to about 15 mol%, or up to about 40 mol% . These compositions have a G '/ G' of at least about 30 Pa, more preferably at least about 40 Pa, at least about 100 Pa, a G '/ G' of about 0.30 to about 0.50, And may have a final HA concentration of about 25 mg / g.

For purposes of this disclosure, "degree of conjugation" as used herein is defined as the molar percentage of a conjugating agent, for example, AA2G, for a repeating unit of hyaluronic acid (eg, HA dimer). Thus, a degree of 10 mol% conjugation means that all 100 HA repeat units contain 10 conjugated AA2G. The degree of conjugation can be calculated using the method illustrated in Example 2 below or other methods known to those skilled in the art.

Example

Example  One: As a crosslinking agent BDD Using HA On gel Bridged AA2G Conjugation

400.6 mg of low molecular weight hyaluronic acid (LMW HA) was hydrated in 1802 mg of 1 wt% NaOH in a syringe for ~ 30 min. 800.7 mg of AA2G was placed in the vial followed by 713.7 mg of BDDE and 1416.8 mg of 10% NaOH. The solution (pH > 12) was reacted in a water bath at 50 [deg.] C for ~20 minutes, and hydrated HA was added. After addition, the mixture was mixed ~ 20 times by passing back and forth between two syringes. The mixed paste was placed in a vial and a 50 占 폚 water bath for? 2.5 hours. 223.5 mg of 12 M HCl was added to 9.05 g PBS, pH 7.4. After ~2.5 hours, HA-AA2G gel was formed. The gel was cut into pieces and a solution of HCl-PBS was added thereto. The gel was neutralized and swollen overnight on an orbital shaker. The gel was resized through a ~ 60 um screen and mixed ~ 20 times by passing back and forth between two syringes. The gel was placed in a 15,000 MWCO RC dialysis bag and dialyzed in PBS, pH 7.4 buffer. Dialysis was performed for ~ 185 hours with frequent changes in PBS buffer. After dialysis, the gel was placed in a syringe and stored in a 4 ° C refrigerator.

Example  2: AA2G Conjugation  decision

The weight of the gel as described in Example 1 was recorded immediately before dialysis and after dialysis. After dialysis, the gel was estimated to be ~ 1 g / ml. Dialysis was stopped at a point where no significant AA2G was produced in 1 L of PBS per 8 hours. AA2G was measured at 260 nm using a UV / Vis spectrophotometer (Nanodrop 2000C, ThermoScientific). The calibration curves of AA2G were calculated using different concentrations of AA2G in 2% HA (A @ 260 nm = 1.4838 [AA2G (mM)])

Weight of HA after dialysis: Starting weight of HA (actual weight before dialysis / theoretical weight)

After dialysis, m mol of AA2G: absorbance at 260 nm after dialysis in the formula (A @ 260 nm = 1.4838 [AA2G (mM)]).

Conjugation at AA2G: (mmol of AA2G / mmol of HA) x 100%

The degree of AA2G conjugation in the gel as described in Example 1 is 14.7 mol%.

Example  3: Gel Rheology  Determination of characteristics

The properties of the gel obtained in Example 1 were measured using a vibrating parallel plate flow system (Anton Paar, Physica MCR 301). The diameter of the used plate was 25 mm. The gap between the plates was set at 1 mm. For each measurement, a frequency sweep was first performed at a constant strain, and then a strain sweep was performed at a fixed frequency. G '(storage modulus) was obtained from the strain sweep curve at 1% strain. The value was 1450Pa for the gel.

Example  4: Variable Conjugation  Degree and gel Rheology  Characteristic As a crosslinking agent  Using BDDE Bridged HA On gel  About AA2G Conjugation

The procedure was similar to that described in Example 1. The degree of conjugation is modified by controlling the cross-linking agent to HA and AA2G mol ratio. The gel properties were measured as described in Example 3. The details are as follows:

400.8 mg of LMW HA was hydrated in 1752.1 mg of 1% NaOH in a syringe for ~ 30 minutes. 800.3 mg of AA2G was placed in the vial followed by 354.1 mg of BDDE and 1402.0 mg of 10% NaOH. The solution (pH > 12) was reacted in a water bath at 50 [deg.] C for ~20 minutes, and hydrated HA was added. After addition, the mixture was mixed ~ 20 times by passing back and forth between two syringes. The mixed paste was placed in a vial and a 50 占 폚 water bath for? 2.5 hours. 140.9 mg of 12 M HCl was added to 9.0053 g PBS, pH 7.4. After ~2.5 hours, HA-AA2G gel was formed. The gel was cut into pieces and a solution of HCl-PBS was added thereto. The gel was neutralized and swollen overnight on an orbital shaker. The gel was resized through a ~ 60 um screen and mixed ~ 20 times by passing back and forth between two syringes. The gel was placed in a 15,000 MWCO RC dialysis bag and dialyzed in PBS, pH 7.4 buffer. Dialysis was performed for ~ 164.5 hours with frequent changes in PBS buffer. After dialysis, the gel was placed in a syringe and stored in a 4 ° C refrigerator. The degree of conjugation is 13%. The gel storage modulus (G ') is 803Pa.

Example  5: As a crosslinking agent BDD Using Bridged HA On gel AA2G Conjugation , Con The degree of jogging was 5.3% G ' Is ~ 300Pa.

400.3 mg of LMW HA was hydrated in 3002.0 mg of 1% NaOH in a syringe for ~ 30 minutes. 800.5 mg of AA2G was placed in the vial followed by 264.3 mg of BDDE and 1100.0 mg of 10% NaOH. The solution (pH > 12) was reacted in a water bath at 50 [deg.] C for ~20 minutes, and hydrated HA was added. After addition, the mixture was mixed ~ 20 times by passing back and forth between two syringes. The mixed paste was placed in a vial and a 50 占 폚 water bath for? 2.5 hours. 104.2 mg of 12 M HCl was added to 8.5128 g PBS, pH 7.4. After ~2.5 hours, a HA-AA2G gel was formed and a solution of HCl-PBS was added thereto. The gel was neutralized and swollen on an orbital shaker over the weekend (~ 55 hours). The gel was resized through a ~ 60 um screen and mixed ~ 20 times by passing back and forth between two syringes. The gel was placed in a 15,000 MWCO RC dialysis bag and dialyzed in PBS, pH 7.4 buffer. Dialysis was performed for ~ 114 hours with frequent changes in PBS buffer. After dialysis, the gel was placed in a syringe and stored in a 4 ° C refrigerator. The degree of conjugation and the gel rheological properties are measured in the procedure as described in Examples 2 and 3. The degree of conjugation is 5.3%. The gel storage rate (G ') is ~ 300Pa.

Example  6: As a crosslinking agent  star- PEG Epoxides  using Bridged HA On gel  AA2G for Conjugation , Conjugation  Of the total, 29.4% G ' Is ~ 235Pa.

200.4 mg of LMW HA was hydrated in 2000 mg of 1% NaOH in a syringe for ~ 30 min. 400 mg of AA2G was placed in the vial followed by 312.7 mg of star-PEG epoxide and 1026.5 mg of 10% NaOH. The solution was reacted in a water bath at < RTI ID = 0.0 > 50 C < / RTI > for ~20 minutes and then added to the hydrated HA. After addition, the mixture was mixed ~ 20 times by passing back and forth between two syringes. The mixed paste was placed in a vial and a 50 占 폚 water bath for? 2.5 hours. 187.4 mg of 12 M HCl was added to 3.034 g PBS, pH 7.4. After ~2.5 hours, a HA-AA2G gel was formed and a solution of HCl-PBS was added thereto. The gel was neutralized and swollen on an orbital shaker over the weekend (~68 hours). The gel was resized through a ~ 60 um screen and mixed ~ 20 times by passing back and forth between two syringes. The gel was placed in a 15,000 MWCO RC dialysis bag and dialyzed in PBS, pH 7.4 buffer. Dialysis was performed for ~95 hours with frequent changes in PBS buffer. After dialysis, the gel was placed in a syringe and stored in a 4 ° C refrigerator. The degree of conjugation and the gel rheological properties are measured in the procedure as described in Examples 2 and 3. The degree of conjugation is 29.4%. The gel storage modulus (G ') is ~ 235 Pa.

Example  7: As a crosslinking agent  star- PEG Epoxides  using Bridged HA On gel  AA2G for Conjugation , Conjugation  Of the total population, 27.8% G ' Is ~ 363Pa.

200.3 mg of LMW HA was hydrated in 2000 mg of 1% NaOH in a syringe for ~ 30 minutes. 400.2 mg of AA2G was placed in the vial followed by 313.4 mg of star-PEG epoxide and 1022.6 mg of 10% NaOH. The solution was added to the hydrated HA. After addition, the mixture was mixed ~ 20 times by passing back and forth between two syringes. The mixed paste was placed in a vial and a 50 占 폚 water bath for? 2.5 hours. 196.5 mg of 12 M HCl was added to 3.016 g PBS, pH 7.4. After ~2.5 hours, a HA-AA2G gel was formed and a solution of HCl-PBS was added thereto. The gel was neutralized and swollen on an orbital shaker overnight (~ 24 hours). The gel was resized through a ~ 60 um screen and mixed ~ 20 times by passing back and forth between two syringes. The gel was placed in a 15,000 MWCO RC dialysis bag and dialyzed in PBS, pH 7.4 buffer. Dialysis was performed for ~ 98.5 hours with frequent changes in PBS buffer. After dialysis, the gel was placed in a syringe and stored in a 4 ° C refrigerator. The degree of conjugation and the gel rheological properties are measured in the procedure as described in Examples 2 and 3. The degree of conjugation is 27.8%. The gel storage modulus (G ') is ~ 363Pa.

Example  8: As a crosslinking agent BDD Using Bridged HMW HA On gel AA2G  Conjugation, Conjugation  Degree is 10 mol% G ' Lt; / RTI >

400.3 mg of LMW HA was hydrated in 2501.3 mg of 4% NaOH in a syringe for ~ 30 minutes. 1200 mg of AA2G was placed in the vial followed by 304.7 mg of BDDE and 1178.6 mg of 16% NaOH. The solution (pH > 12) was reacted in a water bath at 50 [deg.] C for ~20 minutes, transferred to a 20 cc syringe, and hydrated HA was added. After addition, the mixture was mixed ~ 20 times by passing back and forth between two syringes. The mixed paste was placed in a 20 cc vial and a 50 ° C water bath for ~2.5 hours. After ~2.5 hours, HA-AA2G gel was formed. Then, 226.6 mg of 12M HCl was added to 8492.2 mg of 10X PBS, pH 7.4 to obtain a HCl-PBS solution, neutralized by adding HCl-PBS solution, and the gel was swollen. The gel was neutralized and swollen on an orbital shaker for 48 hours. The gel was resized through a ~ 60 um screen and mixed ~ 20 times by passing back and forth between two syringes. The gel was placed in a 20,000 MWCO CE dialysis bag and dialyzed in PBS, pH 7.4 buffer. Dialysis was performed for ~ 114 hours with frequent changes in PBS buffer. After dialysis, the gel was placed in a syringe and stored in a 4 ° C refrigerator. The degree of conjugation and the gel rheological properties are measured in the procedure as described in Examples 2 and 3. Conjugation degree is 10 mol%. The gel storage modulus (G ') is ~240Pa.

Example  9: As a crosslinking agent BDD Using Bridged HMW HA On gel Vitagen Conjugate Sean, Conjugation  Is about 15 mol% G ' Is ~ 365Pa.

398.2 mg of LMW HA was hydrated in 1753.24 mg of 1 wt% NaOH in a syringe for ~ 40 min. HA was inflated by the addition of BDDE (311.7 mg) and HA was inflated for another 80 minutes. The expanded HA / BDDE mixture was pre-reacted at 50 ° C for 20 minutes.

801.9 mg of bitagene was isolated and dissolved in 1459.7 mg of 10 wt% NaOH and mixed with HA pre-reacted with BDDE. The reaction was continued at 50 < 0 > C for another 2.5 hours. After ~2.5 hours, HA-Vitagen gel was formed. Then, 195 mg of 12M HCl was added to 9004.0 mg of 10X PBS, pH 7.4 to obtain a HCl-PBS solution, neutralized by adding HCl-PBS solution, and the gel was swollen. The gel was neutralized and swollen on an orbital shaker for 48 hours. The gel was resized through a ~ 60 um screen and mixed ~ 20 times by passing back and forth between two syringes. The gel was placed in a 20,000 MWCO CE dialysis bag and dialyzed in PBS, pH 7.4 buffer. Dialysis was performed for ~ 120 hours with frequent changes in PBS buffer. After dialysis, the gel was placed in a syringe and stored in a 4 ° C refrigerator. The gel rheological properties are measured in the procedure as described in Example 3. [ The degree of conjugation was determined to be about 15 mol% using a method similar to the AA2G crystal as described in Example 2. The gel storage modulus (G ') is ~ 365Pa.

Example  10: Amidation  Linear through chemistry HA on Vitagen Conjugation

200.3 mg of HMW HA was hydrated in 10 ml water in a 60 cc syringe. 500 mg of Vitagen was dissolved in 0.5 ml of water and the solution was neutralized to pH 4.8. 197.7 mg of EDC and 149 mg of NHS were individually dissolved in 6 ml of water. The solution (rinse and EDC / NHS solution) is added to another 60 cc syringe containing 23.5 ml of water. Two syringes are mixed ~ 20 times by passing back and forth between two syringes. The mixture was stored in one syringe and immersed in a 37 [deg.] C bath for 4 hours. The solution was finally dialyzed against PBS pH 7.4 buffer until no significant bitagens were observed. The degree of conjugation was determined by a method similar to that described in Example 3. The degree of conjugation is about 10 mol%.

Example  11: HA On gel Bridged AA2PG Conjugation

200.4 mg of LMW HA was hydrated in 1000 mg of MES 5.2 buffer in the syringe for ~ 30 minutes. 292 mg of AA2P was placed in the vial and 300 mg of star-PEG amine was added. The solution was allowed to react overnight at room temperature. The gel was hydrated with PBS buffer and dialyzed against PBS buffer to remove unreacted AA2P. Finally, the gel was characterized as described in Examples 2 and 3 to determine the degree of conjugation and the gel rheological properties. The degree of conjugation is about 20 mol%. The gel storage modulus (G ') is about 500Pa.

Example  12

To reduce the appearance of fine lines AA2G With HA / BDD  dermis Filler Formulation

 After dialysis, for any of the gels described in the above examples, a suitable amount of free HA gel was added to the gel to improve or modify gel cohesion and / or scanning ability. For example, the free HA fibers were expanded in phosphate buffer solution to obtain a uniform viscoelastic gel ("glass" HA gel). Dialyzed step The crosslinked gel is added to the HA / BDDE crosslinked gel obtained in Example 1 (e.g., to obtain a composition having from about 1% to about 5% w / w glass HA). The resulting gel is then filled into a ready-to-fill sterile syringe and autoclaved at a temperature and pressure sufficient for sterilization for at least about one minute. After autoclaving, the final HA / AA2G product is packaged and distributed to the physician for use as a dermal filler for deep injection to improve the appearance of fine wrinkles in the orbit or other facial region.

Example  13

Lidocaine  Included HA - AA2G  dermis Filler Formulation

Following the procedure of Example 12, lidocaine chlorhydrate (lidocaine HCl) is added to the mixture after the dialysis step and before addition of the free HA gel. In powder form (lidocaine HCl) can be first solubilized in WFI and filtered through a 0.2 μm filter. A dilute NaOH solution is added to the cohesive HA / AA2G gel to reach a slightly basic pH (for example, a pH of about 7.5 to about 8). The lidocaine HCl solution is then added to the slightly basic gel to reach a final desired concentration, for example, a concentration of about 0.3% (w / w). The resulting pH of the HA / AA2G / lidocaine mixture is then about 7 and the HA concentration is about 24 mg / g. Mechanical mixing is carried out to obtain adequate uniformity in standard reactors with appropriate compounding machines.

Example  14

HA On hydrogel  about Carboxy  Of functional group-containing additives Conjugation

Retinoic acid (AKA, tretinoin), appearance and additives such as alpha-lipoic acid contain carboxyl functional groups (-COOH). These additives are conjugated to HA hydrolysis via esterification using EDC chemistry. An example for conjugation according to the present invention is described as follows:

200 mg of HMW HA was hydrated in 10 ml of pH 4.8 MES buffer in a 60 cc syringe. In another syringe, 200 mg of retinoic acid is dissolved in 5 ml of a water-acetone mixture (water / acetone volume ratio 1: 3). The two syringes are mixed through the syringe connector for about 20 times. 197.7 mg of EDC and 149 mg of NHS were then dissolved individually in 6 ml of water. The syringe containing EDC and NHS is connected to a syringe containing HA and retinoic acid and the reaction is mixed at least 20 times by passing two syringes back and forth. The mixture was stored in one syringe and immersed in a 37 [deg.] C bath for 4 hours. The gel is dialyzed against isopropanol to remove unconjugated retinoic acid and dialyzed against PBS buffer under azeotropic conditions. The gel is packed into a sterile syringe and stored at 4 ° C.

Example  15

HA On hydrogel Hydroxyl  Of functional group-containing additives Conjugation .

Additives such as retinol (AKA, tretinoin), catalase, dimethylaminoethanol and g-tocopherol contain hydroxyl functionality (-OH). These additives are conjugated to HA hydrolysis via esterification using EDC chemistry. A typical example for conjugation is described as follows:

200 mg of HMW HA is hydrated in 10 ml of pH 4.8 2- (N-morpholino) ethane sulfonic acid (MES) buffer in a 60 cc syringe. In another syringe, 200 mg of retinoic acid is dissolved in 5 ml of a water-acetone mixture (water / acetone volume ratio 1: 3). The two syringes are mixed through the syringe connector for about 20 times. 197.7 mg of EDC and 149 mg of NHS were then dissolved individually in 6 ml of water. The syringe containing EDC and NHS is connected to a syringe containing HA and retinol and the reaction is mixed at least 20 times by passing two syringes back and forth. The mixture was stored in one syringe and immersed in a 37 [deg.] C bath for 4 hours. The gel is dialyzed against isopropanol to remove unconjugated retinol and dialyzed against PBS buffer under azeotropic conditions. The gel is packed into a sterile syringe and stored at 4 ° C.

Example  16

After modification HA On hydrogel Hydroxyl  Of functional group-containing additives Conjugation .

This is a two-step process.

Step 1: A crosslinked HA gel, for example a commercial HA dermatophyte filler, such as JUVEDERM®, Allgarden or Restylane® from Irvine, Calif., Medicis Esthetic Medicis Aesthetics, Inc. is treated with EDC / NHS to activate the carboxyl groups of HA.

Step 2: Treat the activated HA hydrogel with an additive containing a hydroxyl group. Additives containing hydroxyl groups are retinol, catalase, dimethylaminoethanol and g-tocopherol hydroxyl functionality (-OH).

A typical example for conjugation of an additive to a crosslinked HA gel is as follows:

2 gm of Juvederm gel are mixed with 200 gm of EDC and 150 mg of NHS at room temperature. Then 200 mg of retinol in a 3 ml acetone-water mixture is added. The mixture is reacted at 37 DEG C for 4 hours. The gel is dialyzed against isopropanol to remove unconjugated retinol and dialyzed against PBS buffer under azeotropic conditions. The gel is packed into a sterile syringe and stored at 4 ° C.

Example  17

HA On hydrogel  Growth factors, Peptides  Or elastin Conjugation

Additives such as epidermal growth factor (EGF), transforming growth factor (TGF) and peptides contain functional amine groups and are conjugated to HA to form advantageous dermal fillers. These additives are conjugated to HA via amidation chemistry. A typical example for conjugation is described as follows:

200.3 mg of HMW HA is hydrated in 10 ml of MES pH 5.4 buffered water. 20 mg of EGF is added to 100 mg of MES solution. To this mixture, 197.7 mg of EDC and 149 mg are added. The resulting reaction mixture was reacted at 37 占 폚 for 4 hours. After the reaction is complete, the gel is dialyzed against isopropanol and dialyzed against PBS buffer under azeotropic conditions. The gel is packed into a sterile syringe and stored at 4 ° C.

The present invention provides a method for improving the viability of transplanted adipose tissue. The method generally includes introducing the composition into the skin of a patient adjacent to the transplanted adipose tissue, wherein the composition is a composition as described elsewhere herein. For example, the composition may comprise vitamin C derivatives covalently conjugated to hyaluronic acid and hyaluronic acid, with a degree of conjugation of about 3 mol% to about 40 mol%. In another aspect of the invention, a method for treating skin comprises introducing into the skin a composition comprising vitamin C that is conjugated to fatty tissue, hyaluronic acid and hyaluronic acid.

Example  18

HA On hydrogel  Growth factors, Peptides  Or elastin Conjugation

To evaluate the mitogenic effect of vitamin C and its derivatives on human adipose tissue derived stem cells (hASC), vitamin C (ascorbic acid) or its derivatives (Vitagen Or AA2G) supplemented with or without hASC were cultured on tissue culture plastic for 4 days in complete MesenPro medium. Growth was assessed by MTT analysis as described by the manufacturer (ATCC, Manassas, Va.). After 4 days, concentrations of ascorbic acid, 0.25, 0.5 and 1 mM were found to enhance the proliferation of the control, which lacks ascorbic acid by 60%, 80% and 96%, respectively (dextrose, The amount of the MTT's purple formazan (the amount of the purple formazan which is solubilized by the detergent) is measured). Using the same concentration of AA2G, 70%, 60% and 50% of the growth of the control group, respectively, was obtained. Similar results were obtained with Vitagen, which showed a 70%, 60% and 30% increase over the control, respectively. In summary, in the presence of growth factors containing medium, vitamin C and its derivatives, AA2G and Vitagen improve hASC proliferation in cell cultures.

Conjugated  Having Vitamin C Bridged HA  Gel

According to a particular embodiment of the present invention which exhibits the Tindle effect and other advantages, a cross-linked HA based gel having conjugated vitamin C using 1,4-butanediol diglycidylether (BDDE) as a cross- The preparation is described in Examples 19 and 20 below. In Example 19, the vitamin C derivative is ascorbic acid 2-glucoside (AA2G), and in Example 20, the vitamin C derivative is ascorbyl 3-aminopropylphosphate (bitagene). These gels have optimal rheological properties, excellent injection capacity and high HA concentration (25 mg / g). Although not constrained by the manipulation of any particular theory, it has been discovered by the inventors that cross-linking BDDE and HA in the presence of AA2G or Vitagen significantly changes the properties of the gel, High crosslink density, high HA concentration, low viscosity and low pressure output. Since AA2G or bitagens are present during crosslinking, the formed gel has these ascorbic acid derivatives bound either alone or as BD crosslinking agents as bridging bridges and HA chains. The microscopic structure of the gel is greatly changed, which results in a gel with very low power output even though it passes through fine needles as much as 30 gauge. In addition, the gel has from about 3 mol% to about 10 mol%, or up to about 15 mol% of the vitamin C conjugated to HA. When the gel is injected, they release active vitamin C from the fibroblast or phosphotase by an endogenous enzyme such as? -Glucosidase. Active vitamin C can trigger skin collagenization and act as a radical scavenger to inhibit gel degradation.

Example  19

Reduced  Having the Tindle effect HA / AA2G Gel Formulation

After adding 1764.0 mg of a 5 wt% NaOH solution, 400.1 mg of LMW HA and 402.3 mg of AA2G mixture in the syringe were hydrated for 60 minutes. In a separate vial, 800.8 mg of AA2G was added followed by 1401.1 mg of 9.1 wt% NaOH solution and 252.6 mg of BDDE. The obtained solution (pH > 12) was reacted in a water bath at 50 캜 for ~ 20 minutes, and transferred to hydrated HA. After addition, the mixture was mixed ~ 20 times by passing back and forth between two syringes. The paste was then transferred to a vial and placed in a 50 ° C water bath for ~ 2.5 hours. After crosslinking, the base was neutralized by the addition of a solution containing 197.0 mg of 12 M HCl and 9.18 g of 10X PBS, pH 7.4, and the gel was swollen on the orbital shaker for 72 hours. The size was changed by applying force to the gel through a ~ 60 um pore size mesh. The gels were resuspended by passing two syringes back and forth, and the gels were mixed ~ 20 times, transferred to a cellulose ester dialysis bag, MWCO ~ 20 kDa, dialyzed against PBS, pH 7.4 buffer for 5 days, Respectively. After dialysis, the gel was provided in a 1 ml COC syringe, air bubbles were removed by centrifugation at 5000 RPM for 5 minutes, and sterilized with water steam. The gel had a final HA concentration of 25 mg / g, about 10 mol% of AA2Gmol% calculated as described in Example 2 and a G 'of about 80 Pa. Other gels were made in a similar manner, which had a G 'value of about 60Pa to about 80Pa.

Example  19A

Reduced  Having the Tindle effect Lidocaine On gel  by HA / AA2G of Formulation

HA / AA2G was produced by lidocaine gel with 0.3% ww lidocaine by adding the amount of lidocaine to the gel of Example 19. A lidocaine solution was prepared by dissolving lidocaine HCl in PBS buffer pH ~ 7.4. The aliquat of the lidocaine solution was added to the gel in dialysis but before sterilization in Example 19. The gel was then thoroughly mixed to obtain a homogenized mixture having a 0.3% lidocaine concentration.

Example  20

Reduced  Having the Tindle effect HA / Vitagen Gel Formulation

401.0 mg of LMW HA was hydrated in a 1 wt% NaOH solution of 2355.0 mg in a syringe for ~ 45 min. 303.8 mg BDDE was added to hydrated HA and mixed 10 times by syringe to syringe blend. The mixture was pre-reacted in a 50 < 0 > C water bath for 15 minutes. 800.1 mg of vitgene were separately dissolved in 950.6 mg of 15 wt% NaOH followed by 510.1 Milli-Q water. The biotagen solution was mixed 30 times back and forth with a pre-heated hydrated HA / BDDE mixture using a syringe to syringe mixture. The mixture was again placed in a 50 ° C water bath and the reaction was allowed to proceed for another 2 hours before adding a solution containing 148.1 mg of 12M HCl and 8523.1 mg of 10X PBS, pH 7.4 to the crosslinking agent. The solution was neutralized by adding HCl-PBS solution, and the gel was swollen. The gel was neutralized and swollen on an orbital shaker for 48 hours. The gel was resized through a ~ 60 um screen and mixed ~ 20 times by passing back and forth between two syringes. The gel was placed in a 20,000 MWCO CE dialysis bag and dialyzed in PBS, pH 7.4 buffer. Dialysis was performed for ~ 197 hours with frequent changes in PBS buffer. After dialysis, the gel was transferred into a 1 ml COC syringe, centrifuged at 5000 RPM for 5 minutes, and sterilized with water steam. The final HA concentration of the gel was 24 mg / g.

Ethyl-3- [3- Dimethylaminopropyl ] Carbodiimide  through Bridged HA  Gel

Hydrochloride ( EDC ) chemistry

The preparation of cross-linked HA based gels is described in Examples 21 and 22 below in accordance with certain embodiments of the present invention that exhibit the tin-mill effect and other advantages. In Example 21, the gel made by EDC chemistry using a crosslinking agent is hexamethylenediamine (HMDA), and in Example 20, 3- [3- (3-aminopropoxy) -2,2-bis 3-amino-propoxymethyl) -propoxy] -propylamine (4-aminamine-4-AA). The crosslinking agent is carried out under mild conditions, for example at room temperature and, for example, at pH 5.4. The reaction conditions can be adjusted to produce highly delineated gels, with optimal rheological properties, good injection capacity and high HA concentration (24 mg / g). HA is crosslinked at very low hydration or reaction concentration and the coupling agent 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) It has been discovered by the inventors that it may be advantageous to have a normal amount of HMDA or 4 AA with -NHS (sulfo-NHS). The gel will have crosslinking points away from each other, and therefore highly crosslinked materials have high damping forces. In contrast, cross-linking of BDDE and HA at such low hydration or reaction concentrations may be impractical due to the relative inefficiency of the cross-linking agent.

Example  21

Reduced  Having the Tindle effect HA / HMDA Gel Formulation

20.0 g of 100 mM MES buffer pH 5.2 was added to a syringe containing 1000.0 mg of LMW HA. A HMDA solution was prepared by dissolving 260.9 mg of HMDA.HCl in 100 mg of MES buffer pH 5.2 of 2010.5 and adding 2 μl of 1 M NaOH to pH 5.2. EDC solution was prepared by dissolving 254.2 mg of EDC in 1188.4 mg of 100 mM MES buffer pH 5.2 and in a separate vial, 44.3 mg of NHS was dissolved in 1341.8 mg of 100 mM MES buffer pH 5.2. For total hydration of HA, 790 占 of HMDA solution was added to hydrated HA for ~ 1 hour. The mixture was homogenized by syringe to syringe mixing 10 times. 490 [mu] l EDC and 490 [mu] l NHS solution were then added to the homogenized paste and mixed 10 times by syringe to syringe mixing again. The mixture was then transferred to the vial, crosslinked for 5 hours at room temperature, and 17.9 mL of 1X PBS buffer pH 7.4 was added. The gel was swollen on the roller for 3 days and then force was applied through a 60 탆 pore size mesh. The size-changed gel was placed in a cellulose ester membrane with MWCO 20 KDa and dialyzed against 1X PBS for 4 days with varying buffer twice a day. The gel was prepared in a 1 ml COC syringe, centrifuged at 5000 RPM for 5 minutes, and sterilized with water steam. The final HA concentration of the gel was 25 mg / g.

Example  22

Reduced  Having the Tindle effect HA /4 AA Gel Formulation

32.55 of 100 mM MES buffer pH 5.2 was added to a syringe containing 1000.4 mg of LMW HA. A 4 AA solution was prepared by dissolving 256.3 mg of 4 AA in 1039.8 mg of 100 mM MES buffer pH 5.2 and adding 380 μL of 6M HCl to pH 5.2. EDC solution was prepared by dissolving 251.2 mg of EDC in 1013.8 mg of 100 mM MES buffer pH 5.2 and in a separate vial, 74.7 mg of NHS was dissolved in 2020.0 mg of 100 mM MES buffer pH 5.2. For total hydration of HA, 260 μl of 4 AA solution was added to hydrated HA for ~ 1 hour. The mixture was homogenized by syringe to syringe mixing 10 times. 277 [mu] l EDC and 273 [mu] l NHS solution were then added to the homogenized paste and mixed 10 times by syringe to syringe mixing again. The mixture was then transferred to the vial, crosslinked for 5 hours at room temperature, and then 6.4 mL of 10X PBS buffer pH 7.4 was added. The gel was swollen on the roller for 3 days and then force was applied through a 60 탆 pore size mesh. The size-changed gel was placed in a cellulose ester membrane with MWCO 20 KDa and dialyzed against 1X PBS for 4 days with varying buffer twice a day. The gel was prepared in a 1 ml COC syringe, centrifuged at 5000 RPM for 5 minutes, and sterilized with water steam. The gel had a final HA concentration of 23 mg / g.

Example  23

Example  19 to 22 Gel Rheology  Determination of characteristics.

The rheological properties of the gel were measured using Anton Paar Physica MCR 301, a vibrating parallel plate flow system. A plate diameter of 25 mm was used at a gap height of 1 mm. Each measurement was performed at a constant temperature of 25 캜. A frequency sweep of 1 Hz to 10 Hz at a constant strain of 2%, and a strain sweep of 1% to 300% at a constant frequency of 5 Hz due to algebraic increase in strain after the algebraic increase of frequency. The storage modulus (G ') and the viscosity (G ") were obtained from the strain sweep at 1% strain.

Example  24

Example  19 to 22 Gel Pressure output  Measure

The force required to extrude the gel through a 30 gauge needle was measured using an Instron 5564 and Bluehill 2 software. The gel was extruded from a 1 ml COC syringe through a 30 G < 2 > TSK needle. The plunger was pushed at a rate of 100 mm / min with respect to 11.35 mm, and the pressure output was recorded.

Example  25

Example  19 to 22 Gel  Bioavailability test

A 50 占 퐇 bolus injection of the gel was implanted into the dermis on the back of the Sprague Dawley rats. The grafts were removed at 1 week and histologically analyzed by hematoxylin and eosin (H & E) staining, CD68 staining for markers for mononuclear inflammatory cells. Three 20X images of CD68 were scored from 0 to 4 based on degree of staining. These values were then averaged to provide a sample score. Four samples were analyzed from each gel.

Example  26

Example  Cytotoxicity test of 19 to 22, ISO  10993-5.

The in vitro cytotoxicity test of the gel was performed by NAMSA according to the Agarose Overlay Method of ISO 10993-5: Part 5: Tests for In Vitro Cytotoxicity. 0.9% NaCl solution, 1 cm long dense polyethylene as negative control, and 1 x 1 cm2 portion of latex as a positive control were administered as well as 0.1 ml of the test article placed on the filter paper in triple wells. Each was placed on an agarose surface directly across a single layer of L929 mouse fibroblasts. After incubation for 24 hours at 37 ° C in 5% CO ₂ , the cultures were macroscopically and microscopically tested for any abnormal cell morphology and cell lysis. The test article was scored from 0 to 4 based on the proximate dissolution zone of the sample. The test materials from Examples 1, 3 and 4 scored as 0 as test articles did not show any evidence of cell lysis or toxicity.

Quantitative analysis of the Tindle effect

In addition, to support visual observations and to perform comparative performance analysis of HA fillers, it was deemed necessary to perform a quantitative analysis of the Tindle effect. There are no quantitative techniques in the literature for the Tindle effect specific to dermal fillers. However, based on existing scientific understanding of skin-light scattering and light interactions, two distinct approaches based on (a) colorimetry and (b) spectroscopy have been used to quantify the tin effects in the skin. Based on these techniques, three distinct quantitative parameters (outlined below) were established to measure the in vivo Tin-D effect.

와 관련됨)에 대해 최대 스코어 5가 주어진다. 시험 샘플을 스코어링하기 위해 맹검 전 스케일에 대해 3명의 독립적 관찰자를 훈련시켰다.a) Tindle effect visual score : The scale has a range of 1 to 5 in increments of 0.5. Score 1 is given at the injection site with normal skin tone and no blue discoloration. A thick, distinctive blue discoloration (typically, Restylane or Juvedim Ultra Plus

The maximum score 5 is given. Three independent observers were trained on the pre-blind scale to score the test sample.

b) Blue component of skin color - "b" : The blue component of the light sent from the skin area injected with various fillers was quantified using a colorimeter (CM2600D, Konica Minolta, New Jersey). This was accomplished by using the "b" component of the Lab color scale.

c) " Blue light %" emitted from the skin : A portable spectrophotometer (CM2600D, Konica Minolta, NJ) was used to quantify the percentage of blue light emitted from the skin over the entire visible light range. This was accomplished by integrating regions under 400 nm to 490 nm and normalizing them under the entire spectrum (400 nm to 700 nm).

Example  27

Gel  Tins Assessment

The gel was injected intradermally via 27G < RTI ID = 0.0 > TSK < / RTI > needles using a linear threading technique into the femurs of 2 month old hairless rats. The gel was implanted deeply to mimic the clinical fine-wrinkle procedure. Tests were carried out on tin at 48 hours after gel implantation. Prior to performing the tin test, the animals were euthanized to improve the contrast of the tin effects due to lack of hemoglobin.

The images of the gels from Examples 19 to 21 after 2 days of transplantation are shown in Fig. Commercial images for the Juvedermain Refinance and Restylane Touch are shown for comparison. The bluish lines (tins) can be seen clearly in the images of the commercial gelsubiderm refine and the resilient touch. The gels from Examples 19, 19A (not shown) and 21 did not exhibit the tin effect

The injection site was scored using a visual score of 1 to 5 in increments of 0.5. The injection site with score 1 did not show skin discoloration, whereas the injection site with score 5 showed severe skin blue discoloration. Spectroscopic analysis was performed on the injection site with the aid of a colorimeter (CM2600D, Konica Minolta, NJ). The blue component of the skin color "b" and the percentage of the blue light emitted from the skin (400 nm to 700 nm) were independently measured. Figures 13 and 14 show the visual tin score and percent of emitted blue light. The gels from Examples 19 and 21 did not exhibit the tin effect and had a lower visual tin score and% of blue light emission value. The tin score and% of the emitted blue light value were higher for the Juvedermill and Restilan touch. Belotero Soft did not show any tines and the values were similar to those of Examples 19 and 21. 13 and 14.

Example  28

By history Gel In vivo  Sustainable evaluation

A 50 [mu] l bolus injection of the gel and commercial gel of the invention was implanted into the dermis on the back of the Sprague Dawley rats. The implants were removed at 1 week and analyzed histologically by hematoxylin and eosin (H & E) staining. The sections were precisely cut at the injection site. Two sections were cut from each tissue sample, and the H & E stained sections were stitched using a stitching scope. The samples were then grouped and scored as follows: none (0%), low (25%), medium (50%) and high (100%) depending on the amount of material left. See FIG.

Example  28A

MRI On by Gel In vivo  Sustainable evaluation

After intradermal injection in female Sprague Dawley rats, the volume and surface area changes of the gels and commercial gels of the present invention over time were evaluated over a period of 40 weeks using Magnetic Resonance Imaging (MRI). The gel was injected at a target volume of 150 占 퐇 per implant. The implant was placed in two opposing areas at two opposing sides towards the tail, two opposing sides from the knee slightly to the two sides, and a midpoint between the head and the tail, slightly to the shoulders with respect to the shoulders. 7 MRI scans were performed on a Tesla 70/30 Bruker Biospec MRI scanner. Images were collected on the day of transplantation (week 0) and at weeks 12, 24, and 40 after transplantation. A plot of absolute volume of gel versus time is shown in Figure 16 below. High specificity gels have a high absolute volume at 40 weeks transplant.

Example  29

Orbital circumference  Compositions of the present invention as used in the treatment of wrinkles

A 40-year-old skinny woman has fine wrinkles in the orbital area and demands dermal filler treatment. Using a 30-gauge needle, the physician introduces 0.6 ml of the HA-based gel according to the invention (as described in Example 19) into the fine wrinkles under her eye and deeply in the tear crease area using the linear threading technique . The gel was not deeply introduced, but no blue discoloration was observed, and the patient was satisfied with the results.

As shown, the compositions of the present invention, e. G., Compositions of Examples 19 and 21, have reduced or no significant tin effects and can be used with certain HA-based commercial gels, e. G., Juvedermill / Surgiderm, 18 and < RTI ID = 0.0 > Velothoros < / RTI > For example, for "fine-wrinkle" formulation Belloterosoft, Example 19 of the present invention not only had a beneficial tin-effect on this commercial gel, but also beneficially had a substantially longer in vivo duration.

Example  30

The appearance of fine wrinkles Improve  ≪ RTI ID = 0.0 >

Additives such as Vitamin A, Vitamin B, Vitamin C, Vitamin D, Vitamin E and derivatives thereof are conjugated to the crosslinked hyaluronic acid gel in a manner to produce various substantially transparent, injectable HA-based gels, alone and in combination . The HA component is at least 90% by weight and is, for example, completely low molecular weight HA, or about 100% low molecular weight HA as defined elsewhere herein. These additives are conjugated to HA hydrogels using any suitable means. Having an HA concentration of at least about 20 mg / g, such as about 23 mg / g, about 24 mg / g, about 25 mg / g, about 30 mg / g, suitable for injection through a fine gauge needle, The conjugated gels are sized and processed to produce injectable pH neutral, cohesive compositions. The gel has a G 'value of at least about 50 Pa, about 60 Pa, about 70 Pa, up to about 80 Pa, and about 100 Pa or less. The gel is packaged and autoclaved using UV light or other suitable means.

Each gel may be applied to the wrinkles of the patient, for example in the orbital area, in the wrinkle area, in the tear area, in the neck area, or in any other face area where dermal filling is beneficial, It is useful for injection into the skin. Despite the undesirable introduction of the gel, discoloration due to the tin effect was not observed and the patient was satisfied with the results.

Finally, although aspects of the present disclosure have been described with reference to various embodiments, those skilled in the art will readily appreciate that the specific disclosed embodiments merely illustrate the subject matter disclosed herein. Accordingly, the subject matter disclosed herein does not in any way limit the specific methods, protocols and / or reagents, etc. As such, those skilled in the art will be able to make numerous and various modifications or variations, or alternatives of the disclosed subject matter may be made in accordance with the teachings herein without departing from the spirit of the specification. Changes in the details may be made without departing from the spirit of the invention as set forth in the appended patent claims. Finally, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the invention, which is defined solely by the claims. In addition, it is intended that all matter contained in the above description or shown in the accompanying drawings be interpreted as illustrative and not in a limiting sense. Therefore, the present invention is not specifically limited as shown and described.

Certain embodiments of the invention are described herein, including the best mode known to the inventors for carrying out the invention. Of course, modifications to these described embodiments will be apparent to those skilled in the art upon reading the foregoing description. The inventors anticipate that those skilled in the art will employ such modifications if appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, the invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto, as permitted by applicable law. Moreover, any combination of the elements described above in all possible variations thereof is not covered by the present invention unless otherwise indicated herein or otherwise clearly contradicted by context.

The groups of alternative elements or embodiments of the invention disclosed herein should not be construed as limitations. Each group member may be referred to and claimed individually or in any combination with any other member of the group or other element found herein. One or more members of the group may be included in or removed from the group for convenience and / or patentability reasons. Whenever such inclusion or deletion occurs, the present specification is deemed to contain a modified group and thus meets the described descriptions of all the Makush groups used in the appended claims.

Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about. &Quot; The term "about" as used herein means that the quantified item, variable, or term includes a range of plus or minus 10% of values exceeding and less than the value of the item, variable or term to which it is referred. Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about. &Quot; The term "about" as used herein means that the quantified item, variable, or term includes a range of plus or minus 10% of values exceeding and less than the value of the item, variable or term to which it is referred. Accordingly, unless otherwise indicated, the numerical parameters set forth in this specification and the appended claims are approximations that may vary depending on the desired properties sought to be obtained by the present invention. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant numbers and by applying ordinary rounding techniques. While the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as concisely as possible. However, any numerical value inherently contains certain errors necessarily resulting from the standard deviation found in its respective test measurements.

The singular terms used in the context of describing the invention and similar references (especially in the context of the following claims), unless otherwise indicated herein or clearly contradicted by context, are intended to include both singular and plural It should be interpreted as covering everything. The citation of the scope of the present specification is intended to serve as a shorthand way of referring specifically to each distinct value falling within its scope. Unless otherwise indicated herein, each individual value is included in the specification as if it were individually recited herein. All methods described herein may be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all embodiments or example language (e.g., "an example ") provided herein is intended only to better describe the present invention and is not to be construed as limiting the scope of the invention otherwise claimed It does not raise. No language in this specification should be construed as indicating any non-claimed element essential to the practice of the invention.

The specific embodiments disclosed herein may be further limited in the appended claims using the language in which they are made or made essentially. When used in the claims, whether an amendment is filed or added, the expanded term "comprising " excludes any elements, steps or components not embodied in the claims. The term " consisting essentially of "encompasses the scope of the claimed subject matter for material and / or steps that do not materially affect the underlying material and / or feature (ies). The embodiments of the invention thus claimed are essentially or clearly described and made possible in the present specification.

All patents, patent publications, and other publications referred to herein and identified herein are, for example, describing compositions and methods described in such publications used in connection with the present invention, and for purposes of disclosure, And is clearly included. These publications are only provided for its disclosure prior to the filing date of the present invention. Nothing herein is to be construed as an admission that the inventors qualify for the invention, or for any other reason, to precede the disclosure. Any reference to the date or representation of the contents of these documents is based on information available to the applicant and does not constitute any concession to the accuracy of the dates or contents of these documents.

Claims (42)

As the dermal filler composition,
A crosslinking component and a crosslinked hyaluronic acid component;
An additive other than the crosslinking component;
The composition is substantially optically clear,
Wherein the composition exhibits a reduced tinned effect when administered into the dermal area of a patient for substantially the same composition but without the additive. The dermal filler composition of claim 1, wherein the additive is a vitamin C derivative. The dermal filler composition of claim 1, wherein the additive is AA2G. The dermal filler composition of claim 1, wherein the additive is Vitagen. The dermal filler composition of claim 1, wherein the hyaluronic acid component is chemically conjugated to the additive. The dermis filler composition of claim 1, wherein the hyaluronic acid component is chemically conjugated to the additive at a degree of conjugation from about 3 mol% to about 40 mol%. The dermal filler composition of claim 1, wherein the hyaluronic acid component is chemically conjugated to the additive at a degree of conjugation from about 3 mol% to about 10 mol%. The dermal filler composition of claim 1, wherein the cross-linking agent is BDDE. The dermis filler composition of claim 1, further comprising an anesthetic. The dermis filler composition of claim 1, further comprising lidocaine. 2. The dermis-peel composition of claim 1 having a G 'value of from about 40 Pa to about 100 Pa. The dermis filler composition of claim 1, having a G 'value of about 100 Pa or less. 2. The dermis filler composition according to claim 1, having a G 'value of at least about 40 Pa. CLAIMS What is claimed is: 1. A method of treating fine wrinkles in a patient's skin,
A composition comprising a hyaluronic acid component, a crosslinking component for crosslinking the hyaluronic acid, and a mixture of additives other than the crosslinking component is introduced into the skin of the patient,
The composition is substantially optically clear,
Wherein the dermis filler composition exhibits a reduced tinned effect for substantially the same composition but without the additive. 15. The method of claim 14, wherein the additive is a vitamin C derivative. 15. The method of claim 14, wherein the additive is AA2G. 15. The method of claim 14, wherein the additive is non-toxic. 15. The method of claim 14, wherein the hyaluronic acid component is chemically conjugated to the additive.  15. The method of claim 14, wherein the hyaluronic acid component is chemically conjugated to the additive at a degree of conjugation of about 3 mol% to about 10 mol%. 15. The method of claim 14, wherein the hyaluronic acid component is chemically conjugated to the additive at a degree of conjugation from about 3 mol% to about 40 mol%. A method of improving the cosmetic appearance of a face,
Comprising administering a substantially optically clear dermal filler that does not exhibit or does not significantly exhibit a tin effect on the dermal area of the patient,
Providing a hyaluronic acid;
Reacting the cross-linking agent with a vitamin C derivative;
Adding the reacted crosslinking agent and a vitamin C derivative to the hyaluronic acid to form a crosslinked hyaluronic acid composition comprising covalently conjugated vitamin C; And
Homogenizing and neutralizing the crosslinked hyaluronic acid composition to obtain an injectable dermatophyte filler composition. 22. The method of claim 21, wherein the additive is a vitamin C derivative. 22. The method of claim 21, wherein the additive is AA2G. 22. The method of claim 21, wherein the additive is non-toxic. 22. The method of claim 21, wherein the hyaluronic acid component is chemically conjugated to the additive. 22. The method of claim 21, wherein the hyaluronic acid component is chemically conjugated to the additive at a degree of conjugation from about 3 mol% to about 40 mol%. 22. The method of claim 21, wherein the hyaluronic acid component is chemically conjugated to the additive at a degree of conjugation from about 3 mol% to about 10 mol%. A method of reducing the appearance of fine wrinkles in a thin skin of a patient,
Comprising administering to said patient a substantially optically clear hyaluronic acid dermal filler composition comprising vitamin C or a vitamin C derivative at a depth of about 1 mm or less. 29. The method of claim 28, wherein the composition is injected at a depth of about 0.8 mm or less. 29. The method of claim 28, wherein the composition is injected at a depth of about 0.6 mm or less. 29. The method of claim 28, wherein the composition is injected at a depth of about 0.4 mm or less. As the dermal filler composition,
A crosslinking component and a crosslinked hyaluronic acid component; And
A vitamin C or vitamin C derivative covalently conjugated to said hyaluronic acid component at a conjugation degree of at least about 3 mol%
Wherein the composition is substantially optically clear and has a G 'value of from about 40 Pa to about 100 Pa. 33. The dermis-peel composition of claim 32 having a hyaluronic acid concentration of from about 18 mg / g to about 30 mg / g. 33. The dermal filler composition of claim 32 having a hyaluronic acid concentration of from about 12 mg / g to about 30 mg / g. 33. The dermal filler composition of claim 32, wherein the degree of conjugation is about 10 mol%. 33. The dermal filler composition of claim 32, wherein the hyaluronic acid is a low molecular weight HA that is at least 90% lower. 33. The dermal filler composition of claim 32, wherein the hyaluronic acid component is a substantially completely low, low molecular weight HA. 1. A injectable composition for reducing the appearance of fine wrinkles in a face,
1,4-butanediol diglycidyl ether (BDDE) -crosslinked low molecular weight hyaluronic acid (HA); And
A vitamin C derivative covalently conjugated with said hyaluronic acid,
Wherein the HA has an average molecular weight of about 300K daltons to about 900K daltons,
The conjugation degree is about 10 mol%
Wherein said composition has a G 'value of from about 60 Pa to about 80 Pa. 39. The injectable composition of claim 38, wherein the HA has an average molecular weight of about 300K daltons to about 500K daltons. 40. The composition of claim 39, wherein the composition has a G ' value of about 80 Pa. 39. The composition of claim 38, wherein the composition is optically clear. 39. The injectable composition of claim 38, having an HA concentration of about 25 mg / g.

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