Gels based on multicomponent hydrogels and uses thereof - Patent Application 20070122997

Abstract

The present disclosure relates to multi-component hydrogels that can be used, for example, as dermal fillers to treat fine lines and wrinkles on the skin. Also disclosed herein are methods for making and using the disclosed hydrogels and related compositions.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 408,718, filed September 21, 2022, which is incorporated herein by reference in its entirety.

[0002] Technical Field The present disclosure relates to hydrogels and compositions comprising the same. The disclosed hydrogels and compositions can be used as dermal fillers or in cosmetic surgery, and they can be effective in treating fine lines and wrinkles on the skin. [Background technology]

[0003] background The following discussion is provided solely to aid the reader in understanding the present disclosure and is not admitted to describe or constitute prior art thereto.

[0004] Water-absorbing gels, or hydrogels, are widely used in the biomedical field, for example, during viscoelastic surgery and as dermal fillers. They are generally prepared by chemical crosslinking of polymers. Various injectable dermal filler products have been developed to treat or correct facial imperfections, such as wrinkles and loss of firmness due to the natural effects of aging. Injectable "dermal fillers" temporarily restore a smoother, more youthful appearance. Ideally, dermal fillers should be long-lasting, soft, smooth, and natural-looking when introduced into or under the skin.

[0005] Glycosaminoglycans (GAGs), or mucopolysaccharides, are long-chain linear polysaccharides composed of repeating disaccharide units (i.e., two sugar units). The repeating disaccharide units consist of uronic and amino sugars. Hyaluronic acid (HA), also known as hyaluronan, is a water-soluble glycosaminoglycan that is a major component of the extracellular matrix and is widely distributed in animal tissues. HA has excellent biocompatibility and does not cause allergic reactions when implanted in patients. Furthermore, HA has the ability to bind large amounts of water, making it an excellent volumizer for soft tissues.

[0006] Alpha hydroxy acids (AHAs) are a class of chemical compounds consisting of carboxylic acids substituted with hydroxyl groups on adjacent carbon atoms. Notable examples are glycolic acid, lactic acid, and citric acid. Polymers made from AHAs are useful as biocompatible materials. For example, poly-L-lactic acid (PLLA) is an immunologically inert, biocompatible, and biodegradable synthetic polymer. Once inside the body, PLLA is eventually degraded and resorbed. Summary of the Invention

[0007] The present application provides hydrogels and compositions comprising a cross-linked glycosaminoglycan (e.g., hyaluronic acid, or "HA"), an uncross-linked glycosaminoglycan (e.g., HA), and a poly(alpha-hydroxy acid) polymer (e.g., poly-L-lactic acid or "PLLA"). The disclosed hydrogels have improved flowability and spreadability, for example, compared to products comprising cross-links between HA and PLLA. The disclosed hydrogels and compositions can be used as dermal fillers or skin boosters, and to treat or reduce the appearance of fine lines (e.g., cheek lines), acne scars, and skin texture. The disclosed hydrogels enhance skin texture as fillers and stimulate the skin to provide immediate improvements in skin texture and texture, with long-term benefits. The disclosed hydrogels and compositions are particularly well-suited for such uses due to their spreadability, providing an immediate lifting effect on the skin and sustained improvement in skin texture, with long-term implantability and stability / shelf life. The hydrogels and compositions disclosed herein improve skin quality attributes, including elasticity, radiance (reducing dry skin and increasing hydration), texture and smoothness (e.g., improving pore size, fine lines), and skin tone over a period of 9 months or more (e.g., up to 12 or 18 months).

[0008] The disclosed hydrogels and compositions are particularly well suited for such uses due to their spreadability, providing an immediate lifting effect to the skin as well as sustained improvement in skin quality, with long-term implantability and stability / shelf life. The hydrogels and compositions disclosed herein improve skin quality attributes, including elasticity, radiance (reducing dry skin and increasing hydration), texture and smoothness (e.g., reducing pore size and fine lines), and skin tone, over a period of 9 months or more, e.g., 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or up to 24 months.

[0009] In one aspect, the present disclosure provides a composition comprising: (a) 10 to 45 mg / ml of a cross-linked glycosaminoglycan hydrogel; (b) 20% to 50% free, uncross-linked glycosaminoglycans by weight of the cross-linked glycosaminoglycan hydrogel; and (c) 10 to 50 mg / ml of a biodegradable poly(α-hydroxy acid) polymer that is not cross-linked to the glycosaminoglycan hydrogel or the free, uncross-linked glycosaminoglycans.

[0010] Another aspect of the present disclosure provides a composition prepared by a process comprising: (a) cross-linking glycosaminoglycans in the presence of 2% to 4% v / v NaOH to obtain a cross-linked glycosaminoglycan hydrogel; (b) filtering the cross-linked glycosaminoglycan hydrogel using a 60-90 micron filter; (c) adding 20% ​​to 50% free, uncross-linked glycosaminoglycans, by weight of the composition, to the filtered cross-linked glycosaminoglycan hydrogel; and (d) adding 10 to 50 mg / ml of a biodegradable poly(α-hydroxy acid) polymer to the composition from (c) under vacuum conditions, wherein the biodegradable poly(α-hydroxy acid) polymer is not cross-linked to the glycosaminoglycan hydrogel. In some embodiments, the process further comprises swelling the cross-linked glycosaminoglycan hydrogel under vacuum conditions between (a) and (b). In some embodiments, the process further comprises precipitating and washing the filtered crosslinked glycosaminoglycan hydrogel from (b) using ethanol. In some embodiments, the process further comprises sterilizing the composition after (d).

[0011] In some embodiments, the composition comprises 10-20 mg / ml, 10-25 mg / ml, 10-30 mg / ml, 10-35 mg / ml, 10-40 mg / ml, 10-45 mg / ml, 15-20 mg / ml, 15-25 mg / ml, 15-30 mg / ml, 15-35 mg / ml, 15-40 mg / ml, or 15-45 mg / ml of the crosslinked glycosaminoglycan hydrogel.

[0012] In some embodiments, the composition comprises 20% to 30%, 20% to 35%, 20% to 40%, or 20% to 45% free uncrosslinked glycosaminoglycans by weight of the crosslinked glycosaminoglycan hydrogel.

[0013] In some embodiments, the composition comprises 10-15 mg / ml, 10-20 mg / ml, 10-25 mg / ml, 10-30 mg / ml, 10-35 mg / ml, 10-40 mg / ml, or 10-50 mg / ml of biodegradable poly(alpha-hydroxy acid) polymer.

[0014] In some embodiments, the cross-linked glycosaminoglycan hydrogel comprises cross-linked hyaluronic acid (HA), cross-linked heparan sulfate (HS), cross-linked heparin (HEP), cross-linked chondroitin sulfate (CS), cross-linked dermatan sulfate (DS), or cross-linked keratan sulfate (KS), or a combination thereof.

[0015] In some embodiments, the free glycosaminoglycans comprise hyaluronic acid (HA), heparan sulfate (HS), heparin (HEP), chondroitin sulfate (CS), dermatan sulfate (DS), or keratan sulfate (KS), or a combination thereof.

[0016] In some embodiments, the biodegradable poly(α-hydroxy acid) polymer comprises poly-L-lactic acid (PLLA), poly-D-lactic acid (PLDA), polyglycolic acid (PLGA), or poly(diol) citrate (PDC), or a combination thereof.

[0017] In some embodiments, the cross-linked glycosaminoglycan hydrogel has a size of about 60-90 microns, and optionally the biodegradable poly(alpha-hydroxy acid) polymer is about 40-63 microns.

[0018] In some embodiments, the cross-linked glycosaminoglycans are cross-linked by one or more multifunctional cross-linkers.

[0019] In some embodiments, the one or more multifunctional crosslinkers are selected from the group consisting of divinyl sulfones, multiepoxides, and diepoxides.

[0020] In some embodiments, the one or more multifunctional crosslinkers are selected from the group consisting of 1,4-butanediol diglycidyl ether (BDDE), 1,2-ethanediol diglycidyl ether (EDDE), and diepoxyoctane.

[0021] In some embodiments, the cross-linked glycosaminoglycans are cross-linked by (i) a non-carbohydrate-based di- or poly-nucleophilic cross-linker, or (ii) a carbohydrate-based di- or poly-nucleophilic cross-linker.

[0022] In some embodiments, the non-carbohydrate-based di- or poly-nucleophilic crosslinker is hexamethylenediamine (HMDA).

[0023] In some embodiments, the carbohydrate-based di- or poly-nucleophilic crosslinker is diaminotrehalose (DATH).

[0024] In some embodiments, the composition is an injectable composition.

[0025] Another aspect of the present disclosure is directed to a method for improving skin quality in a subject in need of such treatment comprising administering to the subject a composition of the present disclosure.

[0026] Another aspect of the present disclosure is directed to a method for preparing a composition, the method comprising: (a) cross-linking glycosaminoglycans in the presence of 2% to 4% v / v NaOH, thereby obtaining a cross-linked glycosaminoglycan hydrogel; (b) filtering the cross-linked glycosaminoglycan hydrogel using a 60 to 90 micron filter; (c) adding 20% ​​to 50% free glycosaminoglycans, by weight of the composition, to the filtered cross-linked glycosaminoglycan hydrogel; and (d) adding 10 to 50 mg / ml of a biodegradable poly(α-hydroxy acid) polymer to the composition of step (c) under vacuum conditions, wherein the biodegradable poly(α-hydroxy acid) polymer is not cross-linked to the glycosaminoglycan hydrogel.

[0027] In some embodiments, the method includes, between (a) and (b), swelling the cross-linked glycosaminoglycan hydrogel under vacuum conditions.

[0028] In some embodiments, the method further comprises precipitating and washing the filtered crosslinked glycosaminoglycan hydrogel from (b) using ethanol.

[0029] In some embodiments, the method further comprises sterilizing the composition after (d).

[0030] In some embodiments, the composition comprises 10-20 mg / ml, 10-25 mg / ml, 10-30 mg / ml, 10-35 mg / ml, 10-40 mg / ml, 10-45 mg / ml, 15-20 mg / ml, 15-25 mg / ml, 15-30 mg / ml, 15-35 mg / ml, 15-40 mg / ml, or 15-45 mg / ml of the crosslinked glycosaminoglycan hydrogel.

[0031] In some embodiments, the composition comprises 20% to 30%, 20% to 35%, 20% to 40%, or 20% to 45% free glycosaminoglycans by weight of the crosslinked glycosaminoglycan hydrogel.

[0032] In some embodiments, the composition comprises 10-15 mg / ml, 10-15 mg / ml, 10-20 mg / ml, 10-25 mg / ml, 10-30 mg / ml, 10-35 mg / ml, 10-40 mg / ml, or 10-45 mg / ml of biodegradable poly(alpha-hydroxy acid) polymer.

[0033] In some embodiments, the cross-linked glycosaminoglycan hydrogel comprises cross-linked hyaluronic acid (HA), cross-linked heparan sulfate (HS), cross-linked heparin (HEP), cross-linked chondroitin sulfate (CS), cross-linked dermatan sulfate (DS), or cross-linked keratan sulfate (KS), or a combination thereof.

[0034] In some embodiments, the free glycosaminoglycans comprise hyaluronic acid (HA), heparan sulfate (HS), heparin (HEP), chondroitin sulfate (CS), dermatan sulfate (DS), or keratan sulfate (KS), or a combination thereof.

[0035] In some embodiments, the biodegradable poly(α-hydroxy acid) polymer comprises poly-L-lactic acid (PLLA), poly-D-lactic acid (PLDA), polyglycolic acid (PLGA), or poly(diol) citrate (PDC), or a combination thereof.

[0036] In some embodiments, the cross-linked glycosaminoglycan hydrogel has a size of about 60-90 microns, and optionally the biodegradable poly(alpha-hydroxy acid) polymer is about 40-63 microns.

[0037] In some embodiments, the cross-linked glycosaminoglycans are cross-linked by one or more multifunctional cross-linkers.

[0038] In some embodiments, the one or more multifunctional crosslinkers are selected from the group consisting of divinyl sulfones, multiepoxides, and diepoxides.

[0039] In some embodiments, the one or more multifunctional crosslinkers are selected from the group consisting of 1,4-butanediol diglycidyl ether (BDDE), 1,2-ethanediol diglycidyl ether (EDDE), and diepoxyoctane.

[0040] In some embodiments, the cross-linked glycosaminoglycans are cross-linked by (i) a non-carbohydrate-based di- or poly-nucleophilic cross-linker, or (ii) a carbohydrate-based di- or poly-nucleophilic cross-linker.

[0041] In some embodiments, the non-carbohydrate-based di- or poly-nucleophilic crosslinker is hexamethylenediamine (HMDA).

[0042] In some embodiments, the carbohydrate-based di- or poly-nucleophilic crosslinker is diaminotrehalose (DATH).

[0043] In some embodiments, the method further comprises formulating the composition as an injectable composition.

[0044] The foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the present disclosure as claimed. Other objects, advantages, and novel features will become readily apparent to those skilled in the art from the following brief description of the drawings and detailed description of the present disclosure. [Brief explanation of the drawings]

[0045] [Figure 1] 1 shows a general representation of an exemplary manufacturing process for the disclosed hydrogels. DETAILED DESCRIPTION OF THE INVENTION

[0046] Detailed Description The present disclosure provides hydrogels and compositions comprising cross-linked glycosaminoglycans (e.g., hyaluronic acid, or "HA"), non-cross-linked glycosaminoglycans (e.g., HA), and poly(α-hydroxy acid) polymers (e.g., poly-L-lactic acid or / "PLLA"). The disclosed hydrogels possess firmness, flowability, and spreadability, making them ideal for cosmetic and dermatological uses, including, but not limited to, treating or reducing the appearance of fine lines and wrinkles. Furthermore, the disclosed hydrogels and related compositions may be stable for more than 18 months; such a shelf life provides further commercial desirability to the disclosed compositions, which can be shipped in ampoules, vials, or pre-filled syringes.

[0047] Generally, the hydrogels and compositions of the present disclosure are designed to deliver a consistent PLLA dose during administration (e.g., injection), something that other hydrogels cannot. In other words, the PPLA particles are uniformly distributed throughout the ampoule / syringe / vial, from the beginning, middle, and end of the ampoule / syringe / vial. Upon administration to a subject, the hydrogels and compositions of the present disclosure improve skin quality characteristics, including improved elasticity, radiance (reduced dry skin and increased moisturization), texture and smoothness (e.g., improved pore size, reduced crepitation), and skin tone, over a period of 9 months or more, for example, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or up to 24 months.

[0048] The disclosed hydrogels are formed using a process that prevents aggregation of poly(α-hydroxy acid) polymers (e.g., PLLA) by preparing crosslinked GAG particles (e.g., crosslinked HA) within a desired size range to provide a more uniformly dispersed hydrogel and by mixing the components under vacuum conditions during certain steps. Furthermore, the addition of non-crosslinked GAG (e.g., "free HA") prevents undesirable stickiness, facilitating the manufacture, transportation, and ultimate use of the hydrogel and hydrogel-containing products. Taken together, the disclosed processes can produce unique and desirable hydrogels suitable for any number of cosmetic and dermatological indications.

[0049] definition As used herein, the singular forms "a," "an," and "the" designate both the singular and the plural, unless expressly stated to designate only the singular.

[0050] It should be understood that all numerical designations are preceded, although not necessarily explicitly stated, by the term "about." The term "about" means that the number being grasped is not limited to the exact numerical value set forth herein, but is intended to refer to a substantial numerical value that is close to the set forth numerical value without departing from the scope of the present invention. As used herein, "about" will be understood by those of ordinary skill in the art and will vary to some extent depending on the context in which it is used. If there are uses of the term that are not clear to those of ordinary skill in the art given the context in which the term is used, "about" will mean plus or minus up to 15%, 10%, 5%, 1%, or 0.1% of the particular term.

[0051] Also, as used herein, "and / or" refers to and includes any and all possible combinations of one or more of the associated listed items, and when interpreted as alternatives ("or"), refers to and includes other combinations.

[0052] As used herein, the terms "administer," "administration," or "administering" refer to (1) providing, administering, dispensing, and / or prescribing, e.g., by or under the direction of a medical professional or a medical professional's authorized representative, and (2) taking, taking, or consuming by a medical professional or a subject. Administration can include, but is not limited to, administration by subcutaneous, intramuscular, subcutaneous tissue, intradermal, or transdermal injection. Administration can be unilateral or bilateral, as needed for a given patient.

[0053] Hydrogels and Compositions One aspect of the present disclosure is directed to a composition comprising 10-45 mg / ml of a crosslinked glycosaminoglycan hydrogel, 20%-50% free glycosaminoglycans by weight of the crosslinked glycosaminoglycan hydrogel, and 10-50 mg / ml of a biodegradable poly(α-hydroxy acid) polymer that is not crosslinked to the glycosaminoglycan hydrogel (i.e., is a free biodegradable poly(α-hydroxy acid) polymer). In some embodiments, the crosslinked glycosaminoglycan hydrogel is a crosslinked hyaluronic acid (HA) hydrogel. In some embodiments, the free glycosaminoglycan is free hyaluronic acid (HA). In some embodiments, the biodegradable poly(α-hydroxy acid) polymer is poly-L-lactic acid (PLLA). Thus, in some embodiments, the disclosed compositions may comprise 10-45 mg / ml of crosslinked HA hydrogel, 20%-50% free HA by weight of the crosslinked HA hydrogel, and 10-50 mg / ml of PLLA that is not crosslinked to the HA hydrogel.

[0054] One aspect of the present disclosure is directed to a composition prepared by a process, the process comprising: (a) cross-linking glycosaminoglycans in the presence of 2% to 4% (e.g., 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, or 4%) v / v sodium hydroxide (NaOH), thereby cross-linking glycosaminoglycans; (b) obtaining a cross-linked glycosaminoglycan hydrogel; (b) filtering the cross-linked glycosaminoglycan hydrogel using a 60-90 micron filter (e.g., a 60, 70, 80, or 90 micron filter); and (c) adding 20% ​​to 50% (e.g., 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 1109%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127 (d) adding 0%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% free glycosaminoglycan to the composition of step (c) under vacuum conditions, and (e) adding 0%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% free glycosaminoglycan to the composition of step (c) under vacuum conditions, and (f) adding 0%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% free glycosaminoglycan to the composition of step (c) under vacuum conditions, and (g) adding 0%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% free glycosaminoglycan to the composition of step (c) under vacuum conditions, and adding a biodegradable poly(α-hydroxy acid) polymer (27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 mg / ml) to the glycosaminoglycan hydrogel, wherein the biodegradable poly(α-hydroxy acid) polymer is not crosslinked to the glycosaminoglycan hydrogel. In some embodiments, the process further comprises swelling the crosslinked glycosaminoglycan hydrogel under vacuum conditions between steps (a) and (b). In some embodiments, the process further comprises precipitating and washing the filtered crosslinked glycosaminoglycan hydrogel from step (b) using ethanol.In some embodiments, the process further comprises sterilizing the composition after step (d). In some embodiments, the glycosaminoglycan is HA. In some embodiments, the biodegradable poly(α-hydroxy acid) polymer is PLLA.

[0055] In some embodiments, crosslinking in step (a) is achieved in the presence of 2.3% to 3.1% (e.g., 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, or 3.1%) v / v NaOH. In some embodiments, crosslinking in step (a) is achieved in the presence of 2.3% to 2.5% (e.g., 2.3%, 2.4%, or 2.5%) v / v NaOH. In some embodiments, crosslinking in step (a) is achieved in the presence of at least 2.4% NaOH.

[0056] In some embodiments, the compositions are used as dermal fillers. The disclosed compositions as dermal fillers exhibit improved fluidity and spreadability when administered as a filler under a patient's skin. When used as a dermal filler, the disclosed compositions also exhibit a more immediate and desirable "lift" due to the increased firmness of the gel. Furthermore, the disclosed hydrogels have low extrusion forces (e.g., less than 12 Newtons (12 N), e.g., 11 N, 10 N, 9 N, 8 N, 7 N, 6 N, or even lower), minimizing clogging even when using fine (e.g., 25-30 gauge, e.g., 25 gauge or 27 gauge) needles. Furthermore, the disclosed hydrogels are ready-to-use, i.e., they do not require reconstitution before administration. HA-based filler products can be tested for firmness (G') and flexibility (xStrain) using rheometry. The G' value reflects the gel strength (stiffness) of the product. xStrain is a measure of the flexibility of the gel (how much strain the gel can withstand and still be reversible). In some embodiments, the disclosed hydrogels exhibit G' values ​​of 20 Pa to 200 Pa (e.g., 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 Pa). In some embodiments, the disclosed hydrogels exhibit G' values ​​of 30 Pa to 100 Pa (e.g., 30, 40, 50, 60, 70, 80, 90, or 100 Pa). In some embodiments, the disclosed hydrogels exhibit G' values ​​of 40 Pa to 80 Pa (e.g., 40, 50, 60, 70, or 80 Pa). In some embodiments, the disclosed hydrogels exhibit xStrain values ​​of 250% to 500% (e.g., 250%, 260%, 270%, 280%, 290%, 300%, 310%, 320%, 330%, 350%, 360%, 370%, 380%, 390%, 400%, 410%, 420%, 430%, 450%, 460%, 470%, 480%, 490%, or 500%).In some embodiments, the disclosed hydrogels exhibit xStrain values ​​of 300% to 400% (e.g., 300%, 310%, 320%, 330%, 350%, 360%, 370%, 380%, 390%, or 400%). In some embodiments, the disclosed hydrogels exhibit xStrain values ​​of 310% to 380% (e.g., 310%, 320%, 330%, 350%, 360%, 370%, or 380%).

[0057] As used herein, the term "vacuum conditions" refers to conditions having an atmospheric pressure of 30 mmHg or less. In some embodiments, the vacuum conditions are achieved by a vacuum homogenizer. In some embodiments, the vacuum conditions also include a temperature of 68-72°C (e.g., 68, 69, 70, 71, or 72°C).

[0058] In some embodiments, the cross-linked glycosaminoglycans and the free glycosaminoglycans are composed of the same type of glycosaminoglycan. In some embodiments, the cross-linked glycosaminoglycans are composed of a first type of glycosaminoglycan, and the free glycosaminoglycans are composed of a second type of glycosaminoglycan, and the first type of glycosaminoglycan and the second type of glycosaminoglycan are different. In some embodiments, the first and second glycosaminoglycans are HA.

[0059] In some embodiments, the glycosaminoglycan is gamma irradiated. In some embodiments, the glycosaminoglycan is not gamma irradiated. In some embodiments, the glycosaminoglycan is heat sterilized.

[0060] In some embodiments, the composition has a concentration of 5-20 mg / ml (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg / ml), 5-15 mg / ml (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 mg / ml), 10-20 mg / ml (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 mg / ml), 10 to 30 mg / ml (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 mg / ml), 10 to 35 mg / ml (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 mg / ml), 10 to 40 mg / ml (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 mg / ml), 15 to 17 mg / ml (e.g., 15, 16, or 17 mg / ml), 15 to 20 mg / ml (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 mg / ml), 15 to 30 mg / ml (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 , 25, 26, 27, 28, 29, or 30 mg / ml), 15-35 mg / ml (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 mg / ml), or 15-40 mg / ml (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 mg / ml) cross-linked glycosaminoglycan (e.g., HA) hydrogel.

[0061] In some embodiments, glycosaminoglycans are crosslinked with one or more crosslinkers, such as multifunctional crosslinkers. The one or more multifunctional crosslinkers are individually selected from the group consisting of divinyl sulfone, multiepoxide, and diepoxide. In certain embodiments, the multifunctional crosslinkers are individually selected from the group consisting of 1,4-butanediol diglycidyl ether (BDDE), 1,2-ethanediol diglycidyl ether (EDDE), and diepoxyoctane. In some embodiments, the crosslinker is 1,4-butanediol diglycidyl ether (BDDE). Thus, in some embodiments, the crosslinks comprise ether bonds. In some embodiments, the concentration of BDDE for crosslinking is 0.001 to 0.05 g (e.g., 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, or 0.05 g) of BDDE per gram of glycosaminoglycan.

[0062] Additionally or alternatively, the crosslinker can comprise or consist of (i) a spacer group and (ii) a linking group formed by the reaction of the functional group of the crosslinker with the carboxylic acid group of the GAG. The spacer group can comprise, for example, hyaluronic acid tetrasaccharide, hyaluronic acid hexasaccharide, trehalose, lactose, maltose, sucrose, cellobiose, or raffinose residues. Thus, crosslinking can be achieved using non-carbohydrate-based di- or poly-nucleophilic crosslinkers, such as hexamethylenediamine (HMDA), or carbohydrate-based di- or poly-nucleophilic crosslinkers, such as diaminotrehalose (DATH), with glycosaminoglycans.

[0063] In some embodiments, the composition comprises 20% to 30% (20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30%), 20% to 35% (20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, or 35%), 20% to 40% (20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, or 35%), or 20% to 40% (20%, 21%, 22%, 23%, 24%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, or 35%) of the crosslinked glycosaminoglycan hydrogel by weight. Contains 5%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40%, or 20%-45% (20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, or 45%) free glycosaminoglycans.

[0064] In some embodiments, the composition contains 10-15 mg / ml (10, 11, 12, 13, 14, or 15 mg / ml), 10-20 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg / ml), 10-25 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 2, 23, 24, or 25 mg / ml), 10-30 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 mg / ml), 10-35 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 mg / ml), 10 to 40 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 mg / ml) ), or 10-45 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 mg / ml) of biodegradable poly(alpha-hydroxy acid) polymer. In some embodiments, the biodegradable poly(alpha-hydroxy acid) polymer is not gamma-irradiated.

[0065] In some embodiments, the cross-linked glycosaminoglycan hydrogel comprises cross-linked hyaluronic acid (HA), cross-linked heparan sulfate (HS), cross-linked heparin (HEP), cross-linked chondroitin sulfate (CS), cross-linked dermatan sulfate (DS), or cross-linked keratan sulfate (KS), or a combination thereof. In some embodiments, the cross-linked glycosaminoglycan hydrogel comprises cross-linked hyaluronic acid (HA).

[0066] In some embodiments, the free glycosaminoglycans comprise hyaluronic acid (HA), heparan sulfate (HS), heparin (HEP), chondroitin sulfate (CS), dermatan sulfate (DS), or keratan sulfate (KS), or a combination thereof. In some embodiments, the free glycosaminoglycans comprise hyaluronic acid (HA).

[0067] In some embodiments, the cross-linked glycosaminoglycan hydrogel comprises a cross-linked hyaluronic acid hydrogel and the free glycosaminoglycan comprises free hyaluronic acid.

[0068] In some embodiments, the biodegradable poly(α-hydroxy acid) polymer comprises poly-L-lactic acid (PLLA), poly-D-lactic acid (PLDA), polyglycolic acid (PLGA), or poly(diol) citrate (PDC), or a combination thereof. In some embodiments, the biodegradable poly(α-hydroxy acid) polymer comprises poly-L-lactic acid (PLLA).

[0069] In certain embodiments, the composition comprises hyaluronic acid (HA) gel crosslinked with 1,4-butanediol diglycidyl ether (BDDE), free HA (i.e., uncrosslinked; approximately 35%), phosphate buffer, approximately 0.7% sodium chloride, and free poly-L-lactic acid (PLLA) (i.e., uncrosslinked).

[0070] In some embodiments, the crosslinked glycosaminoglycan hydrogel has a size of about 60-90 (e.g., about 60, 65, 70, 75, 80, 85, or 90) microns. In some embodiments, the term "about" refers to plus or minus 10% of a given value.

[0071] In some embodiments, the composition further comprises 1 mg / ml to 15 mg / ml (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 mg / ml, or any value therebetween) of a local anesthetic selected from lidocaine, bupivacaine, articaine, etidocaine, or carbocaine. In certain embodiments, the composition comprises 1 mg / ml and 15 mg / ml of lidocaine.

[0072] In some embodiments, the composition further comprises 0.1% to 1.5% (e.g., 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, or 1.5%) sodium chloride.

[0073] In some embodiments, the composition has an osmolality of about 250 mOsm / kg to 350 mOsm / kg (e.g., about 250 mOsm / kg, 275 mOsm / kg, 300 mOsm / kg, 325 mOsm / kg, or 350 mOsm / kg).

[0074] In some embodiments, the composition further comprises 9 mM to 20 mM (e.g., 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mM) phosphate buffer.

[0075] In some embodiments, the composition is a sterile composition. In some embodiments, the composition is an injectable composition. In some embodiments, the composition is packaged as a pre-filled syringe. In some embodiments, the composition can be used to treat fine lines and wrinkles, among other indications.

[0076] The disclosed compositions can be used in methods for treating wrinkles or fine lines in the skin of a subject in need of such treatment, for tightening the skin of a subject in need of such treatment, or for reducing the appearance of wrinkles or fine lines in the skin of a subject. Such methods generally may include administering the disclosed compositions to a subject via injection (e.g., subcutaneous, subcutaneous, intradermal, transdermal, intramuscular, etc.). The injection may utilize a fine needle, such as a 25-30 gauge (G) (e.g., 25, 26, 27, 28, 29, or 30 G) needle. The needle may be about 0.5 inches or longer in length. In some embodiments, the injection may be administered through a cannula.

[0077] Methods for preparing hydrogel compositions Another aspect of the present disclosure is directed to a method for preparing the compositions disclosed herein. In some embodiments, the process for making the composition requires a higher concentration of crosslinker, a higher concentration of sodium chloride, and a higher concentration of phosphate buffer compared to a comparable filler composition. In some embodiments, the process also includes an additional purification / precipitation step before adding the free glycosaminoglycans and free biodegradable poly(α-hydroxy acid) polymers. An exemplary flow diagram illustrating an embodiment of the manufacturing process is provided in the figure.

[0078] Another aspect of the present disclosure is a method for preparing a composition, comprising: (a) cross-linking glycosaminoglycans in the presence of 2% to 4% v / v (e.g., 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, or 4%) NaOH, thereby cross-linking glycosaminoglycans; (b) filtering the crosslinked glycosaminoglycan hydrogel using a 60-90 micron filter; and (c) adding 20% ​​to 50% (e.g., 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 1109%, 1110%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 1 (d) adding 9%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% free glycosaminoglycan to the composition of step (c) under vacuum conditions at 5 to 60 mg / ml (5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50%, or 51%). and adding a biodegradable poly(alpha-hydroxy acid) polymer (4, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 mg / ml) to the glycosaminoglycan hydrogel, wherein the biodegradable poly(alpha-hydroxy acid) polymer is not crosslinked to the glycosaminoglycan hydrogel.

[0079] In some embodiments, crosslinking in step (a) is achieved in the presence of 2.3% to 3.1% (e.g., 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, or 3.1%) v / v NaOH. In some embodiments, crosslinking in step (a) is achieved in the presence of 2.3% to 2.5% (e.g., 2.3%, 2.4%, or 2.5%) v / v NaOH. In some embodiments, crosslinking in step (a) is achieved in the presence of at least 2.4% NaOH.

[0080] In some embodiments, the process further comprises swelling the cross-linked glycosaminoglycan hydrogel under vacuum conditions between steps (a) and (b).

[0081] In some embodiments, the vacuum conditions are achieved by a vacuum homogenizer. In some embodiments, the vacuum conditions also include a temperature of 68-72°C (e.g., 68, 69, 70, 71, or 72°C).

[0082] In some embodiments, the cross-linked glycosaminoglycans and the free glycosaminoglycans are composed of the same type of glycosaminoglycans, hi some embodiments, the cross-linked glycosaminoglycans are composed of a first type of glycosaminoglycans and the free glycosaminoglycans are composed of a second type of glycosaminoglycans, and the first type of glycosaminoglycans and the second type of glycosaminoglycans are different.

[0083] In some embodiments, the glycosaminoglycan is gamma irradiated. In some embodiments, the glycosaminoglycan is not gamma irradiated.

[0084] In some embodiments, the process further comprises precipitating and washing the filtered crosslinked glycosaminoglycan hydrogel from step (b) using ethanol. In some embodiments, the washed crosslinked glycosaminoglycan hydrogel is dried and swollen (rehydrated) prior to step (c).

[0085] In some embodiments, the process further comprises sterilizing the composition after step (d). In some embodiments, sterilization is achieved by autoclaving the composition. In some embodiments, sterilization is achieved by UV treatment.

[0086] In some embodiments, the crosslinking in step (a) is carried out at an ambient temperature of 21° C. to 25° C. (e.g., 21° C., 22° C., 23° C., 24° C., or 25° C.).

[0087] In some embodiments, crosslinking in step (a) is accomplished in 16 to 30 hours (e.g., 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 hours).

[0088] In some embodiments, the composition is at a concentration of 15-17 mg / ml (e.g., 15, 16, or 17 mg / ml), 15-20 mg / ml (e.g., 15, 16, 17, 18, 19, or 20 mg / ml), 15-25 mg / ml (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 mg / ml), 15-30 mg / ml (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 mg / ml). ml), 15-35 mg / ml (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 mg / ml), or 15-40 mg / ml (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 mg / ml) crosslinked glycosaminoglycan hydrogels.

[0089] In some embodiments, the glycosaminoglycans are cross-linked using a cross-linking agent. In some embodiments, the cross-linking agent is 1,4-butanediol diglycidyl ether (BDDE). In some embodiments, the concentration of BDDE for cross-linking is 0.001 to 0.05 g (e.g., 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, or 0.05 g) of BDDE per gram of glycosaminoglycan.

[0090] In some embodiments, the composition comprises 20% to 30% (20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30%), 20% to 35% (20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, or 35%), 20% to 40% (20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30%) of the total weight of the composition. %, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40%, or 20%-45% (20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, or 45%) of free glycosaminoglycan hydrogel.

[0091] In some embodiments, the composition contains 10-15 mg / ml (10, 11, 12, 13, 14, or 15 mg / ml), 10-20 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg / ml), 10-25 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 2, 23, 24, or 25 mg / ml), 10-30 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 mg / ml), 10-35 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 mg / ml), 10 to 40 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 mg / ml) ), or 10-45 mg / ml (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 mg / ml) of biodegradable poly(alpha-hydroxy acid) polymer. In some embodiments, the biodegradable poly(alpha-hydroxy acid) polymer is not gamma-irradiated.

[0092] In some embodiments, the cross-linked glycosaminoglycans and the free glycosaminoglycans are composed of the same type of glycosaminoglycans, hi some embodiments, the cross-linked glycosaminoglycans are composed of a first type of glycosaminoglycans and the free glycosaminoglycans are composed of a second type of glycosaminoglycans, and the first type of glycosaminoglycans and the second type of glycosaminoglycans are different.

[0093] In some embodiments, the cross-linked glycosaminoglycan hydrogel comprises cross-linked hyaluronic acid (HA), cross-linked heparan sulfate (HS), cross-linked heparin (HEP), cross-linked chondroitin sulfate (CS), cross-linked dermatan sulfate (DS), or cross-linked keratan sulfate (KS), or a combination thereof.

[0094] In some embodiments, the free glycosaminoglycans comprise hyaluronic acid (HA), heparan sulfate (HS), heparin (HEP), chondroitin sulfate (CS), dermatan sulfate (DS), or keratan sulfate (KS), or a combination thereof.

[0095] In some embodiments, the biodegradable poly(α-hydroxy acid) polymer comprises poly-L-lactic acid (PLLA), poly-D-lactic acid (PLDA), polyglycolic acid (PLGA), or poly(diol) citrate (PDC), or a combination thereof.

[0096] In some embodiments, the crosslinked glycosaminoglycan hydrogel has a size of about 60-90 (e.g., about 60, 65, 70, 75, 80, 85, or 90) microns. As used herein, the term "about" refers to plus or minus 10% of a given value.

[0097] In some embodiments, the biodegradable poly(alpha-hydroxy acid) polymer has a size of about 30-80 (e.g., about 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80) microns. In some embodiments, the biodegradable poly(alpha-hydroxy acid) polymer has a size of about 40-63 (e.g., about 40, 45, 50, 55, 60, or 63) microns.

[0098] In some embodiments, the composition further comprises 0.1% to 1.5% (e.g., 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, or 1.5%) sodium chloride.

[0099] In some embodiments, the composition has an osmolality of about 250 mOsm / kg to 350 mOsm / kg (e.g., about 250 mOsm / kg, 275 mOsm / kg, 300 mOsm / kg, 325 mOsm / kg, or 350 mOsm / kg).

[0100] In some embodiments, the composition further comprises 9 mM to 20 mM (e.g., 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mM) phosphate buffer.

[0101] In some embodiments, the composition is a sterile composition. In some embodiments, the composition is an injectable composition. In some embodiments, the composition is packaged as a pre-filled syringe. In some embodiments, the composition can be used to treat fine lines and wrinkles, among other indications. [Example]

[0102] An exemplary list of some of the components of the disclosed compositions is shown in Table 1 below.

[0103] Table 1: Exemplary compositions TIFF2025530423000002.tif185160

[0104] Table 2 provides further details about the process steps that can be used and adjusted to prepare the disclosed compositions. These methods are non-limiting and may be adjusted as needed to take into account the desired properties of the resulting composition.

[0105] Table 2: Exemplary methods for preparing compositions TIFF2025530423000003.tif214160

[0106] The present technology is not limited to the specific embodiments described in this application, which are intended as single illustrations of individual aspects of the technology. Many modifications and variations of the present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing description. Such modifications and variations are intended to fall within the scope of the present technology. It is understood that the present technology is not limited to particular methods, reagents, compounds, compositions, or systems, which may, of course, vary. It is also understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

Claims

1. (a) 10-45 mg / ml of a cross-linked glycosaminoglycan hydrogel; (b) 20% to 50% free, non-crosslinked glycosaminoglycans by weight of the crosslinked glycosaminoglycan hydrogel; (c) 10 to 50 mg / ml of a biodegradable poly(α-hydroxy acid) polymer, wherein the biodegradable poly(α-hydroxy acid) polymer is not crosslinked to the glycosaminoglycan hydrogel or the free, uncrosslinked glycosaminoglycan. A composition comprising:

2. 1. A composition prepared by a process comprising: (a) cross-linking glycosaminoglycans in the presence of 2% to 4% v / v NaOH, thereby obtaining a cross-linked glycosaminoglycan hydrogel; (b) filtering the cross-linked glycosaminoglycan hydrogel using a 60-90 micron filter; (c) adding 20% ​​to 50% of free non-crosslinked glycosaminoglycan by weight of the crosslinked glycosaminoglycan hydrogel to the filtered crosslinked glycosaminoglycan hydrogel; (d) adding 10-50 mg / ml of a biodegradable poly(α-hydroxy acid) polymer to the composition from (c) under vacuum conditions, wherein the biodegradable poly(α-hydroxy acid) polymer is not crosslinked to the glycosaminoglycan hydrogel; A composition comprising:

3. 3. The composition of claim 2, wherein the process further comprises swelling the cross-linked glycosaminoglycan hydrogel under vacuum conditions between (a) and (b).

4. 4. The composition of claim 2 or claim 3, wherein the process further comprises precipitating and washing the filtered crosslinked glycosaminoglycan hydrogel from (b) using ethanol.

5. The composition of any one of claims 2 to 4, further comprising sterilizing the composition after (d).

6. 6. The composition of any one of claims 1 to 5, comprising 15 to 20 mg / ml, 15 to 25 mg / ml, 15 to 30 mg / ml, 15 to 35 mg / ml, or 15 to 40 mg / ml of the cross-linked glycosaminoglycan hydrogel.

7. 7. The composition of claim 1, comprising 20% ​​to 30%, 20% to 35%, 20% to 40%, or 20% to 45% free, uncrosslinked glycosaminoglycans by weight of the crosslinked glycosaminoglycan hydrogel.

8. 8. The composition of any one of claims 1 to 7, comprising 10 to 15 mg / ml, 10 to 20 mg / ml, 10 to 25 mg / ml, 10 to 30 mg / ml, 10 to 35 mg / ml, 10 to 40 mg / ml, or 10 to 50 mg / ml of the biodegradable poly(alpha hydroxy acid) polymer.

9. 9. The composition of any one of claims 1 to 8, wherein the cross-linked glycosaminoglycan hydrogel comprises cross-linked hyaluronic acid (HA), cross-linked heparan sulfate (HS), cross-linked heparin (HEP), cross-linked chondroitin sulfate (CS), cross-linked dermatan sulfate (DS), or cross-linked keratan sulfate (KS), or a combination thereof.

10. 10. The composition of any one of claims 1 to 9, wherein the free glycosaminoglycans comprise hyaluronic acid (HA), heparan sulfate (HS), heparin (HEP), chondroitin sulfate (CS), dermatan sulfate (DS), or keratan sulfate (KS), or a combination thereof.

11. 11. The composition of any one of claims 1 to 10, wherein the biodegradable poly(α-hydroxy acid) polymer comprises poly-L-lactic acid (PLLA), poly-D-lactic acid (PLDA), polyglycolic acid (PLGA), or poly(diol) citrate (PDC), or a combination thereof.

12. 12. The composition of any one of claims 1 to 11, wherein the crosslinked glycosaminoglycan hydrogel has a size of about 60 to 90 microns, and optionally the biodegradable poly(alpha-hydroxy acid) polymer is about 40 to 63 microns.

13. The composition of any one of claims 1 to 12, wherein the cross-linked glycosaminoglycans are cross-linked by one or more multifunctional cross-linkers.

14. 14. The composition of claim 13, wherein the one or more multifunctional crosslinkers are selected from the group consisting of divinyl sulfones, multiepoxides, and diepoxides.

15. 14. The composition of claim 13, wherein the one or more multifunctional crosslinkers are selected from the group consisting of 1,4-butanediol diglycidyl ether (BDDE), 1,2-ethanediol diglycidyl ether (EDDE), and diepoxyoctane.

16. 13. The composition of any one of claims 1 to 12, wherein the cross-linked glycosaminoglycans are cross-linked by (i) a non-carbohydrate-based di- or poly-nucleophilic cross-linker, or (ii) a carbohydrate-based di- or poly-nucleophilic cross-linker.

17. 17. The composition of claim 16, wherein the non-carbohydrate-based di- or poly-nucleophilic crosslinker is hexamethylenediamine (HMDA).

18. 17. The composition of claim 16, wherein the carbohydrate-based di- or poly-nucleophilic crosslinker is diaminotrehalose (DATH).

19. The composition according to any one of claims 1 to 18, which is an injectable composition.

20. A method for improving skin quality in a subject in need of such treatment, comprising administering to the subject a composition according to any one of claims 1 to 18.

21. 1. A method for preparing a composition, comprising: (a) cross-linking glycosaminoglycans in the presence of 2% to 4% v / v NaOH, thereby obtaining a cross-linked glycosaminoglycan hydrogel; (b) filtering the cross-linked glycosaminoglycan hydrogel using a 60-90 micron filter; (c) adding 20% ​​to 50% free glycosaminoglycan by weight of the composition to the filtered crosslinked glycosaminoglycan hydrogel; (d) adding 10-50 mg / ml of a biodegradable poly(α-hydroxy acid) polymer to the composition of step (c) under vacuum conditions, wherein the biodegradable poly(α-hydroxy acid) polymer is not crosslinked to the glycosaminoglycan hydrogel; A method comprising:

22. 22. The method of claim 21, further comprising swelling the cross-linked glycosaminoglycan hydrogel under vacuum conditions between (a) and (b).

23. 23. The method of claim 21 or claim 22, further comprising precipitating and washing the filtered crosslinked glycosaminoglycan hydrogel from (b) using ethanol.

24. The method of any one of claims 21 to 23, further comprising sterilizing the composition after (d).

25. 25. The method of any one of claims 21 to 24, wherein the composition comprises 15 to 20 mg / ml, 15 to 25 mg / ml, 15 to 30 mg / ml, 15 to 35 mg / ml, or 15 to 40 mg / ml of the cross-linked glycosaminoglycan hydrogel.

26. 26. The method of any one of claims 21 to 25, wherein the composition comprises 20% to 30%, 20% to 35%, 20% to 40%, or 20% to 45% free glycosaminoglycans by weight of the composition.

27. 27. The method of any one of claims 21 to 26, wherein the composition comprises 10-15 mg / ml, 10-15 mg / ml, 10-20 mg / ml, 10-25 mg / ml, 10-30 mg / ml, 10-35 mg / ml, 10-40 mg / ml, or 10-45 mg / ml of the biodegradable poly(alpha hydroxy acid) polymer.

28. 28. The method of any one of claims 21 to 27, wherein the cross-linked glycosaminoglycan hydrogel comprises cross-linked hyaluronic acid (HA), cross-linked heparan sulfate (HS), cross-linked heparin (HEP), cross-linked chondroitin sulfate (CS), cross-linked dermatan sulfate (DS), or cross-linked keratan sulfate (KS), or a combination thereof.

29. 29. The method of any one of claims 21 to 28, wherein the free glycosaminoglycans comprise hyaluronic acid (HA), heparan sulfate (HS), heparin (HEP), chondroitin sulfate (CS), dermatan sulfate (DS), or keratan sulfate (KS), or a combination thereof.

30. 30. The method of any one of claims 21-29, wherein the biodegradable poly(α-hydroxy acid) polymer comprises poly-L-lactic acid (PLLA), poly-D-lactic acid (PLDA), polyglycolic acid (PLGA), or poly(diol) citrate (PDC), or a combination thereof.

31. 31. The method of any one of claims 21 to 30, wherein the crosslinked glycosaminoglycan hydrogel has a size of about 60 to 90 microns, and optionally the biodegradable poly(alpha-hydroxy acid) polymer is about 40 to 63 microns.

32. 32. The method of any one of claims 21 to 31, wherein the cross-linked glycosaminoglycans are cross-linked by one or more multifunctional cross-linkers.

33. 33. The method of claim 32, wherein the one or more multifunctional crosslinkers are selected from the group consisting of divinyl sulfones, multiepoxides, and diepoxides.

34. 33. The method of claim 32, wherein the one or more multifunctional crosslinkers are selected from the group consisting of 1,4-butanediol diglycidyl ether (BDDE), 1,2-ethanediol diglycidyl ether (EDDE), and diepoxyoctane.

35. 32. The method of any one of claims 21 to 31, wherein the cross-linked glycosaminoglycans are cross-linked by (i) a non-carbohydrate-based di- or poly-nucleophilic cross-linker, or (ii) a carbohydrate-based di- or poly-nucleophilic cross-linker.

36. 36. The method of claim 35, wherein the non-carbohydrate-based di- or poly-nucleophilic crosslinker is hexamethylenediamine (HMDA).

37. 36. The method of claim 35, wherein the carbohydrate-based di- or poly-nucleophilic cross-linker is diaminotrehalose (DATH).

38. 38. The method of any one of claims 21 to 37, further comprising formulating the composition as an injectable composition.

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