Compositions and methods for cold slurries with hyaluronic acid

Abstract

Disclosed herein are compositions comprising an amount of water; hyaluronic acid; and a first excipient, wherein the composition is configured to form a flowable cold slurry when the composition is exposed to a temperature of 0 DEG C or less.

Description

Technical Field

[0001] This application claims priority under 35 U.S.C. §119(c) to U.S. Serial No. 63 / 309,291, filed on February 11, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] The present disclosure generally relates to compositions and methods for making biomaterials that form flowable and / or injectable cold slurries. More particularly, the present invention preferably relates to formulations of biocompatible solutions comprising liquids (e.g., saline, water, or phosphate buffered saline), glycerol, hyaluronic acid, poloxamer, and optional lipids. Background Art

[0003] Cold slurries (e.g., ice slurries) are known in the art as compositions made of sterile ice particles of water, varying amounts of excipients or additives such as antifreeze, hydrotropic molecules, and optionally one or more active pharmaceutical ingredients, as described in U.S. Application Serial No. 15 / 505,042 ("the '042 Application"; Publication No. US2017 / 0274011), the disclosures relating to the formulation of cold slurry compositions being incorporated herein by reference in their entirety. Cold slurries of the prior art may be delivered to a subject, preferably a tissue of a human patient, preferably via injection, for prophylactic, therapeutic, or aesthetic purposes to induce selective or non-selective cryotherapy and / or cryolipolysis. Injectable cold slurries may be used to treat a variety of conditions requiring inhibition of nerve conduction. For example, U.S. Application Serial No. 15 / 505,039 ("the '039 Application"; Publication No. US2017 / 0274078), the disclosure of which is incorporated herein by reference in its entirety for its disclosure relating to reversible inhibition of nerve conduction, discloses the use of a slurry to induce reversible degeneration of nerves (e.g., by Wallerian degeneration) by causing crystallization of lipids in the myelin sheath of the nerves. The '039 Application also discloses the use of an injectable cold slurry to treat a variety of other conditions requiring inhibition of somatic or autonomic nerves, including motor cramps, hypertension, hyperhidrosis, and urinary incontinence.

[0004] Methods for preparing cold slurries are shown in U.S. Application Serial No. 16 / 080,092 (“the '092 Application”; Publication No. US2019 / 0053939), which is incorporated herein by reference in its entirety. However, the method disclosed in the '092 Application requires the point of care to manufacture the cold slurry by installing a medical ice slurry production system. The technology also requires the point of care to take measures to maintain the sterility of the cold slurry during manufacture and prior to application. Alternative methods for preparing cold slurries are disclosed in U.S. Patent No. 11,241,330 and International Publication No. WO2022 / 261494A1 (“'494PCT”). The disclosures regarding the manufacturing methods in WO 2022 / 211904 A1 (“'904PCT”) are incorporated herein by reference. The disclosures regarding cold slurry compositions in the '494PCT are incorporated herein by reference. The disclosure in this application is compatible with the methods and systems disclosed in International Publication No. WO 2017 / 147367A1 (“'367 PCT”) and '904 PCT.

[0005] '904 PCT discloses a method for easily transporting sterile biological materials to the point of care using standard transport technology, wherein the biological material can be converted into a flowable and injectable cold slurry at the point of care without requiring manufacturing equipment available at the point of care and without compromising the sterility of the biological material at the point of care. The disclosure in this application is compatible with the methods and systems disclosed in '904 PCT.

[0006] There is a need for compositions and methods that allow for simple transport, storage, and preparation of flowable and / or injectable cold slurries at the clinical point of care without compromising the sterility of the biological material (e.g., the solution to be converted to the cold slurry) during preparation, without requiring specialized manufacturing equipment available at the point of care, and without compromising the sterility of the biological material at the point of care. The present disclosure meets this need by providing improved cold slurry compositions and preparation methods that allow a biocompatible solution to be received at the point of care in a container that is easily transported and stored, which can be placed in a standard freezer and optionally further physically agitated to convert the biocompatible solution into a therapeutic substance, such as a flowable and / or injectable cold slurry. The present disclosure describes compositions and methods that can provide sufficient and consistent amounts of ice particles after exposure to freezing temperatures and reliably allow injection of the cold slurry through a syringe needle. Summary of the invention

[0007] In certain aspects, the present disclosure provides a composition comprising an amount of water, hyaluronic acid, and a first excipient, wherein the composition is configured to form a flowable cold slurry comprising a plurality of ice crystals when the composition is exposed to a temperature of 0°C or less.

[0008] In certain embodiments, the composition further comprises a water-soluble surfactant. In other embodiments, the water-soluble surfactant is a poloxamer molecule. In certain embodiments, wherein the composition comprises a plurality of poloxamer molecules. In certain embodiments, the composition comprises poloxamer particles, and wherein the poloxamer particles comprise a plurality of poloxamer molecules. In other embodiments, the poloxamer particles are micelles. In certain embodiments, the poloxamer molecules are selected from poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 236, poloxamer 237, poloxamer 238, poloxamer 239, poloxamer 240, poloxamer 241, poloxamer 242, poloxamer 243, poloxamer 244, poloxamer 245, poloxamer 246, poloxamer 247, poloxamer 248, poloxamer 249, poloxamer 250, poloxamer 251, poloxamer 252, poloxamer 253, poloxamer 254, poloxamer 255, poloxamer 256, poloxamer 257, poloxamer 258, poloxamer 259, poloxamer 260, poloxamer 261, poloxamer 262, poloxamer 263, poloxamer 264, poloxamer 26 In some embodiments, the poloxamer is poloxamer 407, and the concentration of poloxamer 407 is about 0.1% (weight / weight) to 10% (weight / weight). In some embodiments, the concentration of poloxamer 407 in the composition is about 0.5% (weight / weight). As used herein, (weight / weight) and (weight / volume) are interchangeable.

[0009] In certain embodiments, the composition further comprises a first excipient selected from the group consisting of salts, ions, lactated Ringer's solution, sugars, biocompatible surfactants, polyols, and combinations thereof.

[0010] In certain embodiments, the first excipient is glycerol. In some embodiments, the concentration of glycerol in the composition is about 12% (w / w) to 25% (w / w). In certain embodiments, the concentration of glycerol in the composition is about 19% (w / w).

[0011] In certain embodiments, the composition further comprises a second excipient. In some embodiments, the second excipient is sodium chloride or sodium phosphate to form saline or phosphate buffered saline.

[0012] In certain embodiments, the composition further comprises a third excipient. In some embodiments, the third excipient is a water-insoluble substance. In some embodiments, the water-insoluble substance is a lipid. In some embodiments, the lipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoylphosphatidyl (DPPC), dicetyl phosphate (DCP), L-α-phosphatidylcholine (soybean PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylglycerol (PG).

[0013] In certain aspects, the compositions disclosed herein are configured to form a plurality of ice crystals when the composition is exposed to a temperature of about -25°C to about -5°C.

[0014] In certain aspects, the compositions disclosed herein are configured to have an injection force of less than about 30 lbf when injected through a 16G needle, a 17G needle, a 18G needle, a 19G needle, a 20G needle, a 22G needle, a 23G needle, or a 24G needle.

[0015] In certain aspects, the compositions disclosed herein are configured to have an injection force of less than about 30 lbf when injected through a 17G needle or an 18G needle.

[0016] In certain aspects, methods are provided for preparing a cold slurry for administration to a patient at a clinical point of care. In certain embodiments, the method comprises: preparing a composition comprising hyaluronic acid and an amount of water; adding a first excipient to the composition, wherein the excipient comprises a water-soluble surfactant; wherein the composition is configured to form a cold slurry comprising a plurality of ice particles when the composition is cooled to a temperature below about 0°C.

[0017] In some aspects, the water-soluble surfactant is a hydrotropic molecule. In some embodiments, the hydrotropic molecule is a poloxamer molecule. In some embodiments, the poloxamer molecule is selected from poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 236, poloxamer 237, poloxamer 238, poloxamer 239, poloxamer 240, poloxamer 241, poloxamer 242, poloxamer 243, poloxamer 244, poloxamer 245, poloxamer 246, poloxamer 247, poloxamer 248, poloxamer 249, poloxamer 250, poloxamer 251, poloxamer 252, poloxamer 253, poloxamer 254, poloxamer 255, poloxamer 256, poloxamer 257, poloxamer 258, poloxamer 259, poloxamer 260, poloxamer 261, poloxamer 262, poloxamer 263, poloxamer 264, poloxamer 265 In some embodiments, the poloxamer is poloxamer 407, and the concentration of poloxamer 407 is about 0.1% (weight / weight) to 10% (weight / weight). In some embodiments, the concentration of poloxamer 407 in the composition is about 0.5% (weight / weight).

[0018] In certain aspects, the methods provided herein further comprise adding a second excipient to the composition, wherein the composition comprising the second excipient is configured to form a cold slurry when the composition is cooled to a temperature below about 0° C. In certain embodiments, the second excipient is selected from salts, ions, lactated Ringer's solution, sugars, biocompatible surfactants, polyols, and combinations thereof.

[0019] In certain embodiments, the second excipient is glycerol. In certain embodiments, the concentration of glycerol in the composition is about 12% (w / w) to 25% (w / w). In certain embodiments, the concentration of glycerol in the composition is about 19% (w / w).

[0020] In certain aspects, the methods provided herein further comprise adding a third excipient to the composition, wherein the composition comprising the second excipient and the third excipient is configured to form a cold slurry when the composition is cooled to a temperature below about 0° C. In certain embodiments, the third excipient is sodium chloride or sodium phosphate to form saline or phosphate buffered saline.

[0021] In certain aspects, the methods provided herein further comprise adding a fourth excipient to the composition, wherein the composition comprising the second excipient, the third excipient, and the fourth excipient is configured to form a cold slurry when the composition is cooled to a temperature below about 0°C.

[0022] In certain embodiments, the fourth excipient is a water-insoluble substance. In some embodiments, the water-insoluble substance is a lipid. In some embodiments, the lipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoylphosphatidyl (DPPC), dicetyl phosphate (DCP), L-α-phosphatidylcholine (soybean PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylglycerol (PG).

[0023] In certain aspects, the methods provided herein further comprise a composition configured to form a plurality of ice crystals when the composition is exposed to a temperature of about -25°C to about -5°C.

[0024] In certain aspects, the methods provided herein include compositions configured to have an injection force of less than about 30 lbf when injected through a 16G needle, 17G needle, 18G needle, 19G needle, 20G needle, 22G needle, 23G needle, or 24G needle. In certain embodiments, the compositions are configured to have an injection force of less than about 30 lbf when injected through a 17G needle or 18G needle.

[0025] In certain aspects, methods are provided for preparing a cold slurry for administration to a patient at a clinical point of care. In certain embodiments, the method comprises: receiving a composition comprising an antifreeze and hyaluronic acid; and cooling the composition to a temperature below about 0° C. to form a cold slurry, wherein the cold slurry comprises a plurality of ice particles. In certain embodiments, the antifreeze is glycerol.

[0026] In some aspects, the composition further comprises an amount of a poloxamer molecule. In some embodiments, the poloxamer molecule is selected from poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 236, poloxamer 237, poloxamer 238, poloxamer 239, poloxamer 240, poloxamer 241, poloxamer 242, poloxamer 243, poloxamer 244, poloxamer 245, poloxamer 246, poloxamer 247, poloxamer 248, poloxamer 249, poloxamer 250, poloxamer 251, poloxamer 252, poloxamer 253, poloxamer 254, poloxamer 255, poloxamer 256, poloxamer 257, poloxamer 258, poloxamer 259, poloxamer 260, poloxamer 261, poloxamer 262, poloxamer 263, poloxamer 264, poloxamer 265 In some embodiments, the poloxamer is poloxamer 407, and the concentration of poloxamer 407 is about 0.1% (weight / weight) to 10% (weight / weight). In some embodiments, the concentration of poloxamer 407 in the composition is about 0.5% (weight / weight).

[0027] In certain aspects, receiving the composition comprises receiving the composition within a container.

[0028] In certain embodiments, the container is a first syringe. In some embodiments, the method further comprises connecting the first syringe to the second syringe and processing the cold slurry by a reciprocating cycle, wherein the reciprocating cycle comprises pushing the cold slurry from the first syringe into the second syringe and pushing the cold slurry from the second syringe into the first syringe.

[0029] In certain aspects, the method further comprises treating the cold slurry through a second, third, or fourth reciprocating cycle.

[0030] In certain aspects, the container is a container configured for surface application.In certain embodiments, the container configured for surface application is a first tube.

[0031] In certain aspects, the method further includes connecting the first tube to the second tube and processing the slurry by a reciprocating cycle, wherein the reciprocating cycle includes pushing the cold slurry from the first tube into the second tube and pushing the cold slurry from the second tube into the first tube.

[0032] In certain aspects, the methods provided herein further comprise monitoring the temperature of the cold slurry.

[0033] In certain aspects, monitoring includes viewing a temperature sensitive indicator on a syringe or container containing the cold slurry, wherein the temperature sensitive indicator is configured to indicate the temperature of the cold slurry.

[0034] In certain embodiments, the temperature sensitive indicator is a temperature sensitive sticker.In certain embodiments, the temperature sensitive indicator provides a visual indication when the cold slurry reaches a predetermined temperature.

[0035] In certain embodiments, the predetermined temperature is about -15°C. In certain embodiments, the predetermined temperature is about -19°C to -11°C, about -18°C to -12°C, about -17°C to -13°C, or about -16°C to -14°C.

[0036] In certain aspects, the temperature sensitive indicator is configured to provide a visual indication when the cold slurry is at a temperature cooler than a predetermined temperature. In certain aspects, the temperature sensitive indicator is configured to provide a visual indication when the cold slurry is hotter than a predetermined temperature.

[0037] In certain aspects, monitoring includes checking a thermometer.

[0038] In certain aspects, monitoring includes viewing a temperature monitoring component embedded within a container containing the cold slurry. In certain embodiments, a temperature component is disposed in the container or along a fluid path.

[0039] In certain aspects, monitoring further comprises listening for an audio indicator configured to indicate when the composition reaches a predetermined temperature.

[0040] In certain aspects, the composition is terminally sterilized. In certain embodiments, the composition is terminally sterilized via autoclaving or steam sterilization. In certain embodiments, autoclaving or steam sterilization comprises subjecting the composition to a temperature of about 118° C. to 121° C. In certain embodiments, the temperature is about 118° C.

[0041] In certain aspects, a cold slurry delivery system is provided. In certain embodiments, the cold slurry delivery system includes a container containing a slurry composition, the container including a sterile barrier and a temperature indicator, wherein the container is configured to allow manual stirring of the slurry composition without breaking the sterile barrier.

[0042] In certain embodiments, the container is a syringe or a tube.

[0043] In certain aspects, the container is configured to be connected to a second container.

[0044] In certain aspects, the container and the second container are configured to be connected using a connector.

[0045] In certain aspects, the container and the second container comprise a first syringe and a second syringe, wherein the first syringe and the second syringe each comprise a male Luer assembly. In certain embodiments, the connector comprises a female Luer assembly.

[0046] In certain embodiments, the first syringe and the second syringe are connected using a female Luer assembly, and wherein the slurry composition can be moved from the first syringe to the second syringe to manually stir the slurry composition.

[0047] In certain aspects, a temperature sensitive indicator is disposed on the container.

[0048] In certain aspects, the temperature sensitive indicator comprises a temperature sensitive sticker.

[0049] In certain aspects, the temperature sensitive indicator provides a visual indication when the slurry composition reaches a predetermined temperature. In certain embodiments, the predetermined temperature is about -15°C. In certain embodiments, the predetermined temperature is about -19°C to -11°C, about -18°C to -12°C, about -17°C to -13°C, or about -16°C to -14°C.

[0050] In certain aspects, the temperature sensitive indicator provides a visual indication when the slurry composition is at a temperature cooler than a predetermined temperature.

[0051] In certain aspects, the temperature sensitive indicator provides a visual indication when the slurry composition is hotter than a predetermined temperature. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] The following figures depict illustrative embodiments of the invention.

[0053] Figure 1 is a table showing exemplary formulations for cold slurry compositions described herein.

[0054] Figure 2 Graphs showing characterization of ice content of cold slurry compositions described herein and cold slurry compositions comprising glycerol and PBS and prepared according to the methods disclosed in, for example, the '367 PCT.

[0055] Figure 3 One embodiment of a composition comprising a plurality of poloxamer micelles, glycerin, and a hyaluronic acid gel is depicted.

[0056] FIG. 4A to FIG. 4B . Figure 4A is an image depicting an exemplary configuration of two syringes to process a slurry composition in a reciprocating process between the two syringes according to certain embodiments described herein. Figure 4B A process flow diagram is shown for preparing an injectable cold slurry using a back-and-forth (BAF) process by subjecting the contents to three BAF cycles, according to certain embodiments described herein.

[0057] FIG. 5A to FIG. 5D . Figure 5A Depicted are exemplary syringe configurations including a temperature sensitive indicator that measures the temperature of the contents of the syringe, wherein the indicator displays a range of temperatures, according to embodiments described herein. Figure 5B Depicted are exemplary syringe configurations including a temperature sensitive indicator displaying a single specific temperature according to embodiments described herein. Figure 5C An exemplary syringe configuration is depicted that includes an external temperature-sensitive indicator that displays "Process Now" when the contents of the syringe reach a predetermined temperature. Figure 5D An exemplary syringe configuration is depicted that includes a plurality of temperature-sensitive indicators that display messages indicating "Wait," "Process Now," and "Discard" when the contents of the syringe reach a series of predetermined temperatures.

[0058] Figure 6 is a graph showing the injection force (lbf) as a function of freezer temperature for a slurry injected through a 17G needle into a syringe containing a composition prepared according to the present disclosure.

[0059] FIG. 7A to FIG. 7B . Fig. 7A is a graph showing the relationship between equilibrium temperature (as a proxy for ice content) and injection temperature for compositions prepared according to the present disclosure. Figure 7B is a graph showing the relationship between injection force (lbf) and injection temperature for compositions prepared in accordance with the present disclosure when injected through a 17G needle.

[0060] Figure 8 Graphs showing the relationship between injection force (lbf) and post-treatment temperature in compositions prepared in accordance with the present disclosure and subjected to one, two, three, or four BAF cycles when injected through an 18G needle.

[0061] Fig. 9 Graph showing injection force (lbf) versus post-treatment temperature for compositions prepared in accordance with the present disclosure subjected to two BAF cycles and injected through either a 17G or 18G needle.

[0062] Fig.10 is a graph showing a characterization of the ice content of the compositions described herein.

[0063] Fig.11 TABLE 1 is a table summarizing the injection force (lbf) and injection reliability of the compositions described herein when prepared according to different processing methods.

[0064] Fig.12 Graph showing the required injection force for a cold slurry composition described herein terminally sterilized using gamma radiation versus a non-gamma irradiated composition. DETAILED DESCRIPTION

[0065] The present disclosure generally relates to compositions and methods for making biomaterials that form flowable and / or injectable cold slurries. More particularly, compositions comprising water, hyaluronic acid, and at least one excipient or additive are disclosed herein. In certain embodiments, at least one excipient or additive is Pluronic TM (Also referred to as "poloxamers"). As used herein, the term "excipient" means any substance that is not itself a therapeutic agent and is used as a diluent, adjuvant and / or carrier for delivering a therapeutic agent (in this case, ice) to a subject or patient, and / or a substance added to a composition to improve its handling characteristics, stability characteristics, or storage characteristics. The terms "excipient" and "additive" are used interchangeably herein. In some embodiments, the solution may also include liposomes, lipids or other lipid structures (e.g., lamellar or non-lamellar structures, bilayer and non-bilayer structures, including lipid nanoparticles, micelles, etc.), water-insoluble substances (i.e., substances that do not dissolve in water), or water-soluble surfactants such as hydrotropic molecules (e.g., polysorbates).

[0066] In some embodiments, the flowable and / or injectable or topically applied composition comprises a significant amount of ice, which provides therapeutic benefits for various applications. For example, therapeutic applications of cold slurries are disclosed in U.S. Application Serial Nos. 16 / 288,073 and 16 / 327,266, the disclosures relating to various therapeutic applications of which are incorporated herein by reference.

[0067] In some embodiments, the final product administered via injection to a human patient or subject (e.g., a human or non-human animal that is not a patient) is a cold slurry comprising ice particles of sterile water and varying amounts of excipients / additives such as hyaluronic acid, poloxamers, and / or antifreeze agents. For example, the percentage of ice particles in the cold slurry can be less than about 10% by weight, about 10% by weight to about 20% by weight, about 20% by weight to about 30% by weight, about 30% by weight to about 40% by weight, about 40% by weight to about 60% by weight, greater than about 60% by weight, etc. of the slurry. The size of the ice particles can be optionally varied by adding, for example, water-soluble surfactants (e.g., hydrotropic molecules), poloxamers (e.g., Pluronic TM F127 or P407) and / or lipid components are controlled to allow fluidity through containers of various sizes (e.g., needle gauge sizes of about 7 to about 43). Containers of various sizes are described in U.S. Serial Application No. 15 / 505,042 (Publication No. US2017 / 0274011), and the disclosures related to containers for injection are incorporated herein by reference. In addition, other methods can be used to control the size of the ice particles to allow fluidity and / or injectability through containers of various sizes (e.g., using a filter or reciprocating the composition between two syringes). In some embodiments, the diameter of most ice particles is smaller than about half of the inner diameter of the injection cavity or container. For example, the diameter of ice particles for a 3 mm catheter can be about 1.5 mm or less.

[0068] In some embodiments, one or more excipients may be included in the cold slurry. The excipient may comprise less than about 10% volume / volume (v / v) of the cold slurry, about 10% v / v to about 20% v / v of the cold slurry, about 20% v / v to about 30% v / v, about 30% v / v to 40% v / v, and greater than about 40% v / v. A variety of added excipients may be used to change the phase transition temperature of the cold slurry (e.g., lower the freezing point), change the ice percentage of the cold slurry, change the viscosity of the cold slurry, prevent agglomeration of ice particles, prevent dendritic ice formation (i.e., crystals with a multi-branched "tree-like" morphology, such as those seen in snowflakes), keep ice particles separate, increase the thermal conductivity of the fluid phase, or improve the overall preventive, therapeutic, or aesthetic efficacy of the flowable and / or injectable cold slurry. In the compositions described herein, such excipients may include hyaluronic acid, poloxamers, polysorbates (or other water-soluble surfactants such as hydrotropes), water-insoluble substances, lipids (including lipid particles), all of which prevent agglomeration of ice particles, prevent dendritic ice formation (i.e., crystals with multi-branched "tree-like" morphology, such as those seen in snowflakes), or keep the ice particles separate so that the cold slurry is flowable and / or injectable when it is removed from the freezer.

[0069] One or more cryoprotectants can be added to sterile water as an excipient to form a cold slurry with a freezing point below 0°C (e.g., about -10°C). Lowering the freezing point of the cold slurry allows it to maintain fluidity and remain injectable while still containing an effective percentage of ice particles. Suitable cryoprotectants include salts (e.g., sodium chloride, sodium sulfobutyl ether beta-cyclodextrin), ions, lactated Ringer's solution, sugars (e.g., glucose, sorbitol, mannitol, hydroxyethyl starch, sucrose, (2-hydroxypropyl)-β-cyclodextrin, or a combination thereof), biocompatible surfactants such as glycerol (also known as glycerol / glycerin / glycerine), other polyols (e.g., polyvinyl alcohol, polyethylene glycol 300, polyethylene glycol 400, propylene glycol), other sugar alcohols or urea, etc. Other exemplary antifreeze agents are disclosed in U.S. Application Serial No. 15 / 505,042 (Publication No. US2017 / 0274011), the disclosure of which is incorporated herein in its entirety with respect to the slurry composition ingredients.

[0070] The present disclosure describes compositions that produce flowable and / or injectable cold slurry when frozen. In some embodiments, the compositions of the present disclosure do not need to be processed or operated to become flowable and / or injectable. However, in other embodiments, operations can be used to further improve fluidity and injectability or improve consistency. In some embodiments, the composition comprises a fluid with a high water content (e.g., about 70% to 80%, about 80% to 90%, or greater than about 90%), a solute (e.g., glycerol) and a suspension of hyaluronic acid for lowering the freezing point. In some embodiments, in order to further improve the fluidity and injectability of the cold slurry, the solution can include one or more of lipids, water-insoluble compounds, or water-soluble surfactants such as hydrotropic compounds (e.g., polysorbates) or poloxamer (e.g., P407). In some embodiments, the solution also includes other excipients, such as sodium chloride or sodium phosphate forming such as saline or phosphate buffered saline.

[0071] The present disclosure provides a variety of compositions. In some embodiments, the composition comprises an effective amount of hyaluronic acid for producing a flowable and / or injectable cold slurry. It is not intended to be bound by any particular theory, it is believed that when the solution is exposed to freezing temperatures (e.g., about -20°C to -15°C, about -15°C to -10°C, about -10°C to -5°C, or about -10°C in some embodiments), the hyaluronic acid in the composition promotes the formation of small ice crystals. In some embodiments, the composition comprising hyaluronic acid is converted into a flowable and / or injectable cold slurry with ice particles when placed in a standard freezer, without the need to apply any mechanical stirring or additional processing to the cold slurry.

[0072] In some embodiments, the composition further comprises an effective amount of a water-soluble surfactant such as a hydrotropic compound (e.g., polysorbate) for producing a flowable and / or injectable ice slurry. In some embodiments, the water-soluble surfactant is a poloxamer (or Pluronic TM) molecules. In some embodiments, any surfactant having a hydrophilic-lipophilic balance (HLB) value greater than 10 is considered water-soluble. In some embodiments, the concentration of the water-soluble surfactant in the composition is about 0.01% (w / w) to 0.5% (w / w), about 0.5% (w / w) to 1% (w / w), about 1% (w / w) to 2% (w / w), about 2% (w / w) to 5% (w / w), about 5% (w / w) to 10% (w / w), or greater than about 10% (w / w). Without intending to be bound by any particular theory, it is believed that when the composition is exposed to freezing temperatures (e.g., about -5°C or lower), the water-soluble surfactant also acts to prevent ice particles from growing too large; large ice particles may prevent the composition from becoming flowable or injectable.

[0073] In some embodiments, the composition also includes one or more of hyaluronic acid, optional hydrotropic molecules and lipids or non-water-soluble particles for producing an effective amount of a flowable and / or injectable cold slurry in the form of an emulsion. In some embodiments, an emulsion is any composition comprising lipids as described herein. In some embodiments, the lipid in the composition is assembled into lipid particles with one or more forms (e.g., lamellar or non-lamellar structure, double-layer and non-double-layer structure, including liposomes, lipid nanoparticles, micelles, etc.) known in the art. The lipid particle morphology of the present disclosure can be determined by any method known in the art, such as by cryo-TEM. In some embodiments, the diameter of the lipid particles in the composition is about 5 μm to about 300 μm. In some embodiments, the diameter of the lipid particles is about 250 μm. In some embodiments, the diameter of the lipid particles in the composition is about 5 μm to 20 μm, or a diameter of about 8 μm to 14 μm. Without intending to be bound by any particular theory, it is believed that when the composition is exposed to freezing temperatures, the lipid or water-insoluble particles prevent the ice particles from growing so large that the composition is no longer flowable or injectable.

[0074] In some embodiments, hyaluronic acid is of natural or synthetic origin. In some embodiments, the molecular weight of hyaluronic acid is about 250kDa to 5,000kDa. In some embodiments, the molecular weight of hyaluronic acid is about 1,000kDa. In some embodiments, the concentration of hyaluronic acid in the composition is about 0.01% (weight / weight) to 2% (weight / weight). In some embodiments, the concentration of hyaluronic acid in the composition is about 0.1% (weight / weight) to 1% (weight / weight). In some embodiments, the concentration of hyaluronic acid in the composition is about 0.5% (weight / weight) to 1.5% (weight / weight). In some embodiments, the concentration of hyaluronic acid in the composition is about 0.05% (weight / weight) to 0.75% (weight / weight). In some embodiments, the concentration of hyaluronic acid in the composition is about 0.5% (weight / weight). In some embodiments, the hyaluronic acid in the composition is in the form of hyaluronic acid gel (i.e., with high viscosity).

[0075] In some embodiments, the excipient is selected from salts, ions, lactated Ringer's solution, sugars, biocompatible surfactants, polyols and combinations thereof. In some embodiments, the excipient is a polyol. In some embodiments, the polyol is glycerol. In some embodiments, the glycerol concentration of the composition is about 12% (weight / weight) to 25% (weight / weight). In some embodiments, the glycerol concentration of the composition is about 20% (weight / weight).

[0076] In some embodiments, the composition comprises a second excipient. In some embodiments, the second excipient is sodium chloride or sodium phosphate forming, for example, saline or phosphate buffered saline.

[0077] In some embodiments, the composition comprises a third excipient. In some embodiments, the third excipient is a water-soluble surfactant. In some embodiments, the third excipient is a hydrotropic compound. In some embodiments, the third excipient is a polysorbate.

[0078] In some embodiments, the composition comprises Pluronic TM(also known as "poloxamers"). In some embodiments, poloxamers form macromolecular assemblies, such as micelles. In some embodiments, the macromolecular assemblies are micelles (or "poloxamer micelles"). In some embodiments, Pluronic or poloxamer is selected from Pluronic L31, Pluronic L35, Pluronic F38, Pluronic L43, Pluronic L44, Pluronic L61, Pluronic F68, Pluronic F77, Pluronic L81, Pluronic P84, Pluronic P85, Pluronic F77, Pluronic F87, Pluronic L92, Pluronic F98, Pluronic L101, Pluronic P103, Pluronic P104, Pluronic P105, Pluronic F108, Pluronic L121, Pluronic P123, Pluronic F127, Poloxamer 101, Poloxamer 105, Poloxamer 108, Poloxamer 122, Poloxamer 123, Poloxamer 124, Poloxamer 181, Poloxamer 183, Poloxamer 188, Poloxamer 212, Poloxamer 215, Poloxamer 217, Poloxamer 231, Poloxamer 234, Poloxamer 235, Poloxamer 237, In some embodiments, the Pluronic or poloxamer is Pluronic F127. In certain embodiments, the poloxamer is poloxamer 407.

[0079] In certain embodiments, the concentration of the poloxamer is about 0% (w / w) to 10% (w / w). In certain embodiments, the concentration of the poloxamer is about 0.1% (w / w) to 10% (w / w). In some embodiments, the concentration of the poloxamer is about 5% (w / w). In certain embodiments, the concentration of the pluronic acid is about 1.5% (w / w). TM Pluronic TM F127, Pluronic TM The concentration of F127 is about 5% (w / w). In some embodiments, the poloxamer is poloxamer 407, wherein the concentration of poloxamer 407 is about 5% (w / w).

[0080] In some embodiments, the composition includes lipids. In some embodiments, the composition includes a plurality of lipids in the form of liposomes formed by phospholipids (e.g., soybean PC). Lipids can be of any type (e.g., phospholipids, cholesterol, conjugated lipids or combinations thereof), or the composition can include any other non-water-soluble substances instead of lipids. Lipids (or lipid particles) or non-water-soluble substances are present in relatively high concentrations, preferably about 6% (weight / weight) to 28% (weight / weight) of the composition. It is not intended to be bound by any particular theory, and it is believed that when the composition is exposed to freezing temperatures (about -25°C to -15°C, about -15°C to -10°C, about -15°C to -5°C, about -10°C to -5°C, or about -10°C in some embodiments), lipids (or lipid particles such as liposomes) or non-water-soluble substances produce emulsions because these substances prevent large ice crystals from forming. This allows the composition to have ice particles while also being flowable and / or injectable. In some embodiments, the composition also includes lipid particles. In some embodiments, the lipid particles are liposomes. In some embodiments, the lipid particles are micelles. In some embodiments, the lipid particles include phospholipids. In some embodiments, the phospholipids are selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoylphosphatidylcholine (DPPC), dicetyl phosphate (DCP), L-α-phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylglycerol (PG), L-α-phosphatidylcholine (soy PC) and combinations thereof. In some embodiments, the lipid is L-α-phosphatidylcholine (soy PC). In some embodiments, the lipid concentration in the composition is about 0% (weight / weight) to 30% (weight / weight).

[0081] In some embodiments, the composition further comprises ethanol. In some embodiments, the concentration of ethanol in the composition is about 0.01% to 0.1%. In some embodiments, the concentration of ethanol in the composition is about 0.07% or less.

[0082] In some embodiments, the composition comprises glycerol, hyaluronic acid, and Pluronic acid in water, saline, or phosphate buffered saline. TM F127. In some embodiments, the composition comprises glycerol, hyaluronic acid and Pluronic acid in saline. TM F127, wherein the molecular weight of the hyaluronic acid is about 1000 kDa. In some embodiments, the composition comprises about 20% (w / w) glycerol, about 0.5% (w / w) hyaluronic acid, and about 5% (w / w) Pluronic acid in water, saline, or phosphate buffered saline. TMIn some embodiments, the composition comprises about 20% (w / w) glycerol, about 0.5% (w / w) hyaluronic acid, and about 5% (w / w) Pluronic acid in water, saline, or phosphate buffered saline. TM F127, wherein the molecular weight of hyaluronic acid is about 1000 kDa.

[0083] In some embodiments, the composition comprises glycerol, PBS, hyaluronic acid, and Pluronic 407. TM In certain embodiments, the composition comprises glycerol at a concentration of about 18.9% (w / w), 1×PBS at a concentration of about 75.6% (w / w), hyaluronic acid having a molecular weight of 1000 kDa and a concentration of about 5% (w / w), and pluronic 407 at a concentration of about 0.5% (w / w). TM F127). See e.g. Figure 1 In certain embodiments, the composition comprises a temperature set point of about -14°C. See id. In certain embodiments, the composition has an ice content wherein the ice content is about 50%. In certain embodiments, the composition is Figure 1 In certain embodiments, the composition comprises a relatively small percentage of hyaluronic acid of about 0.5% (w / w) to 1.5% (w / w), wherein the molecular weight of the hyaluronic acid is about 1,000 kDa. In such embodiments, the composition also comprises glycerol at a concentration of about 15% (w / w) to 25% (w / w), and a small percentage of polysorbate of about 0.25% (w / w) to 1.5% (w / w). This allows the composition to have ice particles after freezing while also being flowable and / or injectable.

[0084] In some embodiments, the composition is filled into a container with a volume less than 10mL and with a shape of the maximum surface area producing a container wall. It is not intended to be bound by any particular theory, it is believed that the ratio of large surface area to volume helps to improve freezing rate, to further prevent the formation of large ice crystals, and therefore improves fluidity and injectability. In some embodiments, the total injection volume of the cold slurry optionally injected into the patient via multiple containers and multiple injections is about 5mL to 10mL, about 10mL to 20mL, about 20mL to 30mL, about 30mL to 40mL, about 40mL to 50mL, about 50mL to 60mL, about 60mL to 70mL or greater than about 70mL. In some embodiments, the total injection volume is about 60mL.

[0085] Methods for producing cold slurries by preparing solutions that prevent the formation of large ice crystals are described in PCT Application Nos. PCT / US20 / 43280 and '494PCT, which are incorporated herein by reference in their entirety. Described herein is an unexpected method for forming cold slurries by producing a composition having hyaluronic acid, water, and at least one excipient, wherein the production of the cold slurry does not require any mechanical manipulation or mechanical stirring of the composition. In some embodiments, the composition further comprises a poloxamer. In some embodiments, the composition comprises a plurality of lipids. In some embodiments, the injectability of the composition is improved by utilizing slight mechanical stirring.

[0086] In some embodiments, the compositions described herein are homogenous mixtures such that the composition medium is uniform throughout the container and the components are evenly distributed. In some embodiments, the addition of poloxamer, hyaluronic acid, and glycerol prevents the formation of large ice crystals, so that a flowable cold slurry can be injected into a subject immediately after removal from a freezer or other cold environment.

[0087] Compositions provided herein can be provided in syringes or other containers.Syringes or other containers can also include visual temperature indicators that can allow visual monitoring of the temperature of the slurry or the approximate temperature of the slurry.Temperature indicators can be temperature sensing labels, stickers, markers, crayons, paints, pellets, etc., including reversible temperature labels that can dynamically track temperature changes.Temperature indicators can be located inside syringes or other containers (e.g., pellets directly placed in the internal solution), on the inner wall of syringes or other containers, on the outer wall of syringes or other containers, or any position that allows visual tracking of the temperature of the contents inside syringes or other containers.In certain embodiments, compositions are provided in syringes, and syringes are placed in freezers.Syringes are taken out from freezers after a period of time (e.g., 24 hours) sufficient to form ice crystals in syringes.After the syringe is taken out from the freezer, for example, a temperature indicator (e.g., an infrared sensor or an external temperature sensitive indicator) is used to monitor the syringe contents.After the syringe contents reach a predetermined temperature, the syringe is connected to a second syringe and subjected to BAF treatment (e.g., three BAF cycles).Then the slurry is injected into a patient or object. In some embodiments, alternative methods of treating the slurry can be used, for example, pushing the slurry through a filter; including a line between two syringe openings around which the slurry must be injected; providing internal components such as magnets, internal blades, or internal forming wires (e.g., springs) in the syringe that provide such treatment. In some embodiments, the BAF treatment or other treatment begins after the contents of the container are removed from the freezer and reach a predetermined temperature. In some embodiments, the BAF treatment or other treatment begins immediately after the container is removed from the freezer before the container reaches a predetermined temperature. Without intending to be bound by any particular theory, it is believed that further mechanical treatment reduces the size of ice crystals in the cold slurry and makes it easier to inject the ice crystals through the needle.

[0088] Reference Figure 2 , two different cold slurry compositions (multiple batches) were characterized with respect to their temperature profiles over time and ice content. The different cold slurry batches were placed on a copper plate heated to 40°C and with thermocouple wires measuring the temperature change of the cold slurry over time. The plotted data shows the temperature change over time (in seconds) for two different cold slurry compositions: 1) a 12.6% (w / w) glycerol and PBS (represented by trace A) C and A M ), and 2) containing 0.5% (w / w) soy-PC, 0.08% (w / w) EtOH, 0.75% (w / w) hyaluronic acid (1,000 kDa), 16% (w / w) glycerol, and PBS (represented by trace B). C and B MFor each cold slurry, the temperature was measured at two different locations: a thermocouple embedded inside the copper plate (Trace A C , B C ), while the other thermocouple is located in the middle of the copper plate and exposed to the outside of the board (Trace A M , B M When a batch of cold slurry is first introduced into the copper board, the thermocouple wire embedded inside the board (Trace A) C , B C ) Initially measure the warm temperature of the heating plate (e.g., trace A C 31°C at time point 0), and then reaches equilibrium at a lower temperature due to the cooling effect of the introduced cold slurry (e.g., trace A C 20°C at about 2 minutes). On the other hand, for the thermocouple wire located in the middle of the plate (Trace A M , B M ), when the cold slurry is initially introduced into the copper plate, the cold slurry immediately contacts the thermocouple wires since the wires are exposed. This results in an initial negative temperature reading in the middle (e.g., trace A) due to the crystallized cold slurry contacting the wires. M =-4°C at time 0), then reaching equilibrium at a warmer temperature as the cold slurry begins to melt on the hot plate (e.g., trace A M 14°C at about 6 minutes). Thermocouple wire exposed to the outside of the board (Trace A) M , B M ) can be used to detect phase transitions during the onset of melting of a crystallized cold slurry. The graph shows that a slurry composition with hyaluronic acid (trace B C and B M ) has a gradual phase transition. The figure also shows that the cold slurry batch with hyaluronic acid composition (trace B C and B M ) reached equilibrium (as measured by the two thermocouple wire positions) in a similar time frame and at similar temperatures of about 10°C to 14°C (depending on the position of the thermocouple (inside / middle)). On the other hand, the cold slurry with a different composition (lacking hyaluronic acid; trace A C and A M ) has a different temperature profile than the composition comprising hyaluronic acid, reaching equilibrium more quickly at a temperature of about 15°C to 17°C (depending on the location of the thermocouple (inside / middle)). Figure 2 It is shown that cold slurries can have different compositions designed to have different temperature profiles or can be designed to perform equivalently.

[0089] Figure 3One embodiment of a composition comprising poloxamer micelles, glycerol, and hyaluronic acid gel is depicted. In this embodiment, the composition comprises pluronic TM (For example, Pluronic TM In some embodiments, the micelles are formed by a hyaluronic acid gel in a water-glycerol and optional ethanol solution (not shown) outside the micelles. In some embodiments, the water droplets are captured in the external medium (i.e., in the hyaluronic acid gel in the water-glycerol and optional ethanol solution (not shown) outside the micelles). Figure 3 The embodiments of the present invention are referred to herein as "micelle dispersions" or "micelle gels". Micellar dispersions or micellar gels contain surfactant micelles that capture water, wherein the micelles are suspended in a hyaluronic acid gel. In some embodiments, the micelles in the composition are in the form of positive micelles (e.g. Figure 3 In some embodiments, a lipid micelle composition comprising hyaluronic acid is provided.

[0090] In some embodiments, the composition of at least part of the crystallization comprising hyaluronic acid, glycerol, water and saline (or PBS) comprises enough flowable and / or injectable ice particles when exposed to freezing temperature (that is, being placed in a freezer) without adding other excipients. In some embodiments, the composition of at least part of the crystallization comprising hyaluronic acid, glycerol and water comprises enough flowable and / or injectable ice particles when exposed to freezing temperature (that is, being placed in a freezer) without adding other excipients. In alternative embodiments, the interpolation of poloxamer, polysorbate or lipid further improves the fluidity and injectability of the composition of the partial crystallization. It is also found that the freezing rate of the material is increased to a faster rate to further improve the fluidity and injectability of the composition. The optimization of the freezing rate includes the material selected for the container (for example, syringe) of the composition, the geometry of the container and the selection of the cold environment or freezer (for example, the humidity of the freezer can be adjusted to improve the fluidity and / or injectability of the resulting cold slurry). It is also found that the injectability or fluidity of the composition of the partial crystallization can be improved by spacing the containers apart from each other when being placed in a freezer.

[0091] In some embodiments, the composition (for example, in the form of a liquid solution) can be packaged and sealed in a container such as a syringe. The syringe can be aseptically filled (for example, using an aseptic procedure), or the syringe can be pre-filled, sealed, and then terminally sterilized (for example, using autoclave or steam sterilization). The composition can also be provided in any other sealed container that can be terminally sterilized, such as a tube for topical ointment, or a larger container for filling a plurality of syringes. In certain embodiments, a pre-filled syringe or other container can be terminally sterilized using gamma radiation or autoclave or steam sterilization at a temperature of about 118 ° C. In certain embodiments, a pre-filled syringe or other container can be terminally sterilized using gamma radiation or autoclave or steam sterilization at a temperature of about 118 ° C to 121 ° C. In a preferred embodiment, when using a hyaluronic acid with a molecular weight of 1,000 kDa, steam is used to terminally sterilize the composition, because the sterilization using radiation will affect the molecular weight of hyaluronic acid. In certain embodiments, the composition is terminally sterilized using hyaluronic acid having a molecular weight of 1,000 kDa, the composition is terminally sterilized using low dose radiation, or radiation is used to irradiate the cold slurry while still frozen.

[0092] In some embodiments, the composition inside the syringe (or other container) is converted into a flowable and / or injectable cold slurry by placing the pre-filled syringe (or other container) in a standard freezer or other cold environment. In certain embodiments, a structure for storing one or more pre-filled syringes (or other containers) in a freezer or other cold environment is provided, wherein the structure is configured to store the first pre-filled syringe (or container) of one or more pre-filled syringes (or other containers) as a predetermined distance from the second pre-filled syringe (or container) of one or more pre-filled syringes (or other containers). In certain embodiments, a structure for storing a plurality of pre-filled syringes (or containers) as a predetermined distance from each other is provided to facilitate the uniform and rapid cooling of cold slurry. See, for example, Example 3. In certain embodiments, the predetermined distance is at least 0.5 inches. In certain embodiments, the predetermined distance is about 0.5 inches. In certain embodiments, the predetermined distance is about 0.5 inches, about 0.75 inches, about 1 inch, about 1.25 inches, about 1.5 inches, about 1.75 inches, about 2 inches, about 2.25 inches, about 2.5 inches, about 2.75 inches, about 3 inches, about 3.25 inches, about 3.5 inches, about 3.75 inches, about 4 inches, or greater than about 4 inches. In some embodiments, the structure is a rack, tray, or box. In some embodiments, liquid nitrogen or other liquid cooling methods can be used to quickly freeze the composition to accelerate the process.

[0093] In some embodiments, after freezing, a syringe or container can be taken out from a freezer, a cold environment or other freezing methods, and can be injected immediately or optionally applied by surface application to apply cold slurry for therapeutic benefit. In some embodiments, slurry can be directly applied to tissue after an invasive surgical method. In some embodiments, a needle can be used to inject cold slurry directly from a syringe. In certain embodiments, the needle is a 16G needle, a 17G needle, a 18G needle, a 19G needle, a 20G needle, a 21G needle, a 22G needle, a 23G needle or a 24G needle. When the container is taken out from the freezer, the cold slurry can also be taken out from the container for surface application, such as by squeezing the container to drive the cold slurry to the target treatment site. In some embodiments, the cold slurry is in a flowable or injectable form immediately after taking it out from the freezer, without any further mechanical operation.

[0094] In certain embodiments, after removal from the freezer, the cold slurry is subjected to mechanical manipulation to improve flowability or injectability prior to injection or, optionally, application by topical application for therapeutic benefit. Figure 4A As shown in , the cold slurry can be provided in a first syringe or container configured to be connected to a second syringe or container, optionally via a female Luer adapter, to facilitate "reciprocating" (BAF) processing of the cold slurry. See, e.g., Example 5. In certain embodiments, as Figure 4A As shown in , a system is provided: the system includes a first sterile syringe or other container and a second sterile syringe or container, wherein the first sterile syringe or other container includes a male Luer assembly and contains cold slurry, and the second sterile syringe or container includes a male Luer assembly and does not contain cold slurry, wherein the first syringe or container and the second syringe or container are connected by a sterile female Luer connector. In certain embodiments, the BAF process consists of: (1) pushing the cold slurry from the first syringe into the second syringe, and (2) pushing the cold slurry from the second syringe back into the first syringe to complete a "reciprocating" cycle. In certain embodiments, the cold slurry is subjected to one, two, three, four or more BAF cycles. See Example 5; Figure 8 In certain embodiments, the cold slurry is subjected to two BAF cycles. In certain embodiments, the cold slurry is subjected to three BAF cycles. An exemplary method comprising three reciprocating cycles is shown in Figure 4B middle. Figure 4B The exemplary method shown in comprises freezing a first syringe for 24 hours, wherein the syringe contains the slurry composition before removing the syringe from the freezer. Next, temperature monitoring (e.g., by using a temperature monitoring device such as Figures 5A to 5DThe temperature of the now cold slurry is observed using a temperature-sensitive sticker depicted in Figure 1 until the cold slurry reaches a predetermined temperature. After reaching the predetermined temperature, the slurry is treated with a second syringe connected to the first syringe using a Luer connector and subjected to three BAF cycles. After three BAF cycles, the slurry is injected. This process can be repeated as needed to prepare the number of syringes required for treatment. Pushing the contents of a syringe from one syringe to another is a well-known technique that does not compromise the sterility of the syringe contents when using sterile components. Optionally, additional elements can be added between the syringes to further split the ice crystals. For example, one of the syringe openings can be covered with a filter or with a wire to split the ice crystals. It should be understood that Figure 4B The method described in can be used with the method and system described in International Publication No. WO 2022 / 055934, wherein a syringe containing a biocompatible composition described herein is transported to a point of care at ambient temperature and then placed in a freezer at the point of care to convert the composition into a cold slurry containing a plurality of ice crystals. The disclosure of the '934 PCT relating to methods for transporting and converting biocompatible compositions is incorporated herein by reference.

[0095] In some embodiments, after being taken out from the freezer, the syringe is placed aside and allowed to warm up to a predetermined temperature for injection or surface application. In certain embodiments, the predetermined temperature is reached after the syringe contents undergo mechanical stirring. In some embodiments, the syringe or other container has an external temperature indicator configured to indicate when a predetermined temperature is reached, for example, by a color change or by displaying a message. In certain embodiments, the temperature indicator is a temperature-sensitive sticker, etc. The temperature indicator can allow visual monitoring of the temperature of the contents of the syringe or the approximate temperature of the contents of the syringe. Temperature indicators, such as temperature-sensitive stickers, etc., can display a range of temperatures, wherein the color change corresponding to an area within a predetermined temperature range indicates that the predetermined temperature has been reached. See, e.g. Figure 5A. The temperature indicator can be a temperature sensing label, sticker, marker, crayon, paint, ball, etc., including a reversible temperature label that can dynamically track temperature changes. The temperature indicator can be located inside the container (e.g., a ball placed directly in the internal solution), on the inner wall of the container, on the outer wall of the container, or anywhere that allows visual tracking of the temperature of the contents inside the container. In some embodiments, the predetermined temperature is about -15°C. In some embodiments, the predetermined temperature is about -19°C to -11°C, about -18°C to -12°C, about -17°C to -13°C, or about -16°C to -14°C. In some embodiments, the color change in the area corresponding to a temperature cooler than the predetermined temperature indicates that the cold slurry is cooler than the predetermined temperature to indicate that the cold slurry should be allowed to heat to the predetermined temperature before injection or application. In some embodiments, the colder temperature is about -20°C. In some embodiments, the colder temperature is about -20°C to -25°C, about -25°C to -30°C, or below about -30°C. In some embodiments, a color change in an area corresponding to a temperature hotter than a predetermined temperature indicates that the cold slurry is hotter than the temperature recommended for injection or application to produce a therapeutic effect. In some embodiments, if a warmer temperature is reached, the syringe can be refrozen. For example, in some embodiments, the warmer temperature is about -10°C to -8°C, about -10°C to -9°C, about -9°C to -8°C, about -10°C to -7°C, about -10°C to -6°C, about -10°C to -5°C, about -10°C to -4°C, about -10°C to -3°C, about -10°C to -2°C, about -10°C to -1°C, about -10°C to 0°C, -9°C to 8°C, about -9°C to -7°C , about -9°C to -6°C, about -9°C to -5°C, about -9°C to -4°C, about -9°C to -3°C, about -9°C to -2°C, about -9°C to -1°C, about -9°C to 0°C, about -8°C to -7°C, about -8°C to -6°C, about -8°C to -5°C, about -8°C to -4°C, about -8°C to -3°C, about -8°C to -2°C, about -8°C to -1°C, about -8°C to 0°C, or hotter than 0°C. In some embodiments, the hotter temperature is at least -10°C to -8°C or hotter. In some embodiments, the hotter temperature is at least -9°C to -8°C or hotter. In some embodiments, the hotter temperature is at least -10°C to -9°C or hotter. In some embodiments, an indicator, such as a temperature sensitive sticker, displays a temperature or a temperature range, indicating when the cold slurry has reached a predetermined temperature. See, e.g. Figure 5B In certain embodiments, the indicator, such as a temperature sensitive sticker or the like, includes an area for displaying a message indicating that a predetermined temperature has been reached. See, e.g. Figure 5CFor example, in some embodiments, the indicator can display a message such as "Processing Now," "Processing," "Stirring," "Starting," or any other suitable message. In some embodiments, an indicator such as a temperature sensitive sticker or the like includes an area for displaying a message indicating that the cold slurry is cooler than a predetermined temperature or hotter than a predetermined temperature. See, e.g. Figure 5D For example, in some embodiments, the indicator may display a message such as "wait", "hold", "not yet reached", "too cold", or any other suitable message to indicate that the cold slurry is colder than the predetermined temperature. In some embodiments, the indicator may display a message such as "discard", "too hot", "throw away", "refrozen", or any other suitable message to indicate that the cold slurry is hotter than the predetermined temperature. In some embodiments, the indicator is provided as Figure 4B The method depicted includes temperature monitoring and reciprocating mechanical stirring.

[0096] In one aspect, a composition is provided wherein the composition requires an injection force of less than about 30 lbs of plunger force to inject the composition through a 16G needle, 17G needle, 18G needle, 19G needle, 20G needle, 21G needle, 22G needle, 23G needle, or 24G needle syringe. In some embodiments, the composition provided herein requires an injection force of less than about 30 lbs of plunger force to inject the composition through a 17G or 18G needle. The compositions disclosed herein according to Figure 2 prepared and tested to verify whether there is a relationship between freezer temperature and injection force. Figure 6 As shown in , colder freezer temperatures require increased injection force to inject cold slurry from a syringe because the composition in the syringe has more ice particles than at warmer temperatures. In order to determine the ice content of cold slurry, a test method and apparatus were designed that allow the ice content to be calculated based on the final equilibrium temperature of the apparatus after a predetermined amount of cold slurry has been dispensed into the apparatus, as described in Example 3. Fig. 7A , as the equilibrium temperature increases, the injection temperature increases. And if Figure 7B As shown in , as the injection temperature decreases, the injection force increases due to the increase in ice content. As the temperature of the cold slurry increases, the injection force decreases. This result was observed for all syringe sizes tested (e.g., 3cc, 6cc) and needle gauges (e.g., 17G, 18G). See, e.g. Figure 6 , Figure 7B , Figures 8 to 9. In addition, the overall reliability of the injection can be increased as the temperature increases, i.e., the number of injection failures (e.g., injections in which the injection force is observed to peak and the slurry fails to expel from the syringe) can be reduced. By monitoring the temperature of the cold slurry, the syringeability can be optimized (reducing the injection force) without reducing the ice content below the therapeutic threshold for a given application. It was found that the reliability of the injection was greatly increased by following the following steps: (1) placing the syringe in a freezer to form a cold slurry containing a plurality of ice crystals; (2) after removing from the freezer, waiting for the cold slurry to reach a predetermined temperature; and (3) subjecting the cold slurry to three BAF cycles between two syringes. These results are shown in > 5mL injection (e.g., 6cc injection). In another aspect, according to the methods described herein, the injection force is reduced by mechanically agitating the cold slurry, e.g., by subjecting the cold slurry to one or more reciprocating cycles. See, e.g. Figure 8 , Figure 10 to Figure 11 ; Example 5.

[0097] Reference Figure 1 , 6cc cold slurry compositions (multiple batches) prepared as described below were characterized with respect to their temperature profile and ice content. Different cold slurry batches were placed on a copper plate heated to 40°C with thermocouple wires measuring the change in the temperature of the cold slurry over time. The data plotted shows the change in the temperature profile according to Figure 1 The prepared solution contained 18.9% (w / w) glycerol, 75.6% (w / w) 1× PBS, 5.0% (w / w) 1000 kDa hyaluronic acid and 0.5% (w / w) Pluronic 407. TM The temperature changes of three batches of cold slurry compositions (F127) over time. The three batches of cold slurry compositions were then subjected to two BAF treatment cycles as described herein. For each cold slurry, the temperature at two different locations was measured: the interior of the embedded copper plate (Trace A C , B C , C C ) and the trace A in the middle of the copper plate that is exposed to the outside of the board M , B M , C M As a batch of cold slurry is introduced into the copper board, the thermocouple wire embedded inside the board (Trace A) C , B C , C C ) Initially measure the warm temperature of the heating plate (e.g., trace A C , B C and C C 35°C to 40°C at time point 0), and then reaches equilibrium at a lower temperature due to the cooling effect of the introduced cold slurry (e.g., trace A C, B C and C C On the other hand, for the thermocouple wire located in the middle of the plate (Trace A) M , B M , C M ), when the cold slurry is first introduced into the copper plate, it immediately contacts the thermocouple wire because the wire is exposed. This results in an initial negative temperature reading at the midpoint (e.g., trace A) due to the crystallized cold slurry contacting the wire. M =-4°C at time point 0), then reaches equilibrium at a warmer temperature as the cold slurry begins to melt on the hot plate (e.g., trace A M , B M and C M about 12°C to 14°C at about 6 minutes). Thermocouple wire exposed to the outside of the plate (Trace A) M , B M , C M ) can be used to detect phase transitions during the onset of melting of a crystallized cold slurry. The figure shows that all tested cold slurry compositions (each as Figure 1 The figure also shows that the cold slurry batches prepared in this way consistently reach equilibrium within a similar time frame and at similar temperatures (as measured by the trace A of the two thermocouple wire positions). M , B M and C M ).therefore, Fig.10 It was demonstrated that cold slurries designed in accordance with the present disclosure produced consistent temperature profiles and ice content when subjected to mechanical agitation.

[0098] Reference Fig.11 , provides the basis for Figure 1 Characterization of slurry compositions prepared and subjected to different processing techniques and injected using 18G or 17G needles. For all tests, cold slurries were contained in 5cc syringes. Reliability was characterized by quantifying the percentage of injections that did not exhibit injection force peaks above a predetermined limit. Fig.11The results presented in the table have a predetermined limit of 40 lbf. First, the injection force of a cold slurry that was frozen at -20°C and not subjected to further processing was measured. The injection reliability using an 18G needle was less than 20%, and the injection reliability using a 17G needle was 90%, meaning that the chance that the cold slurry can be injected through an 18G needle is less than 20%, and the chance of being injected through a 17G needle is 90%. Secondly, the injection force of a cold slurry that was frozen at -20°C and subjected to three BAF cycles after being removed from the freezer, but without waiting and temperature monitoring, was measured. The injection reliability using an 18G needle was 78%, and the injection reliability using a 17G needle was greater than 95%. Finally, the injection force of a cold slurry that was frozen at -20°C, allowed to warm to -15°C (as determined by temperature monitoring), and subjected to three BAF cycles was measured. The injection reliability using an 18G needle and a 17G needle is greater than 95%. Therefore, Fig.11 It was shown that when the slurry was subjected to three BAF cycles after being frozen at -20°C and injected using a 17G needle, an injection reliability of greater than 95% could be achieved. Fig.11 It was also shown that greater than 95% reliability could be achieved using both 18G and 17G needles when the slurry was subjected to three BAF cycles after being frozen at -20°C and allowed to warm to -15°C (as determined by temperature monitoring).

[0099] Reference Fig.12 , presents the results of experiments testing the effect of sterilizing syringes containing slurry compositions using radiation (gamma / e-beam sterilization). The results show that radiation sterilization using gamma / e-beam sterilization results in an increase in the injection force required to expel the cold slurry from the syringe. See Fig.12 Without wishing to be bound by a particular theory, this increase in the injection force required may be due to the effect of the molecular weight of the hyaluronic acid in the composition. In some experiments, syringes containing the slurry composition were sterilized using autoclave (steam sterilization) at different temperatures (e.g., at about 118° C. or at about 121° C.). It was observed that, unlike radiation sterilization, autoclave or steam sterilization did not increase the injection force required to expel the cold slurry from the syringe.

[0100] The compositions described herein can be used for a variety of applications. After the compositions according to some embodiments of the present disclosure are exposed to freezing temperatures so that they form a flowable cold slurry, they can be topically applied to the area for therapeutic treatment. The method of topically applying cold slurry to the ocular surface is described in International Patent Application No. PCT / US21 / 24514, and the disclosures related to the therapeutic use of slurry are incorporated herein by reference as a whole. The compositions described herein can also be used to form a flowable and / or injectable cold slurry that can be injected into the target treatment area for therapeutic effect. The injection method for cold slurry is described in International Patent Application No. US2017 / 0274078, and the disclosures related to the therapeutic use of injection slurry are incorporated herein by reference as a whole.

[0101] The devices, systems, compositions and methods disclosed herein are not limited in scope to the specific embodiments described herein. Indeed, various modifications of the devices, systems and methods besides those described will become apparent to those skilled in the art from the foregoing description.

[0102] Example

[0103] Example 1 - Method for preparing a cold slurry composition comprising hyaluronic acid and poloxamer

[0104] Cold slurry compositions were prepared and tested by varying the following components: glycerol, hyaluronic acid, lipids, and poloxamers (e.g., Pluronic TM F127). A composition having a glycerol content ranging from about 12 wt / wt% to 25 wt / wt% is prepared. The composition further comprises a hyaluronic acid content ranging from about 0.1 wt / wt% to 1 wt / wt% hyaluronic acid, wherein the hyaluronic acid has a molecular weight ranging from 250 kDa to 5000 kDa. The lipid content of the composition ranges from 0 wt / wt% to 30 wt / wt%. The poloxamer content of the composition (e.g., Pluronic TM F127) ranges from 0 wt / wt% to 30 wt / wt%.

[0105] Two basic test methods are used to evaluate the preparation for cold slurry composition. First, the composition is tested to measure the injection force by various needle specifications (e.g., 17G or 18G). Use a force test bench to apply force to the plunger of a syringe with the predetermined speed and peak force required to extrude all materials by a needle (e.g., 17G or 18G needle). Secondly, the device of the heat capacity of the test cold slurry composition is used to characterize the ice content of the cold slurry composition. After the assessment, prepare a glycerol containing 20 w / w%, 0.5 w / w% of 1000kDa hyaluronic acid and 5 w / w% of Pluronic Acid in saline. TMPreparation of F127. Figure 1 .

[0106] Example 2 - Evaluation of Cold Slurry Cooling Technology and Freezer Spacing

[0107] Cold slurry compositions were prepared as described in Example 2. In order to optimize performance, different preparation methods were tested to determine the effect of different preparation methods on injection force, as described below. In this example, experiments were conducted to determine whether the freezing temperature affects the injection force required to eject the cold slurry composition from the syringe. The results showed that lower freezer temperatures required greater injection forces. See Figure 6 . In addition, the cold slurry composition that was rapidly frozen at a temperature of about -60°C required an increased injection force compared to the control. In certain experiments, the cold slurry in multiple syringes was placed in the freezer by spacing the syringes apart to achieve uniform and rapid cooling of the cold slurry. It was observed that if the syringes were placed too close to each other, freezing was adversely affected and larger ice crystals were observed. In contrast, when the cold slurry was injected by syringes that were spaced farther apart in the freezer, a reduced injection force was observed.

[0108] Example 3 - Temperature Monitoring of Injectable Frozen Slurries for Performance Optimization at the Point of Care

[0109] According to the present disclosure, lower injection forces are generally preferred for injecting cold slurry, however, it is desirable that the cold slurry be in the correct temperature range and not too hot to produce a therapeutic effect. It is desirable that the cold slurry maintain a sufficient ice content because the ice content of the cold slurry allows the cold slurry to extract energy from the target tissue and provide a therapeutic effect. If the ice content of the cold slurry is too low, the therapeutic effect may be reduced. Conversely, if the ice content of the cold slurry is too high, the cold slurry may not be injectable. In this embodiment, an experiment was conducted using a copper plate to determine the ice content of the slurry. After a predetermined volume of slurry was dispensed into the device, the ice content was calculated based on the final equilibrium temperature of the device. As Fig. 7A As shown in , the relationship between injection temperature and final equilibrium temperature is inversely related, so that as the injection temperature increases, the ice content of the cold slurry decreases. Fig. 7A .

[0110] Further experiments showed that the injection force decreased as the temperature of the cold slurry increased. Figure 7B. Lower ice content also results in more reliable injections, i.e., the number of failed injections is reduced when the cold slurry has a lower ice content. However, as described above, it is desirable for the cold slurry to maintain sufficient ice content and temperature to maintain a therapeutic effect. Results indicate that injection performance can be optimized by controlling the ice content by targeting a predetermined injection temperature. Therefore, the temperature monitoring device and system are designed to include an indicator that provides an indication when the cold slurry reaches a predetermined temperature, wherein the predetermined temperature is selected to reduce the injection force required to expel the cold slurry from the syringe without causing the ice content of the cold slurry to drop below the therapeutic temperature. See, e.g. FIG. 5A to FIG. 5D ; see also Figure 4B . In addition, other temperature monitoring devices and systems are contemplated that include a plurality of indicators that provide an indication of when the cold slurry is at a temperature that is too cold for injection; when the cold slurry has reached a predetermined temperature, wherein the predetermined temperature is selected to reduce the injection force required to expel the cold slurry from the syringe without reducing the ice content of the cold slurry below the therapeutic temperature; or when the cold slurry has reached a temperature that is too hot to achieve the desired therapeutic effect. See, e.g. Figure 5A , FIG. 5B to FIG. 5C ; see also Figure 4B .

[0111] Example 4 - Method for treating cold slurry between multiple syringes to improve syringeability

[0112] In some experiments, different mechanical treatment methods were tested. In some experiments, a cold slurry composition was prepared by connecting a first syringe to a second syringe using a connector, such as a Luer connector, and pushing the contents of the first syringe "back and forth" into the second syringe. See FIG. 4A to FIG. 4B , Figure 8 For example, in one experiment, a first syringe including a male Luer component and containing a cold slurry composition was connected to a second syringe including a male Luer component and not containing a cold slurry composition using a female-to-female Luer connector connecting the first syringe and the second syringe. FIG. 4A to FIG. 4B The slurry was reciprocated between the first and second syringes, wherein one reciprocating cycle included transferring the cold slurry from the first syringe to the second syringe and then from the second syringe back to the first syringe. It was observed that conditioning the slurry by reciprocatingly pushing the slurry significantly improved the performance in the injection test and reduced the injection force required to expel the cold slurry from the syringe. Figures 8 to 11Without wishing to be bound by a particular theory, it is believed that performing one or more reciprocating cycles reduces the injection force by reducing the size of ice crystals in the cold slurry, dispersing the ice crystals more evenly in the cold slurry, and / or increasing the temperature of the cold slurry by imparting shear forces to the cold slurry composition. Experiments were conducted to compare the injection force required to expel the slurry from the syringe after performing one, two, three, or four reciprocating cycles. Figure 8 The experiment showed that the injection force required to inject the cold slurry through a 6 mL syringe connected to a 17G needle was significantly reduced. Id. The control cold slurry that was not subjected to reciprocating cycles required an injection force of about 50 lbf to 95 lbf. See, e.g. Fig.11 . Cold slurry subjected to a single reciprocating cycle requires a reduced injection force in the range of about 6 lbf to 38 lbf. Id.; see also Figure 8 , Fig. 9 While experiments have shown that the injection temperature of cold slurry is inversely related to the injection force, see Figure 7B , when the data are normalized to temperature, performing one or more reciprocating cycles further reduces the required injection force. Figure 8 . The results showed that increasing the number of consecutive reciprocating cycles to two or three cycles further reduced the injection force. Id. It was observed that after three reciprocating cycles, no additional reduction in injection force was observed in slurries subjected to a fourth reciprocating cycle. Id. Additional experiments showed that using 17G and 18G needles, cold slurries subjected to two reciprocating cycles could consistently be injected using injection pressures below about 30 lbf over the post-treatment temperature range. See Fig. 9 .

[0113] Further experiments included a handling element in a connector (e.g., a Luer connector) between the first and second syringes, and the contents of the first syringe were reciprocally pushed into the second syringe as described above. It was observed that including the handling element in the connector produced neutral results on the required injection force.

[0114] Example 5 - Sterilization Method for Cold Slurry Compositions Containing Hyaluronic Acid

[0115] In certain experiments, various sterilization methods were examined. In some experiments, the syringes containing the slurry composition were sterilized using radiation (gamma ray / electron beam sterilization). The results showed that radiation sterilization using gamma ray / electron beam sterilization resulted in an increase in the injection force required to expel the slurry from the syringe. See Fig.12Without wishing to be bound by a particular theory, this increase in the injection force required may be due to an effect on the molecular weight of the hyaluronic acid in the composition. In some experiments, syringes containing the slurry composition were sterilized using autoclave (steam sterilization) at different temperatures (e.g., at about 118° C. or at about 121° C.). It was observed that, unlike radiation sterilization, autoclave or steam sterilization did not increase the injection force required to expel the slurry from the syringe.

[0116] Example 6 - Method of Producing a Cold Slurry Composition Comprising Hyaluronic Acid and Soy PC

[0117] Procedure for preparing 0.5% Soy PC / HA / PBS / Glycerol or 0.5% Tween80 / HA / PBS / Glycerol for extrusion testing

[0118] 1. Preparation of PBS / Glycerol Solution

[0119] Weigh 20 g of glycerol in a 125 mL narrow-mouth conical flask with a magnetic stirring bar, add 80 mL of 1×PBS, seal the flask mouth with a sealing film to prevent solvent evaporation, and stir the liquid at 500 rpm for 4 hours to obtain PBS / glycerol (41 by weight / weight, or 5 / 1 by volume / volume).

[0120] 2. Preparation of 0.75% HA / PBS / glycerol solution

[0121] A certain amount of HA 1000kD (0.15 g to 0.75 g, to prepare 20 g to 100 g solution) was weighed in a 125 mL narrow-mouth conical flask with a magnetic stirring bar (8 mm × 35 mm), and a certain amount of PBS / glycerol solution (19.85 g to 99.25 g, to prepare 20 g to 100 g solution) was added. The mouth of the flask was sealed with a sealing film to prevent solvent evaporation. The liquid was stirred at 1200 rpm for 12 to 24 hours to obtain 0.75% HA / PBS / glycerol.

[0122] 3. Dissolve soybean PC in ethanol

[0123] 1 g of soybean PC was weighed in a 5 mL glass vial, 0.2 mL of ethanol was added with a pipette, vortexed, and then incubated at 45° C. for 12 to 24 hours to make it a clear solution with a light yellow color.

[0124] 4. Preparation of 0.5% Soybean PC / HA / PBS / Glycerol

[0125] Example of preparing 20 g sample

[0126] Weigh 0.116 g of the above prepared soy PC in ethanol into a 125 mL narrow-mouth conical flask using a disposable pipette. Add 19.884 g of 0.75% HA / PBS / glycerol. Place a magnetic stirring bar (8 mm × 35 mm) and seal the flask mouth with a sealing film to prevent solvent evaporation. Stir the liquid at 1200 rpm for 4 to 24 hours to obtain 0.5% soy PC / HA / PBS / glycerol.

[0127] 5. Preparation of 0.5% Tween 80 / HA / PBS / Glycerol

[0128] Example of preparing 20 g sample

[0129] Weigh 0.1 g of Tween 80 into a 125 mL narrow-mouth conical flask using a disposable pipette. Add 19.9 g of 0.75% HA / PBS / glycerol. Place a magnetic stirring bar (8 mm × 35 mm) and seal the flask with a sealing film to prevent solvent evaporation. Stir the liquid at 1200 rpm for 4 to 24 hours to obtain 0.5% Tween 80 / HA / PBS / glycerol.

[0130] 6. Transfer the above prepared 0.5% soybean PC / HA / PBS / glycerol or 0.5% Tween 80 / HA / PBS / glycerol to a 20 mL glass vial and sonicate for 10 m. Then fill the obtained liquid into 3×3 mL syringes through a 21G needle. Place the syringes in a -20°C freezer.

[0131] After 12 to 24 hours, the -20°C syringe was removed from the freezer and connected to an 18G needle for expulsion testing. The test should be performed within 1 minute of removal from the freezer.

Claims

1. A composition comprising: A certain amount of water; Hyaluronic acid; and A first excipient, wherein the composition is configured to form a flowable cold slurry comprising a plurality of ice crystals when the composition is exposed to a temperature of 0°C or less.

2. The composition according to claim 1, wherein the composition further comprises a water-soluble surfactant.

3. The composition of claim 2, wherein the water-soluble surfactant is a poloxamer molecule.

4. The composition of claim 3, wherein the composition comprises a plurality of poloxamer molecules.

5. The composition of claim 3, wherein the composition comprises poloxamer particles, and wherein the poloxamer particles comprise a plurality of poloxamer molecules.

6. The composition of claim 5, wherein the poloxamer particles are micelles.

7. The composition of claim 3, wherein the poloxamer molecule is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer Poloxamer 235, Poloxamer 237, Poloxamer 238, Poloxamer 282, Poloxamer 284, Poloxamer 288, Poloxamer 331, Poloxamer 333, Poloxamer 334, Poloxamer 335, Poloxamer 338, Poloxamer 401, Poloxamer 402, Poloxamer 403, Poloxamer 407, Poloxamer 105 benzoate, Poloxamer 182 dibenzoate, and combinations thereof.

8. The composition of claim 7, wherein the poloxamer is poloxamer 407, and wherein the concentration of poloxamer 407 is about 0.1% (w / w) to 10% (w / w).

9. The composition of claim 8, wherein the concentration of Poloxamer 407 in the composition is about 0.5% (w / w).

10. The composition of claim 1, wherein the first excipient is selected from the group consisting of salts, ions, lactated Ringer's solution, sugars, biocompatible surfactants, polyols, and combinations thereof.

11. The composition of claim 1, wherein the first excipient is glycerol.

12. The composition of claim 1, wherein the concentration of glycerol in the composition is about 12% (w / w) to 25% (w / w).

13. The composition of claim 12, wherein the concentration of glycerol in the composition is about 19% (w / w).

14. The composition of claim 1, wherein the composition further comprises a second excipient.

15. The composition of claim 14, wherein the second excipient is sodium chloride or sodium phosphate forming saline or phosphate buffered saline.

16. The composition of claim 14, wherein the composition further comprises a third excipient.

17. The composition according to claim 16, wherein the third excipient is a water-insoluble substance.

18. The composition according to claim 17, wherein the water-insoluble substance is a lipid.

19. The composition of claim 18, wherein the lipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoylphosphatidyl (DPPC), dicetyl phosphate (DCP), L-α-phosphatidylcholine (soybean PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylglycerol (PG).

20. The composition of claim 1, wherein the composition is configured to form the plurality of ice crystals when the composition is exposed to a temperature of about -25°C to about -5°C.

21. The composition of claim 1, wherein the composition is configured to have an injection force of less than about 30 lbf when injected through a 16G needle, a 17G needle, a 18G needle, a 19G needle, a 20G needle, a 22G needle, a 23G needle, or a 24G needle.

22. The composition of claim 1, wherein the composition is configured to have an injection force of less than about 30 lbf when injected through a 17G needle or an 18G needle.

23. A method of preparing a cold slurry for administration to a patient at a clinical point of care, the method comprising: preparing a composition comprising hyaluronic acid and an amount of water; adding a first excipient to the composition, wherein the excipient comprises a water-soluble surfactant; as well as wherein the composition is configured to form a cold slurry comprising a plurality of ice particles when the composition is cooled to a temperature below about 0°C.

24. The method of claim 23, wherein the water-soluble surfactant is a hydrotropic molecule.

25. The method of claim 24, wherein the hydrotropic molecule is a poloxamer molecule.

26. The method of claim 25, wherein the poloxamer molecule is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer Poloxamer 235, Poloxamer 237, Poloxamer 238, Poloxamer 282, Poloxamer 284, Poloxamer 288, Poloxamer 331, Poloxamer 333, Poloxamer 334, Poloxamer 335, Poloxamer 338, Poloxamer 401, Poloxamer 402, Poloxamer 403, Poloxamer 407, Poloxamer 105 benzoate, Poloxamer 182 dibenzoate, and combinations thereof.

27. The method of claim 25, wherein the poloxamer is poloxamer 407, and wherein the concentration of poloxamer 407 is about 0.1% (w / w) to 10% (w / w).

28. The method of claim 25, wherein the concentration of Poloxamer 407 in the composition is about 0.5% (w / w).

29. The method of claim 23, further comprising adding a second excipient to the composition, wherein the composition comprising the second excipient is configured to form the cold slurry when the composition is cooled to a temperature below about 0°C.

30. The method of claim 29, wherein the second excipient is selected from the group consisting of salts, ions, lactated Ringer's solution, sugars, biocompatible surfactants, polyols, and combinations thereof.

31. The method of claim 30, wherein the second excipient is glycerol.

32. The method of claim 31, wherein the concentration of glycerol in the composition is about 12% (w / w) to 25% (w / w).

33. The method of claim 31, wherein the concentration of glycerol in the composition is about 19% (w / w).

34. The method of claim 31, further comprising adding a third excipient to the composition, wherein the composition comprising the second excipient and the third excipient is configured to form the cold slurry when the composition is cooled to a temperature below about 0°C.

35. The method of claim 34, wherein the third excipient is sodium chloride or sodium phosphate to form saline or phosphate buffered saline.

36. The method of claim 34, further comprising adding a fourth excipient to the composition, wherein the composition comprising the second excipient, the third excipient, and the fourth excipient is configured to form the cold slurry when the composition is cooled to a temperature below about 0°C.

37. The method of claim 36, wherein the fourth excipient is a water-insoluble substance.

38. The method of claim 37, wherein the water-insoluble substance is a lipid.

39. The method of claim 38, wherein the lipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), egg sphingomyelin (DPSM), dipalmitoylphosphatidyl (DPPC), dicetyl phosphate (DCP), L-α-phosphatidylcholine (soybean PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylglycerol (PG).

40. The method of claim 23, wherein the composition is configured to form a plurality of ice crystals when the composition is exposed to a temperature of about -25°C to about -5°C.

41. The method of claim 23, wherein the composition is configured to have an injection force of less than about 30 lbf when injected through a 16G needle, a 17G needle, a 18G needle, a 19G needle, a 20G needle, a 22G needle, a 23G needle, or a 24G needle.

42. The method of claim 23, wherein the composition is configured to have an injection force of less than about 30 lbf when injected through a 17G needle or an 18G needle.

43. A method of preparing a cold slurry for administration to a patient at a clinical point of care, the method comprising: receiving a composition comprising an antifreeze agent and hyaluronic acid; as well as The composition is cooled to a temperature below about 0°C to form a cold slurry, wherein the cold slurry comprises a plurality of ice particles.

44. The method of claim 43, wherein the antifreeze agent is glycerol.

45. The method of claim 43, wherein the composition further comprises an amount of a poloxamer molecule.

46. ​​The method of claim 45, wherein the poloxamer molecule is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 183, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer Poloxamer 235, Poloxamer 237, Poloxamer 238, Poloxamer 282, Poloxamer 284, Poloxamer 288, Poloxamer 331, Poloxamer 333, Poloxamer 334, Poloxamer 335, Poloxamer 338, Poloxamer 401, Poloxamer 402, Poloxamer 403, Poloxamer 407, Poloxamer 105 benzoate, Poloxamer 182 dibenzoate, and combinations thereof.

47. The method of claim 46, wherein the poloxamer is poloxamer 407, and wherein the concentration of poloxamer 407 is about 0.1% (w / w) to 10% (w / w).

48. The method of claim 46, wherein the concentration of Poloxamer 407 in the composition is about 0.5% (w / w).

49. The method of claim 43, wherein receiving the composition comprises receiving the composition in a container.

50. The method of claim 49, wherein the container is a first syringe.

51. The method according to claim 50 further includes connecting the first syringe to a second syringe and processing the cold slurry through a reciprocating cycle, wherein the reciprocating cycle includes pushing the cold slurry from the first syringe into the second syringe, and pushing the cold slurry from the second syringe into the first syringe.

52. The method of claim 51 further comprising processing the cold slurry through a second, third or fourth reciprocating cycle.

53. The method of claim 49, wherein the container is a container configured for surface application.

54. The method of claim 53, wherein the container configured for surface application is a first tube.

55. The method of claim 54, further comprising connecting the first tube to a second tube and processing the slurry by a reciprocating cycle, wherein the reciprocating cycle comprises pushing the cold slurry from the first tube into the second tube, and pushing the cold slurry from the second tube into the first tube.

56. The method of any one of claims 43 to 55, further comprising monitoring the temperature of the cold slurry.

57. The method of claim 56, wherein the monitoring comprises viewing a temperature sensitive indicator on a syringe or container containing the cold slurry, wherein the temperature sensitive indicator is configured to indicate the temperature of the cold slurry.

58. The method of claim 57, wherein the temperature sensitive indicator is a temperature sensitive sticker.

59. The method of claim 57, wherein the temperature sensitive indicator provides a visual indication when the cold slurry reaches a predetermined temperature.

60. The method of claim 59, wherein the predetermined temperature is about -15°C.

61. The method of claim 59, wherein the predetermined temperature is about -19°C to -11°C, about -18°C to -12°C, about -17°C to -13°C, or about -16°C to -14°C.

62. A method according to any one of claims 59 to 61, wherein the temperature sensitive indicator is configured to provide a visual indication when the cold slurry is at a temperature cooler than the predetermined temperature.

63. A method according to any one of claims 59 to 62, wherein the temperature sensitive indicator is configured to provide a visual indication when the cold slurry is hotter than the predetermined temperature.

64. The method of claim 56, wherein the monitoring comprises checking a thermometer.

65. The method of claim 56, wherein the monitoring comprises viewing a temperature monitoring assembly embedded within a container containing the cold slurry.

66. The method of claim 65, wherein a temperature component is disposed in the container or along the fluid path.

67. The method of any one of claims 56 to 66, wherein the monitoring further comprises listening to an audio indicator configured to indicate when the composition reaches a predetermined temperature.

68. The method of any one of claims 56 to 67, wherein the composition is terminally sterilized.

69. The method of claim 68, wherein the composition is terminally sterilized via autoclaving or steam sterilization.

70. The method of claim 69, wherein the autoclaving or steam sterilization comprises subjecting the composition to a temperature of about 118°C to 121°C.

71. The method of claim 70, wherein the temperature is about 118°C.

72. A cold slurry conveying system, comprising: A container containing a slurry composition, the container comprising a sterile barrier and a temperature indicator, wherein the container is configured to allow manual stirring of the slurry composition without breaching the sterile barrier.

73. The system of claim 72, wherein the container is a syringe or a tube.

74. The system of claim 73, wherein the container is configured to be connected to a second container.

75. The system of claim 74, wherein the container and the second container are configured to be connected using a connector.

76. The system of claim 75, wherein the container and the second container comprise a first syringe and a second syringe, wherein the first syringe and the second syringe each comprise a male Luer assembly.

77. The system of claim 76, wherein the connector comprises a female Luer assembly.

78. The system of claim 77, wherein the first syringe and the second syringe are connected using the female Luer assembly, and wherein the slurry composition is movable from the first syringe to the second syringe to manually stir the slurry composition.

79. The system of any one of claims 72 to 78, further comprising a temperature sensitive indicator disposed on the container.

80. The system of claim 79, wherein the temperature sensitive indicator comprises a temperature sensitive sticker.

81. The system of claim 80, wherein the temperature sensitive indicator provides a visual indication when the slurry composition reaches a predetermined temperature.

82. The system of claim 81, wherein the predetermined temperature is approximately -15°C.

83. The system of claim 81, wherein the predetermined temperature is about -19°C to -11°C, about -18°C to -12°C, about -17°C to -13°C, or about -16°C to -14°C.

84. The system of any one of claims 79 to 83, wherein the temperature sensitive indicator provides a visual indication when the slurry composition is at a temperature cooler than the predetermined temperature.

85. The system of any one of claims 79 to 84, wherein the temperature sensitive indicator provides a visual indication when the slurry composition is hotter than the predetermined temperature.

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