Solid forms of resiquimod and formulations thereof

Abstract

The present disclosure provides solid forms of resiquimod that are useful, for example, as immunomodulatory payload components of certain biomaterials (eg, hydrogels) and / or formulations for administration to a subject.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Application No. 63 / 286,361, filed December 6, 2021, which is incorporated by reference in its entirety herein. [Background technology]

[0002] Surgery is often the first-line treatment for solid tumor cancer, and is generally used in combination with systemic administration of anticancer therapy. However, surgery-induced immunosuppression is associated with the development of postoperative infectious complications and tumor metastasis due to changes in various metabolic and endocrine responses, which ultimately lead to the death of many patients (Hiller, JGet al.Nature Reviews Clinical Oncology, 2018, 15, 205-218).

[0003] Systemic administration of drugs, nutrients, or other substances into the circulatory system affects the entire body. Systemic administration routes include enteral administration (e.g., oral administration resulting in absorption of the drug through the digestive tract) and parenteral administration (e.g., intravenous, intramuscular, and subcutaneous injection). The administration of immunotherapeutic agents usually relies on these systemic administration routes, which may result in undesirable side effects. In some instances, certain promising therapeutic agents are extremely difficult to develop due to the associated toxicities and limitations of current administration methods and systems. Hydrogels are a particularly attractive type of biomaterial and have been used in a wide range of applications, including tissue engineering and regenerative medicine, diagnostics, cell immobilization, and / or drug delivery. However, existing hydrogels have several limitations that limit the practical use of hydrogel-based drug delivery therapies. For example, bulk hydrogels have well-defined dimensions that can make them difficult to extrude through a needle, so many hydrogels are typically formed outside the body and then implanted. Although some hydrogels can be formed in situ in vivo, there may be potential risks and challenges associated with certain crosslinking agents, such as UV radiation and / or crosslinking chemicals. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Hiller,JGet al.Nature Reviews Clinical Oncology,2018,15,205-218 Summary of the Invention [Means for solving the problem]

[0005] The present disclosure provides solid forms of resiquimod, as well as compositions and methods for preparing the solid forms of resiquimod. In some embodiments, the solid forms provided are highly useful as immunomodulatory payload components of certain biomaterials (e.g., hydrogels) and / or formulations, for example, for administration to subjects who have undergone or have undergone tumor resection. In some embodiments, the solid forms provided exhibit certain desirable properties, such as, for example, certain solubility, stability, and / or hygroscopicity.

[0006] In some embodiments, the disclosure provides a method of preparing a formulation comprising providing a solid form of resiquimod. For example, in some embodiments, the disclosure provides a method of preparing a formulation suitable for intraoperative administration comprising providing a solid form of resiquimod.

[0007] In some embodiments, the disclosure provides methods of using the provided formulations of resiquimod. [Brief description of the drawings]

[0008] [Figure 1] 1 is an XRPD pattern of resiquimod form I.

[0009] [Diagram 2] 1 is a DSC thermogram of resiquimod form I.

[0010] [Diagram 3] 1 is a TGA thermogram of resiquimod form I.

[0011] [Figure 4] FIG. 1 is an XRPD pattern of resiquimod form II.

[0012] [Diagram 5] 1 is a DSC thermogram of resiquimod form II.

[0013] [Figure 6] 1 is a TGA thermogram of resiquimod Form II.

[0014] [Figure 7] 1 shows an XRPD pattern of resiquimod form III (top spectrum) and an XRPD pattern of resiquimod form I (bottom spectrum) obtained by drying form III.

[0015] [Figure 8] 1 is a DSC thermogram of a sample of resiquimod Form III after drying.

[0016] [Figure 9] 1 is a TGA thermogram of a sample of resiquimod Form III after drying.

[0017] [Figure 10] FIG. 1 is an XRPD pattern of resiquimod form IV.

[0018] [Figure 11] 1 is a DSC thermogram of resiquimod Form IV.

[0019] [Figure 12] 1 is a TGA thermogram of resiquimod Form IV.

[0020] [Figure 13] The top spectrum is the XRPD pattern of resiquimod form V. Upon grinding and / or drying the sample was converted to resiquimod form I (bottom three spectra).

[0021] [Figure 14] 1 is a DSC thermogram of resiquimod form V.

[0022] [Figure 15] 1 is a TGA thermogram of resiquimod form V.

[0023] [Figure 16] 1 is an XRPD pattern of resiquimod form VI.

[0024] [Figure 17] 1 is a DSC thermogram of resiquimod Form VI.

[0025] [Figure 18] 1 is a TGA thermogram of resiquimod Form VI.

[0026] [Figure 19] FIG. 1 is an XRPD pattern of resiquimod form VII.

[0027] [Figure 20] 1 is a DSC thermogram of resiquimod form VII.

[0028] [Figure 21] 1 is a TGA thermogram of resiquimod form VII.

[0029] [Figure 22] A-B show heat maps showing the gelling properties of an exemplary temperature-responsive polymer combination preparation including P407 at concentrations indicated in % (w / w) and hyaluronic acid (HA) with an average molecular weight of 1.5 MDa at concentrations indicated in % (w / w) in two different buffer systems. The temperature-responsive polymer combination preparation was exposed to a temperature of 37° C. to observe any gel formation. A polymer combination preparation is determined to form a gel if such a polymer combination preparation becomes translucent or opaque and is not flowable when tilted or turned upside down. A corresponds to 10 mM phosphate buffered saline (PBS) at pH 7.4. B corresponds to 0.1 M bicarbonate buffer at pH 8.0.

[0030] [Figure 23] A-B show heat maps showing the gelling properties of an exemplary temperature-responsive polymer combination preparation including P407 at concentrations indicated in % (w / w) and hyaluronic acid (HA) having an average molecular weight of 730 kDa at concentrations indicated in % (w / w) in two different buffer systems. The polymer combination preparation was exposed to a temperature of 37° C. to observe any gel formation. The temperature-responsive polymer combination preparation was exposed to a temperature of 37° C. to observe any gel formation. A polymer combination preparation is determined to form a gel if such polymer combination preparation becomes translucent or opaque and is not flowable when tilted or turned upside down. A corresponds to 10 mM PBS at pH 7.4. B corresponds to 0.1 M bicarbonate buffer at pH 8.0.

[0031] [Figure 24]1 shows a heat map showing the gelation properties of an exemplary temperature-responsive polymer combination preparation comprising P407 at a concentration indicated in % (w / w) and modified chitosan (e.g., carboxymethyl chitosan; CMCH) at a concentration indicated in % (w / w) in 10 mM PBS at pH 7.4. The temperature-responsive polymer combination preparation was exposed to a temperature of 37° C. to observe any gel formation. A polymer combination preparation is determined to form a gel if such a polymer combination preparation becomes translucent or opaque and is not flowable when tilted or turned upside down.

[0032] [Diagram 25] AB show graphical diagrams illustrating the storage modulus of an exemplary temperature-responsive polymer combination formulation compared to a control polymer composition after exposure to a temperature of 37° C. A: Linear scale. B: Logarithmic scale. Abbreviations: "18% P407" = 18% (w / w) P407, "13.5% P407 + 0.65% 1.5 MDa HA (10 mM PBS)" = 13.5% (w / w) P407 + 0.65% (w / w) 1.5 MDa HA in 10 mM PBS at pH 7.4, "13.5% P407 + 0.65% 1.5 MDa HA (0.1 M bicarbonate)" = 13.5% (w / w) P407 + 0.65% (w / w) 1.5 MDa HA in 0.1 M bicarbonate buffer at pH 8, "10% P407 + 1% 1.5 MDa HA (10 mM "13.5%P407+1.3%CMCH" = 13.5% (w / w) P407 + 1.3% (w / w) CMCH in 10 mM PBS pH 7.4, "12.5% ​​Extralink" = 12.5% ​​hyaluronic acid chemically crosslinked with Extralink thiol crosslinker, "1.5% Extralink" = 1.5% hyaluronic acid chemically crosslinked with Extralink thiol crosslinker, "0.5% Extralink" = 0.5% hyaluronic acid chemically crosslinked with Extralink thiol crosslinker.

[0033] [Figure 26]A-D show graphs illustrating the homogeneity of an exemplary thermoresponsive polymer combination preparation in the hydrogel state (when its precursor state was maintained at temperatures between 2-8°C over a period of one month) compared to a control polymer composition, with weekly measurements performed at 37°C (above CGT). Gel homogeneity was determined by measuring the storage modulus of the hydrogel over a period of time. A: Control gel (18% w / w poloxamer 407). B: Thermoresponsive polymer combination preparation of 13.5% w / w poloxamer 407 and 0.65% w / w 1.5 MDa HA in 10 mM PBS pH 7.4. C: Thermoresponsive polymer combination preparation of 10% w / w poloxamer 407 and 1% w / w 1.5 MDa HA in 10 mM PBS pH 7.4. D: Temperature-responsive polymer combination preparation of 13.5% w / w poloxamer 407 and 0.65% w / w 1.5 MDa HA in 0.1 M bicarbonate buffer at pH 8.0.

[0034] [Figure 27]A-E show graphs showing in vivo survival data of animals that underwent tumor resection and received either an exemplary thermoresponsive polymer combination preparation in hydrogel form alone or an exemplary thermoresponsive polymer combination preparation in hydrogel form incorporating resiquimod, compared to a chemically crosslinked control hyaluronic acid hydrogel alone or a chemically crosslinked control hyaluronic acid hydrogel incorporating resiquimod. The x-axis shows time after tumor inoculation. Tumor resection was performed 10 days after tumor inoculation, and the exemplary compositions were administered after tumor resection. A: Control 12.5% ​​(w / v) Extralink® hyaluronic acid (HyStem™) hydrogel with or without resiquimod. B: Thermoresponsive polymer combination preparation of 10% w / w poloxamer 407 and 1% w / w 1.5 MDa HA in 10 mM PBS, pH 7.4, with or without resiquimod. C: Temperature-responsive polymer combination preparation of 13.5% w / w Poloxamer 407 and 0.65% w / w 1.5 MDa HA in 10 mM PBS, pH 7.4, with or without resiquimod. D: Temperature-responsive polymer combination preparation of 13.5% w / w Poloxamer 407 and 0.65% w / w 1.5 MDa HA in 0.1 M bicarbonate buffer, pH 8.0, with or without resiquimod. E: Temperature-responsive polymer combination preparation of 13.5% w / w Poloxamer 407 and 1.3% w / w CMCH in 10 mM PBS, pH 7.4, with or without resiquimod.

[0035] [Figure 28]A-D show graphs depicting in vivo survival data of animals that underwent tumor resection and were administered an exemplary thermoresponsive polymer combination preparation (e.g., a thermoresponsive liquid preparation comprising a combination of 730 kDa or 1.5 MDa hyaluronic acid (HA) and poloxamer (e.g., P407) at different concentrations) either as a sole polymer combination or as a polymer combination incorporating resiquimod. The x-axis indicates time after tumor inoculation. Tumor resection was performed on day 10 after tumor inoculation, and the exemplary compositions were administered after tumor resection. A: A thermoresponsive polymer combination preparation of 10% w / w poloxamer 407 and 2.25% w / w 730 kDa HA in 12.5 mM PBS, pH 8, with or without resiquimod. B: Temperature-responsive polymer combination preparation of 10% w / w poloxamer 407 and 2.25% w / w 730 kDa HA in 25 mM PBS, pH 8, with or without resiquimod. C: Temperature-responsive polymer combination preparation of 12.5% ​​w / w poloxamer 407 and 1.625% 730 kDa HA in 25 mM PBS, pH 8, with or without resiquimod. D: Temperature-responsive polymer combination preparation of 8% w / w poloxamer 407 and 2.25% w / w 730 kDa HA in 25 mM buffered saline, pH 8, with or without resiquimod.

[0036] [Figure 29] FIG. 1 shows a graph illustrating in vivo survival data of animals that underwent tumor resection and were administered an exemplary thermoresponsive polymer combination preparation (e.g., a thermoresponsive liquid preparation comprising a combination of 119 kDa hyaluronic acid (HA) and a poloxamer (e.g., P407)) either as the sole polymer combination or as a polymer combination incorporating resiquimod. Results are shown for a thermoresponsive polymer combination preparation of 10% w / w poloxamer 407 and 4% w / w 119 kDa HA in 25 mM buffered saline (pH 7.4) with or without resiquimod. The x-axis indicates time after tumor inoculation. Tumor resection was performed 10 days after tumor inoculation, and the exemplary composition was administered after tumor resection.

[0037] [Diagram 30] FIG. 1 shows a graph showing in vivo survival data of animals that underwent tumor resection and were administered an exemplary thermoresponsive polymer combination preparation (e.g., a thermoresponsive liquid preparation comprising a combination of 309 kDa hyaluronic acid (HA) and a poloxamer (e.g., P407)) as the sole polymer combination or a polymer combination incorporating resiquimod, or a poloxamer-only control animal cohort. Results are shown for a thermoresponsive polymer combination preparation of 10% w / w poloxamer 407 and 2% w / w 309 kDa HA in 25 mM buffered saline at pH 7.4 with or without resiquimod, and a control preparation of 15% poloxamer 407 biomaterial without active agent. The x-axis shows the time after tumor inoculation. Tumor resection was performed 10 days after tumor inoculation, and the exemplary composition was administered after tumor resection. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0038] definition It should be noted that the concentration of each individual polymer component in the polymer combination preparations described herein is expressed as % (w / w) or weight %, respectively. As used herein, the concentration of a polymer component in a polymer combination preparation, % (w / w), is determined based on the mass or weight of the polymer component relative to (i) the total mass or weight of all individual polymer components present in the polymer combination preparation and (ii) the total mass or weight of the solvent used in the polymer combination preparation.

[0039] The term "about" as used herein in relation to a value refers to a value that is similar in relation to the referenced value. Generally, a person skilled in the art familiar with such a context will recognize the degree of variation associated with "about" in that context. In some embodiments, the term "about" refers to ±10% of the given value.

[0040] As used herein, the terms "administer", "administering", or "administration" generally refer to administration of a composition to a subject to achieve delivery of the composition or agent or payload contained therein to a target site or site to be treated. Those skilled in the art will be aware of the various routes that may be utilized for administration of different agents to a subject, e.g., a human, in the appropriate circumstances. For example, the terms "administer", "administering", or "administration" may refer to implanting, absorbing, ingesting, injecting, inhaling, parenterally administering, or otherwise introducing a composition as described herein, although in the context of administration of a composition comprising a provided polymer combination preparation, administering may refer in some embodiments to implanting, or in some embodiments to injecting.

[0041] As used herein, the term "biocompatible" refers to a material that, for example, when placed in contact with living tissue in vivo, does not cause significant harm to such tissue. The biocompatibility of a material may be evaluated by the ability of such material to pass the biocompatibility tests set forth in International Organization for Standardization (ISO) Standard No. 10993 and / or United States Pharmacopeia (USP) 23 and / or U.S. Food and Drug Administration (FDA) Blue Book Memorandum #G95-1 entitled "Use of International Standard ISO-10993, Biological Evaluation of Medical Devices Part-1: Evaluation and Testing." Typically, these tests measure the toxicity, infectivity, pyrogenicity, irritancy, reactivity, hemolytic activity, carcinogenicity, and / or immunogenicity of the material. In certain embodiments, materials are "biocompatible" if they are not themselves toxic to cells in the in vivo environment of their application. In certain embodiments, substances are "biocompatible" if their addition to cells in vitro results in 20% or less cell death and / or if their administration in vivo does not induce significant severe inflammation or other such adverse effects that are clinically undesirable for the purposes described herein. As will be understood by those skilled in the art, such significant severe inflammation is distinguishable from the transient low-grade inflammation that typically accompanies surgery or the introduction of a foreign body into the body. Furthermore, those skilled in the art reading this disclosure will understand that in some embodiments, the polymer combination preparations described herein and / or their individual polymer components are biocompatible if a degree of immune modulation (e.g., innate immune agonism) over a defined period of time is clinically beneficial and / or desirable, for example, to provide anti-tumor immunity.

[0042] As used herein, the term "biodegradable" refers to a material that, when introduced into a cell, is broken down (e.g., by cellular machinery, e.g., by enzymatic degradation, by hydrolysis, and / or a combination thereof) into components that the cell can either reuse or remove without significant toxic effects to the cell. In certain embodiments, the components produced by the degradation of a biodegradable material are biocompatible and therefore do not induce significant severe inflammation and / or other adverse effects in vivo that are clinically undesirable for the purposes described herein. In some embodiments, biodegradable polymeric materials are broken down into their constituent monomers. In some embodiments, biodegradable polymeric materials can be degraded by organisms, e.g., by enzymatic activity or cellular machinery, in some cases, e.g., by exposure to lysozyme (e.g., having a relatively low pH), or by simple hydrolysis. In some embodiments, the degradation of a biodegradable material (including, e.g., a biodegradable polymeric material) involves hydrolysis of ester bonds. Alternatively or additionally, in some embodiments, degradation of the biodegradable material (including, for example, biodegradable polymeric materials) involves cleavage of urethane bonds. Exemplary biodegradable polymers include, for example, polymers of hydroxy acids such as lactic acid and glycolic acid, including, but not limited to, poly(hydroxyl acid), poly(lactic acid) (PLA), poly(glycolic acid) (PGA), poly(lactic-co-glycolic acid) (PLGA), and copolymers with PEG, polyanhydrides, poly(ortho)esters, polyesters, polyurethanes, poly(butyric acid), poly(valeric acid), poly(caprolactone), poly(hydroxyalkanoates), poly(lactide-caprolactone copolymers), mixtures and copolymers thereof. Many naturally occurring polymers are also biodegradable, including, for example, proteins such as albumin, collagen, gelatin, and prolamines (e.g., zein), as well as polysaccharides such as alginates, cellulose variants, and polyhydroxyalkanoates, such as polyhydroxybutyric acid, mixtures and copolymers thereof.Those of skill in the art will understand or be able to determine when such a polymer is biocompatible and / or when it is a biodegradable variant thereof (e.g., related to the parent polymer by substantially identical structure differing only in the substitution or addition of certain chemical groups, as known in the art).

[0043] The term "cancer" refers to a malignant neoplasm (Stedman's Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990). Of particular interest in the context of some embodiments of the present disclosure are cancers that are treated by cell killing and / or ablative therapy (e.g., surgical resection and / or certain chemotherapeutic drug therapy, such as cytotoxic therapy). In some embodiments, the cancer treated according to the present disclosure is one that has been surgically resected (i.e., at least one tumor has been surgically removed). In some embodiments, the cancer treated according to the present disclosure is one where resection is the standard of care. In some embodiments, the cancer treated according to the present disclosure is one that has metastasized.

[0044] The term "carbohydrate polymer" refers to a polymer that is or includes, for example, one or more carbohydrates having a carbohydrate backbone. For example, in some embodiments, a carbohydrate polymer refers to a polysaccharide or oligosaccharide, or a polymer that contains multiple monosaccharide units covalently linked. The monosaccharide units may all be identical, or in some cases, there may be more than one type of monosaccharide unit present in the carbohydrate polymer. In certain embodiments, the carbohydrate polymer is naturally occurring. In certain embodiments, the carbohydrate polymer is synthetic (i.e., not naturally occurring). In some embodiments, the carbohydrate polymer may include chemical modifications. In some embodiments, the carbohydrate polymer is a linear polymer. In some embodiments, the carbohydrate polymer is a branched polymer.

[0045] As used herein, the term "comparable" refers to two or more agents, entities, situations, sets of conditions, etc. that may not be identical to each other, but are sufficiently similar to allow a comparison between them so that a person skilled in the art will understand that a conclusion can be reasonably drawn based on the observed differences or similarities. In some embodiments, a set of comparable conditions, situations, individuals, or populations is characterized by a number of substantially identical characteristics and one or a few varying characteristics. A person skilled in the art will understand what degree of identity is required for two or more such agents, entities, situations, sets of conditions, etc. to be considered comparable in any given situation in the context. For example, a person skilled in the art will understand that a set of situations, individuals, or populations is comparable to each other when it is characterized by a sufficient number and type of substantially identical characteristics to ensure a reasonable conclusion that differences in results obtained or phenomena observed under or with the different sets of situations, individuals, or populations are caused by or indicate the existence of differences in various of those characteristics. Those skilled in the art will also understand that when the term "comparable" is used in the context of comparing two or more values, such values ​​are comparable to each other such that the difference in the values ​​does not result in a substantial difference in the therapeutic outcome, e.g., induction of anti-tumor immunity and / or incidence of tumor regrowth and / or metastasis. For example, in some embodiments, comparable release rates refer to such release rate values ​​within 15% over 48 hours. In some embodiments, comparable release rates refer to such release rate values ​​within 20% over 48 hours. In some embodiments, comparable release rates refer to such release rate values ​​within 15% over 24 hours.

[0046] Critical gelation temperature: As used herein, the term "critical gelation temperature," abbreviated as "CGT," refers to a threshold temperature at or above which a precursor state of a polymer combination preparation (e.g., as described herein) transitions to a polymer network state (e.g., a hydrogel state) as described herein. In some embodiments, the critical gelation temperature may correspond to a sol-gel transition temperature. In some embodiments, the critical gelation temperature may correspond to a lower critical solution temperature. For a general description of thermoresponsive gels, see Taylor et al., "Thermoresponsive Gels," Gels (2017) 3:4, the contents of which are incorporated herein by reference for purposes described herein. As described in this disclosure, certain embodiments of the polymer combination preparations described herein have been shown to form a polymer network state upon exposure to temperatures of about 35-40°C. One of skill in the art reading this disclosure will understand that such polymer combination preparations do not necessarily have a CGT of about 35-40° C., but rather may have a CGT of less than 35-40° C. For example, in some embodiments, the provided polymer combination preparations may have a CGT of about 20-28° C.

[0047] As used herein, the term "critical gelation weight ratio" refers to a threshold weight ratio of at least two or more polymer components in a provided polymer combination formulation at or above which a precursor state (e.g., as described herein) of such a polymer combination formulation transitions to a polymer network state (e.g., a hydrogel state) as described herein. In some embodiments, such a precursor state-polymer network state transition occurs when both the critical gelation temperature and the critical gelation weight ratio of a provided polymer combination formulation are achieved.

[0048] As used herein, the term "crosslinking" refers to interactions and / or bonds between one entity and another entity to form a network. For example, in some embodiments, the crosslinks present in a polymer network can be or include intramolecular crosslinks, intermolecular crosslinks, or both. In some embodiments, the crosslinks can include interactions and / or bonds between one polymer chain(s) and another polymer chain(s) to form a polymer network. In some embodiments, the crosslinks can be achieved using one or more physical crosslinking techniques, including, for example, one or more environmental triggers and / or physiochemical interactions. Examples of environmental triggers include, but are not limited to, pH, temperature, and / or ionic strength. Non-limiting examples of physiochemical interactions include hydrophobic interactions, charge interactions, hydrogen bonding interactions, stereocomplexation, and / or supramolecular chemistry. In some embodiments, crosslinking can be achieved using one or more covalent crosslinking techniques based on chemical reactions (e.g., the bond between two entities is or includes a covalent bond), for example, in some embodiments, this can include the reaction of an aldehyde and an amine to form a Schiff base, an aldehyde and a hydrazide to form a hydrazine, and / or the Michael reaction of an acrylic acid and either a primary amine or a thiol to form a secondary amine or a sulfide. Examples of such covalent crosslinking techniques include, but are not limited to, small molecule crosslinking and polymer-polymer crosslinking. Various methods for physical and covalent crosslinking of polymer chains are known in the art, for example, as described in Hoare and Kohane, “Hydrogels in drug delivery: Progress and challenges” Polymer (2008) 49:1993-2007, the entire contents of which are incorporated herein by reference for the purposes described herein.

[0049] The term "crosslinker" or "crosslinking agent", used interchangeably herein, refers to an agent that links one entity (e.g., one polymer chain) to another entity (e.g., another polymer chain). In some embodiments, the bond between the two entities (i.e., "crosslink") is or includes a covalent bond. In some embodiments, the bond between the two entities is or includes an ionic bond or ionic interaction. In some embodiments, the crosslinker is a chemical crosslinker, e.g., in some embodiments, this can be or includes a small molecule (e.g., dialdehyde or genipin) to induce covalent bond formation between an aldehyde group and an amino group. In some embodiments, the crosslinker includes a photosensitive functional group. In some embodiments, the crosslinker includes a pH-sensitive functional group. In some embodiments, the crosslinker includes a heat-sensitive functional group.

[0050] The term "hydrogel" has its art-understood meaning and refers to a material formed from a network of hydrophilic polymer chains, sometimes found as a colloidal gel in which the aqueous phase is the dispersion medium. In some embodiments, hydrogels are natural or synthetic polymer networks that are highly absorbent (e.g., capable of absorbing and / or retaining more than 90% water). In some embodiments, hydrogels have a degree of flexibility comparable to that of natural tissues, for example, due to their significant water content.

[0051] The terms "neoplasm" and "tumor" are used interchangeably herein and refer to an abnormal mass of tissue whose proliferation exceeds and is not coordinated with the growth of normal tissue. A neoplasm or tumor can be "benign" or "malignant" depending on the following characteristics: degree of cellular differentiation (including morphology and function), rate of growth, local invasion, and metastasis. "Benign neoplasms" are generally well differentiated, have characteristically slower growth than malignant neoplasms, and remain localized at the site of origin. In addition, benign neoplasms do not have the ability to invade, invade, or metastasize to distant sites. Exemplary benign neoplasms include, but are not limited to, lipomas, chondromas, adenomas, acrochordons, senile hemangiomas, seborrheic keratosis, lentigines, and sebaceous hyperplasia. In some cases, certain "benign" tumors may subsequently give rise to malignant neoplasms, which may arise as a result of further genetic alterations in a subpopulation of the tumor's tumor cells; these tumors are referred to as "pre-malignant neoplasms." One example of a pre-malignant neoplasm is a teratoma. In contrast, "malignant neoplasms" are generally poorly differentiated (anaplastic) and have characteristically rapid growth accompanied by progressive infiltration, invasion, and destruction of surrounding tissue. Moreover, malignant neoplasms generally have the ability to metastasize to distant sites.

[0052] As used herein, the term "poloxamer" refers to a polymer preparation of one or more poloxamers or a polymer preparation comprising one or more poloxamers. In some embodiments, the poloxamer in the polymer preparation may be unconjugated or unmodified, e.g., it is typically a triblock copolymer comprising a hydrophobic chain of polyoxypropylene (polypropylene glycol, PPG) flanked by two hydrophilic chains of polyoxyethylene (polyethylene glycol, PEG). In some embodiments, the polymer preparation of one or more poloxamers or the polymer preparation comprising one or more poloxamers may be unfiltered (e.g., such a polymer preparation may contain impurities and / or relatively low molecular weight polymer molecules compared to a comparable polymer preparation that has been filtered or fractionated or otherwise purified). Examples of poloxamers include, but are not limited to, poloxamer 124 (P124, also known as Pluronic® L44 NF), poloxamer 188 (P188, also known as Pluronic F68 NF), poloxamer 237 (P237, also known as Pluronic F87 NF), poloxamer 338 (P338, also known as Pluronic F108 NF), poloxamer 407 (P407, also known as Pluronic F127 NF), and combinations thereof.

[0053] The term "polymer" is given its ordinary meaning as used in the art, i.e., a molecular structure that includes one or more repeating units (monomers) that are covalently linked. The repeating units may all be identical, or in some cases, there may be more than one type of repeating unit present in a polymer (e.g., a copolymer). In certain embodiments, the polymer is naturally occurring. In certain embodiments, the polymer is synthetic (i.e., not naturally occurring). In some embodiments, the polymer is a linear polymer. In some embodiments, the polymer is a branched polymer. In some embodiments, a polymer for use in accordance with the present disclosure is not a polypeptide. In some embodiments, a polymer for use in accordance with the present disclosure is not a nucleic acid.

[0054] As used herein, the term "polymer combination preparation" refers to a polymeric biomaterial that comprises at least two distinct polymeric components. For example, in many embodiments, the polymeric combination preparation described herein is a polymeric biomaterial that comprises a first polymeric component and a second first polymeric component, where the first polymeric component is or comprises at least one poloxamer, and the second polymeric component is or comprises a non-poloxamer polymer. In some embodiments, the polymeric combination preparation described herein is a polymeric biomaterial in a precursor state that may be useful, for example, for administration to a subject. In some embodiments, the polymeric combination preparation described herein is a polymeric biomaterial in a polymeric network state.

[0055] "Subjects" to which administration is contemplated include, but are not limited to, humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly adults)) and / or non-human animals, e.g., mammals (e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys); domestic animals, e.g., cows, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds (e.g., chickens, ducks, geese, and / or turkeys)). In certain embodiments, the animal is a mammal (e.g., at any stage of development). In some embodiments, the animal (e.g., non-human animal) can be a transgenic or genetically engineered animal. In some embodiments, the subject is a tumor resection subject, e.g., a subject that has recently undergone tumor resection. In some embodiments, the tumor resection subject is a subject who has undergone tumor resection less than 72 hours (including, for example, less than 48 hours, less than 24 hours, less than 12 hours, less than 6 hours, or less than) before receiving the compositions described herein. In some embodiments, the tumor resection subject is a subject who has undergone tumor resection less than 48 hours before receiving the compositions described herein. In some embodiments, the tumor resection subject is a subject who has undergone tumor resection less than 24 hours before receiving the compositions described herein. In some embodiments, the tumor resection subject is a subject who has undergone tumor resection less than 12 hours before receiving the compositions described herein.

[0056] As used interchangeably herein, the terms "extended" or "sustained" generally refer to prolonging an effect and / or process over a desired period of time. For example, in the context of sustained immunomodulation (e.g., in the presence of a composition or preparation described and / or utilized herein), such immunomodulatory effects may be observed for a longer period of time following administration of a particular immunomodulatory payload in the context of a composition including a biomaterial preparation, as otherwise described herein, as compared to that observed by administration of the same payload in the absence of such a biomaterial preparation. In the context of sustained release of one or more agents of interest over a period of time from a composition and / or preparation described herein (e.g., a payload incorporated in a polymer combination preparation described herein and / or a degradation or dissolution product and / or soluble component of a polymer combination preparation described herein that modulates one or more aspects of the immune response, such as, but not limited to, innate immune agonism), such release may occur on a time scale ranging from about 30 minutes to several weeks. In some embodiments, the extent of sustained or sustained release may be characterized in vitro or in vivo. For example, in some embodiments, the release kinetics can be tested in vitro by placing the preparations and / or compositions described herein in an aqueous buffer (e.g., PBS at pH 7.4). In some embodiments, when the preparations and / or compositions described herein are placed in an aqueous buffer (e.g., PBS at pH 7.4), less than 100% or less (e.g., including 90% or less, 80% or less, 70% or less, 50% or less, or less) of the one or more agents of interest (e.g., payloads incorporated into the polymer combination preparations described herein that modulate one or more aspects of the immune response, such as, but not limited to, innate immune agonism, and / or degradation or dissolution products and / or soluble components of the polymer combination preparations described herein) are released from the biomaterial within 3 hours when the preparations and / or compositions described herein are placed in an aqueous buffer (e.g., PBS at pH 7.4). In some embodiments, the release kinetics can be tested in vivo, for example, by implanting the composition into a target site (e.g., mammary fat pad) of an animal subject (e.g., a murine subject).In some embodiments, when the composition is implanted into a target site (e.g., mammary fat pad) of an animal subject (e.g., a murine subject), no more than 70% or less (including, e.g., no more than 60%, no more than 50%, less than 40%, less than 30%, or less) of the one or more agents of interest (e.g., payloads incorporated into the polymer combination preparations described herein and / or degradation or dissolution products and / or soluble components of the polymer combination preparations described herein that modulate one or more aspects of the immune response, including but not limited to innate immune agonism) are released in vivo 8 hours after implantation.

[0057] As used herein, in the context of a temperature-responsive polymer or biomaterial (e.g., a polymeric biomaterial), the term "temperature-responsive" refers to a polymer or biomaterial (e.g., a polymeric biomaterial) that exhibits an instantaneous or intermittent change in one or more of its properties at a critical temperature (e.g., a critical gelation temperature). For example, in some embodiments, one or more of such properties is or includes the solubility of the polymer or biomaterial in a particular solvent. By way of example only, in some embodiments, a temperature-responsive polymer or biomaterial (e.g., a polymeric biomaterial) is characterized by being a homogenous polymer solution or colloid that is stable below a critical temperature (e.g., a critical gelation temperature) and instantaneously forming a polymer network (e.g., a hydrogel) when the critical temperature (e.g., a critical gelation temperature) is reached or exceeded. In some embodiments, a temperature-responsive polymer or biomaterial (e.g., a polymeric biomaterial) may be temperature reversible, where, for example, in some embodiments, a polymer solution may instantaneously form a polymer network at or above a critical gelation temperature, and such resulting polymer network may instantaneously revert to a homogenous polymer solution when the temperature is reduced below the critical gelation temperature.

[0058] The terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset, or inhibiting the progression of a "pathology" as described herein (e.g., a disease, disorder, or condition, including one or more signs or symptoms thereof), e.g., a cancer or tumor. In some embodiments, treatment may be administered after one or more signs or symptoms have developed or been observed. Treatment may be continued after symptoms have resolved, e.g., to delay or prevent recurrence and / or spread.

[0059] As used herein, the term "tumor resection subject" refers to a subject undergoing or having recently undergone tumor resection. In some embodiments, a tumor resection subject is a subject in which at least 70% or more of the total tumor mass (including at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or more (including 100%)) has been removed by surgical resection. In some cases, even when gross visual examination shows that all of the total tumor mass has apparently been removed, those skilled in the art will understand that there may be some residual cancer cells present microscopically at the visible resection margin. In some embodiments, a tumor resection subject may be determined to have negative resection margins (i.e., based on, for example, histological evaluation of the tissue surrounding the tumor resection site, there are no cancer cells microscopically present at the resection margin). In some embodiments, a tumor resection subject may be determined to have positive resection margins (i.e., based on, for example, histological evaluation of the tissue surrounding the tumor resection site, there are cancer cells microscopically present at the resection margin). In some embodiments, the tumor resection subject may have micrometastases and / or dormant disseminated cancer cells that may be encouraged to progress / grow due to a physiological response to surgery. In some embodiments, the tumor resection subject is administered a composition (e.g., as described and / or utilized herein) immediately after the tumor resection is performed (e.g., administered during surgery). In some embodiments, the tumor resection subject is administered a composition (e.g., as described and / or utilized herein) within 24 hours or less (including, for example, within 18 hours, within 12 hours, within 6 hours, within 3 hours, within 2 hours, within 1 hour, within 30 minutes, or within a shorter period) after surgery.

[0060] In some embodiments, the term "tumor site" can be the site where at least a portion of a tumor is present or was present prior to a surgical resection. In some embodiments, the tumor site can still have the entire tumor present. However, in some embodiments, the tumor site can have the entire tumor removed, for example, by tumor resection.

[0061] Resiquimod Resiquimod (i.e., R-848) is an immune response modifier having the following structure: [ka] Resiquimod is an agonist of Toll-like receptor 7 (TLR7) and Toll-like receptor 8 (TLR8) and has been shown to exhibit antiviral and antitumor activity.

[0062] In some embodiments, resiquimod has been shown to be highly useful as an immunomodulatory payload component of certain biomaterials and / or formulations for administration to subjects who have undergone or have undergone tumor resection, for example, see WO2018 / 045058 or WO2019 / 183216, both of which are incorporated herein by reference in their entireties.

[0063] Without wishing to be bound by theory, the present disclosure provides insight that it would be desirable to provide forms (e.g., solid forms) of resiquimod that provide improved solubility, hygroscopicity, stability, and ease of formulation (e.g., particularly for use in the formulations described herein) compared to amorphous resiquimod and / or salt forms of resiquimod. Accordingly, the present disclosure provides several solid forms of resiquimod, as well as methods for preparing and using the same.

[0064] Solid forms of resiquimod In some embodiments, the present disclosure provides a solid form of resiquimod. Resiquimod may exist as an amorphous solid form or as a crystalline solid form, or as a mixture thereof. The crystalline solid form may exist in one or more unique forms, which may be solvates, heterosolvates, hydrates, or nonsolvates, etc. All such forms are contemplated by the present disclosure.

[0065] In some embodiments, the present disclosure provides one or more polymorphic solid forms of resiquimod. As used herein, the term "polymorph" refers to the ability of a compound to exist in one or more different crystal structures. For example, each polymorph may differ in pharmacologic relevant physical properties, such as solubility, stability, and / or water absorption.

[0066] In some embodiments, the present disclosure provides non-solvated polymorphic forms of resiquimod.

[0067] In some embodiments, the present disclosure provides resiquimod as a solvate or heterosolvate.As used herein, the term "solvate" refers to a solid form that contains one or more solvents in stoichiometric or non-stoichiometric amounts incorporated into the crystal structure.For example, a solvate or heterosolvate polymorph may independently contain one or more solvents, such as 0.05, 0.1, 0.2, 0.5, 1.0, 1.5, or 2.0 equivalents incorporated into the crystal lattice.

[0068] In some embodiments, the present disclosure provides resiquimod as a hydrate. As used herein, the term "hydrate" refers to a solvate in which the solvent incorporated into the crystal structure is water.

[0069] As used herein, the term "about" when used in reference to 2θ angle values ​​refers to 2θ=the stated value ±0.2°. In some embodiments, "about" refers to 2θ=the stated value ±0.1°.

[0070] Resiquimod morphology I In some embodiments, the crystalline solid form of resiquimod is resiquimod Form I. In some embodiments, resiquimod Form I is unsolvated.

[0071] In some embodiments, resiquimod Form I is characterized by one or more peaks in its XRPD pattern selected from the peaks at about 8.72, about 12.24, about 16.29, about 17.56, about 19.51, about 21.31, and about 29.15 degrees 2θ. In some embodiments, resiquimod Form I is characterized by two or more peaks in its XRPD pattern selected from the peaks at about 8.72, about 12.24, about 16.29, about 17.56, about 19.51, about 21.31, and about 29.15 degrees 2θ. In some embodiments, resiquimod Form I is characterized in that three or more peaks in its XRPD pattern are selected from the peaks at about 8.72, about 12.24, about 16.29, about 17.56, about 19.51, about 21.31, and about 29.15 degrees 2θ.

[0072] In some embodiments, resiquimod Form I is characterized by peaks in its XRPD pattern at about 8.72, about 12.24, about 16.29, about 17.56, about 19.51, about 21.31, and about 29.15 degrees 2θ. In some embodiments, resiquimod Form I is characterized by an XRPD pattern including substantially all of the peaks selected from the following: [Table 13]

[0073] In some embodiments, Form I of resiquimod is characterized by one or more of the following: (i) an XRPD pattern substantially similar to that shown in Figure 1; (ii) a DSC thermogram substantially similar to that shown in FIG. 2; and (iii) A TGA thermogram substantially similar to that shown in FIG.

[0074] Based on the data provided herein, it will be appreciated that Form I exhibits significantly lower water absorption, making it particularly suitable for storage, handling, and formulation. Additionally, Form I has high solubility in sodium phosphate buffered saline containing 10% by weight poloxamer 407, indicating suitability for use in certain formulations, such as those described herein.

[0075] Morphology of resiquimod II In some embodiments, the crystalline solid form of resiquimod is resiquimod Form II. In some embodiments, resiquimod Form II is a methyl isopropyl ketone solvate.

[0076] In some embodiments, resiquimod Form II is characterized by having one or more peaks in its XRPD pattern selected from the peaks at about 7.75, about 9.65, about 11.23, about 14.38, about 19.90, about 20.80, and about 22.65 degrees 2θ. In some embodiments, resiquimod Form II is characterized by having two or more peaks in its XRPD pattern selected from the peaks at about 7.75, about 9.65, about 11.23, about 14.38, about 19.90, about 20.80, and about 22.65 degrees 2θ. In some embodiments, resiquimod Form II is characterized by having three or more peaks in its XRPD pattern selected from the peaks at about 7.75, about 9.65, about 11.23, about 14.38, about 19.90, about 20.80, and about 22.65 degrees 2θ.

[0077] In some embodiments, resiquimod Form II is characterized by peaks in its XRPD pattern at about 7.75, about 9.65, about 11.23, about 14.38, about 19.90, about 20.80, and about 22.65 degrees 2θ. In some embodiments, resiquimod Form II is characterized by an XRPD pattern including substantially all of the peaks selected from the following: [Table 14]

[0078] In some embodiments, Form II of resiquimod is characterized by one or more of the following: (i) an XRPD pattern substantially similar to that shown in FIG. (ii) a DSC thermogram substantially similar to that shown in FIG. 5; and (iii) A TGA thermogram substantially similar to that shown in FIG.

[0079] Resiquimod morphology III In some embodiments, the crystalline solid form of resiquimod is resiquimod Form III. In some embodiments, resiquimod Form III is unsolvated.

[0080] In some embodiments, Form III of resiquimod is characterized by having one or more peaks in its XRPD pattern selected from the peaks at about 8.69, about 9.18, about 9.48, about 11.97, about 14.41, about 18.53, and about 19.70 degrees 2θ. In some embodiments, Form III of resiquimod is characterized by having two or more peaks in its XRPD pattern selected from the peaks at about 8.69, about 9.18, about 9.48, about 11.97, about 14.41, about 18.53, and about 19.70 degrees 2θ. In some embodiments, Form III of resiquimod is characterized by having three or more peaks in its XRPD pattern selected from the peaks at about 8.69, about 9.18, about 9.48, about 11.97, about 14.41, about 18.53, and about 19.70 degrees 2θ.

[0081] In some embodiments, resiquimod Form III is characterized by peaks in its XRPD pattern at about 8.69, about 9.18, about 9.48, about 11.97, about 14.41, about 18.53, and about 19.70 degrees 2θ. In some embodiments, resiquimod Form III is characterized by an XRPD pattern including substantially all of the peaks selected from the following: [Table 15]

[0082] In some embodiments, resiquimod Form III is characterized by an XRPD pattern substantially similar to that shown in Figure 7 (top).

[0083] Resiquimod morphology IV In some embodiments, the crystalline solid form of resiquimod is resiquimod Form IV. In some embodiments, resiquimod Form IV is a solvate.

[0084] In some embodiments, resiquimod Form IV is characterized by one or more peaks in its XRPD pattern selected from the peaks at about 6.01, about 12.00, about 12.15, about 16.14, about 19.24, about 20.21, about 21.19, about 22.12, and about 24.50 degrees 2θ. In some embodiments, resiquimod Form IV is characterized by two or more peaks in its XRPD pattern selected from the peaks at about 6.01, about 12.00, about 12.15, about 16.14, about 19.24, about 20.21, about 21.19, about 22.12, and about 24.50 degrees 2θ. In some embodiments, resiquimod form IV is characterized in that three or more peaks in its XRPD pattern are selected from the peaks at about 6.01, about 12.00, about 12.15, about 16.14, about 19.24, about 20.21, about 21.19, about 22.12, and about 24.50 degrees 2θ.

[0085] In some embodiments, resiquimod Form IV is characterized by peaks in its XRPD pattern at about 6.01, about 12.00, about 12.15, about 16.14, about 19.24, about 20.21, about 21.19, about 22.12, and about 24.50 degrees 2θ. In some embodiments, resiquimod Form IV is characterized by an XRPD pattern including substantially all of the peaks selected from the following: [Table 16]

[0086] In some embodiments, Form IV of resiquimod is characterized by one or more of the following: (i) an XRPD pattern substantially similar to that shown in FIG. 10; (ii) a DSC thermogram substantially similar to that shown in FIG. 11; and (iii) A TGA thermogram substantially similar to that shown in FIG.

[0087] Resiquimod morphology V In some embodiments, the crystalline solid form of resiquimod is resiquimod form V. In some embodiments, resiquimod form V is unsolvated.

[0088] In some embodiments, resiquimod Form V is characterized by having one or more peaks in its XRPD pattern selected from the peaks at about 8.13, about 10.20, about 10.44, about 16.29, and about 24.56 degrees 2θ. In some embodiments, resiquimod Form V is characterized by having two or more peaks in its XRPD pattern selected from the peaks at about 8.13, about 10.20, about 10.44, about 16.29, and about 24.56 degrees 2θ. In some embodiments, resiquimod Form V is characterized by having three or more peaks in its XRPD pattern selected from the peaks at about 8.13, about 10.20, about 10.44, about 16.29, and about 24.56 degrees 2θ.

[0089] In some embodiments, resiquimod form V is characterized by peaks in its XRPD pattern at about 8.13, about 10.20, about 10.44, about 16.29, and about 24.56 degrees 2θ. In some embodiments, resiquimod form V is characterized by an XRPD pattern including substantially all of the peaks selected from the following: [Table 17]

[0090] In some embodiments, form V of resiquimod is characterized by one or more of the following: (i) an XRPD pattern substantially similar to that shown in Figure 13 (top); (ii) a DSC thermogram substantially similar to that shown in FIG. 14; and (iii) A TGA thermogram substantially similar to that shown in FIG.

[0091] Morphology of resiquimod VI In some embodiments, the crystalline solid form of resiquimod is resiquimod Form VI. In some embodiments, resiquimod Form VI is an anisole solvate.

[0092] In some embodiments, resiquimod Form VI is characterized by having one or more peaks in its XRPD pattern selected from the peaks at about 9.40, about 13.02, about 18.13, about 18.93, about 20.38, about 23.16, and about 27.78 degrees 2θ. In some embodiments, resiquimod Form VI is characterized by having two or more peaks in its XRPD pattern selected from the peaks at about 9.40, about 13.02, about 18.13, about 18.93, about 20.38, about 23.16, and about 27.78 degrees 2θ. In some embodiments, resiquimod Form VI is characterized in that three or more peaks in its XRPD pattern are selected from the peaks at about 9.40, about 13.02, about 18.13, about 18.93, about 20.38, about 23.16, and about 27.78 degrees 2θ.

[0093] In some embodiments, resiquimod Form VI is characterized by peaks in its XRPD pattern at about 9.40, about 13.02, about 18.13, about 18.93, about 20.38, about 23.16, and about 27.78 degrees 2θ. In some embodiments, resiquimod Form VI is characterized by an XRPD pattern including substantially all of the peaks selected from the following: [Table 18]

[0094] In some embodiments, resiquimod Form VI is characterized by one or more of the following: (i) an XRPD pattern substantially similar to that shown in FIG. 16; (ii) a DSC thermogram substantially similar to that shown in FIG. 17; and (iii) A TGA thermogram substantially similar to that shown in FIG.

[0095] Morphology of resiquimod VII In some embodiments, the crystalline solid form of resiquimod is resiquimod Form VII.

[0096] In some embodiments, resiquimod Form VII is characterized by having one or more peaks in its XRPD pattern selected from the peaks at about 6.25, about 9.92, about 10.96, about 16.51, about 18.99, about 23.75, and about 24.24 degrees 2θ. In some embodiments, resiquimod Form VII is characterized by having two or more peaks in its XRPD pattern selected from the peaks at about 6.25, about 9.92, about 10.96, about 16.51, about 18.99, about 23.75, and about 24.24 degrees 2θ. In some embodiments, resiquimod Form VII is characterized in that three or more peaks in its XRPD pattern are selected from the peaks at about 6.25, about 9.92, about 10.96, about 16.51, about 18.99, about 23.75, and about 24.24 degrees 2θ.

[0097] In some embodiments, resiquimod Form VII is characterized by peaks in its XRPD pattern at about 6.25, about 9.92, about 10.96, about 16.51, about 18.99, about 23.75, and about 24.24 degrees 2θ. In some embodiments, resiquimod Form VII is characterized by an XRPD pattern including substantially all of the peaks selected from the following: [Table 19]

[0098] In some embodiments, resiquimod Form VII is characterized by one or more of the following: (i) an XRPD pattern substantially similar to that shown in FIG. 19; (ii) a DSC thermogram substantially similar to that shown in FIG. 20; and (iii) A TGA thermogram substantially similar to that shown in FIG.

[0099] Methods for preparing the provided solid forms In some embodiments, the present disclosure provides methods of preparing a provided solid form of resiquimod (e.g., Form I, Form II, Form III, Form IV, Form V, Form VI, and Form VII of resiquimod).

[0100] In some embodiments, the provided solid form of resiquimod is prepared by dissolving resiquimod (e.g., crystalline or amorphous resiquimod) in a suitable solvent and then allowing the resiquimod to revert to the solid phase. In some embodiments, the solid form of resiquimod is prepared by combining amorphous and / or crystalline resiquimod in a suitable solvent under suitable conditions and isolating the solid form of resiquimod.

[0101] In some embodiments, the suitable solvent is selected from 1-butanol, 1-propanol, 2,2-dimethoxypropane, 2-butanol, 2-methoxyethanol, 2-methyltetrahydrofuran, 2-propanol, acetone, acetonitrile, amyl alcohol, anisole, chloroform, cyclohexane, cyclopentyl methyl ether, dichloromethane, diethyl ether, dioxane, ethanol, ethyl acetate, heptane, hexane, isobutanol, isobutyl acetate, isopropyl acetate, m-xylene, methanol, methyl acetate, methyl ethyl ketone, methyl isobutyl ketone, methyl isopropyl ketone, methyl tert-butyl ether, nitromethane, octane, pentane, petroleum ether, tetrahydrofuran, toluene, water, and xylene, or any combination thereof.

[0102] In some embodiments, a method of preparing a solid form of resiquimod comprises heating a mixture comprising resiquimod and a suitable solvent (e.g., a suitable solvent described herein) to a suitable temperature. In some such embodiments, the suitable temperature is from about 40° C. to about 60° C.

[0103] In some embodiments, a method of preparing a solid form of resiquimod comprises cooling a mixture comprising resiquimod and a suitable solvent (e.g., a suitable solvent described herein) to a suitable temperature. In some such embodiments, the suitable temperature is from about 0° C. to about 10° C.

[0104] In some embodiments, the method of preparing a solid form of resiquimod comprises repeated cycles of heating and cooling, in which a mixture comprising resiquimod and a suitable solvent (e.g., a suitable solvent described herein) is heated to a suitable temperature for a period of time and then cooled to a suitable temperature for a period of time, hi some embodiments, the heating and cooling cycles are repeated, for example, 2, 3, 4, 5, or 6 cycles.

[0105] In some embodiments, a method for preparing a solid form of resiquimod includes slurrying resiquimod in a suitable solvent (e.g., a suitable solvent described herein) at a suitable temperature (e.g., about 40°C to about 60°C).

[0106] In some embodiments, the solid form of resiquimod precipitates from a mixture (e.g., from a solution, suspension, or slurry). In some embodiments, the solid form of resiquimod crystallizes from a solution. In some embodiments, the solid form of resiquimod crystallizes from a solution after seeding the solution (e.g., adding crystals of resiquimod to the solution). In some embodiments, the solid form of resiquimod precipitates or crystallizes from a mixture after partial or total removal of the solvent by methods such as evaporation, distillation, or filtration. In some embodiments, the solid form of resiquimod precipitates or crystallizes from a mixture after addition of a suitable anti-solvent (e.g., water, heptane, hexane, or methyl t-butyl ether). For example, in some embodiments, resiquimod Form I is prepared by dissolving resiquimod in a suitable solvent (e.g., dichloromethane) and adding a suitable anti-solvent (e.g., heptane), thereby forming resiquimod Form I. In some embodiments, the solid form of resiquimod precipitates or crystallizes from the mixture upon cooling to an appropriate temperature (eg, about -20°C, about 0°C, or about 5°C).

[0107] In some embodiments, the method of preparing a solid form of resiquimod includes isolating the solid form of resiquimod. It will be appreciated that the solid form of resiquimod can be isolated by any suitable means. In some embodiments, the solid form of resiquimod (e.g., precipitated or crystallized resiquimod) is separated from the supernatant liquid by filtration. In some embodiments, the solid form of resiquimod (e.g., precipitated or crystallized resiquimod) is separated from the supernatant liquid by decanting the supernatant liquid.

[0108] In some embodiments, the solid form of resiquimod is dried (eg, in air or under reduced pressure, and optionally at elevated temperature).

[0109] In some embodiments, the solid form of resiquimod is prepared by converting one solid form of resiquimod to another solid form of resiquimod.

[0110] Solid form compositions provided The present disclosure also provides compositions comprising one or more solid forms of resiquimod. In some embodiments, the compositions provided comprise crystalline resiquimod (e.g., resiquimod Form I, resiquimod Form II, resiquimod Form III, resiquimod Form IV, resiquimod Form V, resiquimod Form VI, or resiquimod Form VII). In some embodiments, the compositions provided comprise amorphous resiquimod.

[0111] In some embodiments, the compositions provided include crystalline resiquimod and amorphous resiquimod. In some embodiments, the compositions that include crystalline resiquimod are substantially free of amorphous resiquimod. As used herein, the term "substantially free of amorphous resiquimod" means that the composition does not contain a significant amount of the amorphous solid form. In some embodiments, the composition includes at least about 90% crystalline resiquimod by weight. In some embodiments, the composition includes at least about 95% crystalline resiquimod by weight. In some embodiments, the composition includes at least about 97%, about 98%, or about 99% crystalline resiquimod by weight. In some embodiments, the composition includes no more than about 10% amorphous resiquimod by weight. In some embodiments, the composition includes no more than about 5% amorphous resiquimod by weight. In some embodiments, the composition includes no more than about 3%, about 2%, or about 1% amorphous resiquimod by weight.

[0112] In some embodiments, the provided compositions comprising crystalline resiquimod are substantially free of impurities. As used herein, the term "substantially free of impurities" means that the composition does not contain a significant amount of extraneous material. Such extraneous material may include starting materials, residual solvents, or any other impurities that may result from the preparation and / or isolation of crystalline resiquimod. In some embodiments, the provided compositions contain about 10% or less by weight of impurities. In some embodiments, the provided compositions contain about 5% or less by weight of impurities. In some embodiments, the provided compositions contain about 3%, about 2%, or about 1% or less by weight of impurities.

[0113] In some embodiments, the composition comprises a mixture of crystalline solid forms of resiquimod (e.g., a mixture comprising two or more of resiquimod Form I, resiquimod Form II, resiquimod Form III, resiquimod Form IV, resiquimod Form V, resiquimod Form VI, or resiquimod Form VII). In some embodiments, the composition comprises a mixture of resiquimod Form I with one or more other crystalline solid forms of resiquimod (e.g., one or more of resiquimod Form II, resiquimod Form III, resiquimod Form IV, resiquimod Form V, resiquimod Form VI, and resiquimod Form VII).

[0114] Biomaterial Composition The present disclosure also provides certain biomaterial formulations and / or polymer combination compositions comprising resiquimod. In some embodiments, the solid forms provided are useful in the preparation of such compositions.

[0115] Various systems including combinations of biomaterials and immunomodulatory payloads (see, for example, WO2018 / 045058 or WO2019 / 183216) have been reported to be extremely useful, especially when administered to subjects who have undergone or are undergoing tumor resection. The properties of these systems address one or more of the causes of problems associated with certain prior art, including, for example, certain conventional approaches to cancer treatment. For example, the systems can reduce and / or prevent certain adverse events (e.g., skin rash, hepatitis, diarrhea, colitis, hypophysitis, thyroiditis, and adrenal insufficiency) that may be associated with systemic administration of immunotherapeutics. In particular, the systems can reduce or prevent exposure of non-tumor-specific immune cells to systemically administered immunotherapeutic agent(s) and / or to the high doses of such agent(s) that are often required for systemic administration to achieve sufficient concentrations within the tumor, and, in particular, the systems can provide localized immunomodulation (e.g., local agonism of innate immunity) following tumor resection, which can increase efficacy by, among other things, focusing the immunomodulatory effect at the site where it is needed. Additionally or alternatively, such a system providing local immune modulation following resection (e.g., agonism of innate immunity) can, inter alia, break local immune tolerance to cancer and generate systemic anti-tumor immunity, which can, for example, in some embodiments, lead to eradication of disseminated disease.

[0116] The present disclosure provides certain biomaterial formulations that may be particularly useful and / or may provide certain useful effects, e.g., as described herein, and in some embodiments, the solid forms provided are useful in preparing such biomaterial formulations.

[0117] In some embodiments, the present disclosure recognizes the source of problems associated with certain prior art techniques, including, for example, problems associated with certain crosslinked biopolymeric materials. In particular, the present disclosure recognizes that certain crosslinking techniques can produce toxic by-products and / or adversely affect the stability and / or efficacy of an agent(s) (e.g., a therapeutic agent, such as resiquimod) when combined with the biopolymeric material prior to or during crosslinking.

[0118] Additionally or alternatively, the present disclosure recognizes the source of problems associated with techniques that involve preforming (e.g., by crosslinking) biopolymer materials prior to introduction into a subject. For example, the present disclosure recognizes that such preforming produces materials with a defined size and / or structure, which may limit administration options. The present disclosure provides techniques that include specific biomaterial formulations that allow administration by various routes and / or approaches, including by methods such as injection and / or laparoscopic administration, which may be less invasive than implantation. In some such embodiments, formulations with improved administration properties can be administered in a liquid state, and in some embodiments, these formulations can be administered in a preformed gel state characterized by soft space-filling properties. In some such embodiments, the formulations provided are composed of the relevant material in particulate form (e.g., such that the formulation includes a multiplicity of particles characterized by particle size distribution and / or other parameters described herein).

[0119] In particular, in some embodiments, the present disclosure provides temperature-responsive biomaterial formulations that can transition from an injectable state to another state having material properties that provide useful effects, e.g., as described herein, without the introduction of cytotoxic crosslinkers, e.g., ultraviolet radiation and / or small molecule crosslinkers. Some such embodiments thus provide useful techniques for forming gelled materials in situ that have various advantages over alternative techniques and provide solutions to certain problems associated with such alternative techniques as identified herein. For example, the present disclosure recognizes the source of problems associated with various alternative techniques for in situ gelation, as many of these techniques require processing (e.g., exposure to ultraviolet radiation and / or small molecule crosslinkers that may have toxic or other deleterious effects on the recipient and / or drugs (e.g., resiquimod) that may be included in or with the material).

[0120] In some embodiments, the provided temperature-responsive biomaterial formulations (e.g., those described herein) can exhibit one or more immunomodulatory properties. For example, in some embodiments, the provided temperature-responsive biomaterial formulations promote natural immunity when administered to a target site in a subject in need of such promotion (e.g., a tumor resection subject).

[0121] In some embodiments, the present disclosure recognizes that, among other things, certain conventional formulations that are or include poloxamers and that are used to form hydrogels typically use a minimum concentration of 16-20% (w / w) poloxamer (e.g., Poloxamer 407 (P407)). The present disclosure recognizes that such conventional formulations may have certain shortcomings in administration to a subject, including, for example, high solution viscosity that makes them unsuitable for injection, and / or tissue irritation from high concentrations of poloxamer. Moreover, the present disclosure demonstrates that it is possible to develop useful formulations using significantly lower concentration(s) of such poloxamers.

[0122] For example, in some embodiments, the present disclosure recognizes that certain poloxamers (e.g., poloxamer 407 (P407)), which have typically been used at minimum concentrations of 16-20% (w / w) to form hydrogels, when combined with one or more biocompatible polymers, can form useful temperature-responsive biomaterials at concentrations of less than 16% (w / w) (including, for example, less than 14% (w / w), less than 12% (w / w), less than 11% (w / w), less than 10.5% (w / w), less than 10% (w / w), less than 8% (w / w), less than 6% (w / w), or even lower). In some embodiments, such biocompatible polymers can be or include polymers that are not temperature-responsive (e.g., in some embodiments, can be or include hyaluronic acid and / or chitosan or modified chitosan).

[0123] One aspect provided herein relates to a preparation or composition comprising a polymer combination preparation comprising at least a first and a second polymer component, wherein the first polymer component is or comprises a poloxamer and the second polymer component is not a poloxamer, and wherein the first polymer component is present in the polymer combination preparation at a concentration of 12.5% ​​(w / w) or less (e.g., 11% (w / w), 10.5% (w / w), 10% (w / w), 9% (w / w), 8% (w / w), 7% (w / w), 6% (w / w), 5% (w / w), 4% (w / w), or less). In some embodiments, the first polymer component is present in the polymer combination formulation at a concentration of 4% (w / w) to 11% (w / w), or 4% (w / w) to 10.5% (w / w), or 4% (w / w) to 10% (w / w). In some embodiments, the first polymer component is present in the polymer combination formulation at a concentration of 5% (w / w) to 11% (w / w), or 5% (w / w) to 10.5% (w / w), or 5% (w / w) to 10% (w / w). In some embodiments, the first polymer component is present in the polymer combination formulation at a concentration of 6% (w / w) to 11% (w / w), or 6% (w / w) to 10.5% (w / w), or 6% (w / w) to 10% (w / w). In some embodiments, such polymer combination formulations are characterized by transitioning from a precursor state to a polymer network state in response to a gelation trigger, which is or includes one or more of the following: (a) a temperature at or above the critical gelation temperature (CGT) of the polymer combination formulation, (b) a critical gelation weight ratio of the first polymer component to the second polymer component, (c) the total polymer content, (d) the molecular weight of the first and / or second polymer component, or (e) a combination thereof.

[0124] In some embodiments, the crosslinks formed upon transition of the precursor state to the polymer network state do not include covalent crosslinks.

[0125] In many embodiments, such polymer combination preparations are temperature responsive. In some such embodiments, such polymer combination preparations are characterized by transitioning from a precursor state to a polymer network state in response to a temperature equal to or greater than the CGT. For example, in some embodiments, the CGT of the provided polymer combination preparation is 18-39°C. In some embodiments, the CGT of the provided polymer combination preparation is room temperature. In some embodiments, the CGT of the provided polymer combination preparation is 20-25°C. In some embodiments, the CGT of the provided polymer combination preparation is 25-30°C. In some embodiments, the CGT of the provided polymer combination preparation is 30-35°C. In some embodiments, the CGT of the provided polymer combination preparation is the body temperature of the subject.

[0126] While many different poloxamers can be used in the provided polymer combination preparations, in some embodiments, certain poloxamers, such as poloxamer 407 (P407), poloxamer 338 (P338), or poloxamer 188 (P188), are particularly useful in certain polymer combination preparations described herein. For example, in some embodiments, the poloxamer included as the first polymer component in the polymer combination preparations described herein is or includes P407. In some embodiments, the first polymer component (e.g., including P407) is present in the provided polymer combination preparations at a concentration of 4% (w / w) to 12.5% ​​(w / w), or 4% (w / w) to 11% (w / w), or 4% (w / w) to 10.5% (w / w), or 4% (w / w) to 10% (w / w). In some embodiments, the first polymer component (e.g., including P407) is present in the provided polymer combination preparations at a concentration of 5% (w / w) to 12.5% ​​(w / w), or 5% (w / w) to 11% (w / w), or 5% (w / w) to 10.5% (w / w), or 5% (w / w) to 10% (w / w). In some embodiments, the first polymer component (e.g., including P407) is present in the provided polymer combination preparations at a concentration of 6% (w / w) to 12.5% ​​(w / w), or 6% (w / w) to 11% (w / w), or 6% (w / w) to 10.5% (w / w), or 6% (w / w) to 10% (w / w).

[0127] In some embodiments, the polymer combination preparations described herein comprise a total polymer content of at least 6% (w / w), at least 8% (w / w), at least 10% (w / w), at least 12%, or at least 15% (w / w). In some embodiments, the polymer combination preparations described herein comprise a total polymer content of 6% (w / w) to 20% (w / w), or 6% (w / w) to 15% (w / w), or 7% (w / w) to 15% (w / w). In some embodiments, the polymer combination preparations described herein comprise a total polymer content of 8% (w / w) to 20% (w / w), or 8% (w / w) to 15% (w / w), or 10% (w / w) to 15% (w / w).

[0128] In some embodiments, the polymer combination preparations described herein are characterized by a weight ratio of the first polymer component to the second polymer component of 1:1 to 14:1, or 1:1 to 10:1. In some embodiments, the polymer combination preparations described herein are characterized by a weight ratio of the first polymer component to the second polymer component of 1:1 to 1:3, or 1:1 to 1:2. In some embodiments, the polymer combination preparations described herein are characterized by a weight ratio of the first polymer component to the second polymer component of 1:1 to 22:1, or 1:1 to 18:1.

[0129] In some embodiments, the second polymer component in the provided polymer combination preparation is or comprises a carbohydrate polymer. Carbohydrate polymers that may be useful according to the present disclosure include, but are not limited to, hyaluronic acid, chitosan, alginate, and variants and combinations thereof. In some embodiments, the carbohydrate polymer in the provided polymer combination preparation may be present at a concentration of about 5% (w / w) or less. In some embodiments, the carbohydrate polymer in the provided polymer combination preparation may be present at a concentration of 0.5% (w / w) to 10% (w / w), or 0.5% (w / w) to 5% (w / w), or 1% (w / w) to 10% (w / w), or 1% (w / w) to 5% (w / w), or 2% to 10% (w / w).

[0130] In some embodiments, the carbohydrate polymer useful in the particular polymer combination preparation is or comprises hyaluronic acid. In some embodiments, the hyaluronic acid may have an average molecular weight of 50 kDa to 2 MDa. In some embodiments, the hyaluronic acid may have an average molecular weight of 100 kDa to 500 kDa. In some embodiments, the hyaluronic acid may have an average molecular weight of 125 kDa to 375 kDa. In some embodiments, the hyaluronic acid may have an average molecular weight of 100 kDa to 400 kDa. In some embodiments, the hyaluronic acid may have an average molecular weight of 500 kDa to 1.5 MDa. In some embodiments, the molecular weight of the hyaluronic acid is characterized by a weight average molecular weight. In some embodiments, the molecular weight of the hyaluronic acid is characterized by a viscosity average molecular weight, which in some embodiments can be determined by converting the intrinsic viscosity of the hyaluronic acid to an average molecular weight using, for example, the Mark-Houwink equation. In some embodiments, the molecular weight of hyaluronic acid can be measured by size exclusion chromatography-multi-angle laser light scattering (SEC-MALLS).

[0131] In some embodiments, the number average molecular weight (Mn), weight average molecular weight (Mw), and / or dispersity (characterized by the polydispersity index) can be determined by SEC-MALLS.

[0132] In some embodiments, the carbohydrate polymer useful in the particular polymer combination preparation is or includes chitosan acid or modified chitosan. In some embodiments, an exemplary modified chitosan is or includes carboxymethyl chitosan.

[0133] In some embodiments, the preparations or compositions comprising the polymer combination preparations used and / or described herein are in a precursor state. In some embodiments, the preparations or compositions comprising the polymer combination preparations used and / or described herein are in a polymer network state (e.g., having one or more properties described herein).

[0134] In some embodiments, the polymer network state is or includes a viscous solution or colloid. In some embodiments, such a polymer network state may be characterized by a storage modulus of 100 Pa to 500 Pa. In some embodiments, such a polymer network state is or includes a hydrogel. In some embodiments, such a polymer network state may be characterized by a storage modulus of 500 Pa to 10,000 Pa, or 750 Pa to 7500 Pa.

[0135] In some embodiments, the polymer network state of the provided polymer combination preparations is characterized by a storage modulus that is at least 40% lower than the storage modulus of a hydrogel formed from a P407 solution at a concentration of 18% (w / w). In some embodiments, the polymer network state of the provided polymer combination preparations, whose precursor state has been stored at a temperature lower than CGT (e.g., 2-8° C.) for a period of one month or more, is characterized by a storage modulus, measured at, for example, 37° C., that remains substantially the same (e.g., within 20%, within 10%, within 5%, or less) as compared to the storage modulus of a polymer network formed from the precursor state of such freshly prepared provided polymer combination preparations. As will be appreciated by one of skill in the art, the storage modulus of a biomaterial can be affected by biodegradation, chemical degradation (e.g., oxidation), and / or phase separation of the polymeric components in the combination.

[0136] In some embodiments, the polymer combination preparations described and / or used herein have a pH of 5.0-8.5. In some embodiments, the polymer combination preparations described and / or used herein have a pH of 7-8 (e.g., pH 7.4). For example, in some embodiments, the precursor state of the polymer combination preparation is a solution of the polymer combination preparation in a solvent system having a pH of 5.0-8.5 (e.g., in some embodiments, a pH of 7-8). In some embodiments, such a solvent system is a buffered system. In some embodiments, such a buffered system may include one or more salts (e.g., but not limited to, sodium phosphate and / or sodium bicarbonate). In some embodiments, such a solvent system is a buffer system having a buffering capacity greater than 10 mM phosphate buffer. In some embodiments, such a solvent system is a buffer system having a buffering capacity greater than 20 mM phosphate buffer.

[0137] In some embodiments, the preparations or compositions described herein can include a polymer combination preparation (e.g., as described herein) and resiquimod, for example, to treat a disease, disorder, or condition (e.g., cancer). In some embodiments, such polymer combination preparations are characterized in that test animal groups with spontaneous metastasis having the polymer combination preparation in a polymer network state at the tumor resection site have a higher percent survival rate when evaluated 2 or 3 months after administration than comparable test animal groups having the polymer combination preparation without an immunomodulatory payload at the tumor resection site.

[0138] I. Compositions or Preparations Comprising the Provided Polymer Combination Preparations In some embodiments, the present disclosure provides, inter alia, compositions and / or preparations including polymer combination preparations (e.g., those described herein) that are temperature responsive and thus allow for in situ gelation at a target site in the absence of crosslinking treatments (e.g., UV irradiation and / or introduction of chemical crosslinkers) that may be toxic or otherwise adversely affect the recipient and / or payload that may be contained in or with the biomaterial.

[0139] In some embodiments, the present disclosure provides compositions comprising certain polymer combination preparations that are useful for providing sustained release of payloads (e.g., resiquimod) incorporated in the polymer combination preparations. For example, in some embodiments, certain compositions and / or preparations described herein may be highly useful when such compositions incorporating one or more immunomodulatory payloads (e.g., resiquimod) are administered to subjects who have undergone or are undergoing tumor resection. By way of example only, in some embodiments, the compositions or preparations of the present disclosure may comprise at least one innate immune modulating payload (e.g., at least resiquimod). In some embodiments, the compositions or preparations of the present disclosure may comprise at least one innate immune modulating payload and at least one adaptive immune modulating payload. In some embodiments, the compositions or preparations of the present disclosure may comprise at least one innate immune modulating payload, at least one adaptive immune modulating payload, and at least one immunomodulatory cytokine. In some embodiments, the compositions or preparations of the present disclosure may comprise at least one proinflammatory immune response inhibitor.

[0140] In some embodiments, the present disclosure provides compositions comprising certain polymer combination preparations that are sufficient to provide an immunomodulatory response (e.g., provide sufficient innate immune agonism) to achieve a beneficial effect alone without a separate immunomodulatory payload (e.g., resiquimod).

[0141] In some embodiments, the polymer combination preparation described herein is characterized in that it forms a polymer network. Without being bound by any theory, it is noted that in some embodiments, such a network can function as a scaffold or depot for a payload (e.g., an immunomodulatory payload, e.g., resiquimod) within the polymer combination preparation.

[0142] In some embodiments, a biomaterial preparation and a payload drug (e.g., an immunomodulatory payload, For example, a polymer combination preparation comprising resiquimod) can function as a sustained release formulation, in that, for example, the payload is released from the composition more slowly (i.e., over a longer period of time) than would be observed in an otherwise comparable composition lacking the polymer combination preparation (e.g., lacking one or all of the polymer components thereof).

[0143] In some embodiments, the polymer combination preparation used as described herein comprises one or more polymers (e.g., those described herein). In certain embodiments, the polymer combination preparation may comprise one or more positively charged polymers. In some embodiments, the polymer combination preparation used as described herein may comprise one or more negatively charged polymers. In some embodiments, the polymer combination preparation used as described herein may comprise one or more neutral polymers.

[0144] Provided polymer combination formulations In some embodiments, the present disclosure provides, inter alia, a polymer combination preparation comprising at least a first and a second polymer component, wherein the first polymer component is or comprises a poloxamer (e.g., as described herein), the second polymer component is not a poloxamer, and the first polymer component is present in the polymer combination preparation at a concentration of 12.5% ​​(w / w) or less. In some embodiments, such polymer combination preparations are characterized by transitioning from a precursor state to a polymer network state in response to a gelation trigger, the gelation trigger being or comprising one or more of the following: (a) a temperature equal to or greater than the critical gelation temperature (CGT) of the polymer combination preparation, (b) a critical gelation weight ratio of the first polymer component to the second polymer component, (c) a total polymer content, (d) a molecular weight of the first and / or second polymer component, or (e) a combination thereof. The polymer network state of the provided polymer combination preparation has a viscosity substantially greater than that of the precursor state and comprises crosslinks not present in the precursor state. In some embodiments, the precursor state of the provided polymer combination preparation is a liquid state. In some embodiments, the precursor state of the provided polymer combination preparation is an injectable state. In some embodiments, the polymer network state of the provided polymer combination preparation is a more viscous liquid state. In some embodiments, the polymer network state of the provided polymer combination preparation is a hydrogel.

[0145] In some embodiments, the provided polymer combination preparations are temperature responsive, for example, such that their gelation (e.g., transition from a liquid state to a gelled state) can occur upon exposure to a particular temperature. In many such embodiments, exposure to body temperature (e.g., by application to a site) is sufficient to trigger such gelation. In some embodiments, heat can also be applied. By way of example only, in some embodiments, the temperature responsive polymer combination preparations described herein are characterized in that such polymer combination preparations transition from a precursor state (e.g., a liquid state or an injectable state) to a polymer network state (e.g., a more viscous state or a hydrogel) having a viscosity and / or storage modulus substantially greater than that of the precursor state when such polymer combination preparations are exposed to a gelation trigger, which is or includes a temperature at or above the critical gelation temperature (CGT) of the polymer combination preparation. In some embodiments, the CGT of the provided polymer combination preparation is at least 10°C or higher (e.g., at least 10°C, at least 11°C, at least 12°C, at least 13°C, at least 14°C, at least 15°C, at least 16°C, at least 17°C, at least 18°C, at least 19°C, at least 20°C, at least 21°C, at least 22°C, at least 23°C, at least 24°C, at least 25°C, at least 26°C, at least 27°C, at least 28°C, at least 29°C, at least 30°C, at least 31°C, at least 32°C, 33°C, at least 34°C, at least 35°C, at least 36°C, at least 37°C, at least 38°C, at least 39°C, at least 40°C, or higher). In some embodiments, the CGT of the provided polymer combination preparation is about 10°C to about 15°C. In some embodiments, the CGT of the provided polymer combination preparation is about 12°C to about 17°C. In some embodiments, the CGT of the provided polymer combination preparation is from about 14° C. to about 19° C. In some embodiments, the CGT of the provided polymer combination preparation is from about 16° C. to about 21° C.In some embodiments, the CGT of the provided polymer combination preparation is about 18°C ​​to about 23°C. In some embodiments, the CGT of the provided polymer combination preparation is about 20°C to about 25°C. In some embodiments, the CGT of the provided polymer combination preparation is about 22°C to about 27°C. In some embodiments, the CGT of the provided polymer combination preparation is about 24°C to about 29°C. In some embodiments, the CGT of the provided polymer combination preparation is about 26°C to about 31°C. In some embodiments, the CGT of the provided polymer combination preparation is about 28°C to about 33°C. In some embodiments, the CGT of the provided polymer combination preparation is about 30°C to about 35°C. In some embodiments, the CGT of the provided polymer combination preparation is about 32°C to about 37°C. In some embodiments, the CGT of the provided polymer combination preparation is about 34°C to about 39°C. In some embodiments, the CGT of the provided polymer combination preparation is about 35°C to about 39°C. In some embodiments, the CGT of a provided polymer combination preparation is at or near the physiological temperature of a subject (e.g., a human subject) to which such polymer combination preparation is administered.

[0146] In some embodiments, the provided polymer combination preparation is thermoreversible. For example, in some embodiments, the provided polymer combination preparation is characterized by transitioning from a precursor state (e.g., liquid state or injectable state) to a polymer network state (e.g., more viscous state or hydrogel) having a viscosity and / or storage modulus substantially greater than that of the precursor state when the polymer combination preparation is exposed to a temperature equal to or greater than the critical gelation temperature (CGT) of the polymer combination preparation, and the polymer combination preparation can return from the polymer network state to a state (e.g., liquid state or the original state of the provided polymer combination preparation) having a viscosity and / or storage modulus substantially lower than that of the polymer network state.

[0147] In some embodiments, the polymer combination preparation described herein does not include a chemical crosslinker. Those skilled in the art will understand that in some embodiments, the chemical crosslinker is characterized by facilitating the formation of covalent crosslinks between polymer chains. In some embodiments, the chemical crosslinker is or includes a small molecule crosslinker that can be derived from natural sources or can be synthesized. Non-limiting examples of small molecule crosslinkers include genipin, dialdehydes, glutaraldehyde, glyoxal, diisocyanates, glutaric acid, succinic acid, adipic acid, acrylic acid, diacrylates, and the like. In some embodiments, the chemical crosslinker can involve crosslinking using thiols (e.g., EXTRACEL®, HYSTEM®), methacrylates, hexadecyl amides (e.g., HYMOVIS®), and / or tyramines (e.g., CORGEL®). In some embodiments, chemical crosslinkers may involve crosslinking using formaldehyde (e.g., HYLAN-A®), divinyl sulfone (DVS) (e.g., HYLAN-B®), 1,4-butanediol diglycidyl ether (BDDE) (e.g., RESTYLANE®), glutaraldehyde, and / or genipin (see, e.g., Khunmanee et al. “Crosslinking method of hyaluronic-based hydrogel for biomedical applications” J Tissue Eng. 8:1-16 (2017)). Thus, in some embodiments, the crosslinks formed during the transition from the precursor state to the polymer network state include covalent crosslinks.

[0148] In some embodiments, the first polymer component (e.g., a poloxamer described herein) and the second polymer component (e.g., those described herein) are present in the polymer combination formulation at a critical gelling weight ratio of 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 5.5:1, 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, 10:1, 10.5:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the first polymer component (e.g., a poloxamer described herein) and the second polymer component (e.g., those described herein) are present in the polymer combination preparation at a critical gelation weight ratio of 1:1 to 20:1, or 1:1 to 18:1, or 1:1 to 14:1, or 1.5:1 to 14:1, or 2:1 to 13:1, or 1:1 to 10:1, or 2:1 to 20:1, or 2:1 to 18:1, or 2:1 to 10:1. In some embodiments, the first polymer component (e.g., a poloxamer described herein) and the second polymer component (e.g., those described herein) are present in the polymer combination preparation at a critical gelation weight ratio of 1:1 to 10:1. In some embodiments, the first polymer component (e.g., a poloxamer described herein) and the second polymer component (e.g., those described herein) are present in the polymer combination preparation at a critical gelation weight ratio of 2:1 to 10:1. In some embodiments, the first polymer component (e.g., a poloxamer described herein) and the second polymer component (e.g., those described herein) are present in the polymer combination preparation at a critical gelation weight ratio such that the second polymer component can be present in a weight-to-weight amount greater than the weight-to-weight amount of the first polymer component. For example, in some embodiments, the first polymer component (e.g., a poloxamer described herein) and the second polymer component (e.g., those described herein) are present in the polymer combination preparation at a critical gelation weight ratio of 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, etc.In some such embodiments, the concentration of poloxamer may be less than 7% (w / w) or less, for example, 6% (w / w), 5% (w / w), 4% (w / w), or less.

[0149] In some embodiments, the polymer combination preparations provided herein that include at least a first and a second polymer component (e.g., as described herein) can include at least one additional polymer component (e.g., including at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, or more additional polymer components), which in some embodiments can be or can include a polymer component that is biocompatible and / or biodegradable (e.g., as described herein).

[0150] In some embodiments, provided polymer combination preparations comprise a total content of polymer of at least 5% (w / w) or more (including, for example, at least 6% (w / w), at least 7% (w / w), at least 8% (w / w), at least 9% (w / w), at least 10% (w / w), at least 11% (w / w), at least 12% (w / w), at least 13% (w / w), at least 14% (w / w), at least 15% (w / w), at least 16% (w / w), at least 17% (w / w), at least 18% (w / w), at least 19% (w / w), at least 20% (w / w), or more). In some embodiments, the polymer combination preparations provided contain between 5% (w / w) and 20% (w / w), or between 6% (w / w) and 18% (w / w), or between 8% (w / w) and 15% (w / w), or between 9% (w / w) and 12% (w / w) total content of polymer. In some embodiments, the polymer combination preparations described herein contain between 6% (w / w) and 20% (w / w), or between 8% (w / w) and 20% (w / w), or between 10% (w / w) and 15% (w / w) total content of polymer.

[0151] In some embodiments, the first polymer component which is or comprises a poloxamer is present in a provided polymer combination preparation at a concentration of 12.5% ​​(w / w) or less (including, e.g., 12% (w / w) or less, 11.5% (w / w) or less, 11% (w / w) or less, 10.5% (w / w) or less, 10% (w / w) or less, 9.5% (w / w) or less, 9% (w / w) or less, 8% (w / w) or less), 7% (w / w) or less, 6% (w / w) or less, 5% (w / w) or less, or 4% (w / w) or less). In some embodiments, the first polymer component which is or comprises a poloxamer is present in the provided polymer combination preparation at a concentration of 5% (w / w) to 12.5% ​​(w / w), or 8% (w / w) to 12.5% ​​(w / w), or 5% (w / w) to 11% (w / w), or 5% (w / w) to 10% (w / w), or 6% (w / w) to 10% (w / w), or 8% (w / w) to 10% (w / w). In some embodiments, the first polymer component which is or comprises a poloxamer is present in the provided polymer combination preparation at a concentration of 4% (w / w) to 12.5% ​​(w / w), or 4% (w / w) to 11% (w / w), or 4% (w / w) to 10.5% (w / w), or 4% (w / w) to 10% (w / w). In some embodiments, the first polymer component which is or comprises a poloxamer is present in the provided polymer combination preparation at a concentration of 5% (w / w) to 12.5% ​​(w / w), or 5% (w / w) to 11% (w / w), or 5% (w / w) to 10.5% (w / w), or 5% (w / w) to 10% (w / w). In some embodiments, the first polymer component which is or comprises a poloxamer is present in the provided polymer combination preparation at a concentration of 6% (w / w) to 12.5% ​​(w / w), or 6% (w / w) to 11% (w / w), or 6% (w / w) to 10.5% (w / w), or 6% (w / w) to 10% (w / w).

[0152] In some embodiments, the second polymer component may be present in the provided polymer combination preparations at a concentration of 15% (w / w) or less. In some embodiments, the second polymer component may be present in the provided polymer combination preparations at a concentration of 10% (w / w) or less, including, for example, 10% (w / w), 9% (w / w), 8% (w / w), 7% (w / w), 6% (w / w), 5% (w / w), 4% (w / w), 3% (w / w), 2% (w / w), 1% (w / w), 0.5% (w / w), or less. In some embodiments, the second polymer component is at least 0.1% (w / w) (e.g., at least 0.2% (w / w), at least 0.3% (w / w), at least 0.4% (w / w), at least 0.5% (w / w), at least 0.6% (w / w), at least 0.7% (w / w), at least 0.8% (w / w), at least 0.9% (w / w), at least 1% (w / w), at least 1.5% (w / w), at least 2% (w / w), at least 3% (w / w), at least 4% (w / w), at least 5% (w / w), at least 6% (w / w), at least 7% (w / w), at least 8% (w / w), at least 9% (w / w), at least 10% (w / w), at least 15% (w / w), at least 10% (w / w), at least 15% (w / w), at least 20 ... The second polymer component may be present in the provided polymer combination preparations at a concentration of from 0.1% (w / w) to 10% (w / w), or from 0.1% (w / w) to 8% (w / w), or from 0.1% (w / w) to 5% (w / w), or from 1% (w / w) to 5% (w / w). In some embodiments, the second polymer component in the provided polymer combination preparations may be present in a concentration of 0.5% (w / w) to 10% (w / w), or 0.5% (w / w) to 5% (w / w), or 1% (w / w) to 10% (w / w), or 1% (w / w) to 5% (w / w), or 2% to 10% (w / w).

[0153] A. First Polymer Component Comprising One or More Exemplary Poloxamers and Variants Thereof In some embodiments, the polymer combination preparation provided comprises a poloxamer or a variant thereof. Poloxamers are block copolymers that typically contain a hydrophobic chain of polyoxypropylene (e.g., polypropylene glycol (PPG) and / or poly(propylene oxide) (PPO)) flanked by two hydrophilic chains of polyoxyethylene (e.g., polyethylene glycol (PEG) and / or poly(ethylene oxide) (PEO)). Poloxamers are known under the trade names Synperonic, Pluronic, and / or Kolliphor. In general, poloxamers are non-ionic surfactants that, in some embodiments, may have good solubilizing ability, low toxicity, and / or high compatibility with cells, body fluids, and a wide range of chemicals.

[0154] In some embodiments, the poloxamer used according to the present disclosure can be any poloxamer known in the art.For example, as understood by those skilled in the art, poloxamers are generally named with the letter P (after poloxamer) followed by three numbers, the first two numbers multiplied by 100 indicate the approximate molecular mass of the polyoxypropylene chain, and the last number multiplied by 10 indicates the polyoxyethylene content.Just as an example, P407 refers to a poloxamer with a polyoxypropylene molecular mass of 4,000 g / mol and a polyoxyethylene content of 70%. Those skilled in the art will also understand that in the Pluronic and Synperonic trade names, the coding of such poloxamers begins with a letter indicating the physical form at room temperature (e.g., L=liquid, P=paste, F=flake (solid)), followed by two or three numbers, where the first number in the number symbol (two numbers in a three-digit number) multiplied by 300 indicates the approximate molecular weight of the polyoxypropylene chain, and the last number multiplied by 10 indicates the polyoxyethylene content. By way of example only, L61 refers to a liquid preparation of poloxamer having a polyoxypropylene molecular mass of 1,800 g / mol and a polyoxyethylene content of 10%. Furthermore, as will be apparent to those skilled in the art, poloxamer 181 (P181) corresponds to Pluronic L61 and Synperonic PE / L61.

[0155] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 124 (e.g., Pluronic L44 NF), poloxamer 188 (e.g., Pluronic F68 NF), poloxamer 181 (e.g., Pluronic L61), poloxamer 182 (e.g., Pluronic L62), poloxamer 184 (e.g., Pluronic L64), poloxamer 237 (e.g., Pluronic F87 NF), poloxamer 338 (e.g., Pluronic F108 NF), poloxamer 331 (e.g., Pluronic L101), poloxamer 407 (e.g., Pluronic F127 NF), or combinations thereof. In some embodiments, the provided polymer combination preparations may include at least two or more different poloxamers. Additional poloxamers listed in Table 1 of Russo and Villa "Poloxamer Hydrogels for Biomedical Applications" Pharmaceutics (2019) 11(12):671, the contents of which are incorporated herein by reference for purposes described herein, may also be useful in the polymer combination formulations described herein.

[0156] In some embodiments, the poloxamer that may be included in the polymer combination preparation described herein may be or may include poloxamer 407 (P407). In some embodiments, P407 is a poloxamer that is a triblock copolymer with a hydrophobic PPO block sandwiched between two hydrophilic PEO blocks. The approximate length of the two PEO blocks is usually 101 repeat units, while the approximate length of the PPO block is 56 repeat units. In some embodiments, P407 has an average molecular weight of about 12,600 Da, of which about 70% represents PEO. In some embodiments, P407 may readily self-assemble to form micelles, depending on the concentration and ambient temperature. Without being bound to a particular theory, the combination of hydration of the PEO block and dehydration of the hydrophobic PPO block may result in the formation of spherical micelles, and the subsequent packing of the micellar structure results in a 3D cubic lattice that constitutes the main structure of the poloxamer hydrogel. They are also biodegradable, non-toxic, and stable, and therefore suitable for use in controlled release of therapeutic agents. As will be appreciated by those skilled in the art, the concentration of P407 in hydrogel formulations based on poloxamer / water binary mixtures typically ranges from 16-20% w / v, with values ​​of about 18% w / v being most commonly used. See, for example, Pereia et al. "Formulation and Characterization of Poloxamer 407®: Thermoreversible Gel Containing Polymeric Microparticles and Hyaluronic Acid" Quim. Nova, Vol. 36, No. 8, 1121-1125 (2013), the contents of which are incorporated herein by reference in their entirety.

[0157] A variety of cross-linking techniques, including chemical and enzyme-mediated cross-linking techniques, have been used to cross-link P407 alone or in combination with another polymer, at P407 concentrations below the typical range of 16-20% w / v. For example, Ryu et al. “Catechol-functionalized chitosan / pluronic hydrogels for tissue adhesives and hemostatic materials” Biomacromolecules (2011) 12(7):2653-2659, Lee et al. “Thermo-sensitive, injectable, and tissue adhesive sol-gel transition hyaluronic acid / pluronic composite hydrogels prepared from bio-inspired catechol-thiol reaction” Soft Matter (2010) 6:977-983, and Chung et al. “Thermo-sensitive biodegradable hydrogels based on stereocomplexed pluronic multi-block copolymers for controlled protein delivery” J Control Release (2008) 127:22-30, and Lee et al. “Enzyme-mediated cross-linking of pluronic copolymer micelles for injectable and in situ forming hydrogels” Acta Biomater (2011) 7:1468-76, the contents of each of which are incorporated by reference in their entirety. However, in some embodiments, such cross-linking techniques require the use of chemical cross-linkers or enzymes and / or modified P407, which may not be desirable for in vivo administration.In some embodiments, the present disclosure provides insight that, inter alia, certain polymer combination preparations (e.g., those described herein) may be particularly useful for forming temperature-responsive hydrogels in the absence of chemical or enzyme-mediated crosslinking when the concentration of P407 in the polymer combination preparation is 12.5% ​​(w / w) or less (including, for example, 12% (w / w) or less, 11.5% (w / w) or less, 11% (w / w) or less, 10.5% (w / w) or less, 10% (w / w) or less, 9.5% (w / w) or less, 9% (w / w) or less, 8% (w / w) or less). In some embodiments, the concentration of P407 is present in the provided polymer combinations at a concentration of 6% (w / w) to 12.5% ​​(w / w), or 6% (w / w) to 11% (w / w), 5% (w / w) to 12.5% ​​(w / w), or 5% (w / w) to 11% (w / w), or 8% (w / w) to 12.5% ​​(w / w), or 5% (w / w) to 10% (w / w), or 8% (w / w) to 10% (w / w), or 6% (w / w) to 10% (w / w). In some embodiments, the concentration of P407 is present in the provided polymer combination at a concentration of 4% (w / w) to 12.5% ​​(w / w), or 4% (w / w) to 11% (w / w), or 4% (w / w) to 10.5% (w / w), or 4% (w / w) to 10% (w / w). In some embodiments, the concentration of P407 is present in the provided polymer combination at a concentration of 5% (w / w) to 12.5% ​​(w / w), or 5% (w / w) to 11% (w / w), or 5% (w / w) to 10.5% (w / w), or 5% (w / w) to 10% (w / w). In some embodiments, the concentration of P407 is present in the provided polymer combinations at a concentration of between 6% (w / w) and 12.5% ​​(w / w), or between 6% (w / w) and 11% (w / w), or between 6% (w / w) and 10.5% (w / w), or between 6% (w / w) and 10% (w / w).

[0158] In some embodiments, P407 that may be included in the polymer combination preparations described herein may be or may include compendial poloxamer 407. In some embodiments, such compendial poloxamer 407 included in the provided polymer combination preparations has not undergone additional purification steps. In some embodiments, such compendial poloxamer 407 included in the provided polymer combination preparations is unmodified (e.g., in some embodiments, is not genetically modified). In some embodiments, P407 that may be useful in the polymer combination preparations described herein has a sol-gel transition temperature (T ) of at least 18° C. or higher in PBS (including, for example, 18.5° C., 19° C., 19.5° C., 20° C., 20.5° C., 21° C., 21.5° C., 22° C., 22.5° C., 23° C., or 23.5° C.). sol-gel In some embodiments, P407 that may be useful in the polymer combination preparations described herein may have an average molecular weight of 12 kDa or less, e.g., 11.5 kDa or less, 11 kDa or less, 10.5 kDa or less, or less. As will be appreciated by those of skill in the art, the T of P407 in PBS may be sol-gel and / or average molecular weight may vary with purification. For example, in some embodiments, when low molecular weight copolymer molecules and / or impurities are removed from compendial P407, the T of P407 in PBS is sol-gel and / or the average molecular weight may be increased. Alternatively, if high molecular weight copolymer molecules and / or impurities are removed from compendial P407, the T of P407 in PBS may be increased. sol-gel and / or the average molecular weight may be reduced. See, e.g., Fakhari et al. "Thermogelling properties of purified Poloxamer 407" Heliyon (2017) 3(8):e00390, the contents of which are incorporated herein by reference in their entirety.

[0159] In some embodiments, the P407 included in the polymer combination preparations described herein can be unconjugated or unmodified P407 (e.g., P407 that is not covalently conjugated to a moiety such as a polymer or amino acid). Examples of conjugated P407 include, but are not limited to, grafting P407 to a carbohydrate polymer, e.g., chitosan, or thiolated P407. See, e.g., Park et al. "Thermosensitive chitosan-Pluronic hydrogel as an injectable cell delivery carrier for cartilage regeneration" Acta Biomaterialia (2009) 5(6):1956-1965, and Ryu et al. "Catechol-functionalized chitosan / pluronic hydrogels for tissue adhesives and hemostatic materials" Biomacromolecules (2011) 12(7):2653-2659, the contents of each of which are incorporated herein by reference in their entirety.

[0160] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 338.

[0161] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 331.

[0162] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 237.

[0163] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 188.

[0164] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 184.

[0165] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 182.

[0166] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 181.

[0167] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may be or may include poloxamer 124.

[0168] In some embodiments, the poloxamer that may be included in the polymer combination preparations described herein may have a polyoxyethylene content of at least 30% by weight (including, for example, at least 40% by weight, at least 50% by weight, at least 60% by weight, at least 70% by weight, at least 80% by weight, at least 90% by weight, or more). In some embodiments, the poloxamer may have a polyoxyethylene content of 50-90% by weight. In some embodiments, the poloxamer has a polyoxyethylene content of 60-90%. In some embodiments, the poloxamer has a polyoxyethylene content of 70-90%. In some embodiments, the poloxamer has a polyoxyethylene content of about 70%. In some embodiments, the poloxamer has a polyoxyethylene content of about 80%.

[0169] In some embodiments, the poloxamer that can be included in the polymer combination preparations described herein has a poloxamer content of at least 1,500 g / mol or more (e.g., at least 2,000 g / mol, at least 2,500 g / ml, at least 3,000 g / mol, at least 4,000 g / mol, at least 5,000 g / mol, at least 6,000 g / mol, at least 7,000 g / mol, at least 8,000 g / mol, at least 9,000 g / mol, The poloxamer may have an average molecular weight of at least 10,000 g / mol, at least 11,000 g / mol, at least 12,000 g / mol, at least 13,000 g / mol, at least 14,000 g / mol, at least 15,000 g / mol, at least 16,000 g / mol, at least 17,000 g / mol, at least 18,000 g / mol, at least 19,000 g / mol, at least 20,000 g / mol, or more. In some embodiments, the poloxamer may have an average molecular weight of about 1,500 to 20,000 g / mol. In some embodiments, the poloxamer may have an average molecular weight of about 4,000 to 12,000 g / mol. In some embodiments, the poloxamer may have an average molecular weight of about 5,000-15,000 g / mol, or 9,000-15,000 g / mol, or 10,000-15,000 g / mol, or about 11,000-14,000 g / mol, or about 11,500-13,000 g / mol, or about 12,000-13,000 g / mol, or about 6,000-10,000 g / mol, or about 7,000-9,000 g / mol, or about 7,500-8,500 g / mol. In some embodiments, the poloxamer may have an average molecular weight of 9,500-15,000 g / mol. In some embodiments, the poloxamer may have an average molecular weight of 6,000-10,000 g / mol. In some embodiments, the poloxamer may have an average molecular weight of 12,000 to 18,000 g / mol. In some embodiments, the poloxamer may have an average molecular weight of 1,500 to 3,000 g / mol. In some embodiments, the poloxamer may have an average molecular weight of 6,000 to 9,000 g / mol.Those skilled in the art will understand that the average molecular weight described herein can be number average molecular weight, viscosity average molecular weight, or weight average molecular weight. In some embodiments, the polymers described herein (e.g., poloxamers and other polymers described herein) are characterized by weight average molecular weight. In some embodiments, the polymers described herein (e.g., hyaluronic acid described herein) are characterized by viscosity average molecular weight, which in some embodiments can be determined by, for example, converting intrinsic viscosity measurements to average molecular weight using the Mark-Houink formula.

[0170] In some embodiments, poloxamers that can be included in the polymer combination preparations described herein can have polyoxypropylene with an average molecular weight of 1,000 to 5,000 g / mol or 1,500 to 4,500 g / mol.

[0171] In some embodiments, the poloxamer that can be included in the polymer combination preparation described herein can be a poloxamer variant. Examples of poloxamer variants include, but are not limited to, poloxamine (e.g., an amphiphilic block copolymer formed by four arms of poly(ethylene oxide)-poly(propylene oxide) (PEO-PPO) blocks bound to a central ethylenediamine moiety), acrylate-modified poloxamer, thiol-modified poloxamer, and combinations thereof. See, for example, Niu et al., J. Controlled Release, 2009, 137:49-56, and Alvarex-Lorenzo et al. "Poloxamine-based nanomaterials for drug delivery" Frontiers in Bioscience (2010), the contents of each of which are incorporated herein by reference at least for their disclosure regarding modified poloxamers.

[0172] B. Second Polymer Component Comprising One or More Exemplary Polymers That Are Not PoloxamersIn some embodiments, the polymer combination preparations described herein may include at least two polymer components (e.g., at least three, at least four, at least five, or more polymer components). In some embodiments, the second polymer component of the provided polymer combination preparations that include a poloxamer as the first polymer component at a concentration of 12.5% ​​(w / w) or less may be or include at least one (e.g., at least two, at least three, at least four, or more) biocompatible and / or biodegradable polymer component.Examples of such biocompatible and / or biodegradable polymeric components include, but are not limited to, immunomodulatory polymers, carbohydrate polymers (e.g., carbohydrates, e.g., polymers that are or include a carbohydrate backbone (e.g., including but not limited to, chitosan, alginate, hyaluronic acid, and / or variants thereof)), polyacrylic acid, silica gel, polyethyleneimine (PEI), polyphosphazene, and / or variants thereof, cellulose, chitin, chondroitin sulfate, collagen, dextran, gelatin, ethylene vinyl acetate (EVA), fibrin, polylactic acid-co-glycolic acid (PLGA), polylactic acid (PLA), polyglycolic acid (PGA), polyethylene glycol (PEG), PEG diacrylate (PEGDA), disulfide-containing PEGDA (PEGSSDA), PEG dimethacrylate (PEGDMA), polydioxanone (PDO), polyhydroxybutyric acid (PHB), poly(2-hydroxyethyl methacrylate) (pHEMA), polycarboxybetaine (PC B), polysulfobetaine (PSB), polycaprolactone (PCL), poly(β-amino ester) (PBAE), poly(ester amide), poly(propylene glycol) (PPG), poly(aspartic acid), poly(glutamic acid), poly(propylene fumarate) (PPF), poly(sebacic anhydride) (PSA), poly(trimethylene carbonate) (PTMC), poly(desaminotyrosyltyrosine alkyl ester carbonate) (PDTE), poly[bis(trifluoroethoxy)phosphazene], poly(phenylene glycol), ... xymethylene, single-walled carbon nanotubes, polyanhydrides, poly(N-vinyl-2-pyrrolidone) (PVP), poly(vinyl alcohol) (PVA), poly(acrylic acid) (PAA), poly(methacrylic acid) (PMA), polyacetals, poly(alpha esters), poly(ortho esters), polyphosphoesters, polyurethanes, polycarbonates, polyamides, polyhydroxyalkanoates, polyglycerols, polyglucuronic acids, starches, variants thereof, and / or combinations thereof.

[0173] In some embodiments, the second polymer component of the provided polymer combination preparation is or comprises a non-ionic polymer component. Examples of such non-ionic polymer components include, but are not limited to, polyvinyl alcohol (PVA), polyethylene oxide (PEO), and combinations thereof. In some embodiments, the second polymer component of the provided polymer combination preparation is or comprises a cationic polymer component, such as, but not limited to, chitosan, amino group-containing polymer, collagen, gelatin, and combinations thereof. In some embodiments, the second polymer component of the provided polymer combination preparation is or comprises an anionic polymer component, examples of which include, but are not limited to, alginate, gellan gum, pectin, xanthan gum, carboxymethyl cellulose (CMC), polyacrylic acid, polyaspartic acid, and combinations thereof.

[0174] In some embodiments, the second polymer component of the provided polymer combination preparations is or includes an immunomodulatory polymer, e.g., a polymer that modulates one or more aspects of the immune response (e.g., a polymer that induces innate immune agonism). In some embodiments, the immunomodulatory polymer can be or include a polymer agonist of innate immunity, such as those described in International Patent Application No. PCT / US20 / 31169, filed May 1, 2020 (published as WO2020 / 223698A1). In some embodiments, the immunomodulatory polymer can be or include a carbohydrate polymer (e.g., a carbohydrate, e.g., a polymer that is or includes a carbohydrate backbone (e.g., including but not limited to chitosan, alginate, hyaluronic acid, and / or variants thereof)).

[0175] In some embodiments, provided polymer combination preparations include at least one poloxamer at a concentration of 12.5% ​​or less (e.g., 11% (w / w), 10.5% (w / w), 10% (w / w), 9% (w / w), 8% (w / w), 7% (w / w), 6% (w / w), or less), and a second polymer component, which can be or can include a carbohydrate polymer (e.g., a carbohydrate, e.g., a polymer that is or includes a carbohydrate backbone (e.g., including but not limited to, hyaluronic acid, chitosan, and / or variants thereof)).

[0176] (i) Exemplary Hyaluronic Acid and Its Variants In some embodiments, the carbohydrate polymer included in the provided polymer combination preparations containing poloxamer is or includes hyaluronic acid or a variant thereof. Hyaluronic acid (HA), also known as hyaluronan or hyaluronate, is a non-sulfated member of a class of polymers known as glycosaminoglycans (GAGs) that are widely distributed in body tissues. HA is found as a component of the extracellular matrix of tissues forming a pericellular coat on the cell surface. In some embodiments, HA is a soluble form of hyaluronic acid (HA), also known as hyaluronan or hyaluronate. 14 H 21 NO 11 ) n (which in some embodiments may be present as a salt, e.g., sodium, potassium, and / or calcium salt), where n may vary depending on the source, isolation technique, and / or measurement method.

[0177] In some embodiments, HA that may be useful according to the present disclosure may be isolated or derived from a number of natural sources. For example, in some embodiments, HA may be isolated or derived from, for example, human umbilical cord, cockscomb, and / or vertebrate connective tissue matrix. In some embodiments, HA may be isolated or derived from capsular components of bacteria, such as Streptococcus. See, for example, Kendall et al, (1937), Biochem. Biophys. Acta, 279, 401-405. In some embodiments, HA and / or variants thereof may be produced by microbial fermentation. In some embodiments, HA and / or variants thereof may be recombinant HA or variants thereof, produced, for example, using gram-positive and / or gram-negative bacteria as hosts, including, but not limited to, Bacillus, Lactococcus lactis, Agrobacterium, and / or Escherichia coli.

[0178] As noted in International Patent Application No. PCT / US20 / 31169, filed May 1, 2020 (published as WO2020 / 223698A1), the biological activity of HA varies depending on its molecular weight. For example, HA with a high molecular weight (high molecular weight HA) may have anti-inflammatory or immunosuppressive activity, whereas HA with a low molecular weight (low molecular weight HA) may exhibit pro-inflammatory or immunostimulatory effects. For example, Gao et al. “A low molecular weight hyaluronic acid derivative accelerates excisional wound healing by modulating pro-inflammation, promoting epithelialization and neovascularization, and remodeling collagen” Int. J. Mol Sci (2019) 20:3722, Cyphert et al. “Size Matters: Molecular Weight Specificity of Hyaluronan Effects in Cell Biology.” Int. J. Cell Biol.(2015) 2015:563818, Dicker et al. “Hyaluronan:A simple polysaccharide with diverse biological functions” Acta Biomater.(2014) 10:1558-1570, Aya and Stern “Hyaluronan in wound healing:Rediscovering a major player.”Wound Repair Regen.(2014) 22:579-593, and Frenkel “The role of hyaluronan in See, “wound healing” Int. Wound J. (2014) 11:159-163, the entire contents of each of which are incorporated by reference in their entirety for the purposes described herein.Thus, in some embodiments, the HA or variant thereof that may be included in the provided polymer combination preparation may have a low molecular weight, for example, an average molecular weight of 500 kDa or less (e.g., 450 kDa, 400 kDa, 350 kDa, 300 kDa, 250 kDa, 200 kDa, 150 kDa, 100 kDa, 50 kDa, or less). In some embodiments, the HA or variant thereof that may be included in the provided polymer combination preparation may have an average molecular weight of about 100 kDa to about 200 kDa. In some embodiments, the HA or variant thereof that may be included in the provided polymer combination preparation may have an average molecular weight of about 100 kDa to about 150 kDa. In some embodiments, the HA or variant thereof that may be included in the provided polymer combination preparation may have an average molecular weight of about 250 kDa to about 350 kDa. In some embodiments, the HA or variant thereof that may be included in the provided polymer combination preparations may have an average molecular weight of about 300 kDa to about 400 kDa. In some embodiments, the polymer combination preparations described herein, which may include a poloxamer (e.g., as described herein) and a low molecular weight HA or variant thereof in the absence of an immunomodulatory payload, may be useful for inducing innate immune agonism.

[0179] In some embodiments, HA or variants that may be included in the provided polymer combination preparations may have a high molecular weight, for example, an average molecular weight of more than 500 kDa or more (e.g., including 550 kDa, 600 kDa, 650 kDa, 700 kDa, 750 kDa, 800 kDa, 850 kDa, 900 kDa, 950 kDa, 1 MDa, 1.1 MDa, 1.2 MDa, 1.3 MDa, 1.4 MDa, 1.5 MDa, 1.6 MDa, 1.7 MDa, 1.8 MDa, 1.9 MDa, 2 MDa, 2.5 MDa, 3 MDa, 3.5 MDa, 4 MDa, 4.5 MDa, or more). In some embodiments, HA or variants thereof that may be useful according to the present disclosure may have an average molecular weight of about 600 kDa to about 900 kDa. In some embodiments, HA or variants thereof that may be useful according to the present disclosure may have an average molecular weight of about 700 kDa to about 900 kDa. In some embodiments, HA or variants thereof that may be included in the provided polymer combination preparations may have an average molecular weight of about 500 kDa to about 800 kDa. In some embodiments, HA or variants thereof that may be included in the provided polymer combination preparations may have an average molecular weight of about 600 kDa to about 800 kDa. In some embodiments, HA or variants thereof that may be included in the provided polymer combination preparations may have an average molecular weight of about 700 kDa to about 800 kDa. In some embodiments, HA or variants thereof that may be useful according to the present disclosure may have an average molecular weight of about 1 MDa to about 3 MDa. In some embodiments, the polymer combination preparations described herein that may include a poloxamer (e.g., as described herein) and a high molecular weight HA or variant thereof in the absence of an immunomodulatory payload may be useful for resolving inflammation (e.g., immunosuppressive inflammation).

[0180] In some embodiments, the polymer combination preparation provided comprises a hyaluronic acid variant. In some embodiments, the hyaluronic acid variant is water-soluble. In some embodiments, the hyaluronic acid variant can be a chemically modified hyaluronic acid, for example, in some embodiments, the hyaluronic acid is esterified. Examples of chemical modifications to hyaluronic acid include, but are not limited to, the addition of thiol, haloacetate, butanediol, diglycidyl, ether, dihydrazide, aldehyde, glycan, and / or tyramine functional groups. Additional hyaluronic acid modifications and hyaluronic acid variants are known in the art. See, e.g., Highley et al., "Recent advances in hyaluronic acid hydrogels for biomedical applications" Curr Opin Biotechnol (2016) Aug 40:35-40; Burdick & Prestwich, "Hyaluronic acid hydrogels for biomedical applications" Advanced Materials (2011); Prestwhich, "Hyaluronic acid-based clinical biomaterials derived for cell and molecule delivery in regenerative medicine" J. Control Release (2011) Oct 30;155(2):193-199 (each of which is incorporated by reference in their entirety for purposes described herein).

[0181] In some embodiments, the provided polymer combination preparations include at least one poloxamer present at a concentration of 12.5% ​​(w / w) or less, and a second polymer component that may be or may include hyaluronic acid or a variant thereof. In some such embodiments, the HA or a variant thereof may be present in the provided polymer combination preparations at a concentration of about 10% (w / w) or less, including, for example, 9% (w / w), 8% (w / w), 7% (w / w), 6% (w / w), 5% (w / w), 4% (w / w), 3% (w / w), 2% (w / w), or 1% (w / w), or less. In some embodiments, HA or a variant thereof may be present in provided polymer combination preparations at a concentration of about 0.5% (w / w) to about 5% (w / w), e.g., 0.5% (w / w), 0.6% (w / w), 0.7% (w / w), 0.8% (w / w), 0.9% (w / w), 1% (w / w), 1.5% (w / w), 2% (w / w), 2.5% (w / w), 3% (w / w), 3.5% (w / w), 4% (w / w), 4.5% (w / w), or 5% (w / w). In some embodiments, HA or a variant thereof having a low molecular weight (e.g., as described herein) may be present in the provided polymer combination preparation at a concentration of at least about 1.5% (w / w) or more (e.g., including at least 2% (w / w), at least 2.5% (w / w), at least 3% (w / w), at least 4% (w / w), at least 5% (w / w), at least 6% (w / w), at least 7% (w / w), at least 8% (w / w), at least 9% (w / w), or more). In some embodiments, HA or a variant thereof having a low molecular weight (e.g., as described herein) may be present in the provided polymer combination preparation at a concentration of about 1.5% (w / w) to about 5% (w / w). In some embodiments, HA or a variant thereof having a low molecular weight (e.g., as described herein) may be present in the provided polymer combination preparation at a concentration of about 0.5% (w / w) to about 10% (w / w).In some embodiments, HA or a variant thereof having a low molecular weight (e.g., as described herein) may be present in the provided polymer combination preparation at a concentration of about 1% (w / w) to about 10% (w / w) or about 1.5% (w / w) to about 10% (w / w). In some embodiments, HA or a variant thereof having a low molecular weight (e.g., as described herein) may be present in the provided polymer combination preparation at a concentration of about 0.7% (w / w) to about 4% (w / w) or about 1.5% (w / w) to about 4% (w / w). In some embodiments, HA or a variant thereof having a low molecular weight (e.g., as described herein) may be present in the provided polymer combination preparation at a concentration of about 3% (w / w) to about 7% (w / w). In some embodiments, HA or a variant thereof having a high molecular weight (e.g., as described herein) may be present in a provided polymer combination preparation at a concentration of 2% (w / w) or less, including, for example, 1.5% (w / w), 1.25% (w / w), 1% (w / w), or less. In some embodiments, HA or a variant thereof having a high molecular weight (e.g., as described herein) may be present in a provided polymer combination preparation at a concentration of about 0.5% (w / w) to about 3% (w / w).

[0182] (ii) Exemplary Chitosan and Its Variants In some embodiments, the carbohydrate polymer included in the provided polymer combination preparations with poloxamers (e.g., as described herein) can be or include chitosan or a variant thereof. Examples of chitosan and / or its variants that can be included in the polymer combination preparations described herein include, but are not limited to, chitosan, chitosan salts (e.g., chitosan HCl, chitosan chloride, chitosan lactate, chitosan acetate, chitosan glutamate), alkyl chitosan, aromatic chitosan, carboxyalkyl chitosan (e.g., carboxymethyl chitosan), hydroxyalkyl chitosan (e.g., hydroxypropyl chitosan, hydroxyethyl chitosan), aminoalkyl chitosan, acylated chitosan, phosphorylated chitosan, thiolated chitosan, quaternary ammonium chitosan (e.g., N-(2- hydroxyl)propyl-3-trimethylammonium chitosan chloride), guanidyl chitosan, chitosan oligosaccharide, glycated chitosan (e.g., N-dihydrogalactochitosan), chitosan poly(sulfonamide), chitosan-phenylsuccinic acid (e.g., the product formed by the reaction of phenylsuccinic anhydride or a variant thereof (e.g., 2-phenylsuccinic anhydride, 2-phenylsuccinic acid derivatives, 2-O-acetyl-L-malic anhydride, etc.) and chitosan) (e.g., chitosan phenylsuccinic acid hemiamide, which is a ring-opened amido carboxylic acid derivative), and variants or combinations thereof. In some embodiments, the carbohydrate polymer included in the provided polymer combination preparations with poloxamer (e.g., as described herein) can be or can include a carboxyalkyl chitosan (e.g., carboxymethyl chitosan).

[0183] One of skill in the art will appreciate that in some cases, chitosan and / or variants thereof may be produced by deacetylation of chitin. In some embodiments, chitosan or variants thereof included in a polymer combination preparation with poloxamer (e.g., as described herein) are characterized by a degree of deacetylation (i.e., percent of acetyl groups removed) of at least 70% or more (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more (including up to 100%)). In some embodiments, chitosan or variants thereof are characterized by a degree of deacetylation of 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, or less. Combinations of the above ranges are also possible. For example, chitosan or variants thereof may be characterized by a degree of deacetylation of 80%-95%, 70%-95%, or 75%-90%. As will be appreciated by one of skill in the art, the degree of deacetylation (% DA) can be determined by a variety of methods known in the art, such as, in some cases, by NMR spectroscopy.

[0184] In some embodiments, the chitosan or variant thereof included in the polymer combination preparation with poloxamer (e.g., as described herein) has a molecular weight of at least 5 kDa or more (e.g., at least 10 kDa or more (e.g., at least 20 kDa, at least 30 kDa, at least 40 kDa, at least 50 kDa, at least 60 kDa, at least 70 kDa, at least 80 kDa, at least 90 kDa, at least 100 kDa, at least 110 kDa, at least 120 kDa, at least 130 kDa, at least 140 kDa, at least 150 kDa, at least 160 kDa, at least 170 kDa, at least 180 kDa, at least 190 kDa, at least 200 kDa, at least 210 kDa, at least 220 kDa, at least 230 kDa, at least 240 kDa, at least 250 kDa, at least 260 kDa, at least 270 kDa, at least 280 kDa, at least 290 kDa, at least 300 kDa, at least 310 kDa, at least 320 kDa, at least 330 kDa, at least 340 kDa, at least 350 kDa, at least 360 kDa, at least 370 kDa, at least 380 kDa, at least 390 kDa, at least 400 kDa, at least 400 kDa, at least 410 kDa, at least 420 kDa, at least 430 kDa, at least 440 kDa, at least 450 kDa, a, including at least 160 kDa, at least 170 kDa, at least 180 kDa, at least 190 kDa, at least 200 kDa, at least 210 kDa, at least 220 kDa, at least 230 kDa, at least 240 kDa, at least 250 kDa, at least 260 kDa, at least 270 kDa, at least 280 kDa, at least 290 kDa, at least 300 kDa, at least 350 kDa, at least 400 kDa, at least 500 kDa, at least 600 kDa, at least 700 kDa, or more. In some embodiments, the chitosan or variant thereof included in the polymer combination preparation with poloxamer (e.g., as described herein) can have an average molecular weight of 750 kDa or less (including, for example, 700 kDa or less, 600 kDa or less, 500 kDa or less, 400 kDa or less, 300 kDa or less, 200 kDa or less, 100 kDa or less, 50 kDa or less, or less). Combinations of the above ranges are also possible. For example, in some embodiments, the chitosan or variant thereof included in the polymer combination preparation with poloxamer (e.g., as described herein) is characterized by an average molecular weight of 10 kDa to 700 kDa, or 20 kDa to 700 kDa, or 30 kDa to 500 kDa, or 150 kDa to 600 kDa, or 150 kDa to 400 kDa, or 50 kDa to 150 kDa, or 10 kDa to 50 kDa.In some embodiments, the chitosan or variant thereof included in the polymer combination preparation with poloxamer (e.g., as described herein) is characterized by an average molecular weight of 20 kDa to 700 kDa or 30 kDa to 500 kDa. As noted herein, the average molecular weight can be a number average molecular weight, a weight average molecular weight, or a peak average molecular weight.

[0185] In some embodiments, the chitosan or variant thereof contained in the polymer combination preparation with poloxamer (e.g., as described herein) is characterized by a molecular weight distribution in the range of 10 kDa to 700 kDa, or 20 kDa, or 700 kDa, or 30 kDa to 500 kDa, or 150 kDa to 600 kDa, or 150 kDa to 400 kDa, or 50 kDa to 150 kDa, or 10 kDa to 50 kDa. In some embodiments, the chitosan or variant thereof contained in the polymer combination preparation with poloxamer (e.g., as described herein) is characterized by a molecular weight distribution in the range of 20 kDa to 700 kDa, or 30 kDa to 500 kDa.

[0186] In some embodiments, the chitosan or variant thereof included in a polymer combination preparation with a poloxamer (e.g., as described herein) may be characterized by a viscosity of 3,500 mPa·s or less (including, for example, 3,000 mPa·s or less, 2,500 mPa·s or less, 2,000 mPa·s or less, 1,500 mPa·s or less, 1,000 mPa·s or less, 500 mPa·s or less, 250 mPa·s or less, 200 mPa·s or less, 150 mPa·s or less, 100 mPa·s or less, 75 mPa·s or less, 50 mPa·s or less, 25 mPa·s or less, 20 mPa·s or less, 15 mPa·s or less, 10 mPa·s or less, or less). In some embodiments, the chitosan or variants thereof may be characterized by a viscosity of at least 5 mPa·s or more (including, for example, at least 10 mPa·s, at least 20 mPa·s, at least 30 mPa·s, at least 40 mPa·s, at least 50 mPa·s, at least 60 mPa·s, at least 70 mPa·s, at least 80 mPa·s, at least 90 mPa·s, at least 100 mPa·s, at least 125 mPa·s, at least 150 mPa·s, at least 175 mPa·s, at least 250 mPa·s, at least 500 mPa·s, at least 1,000 mPa·s, at least 1,500 mPa·s, at least 2,000 mPa·s, at least 2,500 mPa·s or more). Combinations of the above ranges are also possible. For example, in some embodiments, such viscous polymer solutions of or including chitosan or variants thereof may be characterized by a viscosity of 5 mPa·s to 3,000 mPa·s, or 5 mPa·s to 300 mPa·s, or 5 mPa·s to 200 mPa·s, or 20 mPa·s to 200 mPa·s, or 5 mPa·s to 20 mPa·s. In some embodiments, the viscosity of chitosan or variants thereof described herein is measured at 1% in 1% acetic acid at 20° C.

[0187] In some embodiments, the poloxamer-containing polymer combination preparation (e.g., as described herein) comprises at least one or more (e.g., one, two, three, or more) chitosan and / or variants thereof (e.g., including modified chitosan and / or chitosan or a salt of modified chitosan, e.g., chloride or glutamate). For example, in some embodiments, the chitosan and / or variants thereof (e.g., including modified chitosan and / or chitosan or a salt of modified chitosan, e.g., chloride or glutamate) may be characterized by a degree of deacetylation of 70%-95%, or 75%-90%, or 80%-95%, or greater than 90%. In some embodiments, chitosan and / or variants thereof (including, e.g., modified chitosan and / or a salt of chitosan or modified chitosan, e.g., chloride or glutamate) may be characterized by an average molecular weight (e.g., measured as chitosan or a chitosan salt, e.g., chitosan acetate) of 10 kDa to 700 kDa, 20 kDa to 600 kDa, 30 kDa to 500 kDa, 150 kDa to 400 kDa, or 200 kDa to 600 kDa. In some embodiments, chitosan and / or variants thereof (including, for example, modified chitosan and / or a salt of chitosan or modified chitosan, such as the chloride salt or glutamate salt) may be characterized by a molecular weight distribution (e.g., measured as chitosan or a chitosan salt, such as chitosan acetate) in the range of 10 kDa to 700 kDa, 20 kDa to 600 kDa, 30 kDa to 500 kDa, 150 kDa to 400 kDa, or 200 kDa to 600 kDa. In some embodiments, chitosan and / or variants thereof (including, for example, a salt thereof, such as the chloride salt or glutamate salt) may be characterized by a viscosity in the range of 5 to 3,000 mPa·s, or 5 to 300 mPa·s, or 20 to 200 mPa·s.In some embodiments, such chitosan and / or variants thereof (e.g., including salts thereof, such as the chloride salt or glutamate salt) may be or include PROTASAN™ UltraPure chitosan chloride and / or chitosan glutamate (e.g., available from NovoMatrix®, a business unit of FMC Health and Nutrition (now part of Du Pont; Product Numbers: CL 113, CL 114, CL 213, CL 214, G 113, G 213, G 214)). In some embodiments, such chitosan and / or variants thereof (including, for example, salts thereof, such as, for example, chloride salts or glutamate salts) may be or include chitosan, chitosan oligomers, and / or variants thereof (including, for example, chitosan HCl, carboxymethyl chitosan, chitosan lactate, chitosan acetate), obtained, for example, from Heppe Medical Chitosan GMBH (e.g., Chitoceuticals® or Chitoscience®).

[0188] In some embodiments, the chitosan or variant thereof included in the poloxamer-containing polymer combination preparation (e.g., as described herein) is or includes a carboxyalkylchitosan (e.g., carboxymethylchitosan) characterized by at least one or all of the following properties: (1) a degree of deacetylation of 80%-95%, (ii) an average molecular weight of 30 kDa-500 kDa, or a molecular weight distribution of 30 kDa-500 kDa, and (iii) a viscosity in the range of 5-300 mPa·s.

[0189] In some embodiments, the chitosan or variant thereof included in the polymer combination preparation with poloxamer (e.g., as described herein) is or includes a variant of chitosan (e.g., as described herein). In some embodiments, such a variant of chitosan may include chemical modification(s) of one or more chemical moieties of the chitosan chain, such as, for example, hydroxyl and / or amino groups. In some embodiments, such a variant of chitosan is or includes a modified chitosan, such as, for example, but not limited to, a glycated chitosan (e.g., a chitosan modified by the addition of one or more mono- or oligosaccharide side chains to one or more of its free amino groups). Exemplary saccharified chitosans that are useful herein include, for example, but are not limited to, those described in US 5,747,475, US 6,756,363, WO 2013 / 109732, US 2018 / 0312611, and US 2019 / 0002594, the contents of each of which are incorporated herein by reference for purposes described herein.

[0190] In some embodiments, the chitosan or variant thereof included in the poloxamer-containing polymer combination preparation (e.g., as described herein) is or includes chitosan conjugated to a polymer (e.g., a hydrophilic polymer such as polyethylene glycol) that increases the solubility of chitosan in an aqueous environment.

[0191] In some embodiments, the chitosan or its variants contained in the polymer combination preparation with poloxamer (e.g., as described herein) is or includes thiolated chitosan.Various modifications to chitosan are known in the art, including, but not limited to, carboxylation, pegylation, galactosylation (or other glycosylation), and / or thiolation, as described, for example, in Ahmadi et al.Res Pharm Sci.,10(1):1-16(2015) (the contents of which are incorporated herein by reference for the purposes described herein).Those skilled in the art who read this disclosure will understand that other modified chitosans may be useful for the particular application in which the method is performed.

[0192] In some embodiments, the provided polymer combination preparations include at least one poloxamer present at a concentration of 12.5% ​​or less, and a second polymer component that may be or may include chitosan or a variant thereof. In some such embodiments, chitosan or a variant thereof may be present in the provided polymer combination preparations at a concentration of about 10% (w / w) or less, including, for example, 9% (w / w), 8% (w / w), 7% (w / w), 6% (w / w), 5% (w / w), 4% (w / w), 3% (w / w), 2% (w / w), 1% (w / w), 0.5% (w / w), 0.4% (w / w), 0.3% (w / w), 0.2% (w / w), 0.1% (w / w), or less. In some embodiments, chitosan or a variant thereof may be present in the provided polymer combination preparations at a concentration of 0.1% (w / w) to 10% (w / w), or 0.1% (w / w) to 8% (w / w), or 0.1% (w / w) to 5% (w / w), or 1% (w / w) to 5% (w / w), or about 1% (w / w) to about 3% (w / w).

[0193] C. Payload (e.g., Therapeutic Agent) Provided biomaterial compositions include resiquimod (e.g., as a payload or therapeutic agent). In some embodiments, provided biomaterial compositions include resiquimod at concentrations of 0.005 mg / mL, 0.01 mg / mL, 0.02 mg / mL, 0.05 mg / mL, 0.10 mg / mL, 0.12 mg / mL, 0.14 mg / mL, 0.16 mg / mL, 0.18 mg / mL, 0.20 mg / mL, 0.22 mg / mL, 0.25 mg / mL, 0.30 mg / mL, 0.35 mg / mL, 0.40 mg / mL, 0.45 mg / mL, or 0.50 mg / mL. L, 0.50mg / mL, 0.55mg / mL, 0.60mg / mL, 0.65mg / mL, 0.70mg / mL, 0.75mg / mL, 0.80mg / mL, 0.85mg / mL, 0.90mg / mL, 0.95mg / mL, 1.00mg / mL, 1.05mg / mL, 1.10mg / mL, 1.15mg / mL, 1.20mg / mL, 1.25mg / mL, 1.50mg / mL, or 1.80mg / mL. In some embodiments, provided biomaterial compositions include resiquimod at a concentration of 0.005 mg / mL to 1.80 mg / mL, 0.01 mg / mL to 1.80 mg / mL, 0.01 mg / mL to 0.50 mg / mL, 0.005 mg / mL to 1.00 mg / mL, 0.05 mg / mL to 1.00 mg / mL, 0.05 mg / mL to 0.50 mg / mL, 0.05 mg / mL to 0.30 mg / mL, 0.05 mg / mL to 0.20 mg / mL, 0.10 mg / mL to 0.25 mg / mL, 0.10 mg / mL to 0.20 mg / mL, 0.12 mg / mL to 0.18 mg / mL, or 0.14 mg / mL to 0.20 mg / mL.

[0194] In some embodiments, provided biomaterial compositions include resiquimod as the sole immunomodulatory payload.

[0195] In some embodiments, provided biomaterial compositions can further comprise and / or be administered in combination with one or more additional payloads (e.g., one or more additional therapeutic agents, e.g., immunomodulatory agents). Exemplary immunomodulatory agents include those described in WO2018 / 045058, WO2019 / 183216, and PCT / US21 / 46392, the contents of each of which are incorporated herein by reference.

[0196] D. Solvent Systems In some embodiments, the polymer combination preparation, or the individual components of the polymer combination preparation, are prepared in or present in a suitable solvent system. For example, in some embodiments, such a solvent system has a pH in the range of 4.5 to 8.5. In certain embodiments, such a solvent system has a pH in the range of 4.5 to 7. In certain embodiments, the polymer combination preparation, or the individual components of the polymer combination preparation, are prepared in or present in a suitable solvent system having a pH of 7 to 9. In certain embodiments, the polymer combination preparation, or the individual components of the polymer combination preparation, are prepared in or present in a suitable solvent system having a pH of 7 to 7.5 (e.g., pH 7.4). In certain embodiments, the polymer combination preparation, or the individual components of the polymer combination preparation, are prepared in or present in a suitable solvent system having a pH of 7.5 to 8.5. In certain embodiments, the polymer combination formulation, or the individual components of the polymer combination formulation, are prepared in or are present in a suitable solvent system having a pH of 8.

[0197] In certain embodiments, the polymer combination preparation, or the individual components of such polymer combination preparation, are prepared in or present in water. In some embodiments, the polymer combination preparation, or the individual components of such polymer combination preparation, are prepared in or present in an aqueous buffer system. In some embodiments, such an aqueous buffer system may include one or more salts (such as, but not limited to, sodium phosphate and / or sodium bicarbonate). In some embodiments, such a solvent system is an aqueous buffer system having a buffer capacity higher than 10 mM phosphate buffer. In some embodiments, such a solvent system is an aqueous buffer system having a buffer capacity higher than 20 mM phosphate buffer. In certain embodiments, the polymer combination preparation, or the individual components of such polymer combination preparation, are prepared in or present in a phosphate buffer, such as phosphate buffered saline (PBS). In certain embodiments, the polymer combination preparation, or the individual components of such polymer combination preparation, are prepared in or present in a bicarbonate buffer. In some embodiments, the polymer combination preparation and / or its individual components are prepared in or present in an aqueous buffer system having a concentration range of 1 mM to 500 mM, or 5 mM to 250 mM, or 10 mM to 150 mM, or 1 mM to 50 mM, or 5 mM to 50 mM, or 5 mM to 100 mM, or 50 mM to 100 mM. In certain embodiments, a suitable aqueous buffer (e.g., phosphate buffer) is prepared at a concentration of 10 mM to 50 mM. In certain embodiments, a suitable aqueous buffer (e.g., phosphate buffer) is prepared at a concentration of 10 mM to 30 mM. In certain embodiments, a suitable aqueous buffer (e.g., bicarbonate buffer) is prepared at a concentration of 100 mM to 200 mM. In certain embodiments, the polymer combination preparation or its individual components are prepared or present in sodium phosphate buffer at a concentration of 10 mM to 50 mM or 10 mM to 30 mM.In some embodiments, the aqueous buffer system may include 0.9% saline, hi some embodiments, the aqueous buffer system may include 0.5% saline to 1.5% saline by weight, or 0.5% saline to 1.0% saline by weight.

[0198] It will be appreciated that the concentration of the aqueous buffer system may vary when combined with one or more polymeric components, payloads, and / or other components. Generally, when a concentration of an aqueous buffer system is specified throughout this disclosure, the concentration refers to the concentration prior to combination with one or more polymeric components, payloads, and / or other components.

[0199] E. Optional Additives In some embodiments, the polymer combination preparation may include one or more additives. In some embodiments, such additives may be or include thickening agents. As will be understood by those skilled in the art, such thickening agents may improve the suspension of components or emulsification to increase the stability of the combination. In some embodiments, such thickening agents may be useful to prevent, reduce, or delay phase separation of individual polymer components in the polymer combination preparation. Examples of thickening agents include, but are not limited to, cellulose derivatives, starches, pectins, xanthan, and / or any combination thereof.

[0200] II. CERTAIN SPECIFIC CHARACTERISTICS AND / OR CHARACTERISTICS OF PROVIDED POLYMER COMBINATION PREPARATIONS OR COMPOSITIONS COMPRISING THE SAME The provided polymer combination preparation or composition comprising the same may be characterized by one or more (e.g., one, two, three or more) of certain properties and / or characteristics described herein. Those skilled in the art who read this disclosure will understand that the provided polymer combination preparation or composition comprising the same may be configured to provide suitable material properties and / or characteristics for a particular application. For example, in some embodiments, suitable material properties and / or characteristics for a particular application may be determined based on, for example, the properties of the tissue surrounding the tumor, the route of administration, the site of administration, and / or the desired duration of immune modulation for which the method is performed.

[0201] A. Immunomodulatory properties In some embodiments, the provided polymer combination preparations may be non-immunomodulatory. In some such embodiments, the provided polymer combination preparations and / or compositions comprising same may include an immunomodulatory payload (e.g., resiquimod) such that the resulting composition or preparation is immunomodulatory.

[0202] In some embodiments, the provided polymer combination preparations comprising poloxamers may include a second or additional polymer component such that the resulting polymer combination preparation may itself be immunomodulatory in the absence of an immunomodulatory agent payload.

[0203] In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may indirectly or directly activate one or more pattern recognition receptors of one or more types of cells of the innate immune system, such as, for example, dendritic cells, macrophages, monocytes, neutrophils, and / or natural killer (NK) cells, such that at least one or more innate immune responses are elicited (e.g., as described herein). Examples of such pattern recognition receptors are or include C-type lectin receptors (CLRs), nucleotide-binding oligomerization domain-like receptors (NOD-like receptors or NLRs), retinoic acid-inducible gene I-like receptors (RLRs), and / or Toll-like receptors (TLRs). In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly activate at least one or more C-type lectin receptors (CLRs) of a number of different cells of the innate immune system (e.g., dendritic cells, macrophages, etc.), including, for example, the mannose receptor and / or the asialoglycoprotein receptor family (e.g., dectin-1, dectin-2, macrophage-inducible C-type lectin (Mincle), dendritic cell-specific ICAM3-binding nonintegrin (DC-SIGN), and DC NK lectin group receptor 1 (DNGR-1)). In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly activate at least one or more NOD-like receptors (NLRs) on different types of leukocytes (e.g., lymphocytes, macrophages, dendritic cells), including, for example, NLRA (e.g., CIITA), NLRB (e.g., NAIP), NLRC (e.g., NOD1, NOD2, NLRC3, NLRC4, NLRC5, NLRX1), and / or NLRP (e.g., NLRP1, NLRP2, NLRP3, NLRP4, NLRP5, NLRP6, NLRP7, NLRP8, NLRP9, NLRP10, NLRP11, NLRP12, NLRP13, NLRP14).In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly activate at least one or more RIG-I-like receptors (RLRs) of, for example, myeloid cells, including, for example, RIG-I, MDA5, and / or LGP2. In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly activate at least one or more Toll-like receptors (TLRs) of different types of leukocytes (e.g., dendritic cells, myeloid dendritic cells, monocytes, macrophages, and / or neutrophils), including, for example, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, and / or TLR10.

[0204] In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may indirectly or directly activate or induce (e.g., increase the level and / or activity of) inflammasomes, for example, in myeloid cells, such that at least one or more innate immune responses (and / or one or more characteristics of an innate immune response) are elicited (e.g., as described herein). In some embodiments, inflammasomes are multiprotein complexes that typically activate one or more inflammatory responses, for example, promoting the maturation and / or secretion of one or more inflammatory cytokines, for example, interleukin 1β and / or interleukin 18. In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may indirectly or directly activate or induce (e.g., increase the level and / or activity of) inflammasomes comprising Absent in Melanoma 2 (AIM2)-like receptors ("AIM2 inflammasomes"). In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may indirectly or directly activate or induce (e.g., increase the level and / or activity of) inflammasomes comprising one or more NLRs, including, for example, NLRP1 (e.g., NALP1b), NLRP3 (e.g., NALP3), and / or NLRC4 (e.g., IPAF).

[0205] In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly activate one or more components involved in the cGAS-STING pathway (e.g., the cGAS-STING pathway and / or its components as described in Chen et al., "Regulation and function of the cGAS-STING pathway of cytosolic DNA sensing" Nature Immunology (2016) 17:1142-1149, which is incorporated by reference in its entirety for purposes described herein) such that innate immunity is elicited. In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly induce the activity and / or levels of NFκB and / or other components associated with the NFκB pathway (e.g., NFκB activation during an innate immune response as described in Dev et al., "NF-κB and innate immunity" Curr. Top. Microbiol. Immunol. (2011) 349:115-43, which is incorporated by reference in its entirety for purposes described herein). In some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly cause the generation of reactive oxygen species, for example, during an innate immune response.

[0206] As will be apparent to one of skill in the art upon reading this disclosure, in some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly activate one or more of the components and / or pathways (e.g., those described herein) associated with the activation of innate immunity. For example, in some embodiments, the provided polymer combination preparations and / or compositions or preparations comprising the provided polymer combination preparations may directly or indirectly activate one or more pattern recognition receptors (e.g., those described herein) of one or more types of cells of the innate immune system, and may also activate or induce (e.g., increase the level and / or activity of) inflammasomes, for example, in myeloid cells.

[0207] B. Viscosity In some embodiments, the polymer combination formulations described herein (e.g., in a precursor state or in a polymer network state such as a viscous solution) have a viscosity of 25,000 mPa·s or less or less (e.g., 24,000 mPa·s or less, 23,000 mPa·s or less, 22,000 mPa·s or less, 21,000 mPa·s or less, 20,000 mPa·s or less, 19,000 mPa·s or less, 18,000 mPa·s or less, 17,000 mPa·s or less, 16,000 mPa·s or less, 15,000 mPa·s or less, 14,000 mPa·s or less, 13,000 mPa·s or less, 12,000 mPa·s or less, 11,000 mPa·s or less, 10,000 mPa·s or less). s or less, 9,000 mPa·s or less, 8,000 mPa·s or less, 7,000 mPa·s or less, 6,000 mPa·s or less, 5,000 mPa·s or less, 4,000 mPa·s or less, 3,500 mPa·s or less, 3,000 mPa·s or less, 2,500 mPa·s or less, 2,000 mPa·s or less, 1,500 mPa·s or less, 1,000 mPa·s or less, 500 mPa·s or less, 250 mPa·s or less, 200 mPa·s or less, 150 mPa·s or less, 100 mPa·s or less, 75 mPa·s or less, 50 mPa·s or less, 25 mPa·s or less, 20 mPa·s or less, 15 mPa·s or less, 10 mPa·s or less, or less.In some embodiments, the polymer combination formulations described herein (e.g., in a precursor state or in a polymer network state such as a viscous solution) have a viscosity of at least 5 mPa·s or more (e.g., at least 10 mPa·s, at least 20 mPa·s, at least 30 mPa·s, at least 40 mPa·s, at least 50 mPa·s, at least 60 mPa·s, at least 70 mPa·s, at least 80 mPa·s, at least 90 mPa·s, at least 100 mPa·s, at least 125 mPa·s, at least 150 mPa·s, at least 175 mPa·s, at least 250 mPa·s, at least 500 mPa·s, at least 1,000 mPa·s, at least 1,500 mPa·s, at least 2,000 mPa·s, at least 2,500 mPa·s, at least 3,000 mPa·s, at least 4,000mPa·s, at least 5,000mPa·s, at least 6,000mPa·s, at least 7,000mPa·s, at least 8,000mPa·s, at least 9,000mPa·s, at least 10,000mPa·s, at least 11,000mPa·s, at least 12,000mPa·s, at least 13,000mPa·s, at least 14,000mPa·s, at least 15 The composition may be characterized by a viscosity of at least 1,000 mPa·s, including at least 16,000 mPa·s, at least 17,000 mPa·s, at least 18,000 mPa·s, at least 19,000 mPa·s, at least 20,000 mPa·s, at least 21,000 mPa·s, at least 22,000 mPa·s, at least 23,000 mPa·s, at least 24,000 mPa·s, or more. Combinations of the above ranges are also possible. For example, in some embodiments, the polymer combination preparations described herein (e.g., in a precursor state or in a polymer network state such as a viscous solution) may be characterized by a viscosity of 5 mPa·s to 10,000 mPa·s, or 10 mPa·s to 5,000 mPa·s, or 5 mPa·s to 200 mPa·s, or 20 mPa·s to 100 mPa·s, or 5 mPa·s to 20 mPa·s, or 3 mPa·s to 15 mPa·s.In some embodiments, the polymer combination preparations described herein (e.g., in a precursor state or in a polymer network state, such as, for example, a viscous solution) can be a viscous solution having a viscosity comparable to that of honey (e.g., having mPa·s and / or centipoise comparable to that of honey, e.g., about 2,000-10,000 mPa·s). In some embodiments, the polymer combination preparations described herein (e.g., in a precursor state or in a polymer network state, such as, for example, a viscous solution) can be a viscous solution having a viscosity comparable to that of natural syrup (e.g., syrup derived from sap, syrup derived from molasses, etc.) (e.g., having mPa·s and / or centipoise comparable to that of natural syrup, e.g., about 15,000-20,000 mPa·s). In some embodiments, the polymer combination preparations described herein (e.g., in a precursor state or in a polymer network state, such as, for example, a viscous solution) can be viscous solutions having a viscosity comparable to ketchup (e.g., having mPa·s and / or centipoise comparable to ketchup, e.g., tomato ketchup, e.g., about 5,000-20,000 mPa·s). One of skill in the art reading this disclosure will understand that in some cases, the viscosity of the polymer combination preparations described herein can be selected or adjusted based on, for example, the route of administration (e.g., injection vs. implant), the injection volume and / or injection time, and / or the duration of the effect of natural immune stimulation. As will also be understood by one of skill in the art, the viscosity of the polymer depends, for example, on the temperature and the concentration of the polymer in the experimental sample. In some embodiments, the viscosity of the polymer combination preparations described herein can be, for example, 1000 s. -1 The shear rate may be measured at 20°C.

[0208] In some embodiments, a polymer combination preparation comprising a poloxamer (e.g., as described herein) (e.g., in a precursor state or in a polymer network state such as, for example, a viscous solution) may be characterized by a viscosity of 3,500 mPa·s or less (including, for example, 3,000 mPa·s or less, 2,500 mPa·s or less, 2,000 mPa·s or less, 1,500 mPa·s or less, 1,000 mPa·s or less, 500 mPa·s or less, 250 mPa·s or less, 200 mPa·s or less, 150 mPa·s or less, 100 mPa·s or less, 75 mPa·s or less, 50 mPa·s or less, 25 mPa·s or less, 20 mPa·s or less, 15 mPa·s or less, 10 mPa·s or less, or less). In some embodiments, a polymer combination preparation (e.g., as described herein) comprising a poloxamer (e.g., in a precursor state or in a polymer network state, e.g., a viscous solution) can be characterized by a viscosity of at least 5 mPa·s or more (e.g., including at least 10 mPa·s, at least 20 mPa·s, at least 30 mPa·s, at least 40 mPa·s, at least 50 mPa·s, at least 60 mPa·s, at least 70 mPa·s, at least 80 mPa·s, at least 90 mPa·s, at least 100 mPa·s, at least 125 mPa·s, at least 150 mPa·s, at least 175 mPa·s, at least 250 mPa·s, at least 500 mPa·s, at least 1,000 mPa·s, at least 1,500 mPa·s, at least 2,000 mPa·s, at least 2,500 mPa·s, or more). Combinations of the above ranges are also possible. For example, in some embodiments, such viscous polymer solutions (e.g., in a precursor state or in a polymer network state, e.g., a viscous solution) can be characterized by a viscosity of 5 mPa·s to 3,000 mPa·s, or 5 mPa·s to 300 mPa·s, 5 mPa·s to 200 mPa·s, or 20 mPa·s to 200 mPa·s, or 5 mPa·s to 20 mPa·s. In some embodiments, the viscosity of the polymer combination preparations described herein can be, for example, greater than 1000 s. -1 The shear rate may be measured at 20°C.

[0209] In particular, the present disclosure recognizes that hydrogel technologies, including certain crosslinking techniques (e.g., certain chemical crosslinking techniques, ultraviolet light, etc.), may generate toxic by-products and / or adversely affect the stability or efficacy of agents (e.g., therapeutic agents) that may be combined with the polymer combination preparation.

[0210] Alternatively or additionally, the present disclosure recognizes that, in some embodiments, certain advantages may be achieved by administering the component(s) of the polymer combination preparation to form during and / or upon administration of the immunomodulatory composition described herein, as compared to preforming (e.g., by crosslinking) the polymer biomaterial prior to introduction into the subject. For example, administration of a preformed biomaterial requires a corresponding incision and / or surgical intervention to facilitate administration. For example, in some embodiments, the present disclosure recognizes that such preforming produces a material with a defined size and / or structure, which may limit administration options, as the dimensions of the preformed material may differ from the dimensions of the target site (e.g., resection cavity). In some embodiments, a hydrogel may form during and / or upon administration. In some embodiments, the polymer combination preparation administered to the target site may include a preformed hydrogel of the polymer combination preparation.

[0211] In some embodiments, the present disclosure recognizes that the polymer combination preparations useful for administration to the target site described herein may be viscous solutions. For example, in some embodiments, a liquid polymer combination preparation may be introduced to a target site such that upon administration to the target site, an immunomodulatory composition described herein in the form of a viscous solution (e.g., a solution having a viscosity of about 5,000-15,000 centipoise at body temperature, e.g., a solution having a viscosity of about 10,000 centipoise at body temperature) is formed.

[0212] In some embodiments, the present disclosure recognizes that the polymer combination preparations useful for administration to a target site described herein may be viscous solutions that may be substantially retained at the target site for a period of time when administered. In some embodiments, such viscous liquid polymer combination preparations have a viscosity that is low enough to be injectable (e.g., through a syringe tip or catheter and / or syringe needle) but high enough to be substantially retained at the target site for a period of time when administered. In some embodiments, such viscous liquid polymer combination preparations may have a viscosity of about 500-10,000 centipoise at room temperature. In some embodiments, such viscous liquid polymer combination preparations may have a viscosity of about 500-3,000 centipoise at room temperature. In some embodiments, such viscous liquid polymer combination preparations may have a viscosity of about 1,000-8,000 centipoise at room temperature. In some embodiments, such viscous liquid polymer combination preparations may have a viscosity of about 2,000-6,000 centipoise at room temperature. In some embodiments, such viscous liquid polymer combination preparations may have a viscosity of about 3,000-7,000 centipoise at room temperature. In some embodiments, such viscous liquid polymer combination preparations may have a viscosity of about 4,000-8,000 centipoise at room temperature. In some embodiments, such viscous liquid polymer combination preparations may have a viscosity of about 5,000-9,000 centipoise at room temperature. In some embodiments, such viscous liquid polymer combination preparations may have a viscosity of about 6,000-10,000 centipoise at room temperature.

[0213] In some embodiments, the present disclosure recognizes that there may be viscosity requirements and / or limitations on the injectability of liquid polymer combination preparations. For example, in some embodiments, the injectable polymer combination preparation may be characterized by a viscosity suitable for loading and controlling release through a range of gauge needles (e.g., 14-20 gauge needles, e.g., 16-18 gauge needles). Alternatively, in some embodiments, the injectable polymer combination preparation may be characterized by a viscosity suitable for loading and controlling release through a range of diameter syringe tips (i.e., without a needle attached or with a catheter). In some embodiments, the polymer combination preparation included in the immunomodulatory composition (e.g., as described herein) loaded into a syringe may further include a plasticizer.

[0214] The present disclosure provides techniques including specific polymer combination preparations and administration methods that allow for interventions that may be less invasive than implantation and / or less toxic than systemic administration. In some such embodiments, preparations with improved administration properties may be administered in a liquid state. In some embodiments, they may be administered in a preformed gel state characterized by soft space-filling properties. In some embodiments, they may be administered subcutaneously. In some embodiments, they may function as a proximal depot for sustained release of an immunomodulatory payload (e.g., resiquimod). In some embodiments, they may allow for reprogramming of tissues (e.g., tumors and / or, e.g., sentinel and / or draining lymph nodes). In some embodiments, they may be administered prior to or at the same time as tumor resection surgery. In some embodiments, they may be administered ipsilaterally when compared to the tumor resection site and / or primary tumor site. In some embodiments, they may be administered contralaterally when compared to the tumor resection site and / or primary tumor site. In some embodiments, they may be administered to patients with metastatic, disseminated and / or recurrent cancer. In some such embodiments, the provided preparation is comprised of a suitable material in particulate form (e.g., such that the preparation comprises a plurality of particles characterized by a size distribution and / or other parameters, e.g., as described herein).

[0215] C. Storage modulus: Polymer network state In some embodiments, when the polymer combination preparations described herein are in a polymer network state, such polymer network state can have a viscosity of at least 100 Pa, at least 200 Pa, at least 300 Pa, at least 400 Pa, at least 500 Pa, at least 600 Pa, at least 700 Pa, at least 800 Pa, at least 900 Pa, at least 1,000 Pa, at least 1,100 Pa, at least 1,200 Pa, at least 1,300 Pa, at least 1,400 Pa, at least 1,500 Pa, at least 1,600 Pa, at least 1,700 Pa, at least 1,800 Pa, at least 1,900 Pa, at least 2,000 Pa, at least 2,100 Pa, The elastic modulus may be characterized by a storage modulus of at least 2,200 Pa, at least 2,300 Pa, at least 2,400 Pa, at least 2,500 Pa, at least 2,600 Pa, at least 2,700 Pa, at least 2,800 Pa, at least 2,900 Pa, at least 3,000 Pa, at least 3,500 Pa, at least 4,000 Pa, at least 4,500 Pa, at least 5,000 Pa, at least 6,000 Pa, at least 7,000 Pa, at least 8,000 Pa, at least 9,000 Pa, at least 10,000 Pa, at least 11,000 Pa, at least 12,000 Pa, at least 13,000 Pa, at least 14,000 Pa, at least 15,000 Pa, or more. In some embodiments, such polymer network state of the provided polymer combination preparations may be characterized by a storage modulus of 15 kPa or less, 14 kPa or less, 13 kPa or less, 12 kPa or less, 11 kPa or less, 10 kPa or less, 9 kPa or less, 8 kPa or less, 7 kPa or less, 6 kPa or less, or less. Combinations of the above ranges are also possible. For example, in some embodiments, such polymer network state of the provided polymer combination preparations may be characterized by a storage modulus of 100 Pa to 15 kPa, or 100 Pa to 10 kPa, or 100 Pa to 7.5 kPa, or 200 Pa to 5,000 Pa, or 300 Pa to 2,500 Pa, or 500 Pa to 2,500 Pa, or 100 Pa to 500 Pa.In some embodiments, the polymer network state of the provided polymer combination preparations may be characterized by a storage modulus of 1,000 Pa to 10,000 Pa, or 2,000 Pa to 10,000 Pa, or 3,000 Pa to 10,000 Pa, or 4,000 Pa to 10,000 Pa, or 5,000 Pa to 10,000 Pa, or 6,000 Pa to 10,000 Pa. One of ordinary skill in the art will appreciate that a variety of rheological characterization methods (e.g., as described in Weng et al., "Rheological Characterization of in situ Crosslinkable Hydrogels Formulated from Oxidized Dextran and N-Carboxyethyl Chitosan" Biomacromolecules, 8:1109-1115 (2007)) can be used to measure the storage modulus of a material, and in some cases, the storage modulus of a material can be measured by a rheometer and / or dynamic mechanical analysis (DMA). Those skilled in the art will also understand that rheological characterization may vary with ambient conditions, e.g., temperature and / or pH. Thus, in some embodiments, the provided polymer combination preparations are characterized by a storage modulus (e.g., as described herein) measured at the body temperature of a subject (e.g., a human subject at 37° C.), e.g., at pH 5-8 or physiological pH (e.g., pH 7). As will be apparent to those skilled in the art upon reading the disclosure provided herein, the storage modulus of the provided polymer combination preparations, e.g., in particulate form, refers to the bulk storage modulus of the particle population.

[0216] In some embodiments, the polymer network state of the polymer combination preparations provided herein may be characterized by a storage modulus less than that of an 18% (w / w) poloxamer hydrogel. For example, in some embodiments, the polymer network state of the polymer combination preparations provided herein may be characterized by a storage modulus measured at 37° C. that is reduced by at least 10% or more (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, or more) compared to that of an 18% (w / w) poloxamer hydrogel.

[0217] In some embodiments, the polymer network state of the polymer combination preparations provided herein may be characterized by a storage modulus (e.g., as described herein) that remains substantially the same (e.g., within 20% or within 10% or within 5%) when stored at a suitable temperature for a period of time. For example, in some embodiments, the polymer network state of the polymer combination preparations provided herein may be characterized by a storage modulus (e.g., as described herein) measured at 37°C that remains substantially the same (e.g., within 20% or within 10% or within 5%) when stored at a temperature between 4°C and 10°C (e.g., 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, or 10°C) for a period of time, for example, at least 1 week or more (e.g., at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, or longer). In some embodiments, the polymer network state of the polymer combination preparations provided herein may be characterized by a storage modulus measured at 37° C. (e.g., as described herein) that remains substantially the same (e.g., within 20%, or within 10%, or within 5%) when stored at room temperature (e.g., 20° C.-25° C.) for a period of time, such as at least one week or more (e.g., at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, or longer).

[0218] D. Phase angle: polymer network state In some embodiments, the polymer network state of the provided polymer combination preparation may be characterized by a phase angle indicative of a viscoelastic material. For example, in some embodiments, the polymer network state of the provided polymer combination preparation may be characterized by a phase angle of 1° to 50°, or 2° to 45°, or 3° to 40°, or 3° to 35°, or 3° to 30°, or 3° to 25°, or 5° to 30°, or 10° to 30°, or 15° to 25°, or 20° to 35°. In some embodiments, the polymer network state of the provided polymer combination preparation may be characterized by a phase angle of 10° to 30° or 15° to 25°. In some embodiments, the polymer network state of the provided polymer combination preparation may be characterized by a phase angle of 5° to 15° or 10° to 20°. As will be understood by those skilled in the art, the phase angle of a polymeric biomaterial can be determined by dynamic mechanical analysis, such as frequency sweep analysis, including, for example, the determination of the shear storage modulus and shear loss modulus of a sample. Those skilled in the art will understand that the storage modulus or elastic modulus of a material can be determined based on its stored energy and represents the elastic properties of the material, while the loss modulus or viscous modulus can be determined based on the energy dissipated as heat and represents the viscous properties of the material. The phase angle (δ) is the arctangent of the ratio of the storage modulus to the loss modulus, and its value indicates whether the material is more elastic or more viscous. Typically, a phase angle of more than 45° indicates that the viscous properties dominate and the material behaves more like a solution. As the phase angle approaches 0°, the elastic (solid or gel-like) properties dominate. For example, a material with a large storage modulus and a small phase angle indicates a stronger (more elastic) gel than one with a smaller storage modulus and phase angle. In some embodiments, the phase angle of a provided polymer combination preparation (e.g., as described herein) in a polymer network state can be determined by a frequency sweep analysis performed at a temperature corresponding to the body temperature of the subject being treated. In some embodiments, the frequency sweep analysis can be performed over a frequency range of 0.1-10 Hz while applying a constant 0.4% strain.

[0219] E. Dissolution / degradation rate The polymer combination preparations described herein are generally biocompatible. In some embodiments, at least one polymer component in the provided polymer combination preparations may be biodegradable in vivo. In some embodiments, at least one polymer component in the provided polymer combination preparations may exhibit resistance to biodegradation (e.g., by enzymatic and / or oxidative mechanisms). In some embodiments, at least one polymer component in the provided polymer combination preparations may be chemically oxidized. Thus, in some embodiments, the polymer combination preparations are capable of being chemically and / or biologically degraded in a physiological environment, e.g., in the subject's body, e.g., at the subject's target site. Those skilled in the art reading this disclosure will understand that the degradation rate of the provided polymer combination preparations may vary based on, for example, the type of poloxamer and / or the second polymer (e.g., in some embodiments, carbohydrate polymers such as hyaluronic acid and / or chitosan described herein) and their material properties and / or concentrations (e.g., as described herein). For example, the half-life (time for 50% of the polymer combination preparation to degrade into monomers and / or other non-polymeric moieties) of the provided polymer combination preparations may be on the order of days, weeks, months, or years. In some embodiments, the polymer combination preparations described herein may be biologically degraded, for example, by enzymatic activity or cellular mechanisms, for example, by exposure to lysozyme (e.g., having a relatively low pH), or by simple hydrolysis. In some examples, the provided polymer combination preparations may be degraded into monomers (e.g., polymer monomers) and / or non-polymeric moieties that are non-toxic to cells. As will be understood by those skilled in the art, the provided polymer combination preparations have a longer residence time at the target site (e.g., tumor resection site) upon administration if such provided polymer combination preparations have a slower degradation rate in vivo.

[0220] In some embodiments, the polymer combination preparations provided herein remain substantially homogenous (e.g., have no detectable phase separation) when stored at a temperature between 4° C. and 10° C. (e.g., 4° C., 5° C., 6° C., 7° C., 8° C., 9° C., or 10° C.) for a period of time, including at least one week or more (e.g., at least two weeks, at least three weeks, at least four weeks, at least five weeks, at least six weeks, at least seven weeks, at least two months, at least three months, at least four months, at least five months, at least six months, or longer). In some embodiments, the polymer combination preparations provided herein remain substantially homogenous (e.g., have no detectable phase separation) when stored at room temperature for a period of time, including at least one week or more (e.g., at least two weeks, at least three weeks, at least four weeks, at least five weeks, at least six weeks, at least seven weeks, at least two months, at least three months, at least four months, at least five months, at least six months, or longer).

[0221] In some embodiments, the polymer combination preparations provided herein may be characterized in that, when stored at a temperature between 4° C. and 10° C. (e.g., 4° C., 5° C., 6° C., 7° C., 8° C., 9° C., or 10° C.) for a period of time, such as at least one week or more (e.g., at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, or longer), 20% or less or less of the polymer combination preparation is degraded (e.g., by biodegradation or chemical degradation). In some embodiments, the polymer combination preparations provided herein may be characterized in that when stored at room temperature for a period of time, e.g., at least one week or more (including, e.g., at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, or longer), 20% or less or less of the polymer combination preparation is degraded (e.g., by biodegradation or chemical degradation).

[0222] In some embodiments, the provided polymer combination preparations are characterized in that when assessed in vivo by administration to a target site (e.g., tumor resection site) of a test subject (e.g., as described herein), at least 10% or more of such provided polymer combination preparations (including, for example, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more) remain at the target site in vivo two or more days after administration. In some embodiments, no more than 90%, no more than 80%, no more than 70%, no more than 60%, no more than 50%, no more than 40%, no more than 30%, no more than 20%, or less of such provided polymer combination preparations remain at the target site in vivo two or more days after administration. Combinations of the above are also possible. For example, in some embodiments, provided polymer combination preparations are characterized in that, when assessed in vivo by administration to a target site (e.g., a tumor resection site) in a test subject (e.g., as described herein), between 30% and 80%, or between 40% and 70% of such provided polymer combination preparations remain at the target site in vivo two or more days after administration.

[0223] In some embodiments, the provided polymer combination preparations are characterized in that when assessed in vivo by administration to a target site (e.g., tumor resection site) of a test subject (e.g., as described herein), at least 10% or more of such provided polymer combination preparations (including, for example, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more) remain at the target site in vivo 3 days or more after administration. In some embodiments, no more than 90%, no more than 80%, no more than 70%, no more than 60%, no more than 50%, no more than 40%, no more than 30%, no more than 20%, or less of such provided polymer combination preparations remain at the target site in vivo 3 days or more after administration. Combinations of the above are also possible. For example, in some embodiments, provided polymer combination preparations are characterized in that, when assessed in vivo by administration to a target site (e.g., a tumor resection site) in a test subject (e.g., as described herein), between 30% and 80%, or between 40% and 70% of such provided polymer combination preparations remain at the target site in vivo 3 days or more after administration.

[0224] In some embodiments, the provided polymer combination preparations are characterized in that when assessed in vivo by administration to a target site (e.g., a tumor resection site) of a test subject (e.g., as described herein), at least 10% or more of such provided polymer combination preparations (including, for example, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more) remain at the target site in vivo 5 days or more after administration. In some embodiments, no more than 90%, no more than 80%, no more than 70%, no more than 60%, no more than 50%, no more than 40%, no more than 30%, no more than 20%, or less of such provided polymer combination preparations remain at the target site in vivo 5 days or more after administration. Combinations of the above are also possible. For example, in some embodiments, provided polymer combination preparations are characterized in that, when assessed in vivo by administration to a target site (e.g., a tumor resection site) in a test subject (e.g., as described herein), between 30% and 80%, or between 40% and 70% of such provided polymer combination preparations remain at the target site in vivo 5 days or more after administration.

[0225] In some embodiments, the provided polymer combination preparations are characterized in that when assessed in vivo by administration to a target site (e.g., tumor resection site) of a test subject (e.g., as described herein), at least 10% or more of such provided polymer combination preparations (including, for example, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more) remain at the target site in vivo 7 days or more after administration. In some embodiments, no more than 90%, no more than 80%, no more than 70%, no more than 60%, no more than 50%, no more than 40%, no more than 30%, no more than 20%, or less of such provided polymer combination preparations remain at the target site in vivo 7 days or more after administration. Combinations of the above are also possible. For example, in some embodiments, provided polymer combination preparations are characterized in that, when assessed in vivo by administration to a target site (e.g., a tumor resection site) in a test subject (e.g., as described herein), between 30% and 80%, or between 40% and 70% of such provided polymer combination preparations remain at the target site in vivo 7 days or more after administration.

[0226] In some embodiments, the provided polymer combination preparations are characterized in that when assessed in vivo by administration to a target site (e.g., a tumor resection site) of a test subject (e.g., as described herein), at least 10% or more of such provided polymer combination preparations (including, for example, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more) remain at the target site in vivo 14 days or more after administration. In some embodiments, no more than 90%, no more than 80%, no more than 70%, no more than 60%, no more than 50%, no more than 40%, no more than 30%, no more than 20%, or less than or equal to the amount of such provided polymer combination preparations remain at the target site in vivo 14 days or more after administration. Combinations of the above are also possible. For example, in some embodiments, provided polymer combination preparations are characterized in that, when assessed in vivo by administration to a target site (e.g., a tumor resection site) in a test subject (e.g., as described herein), between 30% and 80%, or between 40% and 70% of such provided polymer combination preparations remain at the target site in vivo 14 days or more after administration.

[0227] In some embodiments, the provided polymer combination preparations are characterized in that when assessed in vivo by administration to a target site (e.g., a tumor resection site) in a test subject (e.g., as described herein), 10% or less or less (including, e.g., 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4% or less, 3% or less, 2% or less, 1% or less, or less) of such provided polymer combination preparation remains at the target site in vivo 10 days or more after administration.

[0228] In some embodiments in which the provided polymer combination preparations are immunomodulatory (e.g., act as polymer biomaterial agonists of innate immunity as described in PCT / US20 / 31169, filed May 1, 2020 (published as WO2020 / 223698A1)), the provided polymer combination preparations are characterized in that, when evaluated in vivo by administration to a target site (e.g., a tumor resection site) in a test subject (e.g., as described herein), such provided polymer combination preparations dissolve or degrade at a rate such that one or more aspects of the immune response are stimulated. For example, in some embodiments, such provided polymer combination preparations dissolve or degrade at a rate such that innate immunity is stimulated in one or more aspects (e.g., activation of pattern recognition receptors, inflammasomes, and / or the cGAS-STING pathway; and / or production of inflammatory cytokines, and / or upregulation of antigen presentation machinery and / or costimulatory molecules) for at least 2 days or longer (including, for example, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 9 days, at least 10 days, or longer). In some embodiments, such provided polymer combination preparations dissolve or degrade at a rate such that innate immunity is stimulated in one or more aspects (e.g., including but not limited to, activation of pattern recognition receptors, inflammasomes, and / or cGAS-STING pathway; and / or production of inflammatory cytokines, and / or upregulation of antigen presentation machinery and / or costimulatory molecules, e.g., as described herein) for 15 days or less or shorter (e.g., including 10 days or less, 9 days or less, 8 days or less, 7 days or less, 6 days or less, 5 days or less, 4 days or less, 3 days or less, or shorter).

[0229] F. Payload Release Rate In some embodiments, the polymer combination preparations described herein may be useful for delivering one or more payloads (e.g., resiquimod). For example, in some embodiments, one or more payloads may be dispersed in the polymer combination preparation such that when administered at a target site (e.g., a tumor resection site), the polymer combination preparation extends the release of the therapeutic agent at the target site compared to administration of the same therapeutic agent in solution. In certain embodiments, such polymer combination preparations may extend the release of the therapeutic agent at the target site (e.g., a tumor resection site) by at least 5 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 60 minutes, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 18 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, or 4 weeks compared to administration of the same therapeutic agent in solution. In some embodiments, such polymer combination preparations may extend the release of a therapeutic agent, such that when assessed at a particular time point after administration, more of the therapeutic agent is present at the target administration site (e.g., tumor resection site) than is observed when the therapeutic agent is administered in solution. For example, in some embodiments, when assessed 24 hours after administration, the amount of therapeutic agent released and present at the target administration site (e.g., tumor resection site) is at least 30% (including, for example, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or more) greater than when the therapeutic agent is administered in solution. In some embodiments, when assessed 48 hours after administration, the amount of therapeutic agent released and present at the target administration site (e.g., tumor resection site) is at least 30% (including, for example, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or more) greater than when the therapeutic agent is administered in solution.In some embodiments, when assessed 3 days after administration, the amount of therapeutic agent released and present at the target site of administration (e.g., tumor resection site) is at least 30% (including at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or more) greater than that observed when the therapeutic agent is administered in solution. In some embodiments, when assessed 5 days after administration, the amount of therapeutic agent released and present at the target site of administration (e.g., tumor resection site) is at least 30% (including at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or more) greater than that observed when the therapeutic agent is administered in solution.

[0230] G. In Vivo Efficacy In some embodiments, at least one therapeutic agent (e.g., resiquimod) may be incorporated into the polymer combination preparation and / or composition comprising the same described herein. In some embodiments, such polymer combination preparations are characterized in that a test animal group with spontaneous metastasis having the polymer combination preparation in a polymer network state at the tumor resection site has a higher survival rate when evaluated 2 months after administration than a comparable test animal group having the same polymer combination preparation at the tumor resection site without an immunomodulatory payload. In some such embodiments, the increase in survival observed in a test animal group with spontaneous metastasis having a provided polymer combination preparation (incorporated with an immunomodulatory payload) at the tumor resection site is at least 30% or more (including at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more) compared to the survival rate of a comparable test animal group having the same polymer combination preparation at the tumor resection site without an immunomodulatory payload when evaluated 2 months after administration.

[0231] III. EXEMPLARY EMBODIMENTS OF PROVIDED POLYMER COMBINATION PREPARATIONS In some embodiments, the polymer combination preparations described herein are prepared in a phosphate or carbonate buffer at a pH of 7-8. In some embodiments, the phosphate buffer may have a concentration of 10-50 mM (including, for example, 10 mM, 20 mM, 30 mM, 40 mM, or 50 mM). In some embodiments, the bicarbonate buffer may have a concentration of 25-200 mM (including, for example, 25 mM, 50 mM, 75 mM, 100 mM, 125 mM, 150 mM, 175 mM, or 200 mM).

[0232] In some embodiments, the polymer combination preparations described herein are temperature responsive and have a critical gelation temperature of about 10-30°C. In some embodiments, such polymer combination preparations described herein may have a critical gelation temperature near room temperature, e.g., 10-15°C. In some embodiments, such polymer combination preparations described herein may have a critical gelation temperature near room temperature, e.g., 15-20°C. In some embodiments, such polymer combination preparations described herein may have a critical gelation temperature near room temperature, e.g., 20-25°C. In some embodiments, such polymer combination preparations described herein may have a critical gelation temperature of about 25-28°C. In some embodiments, such polymer combination preparations described herein may have a critical gelation temperature of about 28-32°C. In some embodiments, such polymer combination preparations described herein may have a critical gelation temperature of about 32-34°C. In some embodiments, such polymer combination preparations described herein may have a critical gelling temperature of about 34-37°C.

[0233] In certain embodiments, the polymer combination preparation comprises 5-12.5% ​​(w / w) or 6-10% (w / w) poloxamer 407 and 0.5-3% (w / w) hyaluronic acid having an average molecular weight of 1-2 MDa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) have a viscosity of about 300 Pa to about 4,600 Pa or about 300 Pa to about 6,500 Pa (e.g., about 400-800 Pa, about 600-1,000 Pa, about 800-1,200 Pa, about 1,000-1,400 Pa, about 1,200-1,600 Pa, about 1,400-1,800 Pa, about 1,600-2,000 Pa, about 1,800-2,200 Pa, about 2,000-2,400 Pa, about 2,200-2,600 Pa, about 2,400-2,800 Pa, about 2,600-3,000 Pa, about The polymer may be characterized by a storage modulus that may be in the range of about 2,800-3,200 Pa, about 3,000-3,400 Pa, about 3,200-3,600 Pa, about 3,400-3,800 Pa, about 3,600-4,000 Pa, about 3,800-4,200 Pa, about 4,000-4,400 Pa, about 4,200-4,600 Pa, about 4,400-4,800 Pa, about 4,600-5,000 Pa, about 4,800-5,200 Pa, about 5,000-5,400 Pa, about 5,200-5,600 Pa, about 5,400-5,800 Pa, about 5,600-6,000 Pa, or about 5,800-6,500 Pa). In some embodiments, such polymer combination preparations (eg, upon transition to a polymer network state) may be characterized by a phase angle of about 2-20°.

[0234] In certain embodiments, the polymer combination preparation comprises 5-12.5% ​​(w / w), 7-12.5% ​​(w / w), 7-11.5% (w / w), 6-11.5% (w / w), 5-11.5% (w / w), 5-11% (w / w), 5-10.5% (w / w), 6-10.5% (w / w), 6-10% (w / w), 7-11% (w / w), or 8-11% (w / w) of poloxamer 407. with 0.5-3% (w / w) hyaluronic acid, 0.5-2% (w / w) hyaluronic acid, 1-2% (w / w) hyaluronic acid, 1-3% (w / w) hyaluronic acid, 1-4% (w / w) hyaluronic acid, 2-5% (w / w) hyaluronic acid, 3-6% (w / w) hyaluronic acid, or 4-7% (w / w) hyaluronic acid having an average molecular weight of 500 kDa to 900 kDa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 100 Pa to about 7,600 Pa, 100 Pa to about 15,000 Pa, or 500 Pa to about 18,000 Pa.In some embodiments, such polymer combination preparations (e.g., at the transition to a polymer network state) have a viscosity of about 300 Pa to about 8,000 Pa, (e.g., about 400 to 800 Pa, about 600 to 1,000 Pa, about 800 to 1,200 Pa, about 1,000 to 1,400 Pa, about 1,200 to 1,600 Pa, about 1,400 to 1,800 Pa, about 1,600 to 2,000 Pa, or about 1,500 to 2,000 Pa). ,000Pa, approx. 1,800~2,200Pa, approx. 2,000~2,400Pa, approx. 2,200~2,600Pa, approx. 2,400~2,800Pa, approx. 2,600~3,000Pa, approx. 2,800~3,200Pa, approx. 3,000~3,400Pa, approx. 3,200~3,600Pa, approx. 3,400~3,800Pa, approx. 3,600~4,000Pa, approx. 3,800~4 ,200Pa, approx. 4,000~4,400Pa, approx. 4,200~4,600Pa, approx. 4,400~4,800Pa, approx. 4,600~5,000Pa, approx. 4,800~5,200Pa, approx. 5,000~5,400Pa, approx. 5,200~5,600Pa, approx. 5,400~5,800Pa, approx. 5,600~6,000Pa, approx. 5,800~6,200Pa, approx. 5,800~ The polymer combination preparations may be characterized by a storage modulus that may range from about 6,400 Pa, about 6,000-6,400 Pa, about 6,200-6,600 Pa, about 6,400-6,800 Pa, about 6,600-7,000 Pa, about 6,800-7,200 Pa, about 7,000-7,400 Pa, about 7,200-7,600 Pa, about 7,400-7,800 Pa, about 7,600-8,000 Pa). In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a phase angle of about 2-20°.

[0235] In certain embodiments, the polymer combination preparation comprising high molecular weight hyaluronic acid comprises the formulations described in Table 10 of Example 5.

[0236] In certain embodiments, the polymer combination preparation comprises 5-12.5% ​​(w / w), 8-12.5% ​​(w / w), 6-11.5% (w / w), 6-11% (w / w), 7-11% (w / w), 8-11% (w / w), 6-10.5% (w / w), or 6-10% (w / w) of poloxamer 407 with 1-4% (w / w) of hyaluronic acid having an average molecular weight of 100 kDa to 500 kDa, 2-5% (w / w) of hyaluronic acid, or 1-10% (w / w) of hyaluronic acid, or 1.5-10% (w / w) of hyaluronic acid, or 3-6% (w / w) of hyaluronic acid, or 4-7% (w / w) of hyaluronic acid. In certain embodiments, the polymer combination preparation is 10.9% (w / w), 10.8% (w / w), 10.7% (w / w), 10.6% (w / w), 10.5% (w / w), 10.4% (w / w), 10.3% (w / w), 10.2% (w / w), 10.1% (w / w), 10.0% (w / w), 9.9% (w / w), 9.8% (w / w), 9.7% (w / w), 9.6% (w / w), 9.5 ...8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% (w / w), 10.8% % (w / w), or 9.0% (w / w) poloxamer 407, together with 1-4% (w / w) hyaluronic acid having an average molecular weight of 100 kDa to 500 kDa, or 2-5% (w / w) hyaluronic acid, or 1-10% (w / w) hyaluronic acid, or 1.5-10% (w / w) hyaluronic acid, or 3-6% (w / w) hyaluronic acid, or 4-7% (w / w) hyaluronic acid. In certain embodiments, the polymer combination preparation comprises 5-12.5% ​​(w / w), 8-12.5% ​​(w / w), 8-11% (w / w), 6-11% (w / w), 6-10.5% (w / w), or 6-10% (w / w) of poloxamer 407 and 0.5-10% (w / w) of hyaluronic acid having an average molecular weight of 100 kDa to 300 kDa or 1.5-10% (w / w) of hyaluronic acid or 2-6% (w / w) of hyaluronic acid or 4-9% (w / w) of hyaluronic acid.In certain embodiments, the polymer combination preparation is 10.9% (w / w), 10.8% (w / w), 10.7% (w / w), 10.6% (w / w), 10.5% (w / w), 10.4% (w / w), 10.3% (w / w), 10.2% (w / w), 10.1% (w / w), 10.0% (w / w), 9.9% (w / w), 9.8% (w / w), 9.7% (w / w), 9.6% (w / w), 9.5% (w / w), or 9.0% (w / w) of poloxamer 407, and 0.5-10% (w / w) of hyaluronic acid having an average molecular weight of 100 kDa to 300 kDa, or 1.5-10% (w / w) of hyaluronic acid, or 2-6% (w / w) of hyaluronic acid, or 4-9% (w / w) of hyaluronic acid. In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) of poloxamer 407 and 2-6% (w / w) of hyaluronic acid having an average molecular weight of 100 kDa to 200 kDa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 400 Pa to about 3,400 Pa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a phase angle of about 2-35° or 2-20°. In certain embodiments, the polymer combination preparations include 5-11% (w / w), 6-10.5% (w / w), or 6-10% (w / w) poloxamer 407 and 1-10% (w / w) or 1.5-10% (w / w) hyaluronic acid having an average molecular weight of 100 kDa to 200 kDa.In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) have a viscosity of about 400 Pa to about 5,000 Pa or 300 Pa to about 6,500 Pa (e.g., about 400-800 Pa, about 600-1,000 Pa, about 800-1,200 Pa, about 1,000-1,400 Pa, about 1,200-1,600 Pa, about 1,400-1,800 Pa, about 1,600-2,000 Pa, about 1,800-2,200 Pa, about 2,000-2,400 Pa, about 2,200-2,600 Pa, about 2,400-2,800 Pa, about 2,600-3,000 Pa, about 2,800-3,200 Pa, about 3,000-4,000 Pa, about 4,000-5,000 Pa, about 5,000-6,000 Pa, about 6,000-7,000 Pa, about 7,000-8,000 Pa, about 8,000-9,000 Pa, about 9,000-10,000 Pa, about 10,000-11,000 Pa, about 12,000-13,000 Pa, about 14,000-15,000 Pa, about 15,000-16,000 Pa, about 16,000-20,000 Pa, about 17,000-20,000 Pa, about 18,000-20,000 Pa, about 20,000-21,000 Pa, about 21,000-22,000 Pa, about 3,400Pa, approx. 3,200~3,600Pa, approx. 3,400~3,800Pa, approx. 3,600~4,000Pa, approx. 3,800~4,200Pa, approx. 4,000~4,400Pa, approx. 4,200~4,600Pa, approx. 4,400~4,800Pa, approx. 4,600~5,000Pa, approx. 4,800~5,200Pa , about 5,000-5,400 Pa, about 5,200-5,600 Pa, about 5,400-5,800 Pa, about 5,600-6,000 Pa, about 5,800-6,200 Pa, about 5,800-6,400 Pa, about 6,000-6,400 Pa, about 6,200-6,500 Pa). In some embodiments, such polymer combination formulations (e.g., upon transition to a polymer network state) may be characterized by a phase angle of about 20-35°.

[0237] In certain embodiments, the polymer combination preparation comprising low molecular weight hyaluronic acid comprises the formulation described in Table 9 of Example 5.

[0238] In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) poloxamer 407 or 6-10% (w / w) poloxamer 407 and 1-10% (w / w) or 1.5-9% (w / w) or 1-5% (w / w) or 5-10% (w / w) hyaluronic acid having an average molecular weight of 70 kDa to 200 kDa or 80 kDa to 150 kDa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 200 Pa to about 6,500 Pa or from about 200 Pa to about 5,900 Pa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a phase angle of about 2-35° or 2-20°. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) have a viscosity of about 400 Pa to about 6,500 Pa or 400 Pa to about 4,600 Pa (e.g., about 400-800 Pa, about 600-1,000 Pa, about 800-1,200 Pa, about 1,000-1,400 Pa, about 1,200-1,600 Pa, about 1,400-1,800 Pa, about 1,600-2,000 Pa, about 1,800-2,200 Pa, about 2,000-2,400 Pa, about 2,200-2,600 Pa, about 2,400-2,800 Pa, about 2,600-3,000 Pa, about 2,800-3,200 Pa, about 3,000-4,000 Pa, or about 4,500 Pa). 3,400Pa, approx. 3,200~3,600Pa, approx. 3,400~3,800Pa, approx. 3,600~4,000Pa, approx. 3,800~4,200Pa, approx. 4,000~4,400Pa, approx. 4,200~4,600Pa, approx. 4,400~4,800Pa, approx. 4,600~5,000Pa, approx. 4,800~5,200Pa , about 5,000-5,400 Pa, about 5,200-5,600 Pa, about 5,400-5,800 Pa, about 5,600-6,000 Pa, about 5,800-6,200 Pa, about 5,800-6,400 Pa, about 6,000-6,400 Pa, about 6,200-6,500 Pa).In some embodiments, such polymer combination formulations (eg, upon transition to a polymer network state) can be characterized by a phase angle of about 2-32° or about 15-35°.

[0239] In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) poloxamer 338 and 1-3% (w / w) hyaluronic acid having an average molecular weight of 1-2 MDa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 980 Pa to about 1,300 Pa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a phase angle of about 2-35° or 2-20°.

[0240] In certain embodiments, the polymer combination preparation comprises 5-12.5% ​​(w / w) or 8-11.5% (w / w) or 8-11% (w / w) of poloxamer 338 with 1-4% (w / w) of hyaluronic acid having an average molecular weight of 500 kDa to 900 kDa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 1,400 Pa to about 2,700 Pa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a phase angle of about 2-35° or 2-20°.

[0241] In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) poloxamer 338 and 1-5% (w / w) hyaluronic acid having an average molecular weight of 100 kDa to 350 kDa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 500 Pa to about 1,350 Pa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a phase angle of about 2-35° or 2-20°.

[0242] In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) poloxamer 407 and 2.5-5% (w / w) modified chitosan (e.g., carboxymethyl chitosan). In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 1,000 Pa to about 5,000 Pa. In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a phase angle of about 2-35° or 2-20°.

[0243] In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) poloxamer 407, 0.5-5% (w / w) hyaluronic acid having an average molecular weight of 500 kDa to 900 kDa, and 0.1-1.5% modified chitosan (e.g., carboxymethyl chitosan). In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 400 Pa to about 3,400 Pa (e.g., about 400-800 Pa, about 600-1,000 Pa, about 800-1,200 Pa, about 1,000-1,400 Pa, about 1,200-1,600 Pa, about 1,400-1,800 Pa, about 1,600-2,000 Pa, about 1,800-2,200 Pa, about 2,000-2,400 Pa, about 2,200-2,600 Pa, about 2,400-2,800 Pa, about 2,600-3,000 Pa, about 2,800-3,200 Pa, about 3,000-3,400 Pa). In some embodiments, such polymer combination formulations (eg, upon transition to a polymer network state) can be characterized by a phase angle of about 2-35° or 2-20°.

[0244] In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) or 6-11% (w / w) or 6-10.5% (w / w) or 6-10% (w / w) of poloxamer 407, 0.5%-10% (w / w) or 1-10% (w / w) or 1-5% (w / w) of hyaluronic acid having an average molecular weight of 80 kDa to 150 kDa, and 0.1-5% (w / w) or 0.2-5% (w / w) or 0.1-3% (w / w) of modified chitosan (e.g., carboxymethyl chitosan and / or chitosan-phenylsuccinate). In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 400 Pa to about 3,400 Pa (e.g., about 400-800 Pa, about 600-1,000 Pa, about 800-1,200 Pa, about 1,000-1,400 Pa, about 1,200-1,600 Pa, about 1,400-1,800 Pa, about 1,600-2,000 Pa, about 1,800-2,200 Pa, about 2,000-2,400 Pa, about 2,200-2,600 Pa, about 2,400-2,800 Pa, about 2,600-3,000 Pa, about 2,800-3,200 Pa, about 3,000-3,400 Pa). In some embodiments, such polymer combination formulations (eg, upon transition to a polymer network state) can be characterized by a phase angle of about 2-35° or 2-20°.

[0245] In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) poloxamer 407, 1-5% (w / w) hyaluronic acid having an average molecular weight of 500 kDa to 900 kDa, and 0.2-4% modified chitosan (e.g., carboxymethyl chitosan). In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 400 Pa to about 3,400 Pa (e.g., about 400-800 Pa, about 600-1,000 Pa, about 800-1,200 Pa, about 1,000-1,400 Pa, about 1,200-1,600 Pa, about 1,400-1,800 Pa, about 1,600-2,000 Pa, about 1,800-2,200 Pa, about 2,000-2,400 Pa, about 2,200-2,600 Pa, about 2,400-2,800 Pa, about 2,600-3,000 Pa, about 2,800-3,200 Pa, about 3,000-3,400 Pa). In some embodiments, such polymer combination formulations (eg, upon transition to a polymer network state) can be characterized by a phase angle of about 2-35° or 2-20°.

[0246] In certain embodiments, the polymer combination preparation comprises 8-12.5% ​​(w / w) or 8-11% (w / w) poloxamer 407, 1-5% (w / w) hyaluronic acid having an average molecular weight of 100 kDa to 500 kDa, and 0.2-4% modified chitosan (e.g., carboxymethyl chitosan). In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 400 Pa to about 3,400 Pa (e.g., about 400-800 Pa, about 600-1,000 Pa, about 800-1,200 Pa, about 1,000-1,400 Pa, about 1,200-1,600 Pa, about 1,400-1,800 Pa, about 1,600-2,000 Pa, about 1,800-2,200 Pa, about 2,000-2,400 Pa, about 2,200-2,600 Pa, about 2,400-2,800 Pa, about 2,600-3,000 Pa, about 2,800-3,200 Pa, about 3,000-3,400 Pa). In some embodiments, such polymer combination formulations (eg, upon transition to a polymer network state) can be characterized by a phase angle of about 2-35° or 2-20°.

[0247] In certain embodiments, the polymer combination preparation comprises 3.5-5.5% (w / w) or 4-5% (w / w) of poloxamer 407 and 1.5-3.5% (w / w) of high molecular weight hyaluronic acid (e.g., hyaluronic acid having an average molecular weight of greater than 500 kDa, e.g., 600-1500 kDa or 700-1500 kDa). In some embodiments, such polymer combination preparations (e.g., upon transition to a polymer network state) may be characterized by a storage modulus that may range from about 200 Pa to about 10,000 Pa, 500 Pa to about 9,000 Pa, or about 1,000 Pa to about 8,000 Pa, or 1,000 Pa to about 6,000 Pa.

[0248] In some embodiments, the polymer combination preparation further comprises resiquimod. In some embodiments, the polymer combination preparation further comprises resiquimod at 0.02 mg / mL, 0.05 mg / mL, 0.10 mg / mL, 0.12 mg / mL, 0.14 mg / mL, 0.16 mg / mL, 0.18 mg / mL, 0.20 mg / mL, 0.22 mg / mL, 0.25 mg / mL, 0.30 mg / mL, 0.35 mg / mL, 0.40 mg / mL, 0.45 mg / mL, 0.50 mg / mL, 0.60 mg / mL, 0.70 mg / mL, 0.80 mg / mL, 0.90 mg / mL, 0.10 mg / mL, 0.12 mg / mL, 0.14 mg / mL, 0.16 mg / mL, 0.18 mg / mL, 0.20 mg / mL, 0.22 mg / mL, 0.25 mg / mL, 0.30 mg / mL, 0.35 mg / mL, 0.40 mg / mL, 0.45 mg / mL, 0.50 mg / mL, 0.70 mg / mL, 0.75 mg / mL, 0.90 mg / mL, 0.9 ... / mL, 0.55mg / mL, 0.60mg / mL, 0.65mg / mL, 0.70mg / mL, 0.75mg / mL, 0.80mg / mL, 0.85mg / mL, 0.90mg / mL, 0.95mg / mL, 1.00mg / mL, 1.05mg / mL, 1.10mg / mL, 1.15mg / mL, 1.20mg / mL, 1.25mg / mL, 1.50mg / mL, or 1.80mg / mL. In some embodiments, biomaterial compositions provided include resiquimod at a concentration of 0.01 mg / mL to 1.80 mg / mL, 0.01 mg / mL to 0.50 mg / mL, 0.05 mg / mL to 1.00 mg / mL, 0.05 mg / mL to 0.50 mg / mL, 0.05 mg / mL to 0.30 mg / mL, 0.05 mg / mL to 0.20 mg / mL, 0.10 mg / mL to 0.25 mg / mL, 0.10 mg / mL to 0.20 mg / mL, 0.12 mg / mL to 0.18 mg / mL, or 0.14 mg / mL to 0.20 mg / mL. In some embodiments, such polymer combination preparations include resiquimod as the sole immunomodulatory payload at a concentration of 0.01 mg / mL to 0.50 mg / mL, 0.05 mg / mL to 0.50 mg / mL, 0.05 mg / mL to 0.30 mg / mL, 0.05 mg / mL to 0.20 mg / mL, 0.10 mg / mL to 0.25 mg / mL, 0.10 mg / mL to 0.20 mg / mL, 0.12 mg / mL to 0.18 mg / mL, or 0.14 mg / mL to 0.20 mg / mL. In some embodiments, such polymer combination preparations further include resiquimod and an additional payload (e.g., an additional immunomodulatory payload).

[0249] Methods for preparing biomaterial compositions are provided. In some embodiments, the present disclosure provides methods of preparing the provided biomaterial compositions (e.g., polymer combination preparations and compositions thereof). In some embodiments, the provided methods of preparing the provided biomaterial compositions use solid forms of resiquimod and compositions thereof described herein. In some embodiments, resiquimod is provided and / or used in accordance with the present disclosure in a form such as a solid form. In some embodiments, resiquimod is provided and / or used in accordance with the present disclosure as an amorphous form, a crystalline form, or a mixture thereof. In some embodiments, resiquimod is provided and / or used in accordance with the present disclosure as a composition comprising one or more solid forms of resiquimod described herein.

[0250] In some embodiments, the disclosure provides methods that include: (i) providing at least one solid form of resiquimod (e.g., resiquimod Form I, resiquimod Form II, resiquimod Form III, resiquimod Form IV, resiquimod Form V, resiquimod Form VI, or resiquimod Form VII); and (ii) combining the at least one solid form of resiquimod with a poloxamer and a second polymer component (e.g., hyaluronic acid and / or chitosan) in a suitable buffer.

[0251] In some embodiments, the disclosure provides a method comprising: (i) providing at least one solid form of resiquimod (e.g., resiquimod Form I, resiquimod Form II, resiquimod Form III, resiquimod Form IV, resiquimod Form V, resiquimod Form VI, or resiquimod Form VII); (ii) combining the at least one solid form of resiquimod with a poloxamer in a suitable buffer; and (iii) adding a second polymer component (e.g., hyaluronic acid and / or chitosan).

[0252] In some embodiments, the disclosure provides a method comprising: (i) providing a polymer combination preparation, e.g., as described herein; and (ii) combining the polymer combination preparation with at least one solid form of resiquimod (e.g., resiquimod Form I, resiquimod Form II, resiquimod Form III, resiquimod Form IV, resiquimod Form V, resiquimod Form VI, or resiquimod Form VII).

[0253] In some embodiments, the disclosure provides a method comprising: (i) mixing a suitable amount of a poloxamer and at least a second polymer component (e.g., hyaluronic acid and / or chitosan) in a suitable buffer to provide a polymer combination preparation, e.g., as described herein; and (ii) combining the polymer combination preparation with at least one solid form of resiquimod (e.g., resiquimod Form I, resiquimod Form II, resiquimod Form III, resiquimod Form IV, resiquimod Form V, resiquimod Form VI, or resiquimod Form VII).

[0254] In some embodiments, the polymer combination preparations described herein can be prepared by mixing an appropriate amount of poloxamer and at least a second polymer component (e.g., hyaluronic acid and / or chitosan) in a suitable buffer. The poloxamer and at least a second polymer component (e.g., hyaluronic acid and / or chitosan) can be, independently, a solid particle preparation or a liquid preparation. In some embodiments, a payload (e.g., resiquimod and optionally further payloads) can be added to such a polymer mixture solution. In some embodiments, the polymer mixture solution can be mixed at a low speed (e.g., a speed of less than 100 rpm) until a homogenous polymer solution is formed. To induce gel formation, such a homogenous polymer solution can be exposed to a temperature at or above the critical gelation temperature for a time sufficient for gel formation (e.g., 10-15 minutes).

[0255] In some embodiments, the present disclosure recognizes that, inter alia, mixing a solid particle preparation of hyaluronic acid (HA) with at least a second polymer preparation (e.g., a poloxamer), which may be a solid particle preparation or a liquid preparation, may facilitate the formation of a homogenous polymer solution compared to mixing a liquid preparation of HA and at least a second polymer.

[0256] Thus, one aspect provided herein relates to a method for preparing a homogenous polymer combination of a hyaluronic acid (HA) polymer preparation and a second polymer preparation, the method comprising combining the HA polymer preparation and the second polymer preparation when the HA polymer preparation is in solid particle form. In some embodiments, the solid particle preparation of HA polymer comprises a HA polymer in powder form. Those skilled in the art who read this disclosure will understand that HA polymers tend to be hygroscopic, and in some embodiments, the HA polymer in the solid particle preparation can be or comprise a hydrated HA polymer.

[0257] In some embodiments, the HA polymer preparation in solid particulate form may be combined with at least a second polymer preparation (e.g., a poloxamer) in solid particulate form (e.g., in some embodiments, a powder) and then dissolved together simultaneously in a liquid solution (e.g., a buffer solution). In some embodiments, the HA polymer preparation in solid particulate form may be combined with at least a second polymer preparation (e.g., a poloxamer) in liquid form, which in some embodiments may be a solution of the second polymer in a solvent system (e.g., as described herein).

[0258] In some embodiments, the HA and the second and optional additional polymer preparation(s) are combined under sufficient conditions and for sufficient time to produce a homogenous polymer mixture. In some embodiments, such homogenous polymer mixtures are characterized by no detectable phase separation being observed after maintaining the homogenous polymer mixture at a temperature below the critical gelation temperature (e.g., in some embodiments, at 2-8° C., or in some embodiments, at ambient temperature) for at least 1 hour or more (e.g., at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 12 hours, at least 18 hours, at least 24 hours, or longer). In some embodiments, such homogenous polymer mixtures are characterized by no detectable phase separation being observed after maintaining the homogenous polymer mixture at a temperature below the critical gelation temperature (e.g., in some embodiments, at 2-8° C., or in some embodiments, at ambient temperature) for at least 1 week or more (e.g., at least 2 weeks, at least 3 weeks, at least 4 weeks, or longer). In some embodiments, such homogenous polymer mixtures produced are characterized in that no detectable phase separation is observed after maintaining the homogenous polymer mixtures produced at a temperature below the critical gelation temperature (e.g., in some embodiments, at 2-8° C., or in some embodiments, at ambient temperature) for at least one month or more (e.g., including at least two months, at least three months, or longer).

[0259] In some embodiments, the HA and the second polymer preparation (e.g., poloxamer) and optionally the additional polymer preparation(s) are combined by mixing them at ambient temperature and / or at low shear rate. In some embodiments, mixing can be performed by mechanical stirring. As will be understood by those skilled in the art, the shear rate is usually determined by the size and rpm of the stirring unit (e.g., impeller or stirrer, e.g., magnetic stirrer), and the highest shear rate is usually the speed at the tip of the stirring unit (e.g., impeller or stirrer). In some embodiments, a cylindrical stirrer can be used to induce radial flow. In some embodiments, at least two-blade (e.g., 2, 3, or 4-blade) impellers can be used to induce axial or radial flow depending on the blade geometry. In axial flow, the movement is parallel to the axis (up and down), and in radial flow, the movement is perpendicular to the axis. In some embodiments, the HA and the second polymer preparation are combined by mixing them at ambient temperature and at a speed of less than 100 rpm.

[0260] In some embodiments, the HA and the second polymer preparation (e.g., poloxamer) and optionally the additional polymer preparation(s) are mixed for at least 5 hours (including, for example, at least 10 hours, at least 15 hours, at least 20 hours, at least 25 hours, at least 30 hours, or longer). In some embodiments, the HA and the second polymer preparation and optionally the additional polymer preparation(s) are mixed for 5-30 hours or 10-24 hours.

[0261] In some embodiments, the HA and second polymer preparation (e.g., poloxamer) and optionally additional polymer(s) may be mixed at a temperature of 2-8° C., for example, in some embodiments, for at least 5 hours (including, for example, at least 10 hours, at least 15 hours, at least 20 hours, at least 25 hours, at least 30 hours, or longer).

[0262] In some embodiments, the HA and second polymer preparation (e.g., poloxamer) and optionally additional polymer(s) (e.g., CMCH) are mixed at a temperature of 2-8° C. and then quickly brought to a temperature equal to or greater than the respective CGT (e.g., a suitable CGT as described herein) to reach a polymer network state, e.g., to prevent phase separation. In some such embodiments, the resulting polymer network can be stored at a temperature equal to or greater than the respective CGT (e.g., a suitable CGT as described herein), e.g., in some embodiments, at ambient temperature, until ready for delivery. In some embodiments, the resulting polymer network can be delivered at a temperature below the respective CGT (e.g., a suitable CGT as described herein) to provide it as a solution and / or liquid preparation.

[0263] In some embodiments, the payload (e.g., resiquimod and optionally additional payload) may be incorporated into a homogenous mixture of HA and the second polymer preparation. In some embodiments, the payload (e.g., resiquimod and optionally additional payload) may be added by combining the HA and the second polymer preparation with the payload. In some embodiments, the combined payload (e.g., resiquimod and optionally additional payload) may be a solid particle preparation. In some embodiments, the combined payload (e.g., resiquimod and optionally additional payload) may be a liquid preparation (e.g., a liquid preparation of a payload prepared from a solid form of resiquimod described herein).

[0264] In some embodiments, the homogenous polymer mixture produced (with or without a payload) may be exposed to a gelation temperature at or above the critical gelation temperature of the polymer mixture for a sufficient time such that a hydrogel is formed. In some embodiments, the homogenous polymer mixture produced (with or without a payload) may be exposed to a gelation temperature of about 35-39° C. In some embodiments, the homogenous polymer mixture produced (with or without a payload) may be exposed to a gelation temperature of about 37° C. In some embodiments, the homogenous polymer mixture produced (with or without a payload) is exposed to a gelation temperature for 5 minutes to 30 minutes.

[0265] Uses of provided biomaterial compositions The preparations and / or compositions described herein may be useful in a variety of medical applications, including, but not limited to, immunomodulation and / or drug delivery. Thus, in some embodiments, the preparations and / or compositions described herein may be formulated as pharmaceutical compositions for administration to a subject in need thereof. Thus, in one aspect, methods are provided herein that include administering to a subject in need thereof the preparations or compositions described and / or used herein or pharmaceutical compositions comprising same.

[0266] In some embodiments, the provided polymer combination preparations and / or compositions can be routinely formulated as pharmaceutical compositions for administration to a subject in need thereof (e.g., as described herein). In some embodiments, such pharmaceutical compositions can include pharma- ceutically acceptable carriers or excipients, which, as used herein, include any solvent, dispersion medium, diluent, or other liquid vehicle, dispersion or suspension aid, surfactant, isotonicity agent, thickening or emulsifying agent, preservative, solid binder, lubricant, and the like, suitable for the particular dosage form desired. Remington's The Science and Practice of Pharmacy, 21st Edition, AR Gennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006, incorporated herein by reference) discloses various excipients used in formulating pharmaceutical compositions and known techniques for their preparation. Suitable pharma- ceutically acceptable carriers include, but are not limited to, water, salt solutions (e.g., NaCl), saline, buffered saline, glycerol, sugars such as mannitol, lactose, trehalose, sucrose, or others, dextrose, fatty acid esters, and the like, and combinations thereof.

[0267] Pharmaceutical compositions may be mixed with auxiliary agents (e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring agents, flavoring agents, and / or aromatic agents) that do not adversely react with the active compounds or interfere with their activity, as needed. In some embodiments, pharmaceutical compositions may be sterile. Suitable pharmaceutical compositions may contain minor amounts of wetting agents or emulsifiers or pH buffering agents, as needed. Pharmaceutical compositions may be liquid solutions, suspensions, or emulsions.

[0268] The pharmaceutical composition may be formulated according to routine procedures as a pharmaceutical composition adapted for administration to humans. The preparation of the pharmaceutical composition must be suitable for the mode of administration. For example, in some embodiments, the pharmaceutical composition for injection may usually comprise a sterile aqueous isotonic buffer. If necessary, the pharmaceutical composition may also comprise a local anesthetic to ease pain at the injection site. In some embodiments, the components of the pharmaceutical composition (e.g., as described herein) are supplied separately or mixed together in a single-use form, for example, as a dry lyophilized powder or a moisture-free concentrate in a hermetically sealed container, such as an ampoule or sachet, or in a sterile syringe indicating the amount of the composition comprising the polymer combination preparation (e.g., as described herein). When the pharmaceutical composition is administered by injection, in some embodiments, the dry lyophilized powder composition comprising the polymer combination preparation (e.g., as described herein) may be reconstituted with an aqueous buffer and then injected into the target site of the subject that needs to be injected. In some embodiments, liquid compositions including a polymer combination preparation (e.g., those described herein) may be provided in a syringe for injection and / or administration via a robotic surgery system (e.g., the da Vinci System).

[0269] In some embodiments, a liquid composition including a polymer combination preparation (e.g., as described herein) may be provided in a syringe for administration with or without a needle, cannula, or trocar.

[0270] In some embodiments, a liquid composition including a polymer combination preparation (eg, those described herein) may be administered by nebulization.

[0271] In some embodiments, administration of liquid compositions including polymer combination preparations (eg, those described herein) can be gas-assisted for use in minimally invasive surgery.

[0272] In some embodiments, administration of a liquid composition comprising a polymer combination formulation (e.g., those described herein) may be accomplished by using a multi-barrel syringe, where each barrel contains a separate polymer component formulation and the multiple polymer component formulations are combined when a shared plunger is depressed.

[0273] Although the description of pharmaceutical compositions provided herein is directed primarily to pharmaceutical compositions suitable for ethical administration to humans, it will be understood by those skilled in the art that such compositions are generally suitable for administration to any type of animal or in vitro or ex vivo cell. Modifications of pharmaceutical compositions suitable for administration to humans to make them suitable for administration to a variety of animals or in vitro or ex vivo cells are well understood, and such modifications can be designed and / or performed by those skilled in the art, e.g., veterinary pharmacologists, with no more than routine experimentation, if any.

[0274] The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the field of pharmacology. For example, such preparation methods include mixing the components of the provided polymer combination preparation and resiquimod with a diluent or other excipient and / or one or more other auxiliary ingredients, and then forming and / or packaging the product into a desired single-use or multi-use unit, as needed and / or desired. Alternatively, such preparation methods may also include preforming a polymer network biomaterial from the components of the polymer combination preparation described herein before forming and / or packaging the product into a desired single-use or multi-use unit.

[0275] The pharmaceutical compositions according to the present disclosure can be prepared, packaged, and / or sold in bulk as a single-use unit and / or as a plurality of single-use units. As used herein, a "single-use unit" is a discrete amount of the pharmaceutical composition described herein. For example, a single-use unit of a pharmaceutical composition comprises a predetermined amount of the composition and / or polymer combination preparation described herein, which in some embodiments can be or include a preformed polymer network of the polymer combination preparation (e.g., as described herein), or in some embodiments can be or include a liquid or colloidal mixture of the individual components of the polymer combination preparation (e.g., as described herein).

[0276] The relative amounts of the individual components of the polymer combination preparation and resiquimod provided in the pharmaceutical compositions described herein (e.g., as a preformed polymer network biomaterial or as a precursor component(s) of such polymer network biomaterial), and optionally any additional agents, e.g., pharma- ceutically acceptable excipients and / or any additional components, may vary depending, for example, on the desired material properties of the polymer biomaterial, the size of the target site, the injection volume, the health and medical condition of the subject to be treated, and / or the type of cancer, and may also depend on the route by which such pharmaceutical composition is administered. In some embodiments, the polymer combination preparation and resiquimod may be provided in the pharmaceutical composition in an effective amount to provide a desired therapeutic effect (e.g., but not limited to, eliciting at least one or more aspects of anti-tumor immunity, e.g., eliciting innate immunity). In some embodiments, the polymer combination preparation and resiquimod may be provided in the pharmaceutical composition in an effective amount to treat cancer. In some embodiments, the polymer combination preparation and resiquimod may be provided in the pharmaceutical composition in an effective amount to inhibit or reduce the risk or incidence of tumor recurrence and / or metastasis. In certain embodiments, the effective amount is a therapeutically effective amount of the polymer combination preparation and resiquimod. In certain embodiments, the effective amount is a prophylactically effective amount of the polymer combination preparation and resiquimod.

[0277] In certain embodiments, the pharmaceutical compositions consist essentially of or consist of the polymer combination preparation (e.g., as described herein) and resiquimod, to the extent that such compositions may contain one or more substances / drugs other than the polymer combination preparation and resiquimod, and such other substance(s) / drug(s) individually or together do not substantially alter the relevant immunomodulatory property(s) of the polymer combination preparation and resiquimod, e.g., the innate immunomodulatory property(s) thereof.

[0278] In certain embodiments, the pharmaceutical composition does not include cells. In certain embodiments, the pharmaceutical composition does not include adoptively transferred cells. In certain embodiments, the pharmaceutical composition does not include T cells. In certain embodiments, the pharmaceutical composition does not include tumor antigens. In certain embodiments, the pharmaceutical composition does not include ex vivo loaded tumor antigens.

[0279] In certain embodiments, the pharmaceutical composition is in a liquid form (e.g., a solution or colloid). In certain embodiments, the pharmaceutical composition is in a solid form (e.g., a gel form). In certain embodiments, the transition from liquid form to solid form can occur outside the subject's body by sufficient crosslinking, such that the resulting material has a storage modulus consistent with the solid form that allows it to be physically manipulated and implanted in a surgical procedure. Thus, in some embodiments, the pharmaceutical composition in solid form may be suitable for carrying out the intended use (e.g., surgical implantation) of the present disclosure. In certain embodiments, the transition from liquid form to solid form can occur in situ (e.g., within the subject's body) by thermal crosslinking, such that the resulting material has a storage modulus consistent with the solid form. In certain embodiments, the pharmaceutical composition is a suspension.

[0280] In some embodiments, the preparations or compositions described and / or used herein, or pharmaceutical compositions comprising the same, may be useful for the treatment of cancer. In some such embodiments, the subject to which the treatment is administered is a subject suffering from cancer. In some embodiments, the subject to which the treatment is administered is a subject suffering from or susceptible to recurrent or disseminated cancer. In some embodiments, the subject to which the treatment is administered is a tumor resection subject.

[0281] In many embodiments, the polymer combination preparations described herein and compositions comprising same are biocompatible and useful for various medical applications, for example, in some embodiments, as drug delivery carriers or formulations (e.g., sustained release drug delivery compositions). For example, in some embodiments, the polymer combination preparations described herein and compositions comprising same are useful for treating a disease, disorder, or condition. In some embodiments, the polymer compositions described herein and compositions comprising same are useful for treating cancer. In some embodiments, the polymer combination preparations described herein and compositions comprising same are useful for delaying the onset of cancer, slowing its progression, or ameliorating one or more symptoms thereof. In some embodiments, the polymer combination preparations described herein and compositions comprising same are useful for reducing or inhibiting the regrowth of primary tumors. In some embodiments, the polymer combination preparations described herein and compositions comprising same reduce or inhibit the occurrence of tumor recurrence and / or metastasis. In some embodiments, the polymer combination preparations described herein and compositions comprising same are useful for inducing anti-tumor immunity.

[0282] Accordingly, some aspects provided herein relate to a method of administering a composition comprising a polymer combination preparation described herein to a target site of a subject in need of administration. In some embodiments, the subject to which such a composition is administered may have a tumor. In some such embodiments, the method comprises intratumoral or peritumoral administration of a composition comprising a polymer combination preparation described herein. In some embodiments, the subject to which such a composition is administered may have or may have had tumor removal (e.g., by surgical tumor resection). In some embodiments, the subject to which such a composition is administered may have tumor recurrence and / or metastasis. In some such embodiments, the method comprises intraoperative administration of a composition comprising a polymer combination preparation described herein at the site of tumor resection of the subject.

[0283] In some embodiments, the composition administered to a subject in need thereof comprises a polymer combination preparation and resiquimod. In some embodiments, such provided compositions for use in the methods of the present disclosure can be formulated as pharmaceutical compositions as described herein.

[0284] In some embodiments, the method includes administering a provided preparation or composition, or a pharmaceutical composition comprising the same, to a target site of a tumor resection subject. In some embodiments, the preparation or composition, or a pharmaceutical composition comprising the same, is administered to the tumor resection site.

[0285] In some embodiments, administration can be by implantation. For example, in some embodiments, preparations or compositions that include polymer combination preparations in a polymer network state (e.g., a hydrogel) can be administered by implantation.

[0286] In some embodiments, administration can be by injection. In some embodiments, injection can be by a robotic arm. For example, in some embodiments, a preparation that includes a polymer combination preparation in a precursor state (e.g., a liquid state or an injectable state) is administered by injection, and upon administration, the precursor state transitions to a polymer network state (e.g., a more viscous solution or a colloidal state or a hydrogel).

[0287] In some embodiments, administration can occur simultaneously with or after laparoscopy. In some embodiments, administration can occur simultaneously with or after minimally invasive surgery (MIS), such as robotic-assisted MIS, robotic surgery, and / or laparoscopic surgery for tumor resection.

[0288] In certain embodiments, the methods provided herein include administering a provided composition to a target site in a subject in need of administration after removal of a tumor, for example, after removal of 50% or more of the tumor mass in the subject (including, for example, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more of the tumor mass in the subject). In certain embodiments, the methods provided herein include administering a provided composition to a target site in a subject in need of administration after removal of 50% or more of the tumor mass in the subject (including, for example, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more of the tumor mass in the subject). In some embodiments, the methods provided herein include performing a tumor resection to remove the tumor in the subject prior to administration of a provided composition.

[0289] In some embodiments, the compositions described and / or utilized herein are administered to a target site of a tumor resection subject immediately after the tumor is removed by surgical tumor resection. In some embodiments, the compositions described and / or utilized herein are administered intraoperatively to a target site of a tumor resection subject. In some embodiments, the compositions described and / or utilized herein are administered postoperatively to a target site of a tumor resection subject within 24 hours or less (including, for example, within 18 hours, within 12 hours, within 6 hours, within 3 hours, within 2 hours, within 1 hour, within 30 minutes, or less) after the tumor is removed by surgical tumor resection. In some embodiments, the compositions described and / or utilized herein are administered one or more times postoperatively to one or more target sites at one or more time points within 12 months or less of the surgical intervention (including, for example, within 11 months, within 10 months, within 9 months, within 8 months, within 7 months, within 6 months, within 5 months, within 4 months, within 3 months, within 2 months, or within 1 month of the surgical intervention). In some embodiments, the compositions described and / or utilized herein are administered one or more times post-operatively to one or more target sites at one or more time points within 31 days (including, for example, within 30 days, within 29 days, within 28 days, within 27 days, within 26 days, within 25 days, within 24 days, within 23 days, within 22 days, within 21 days, within 20 days, within 19 days, within 18 days, within 17 days, within 16 days, within 15 days, within 14 days, within 13 days, within 12 days, within 11 days, within 10 days, within 9 days, within 8 days, within 7 days, within 6 days, within 5 days, within 4 days, within 3 days, within 2 days, or within 1 day) of the surgical intervention).

[0290] In some embodiments, the target site of administration is or includes a tumor resection site. In some embodiments, such tumor resection sites may be characterized by a lack of total residual tumor antigen. In some embodiments, such tumor resection sites may be characterized by negative resection margins (i.e., no cancer cells are visible microscopically at the resection margins, e.g., based on histological evaluation of tissue surrounding the tumor resection site). In some embodiments, such tumor resection sites may be characterized by positive resection margins (i.e., cancer cells are visible microscopically at the resection margins, e.g., based on histological evaluation of tissue surrounding the tumor resection site). In some embodiments, such tumor resection sites may be characterized by the presence of total residual tumor antigen. In some embodiments, the target site of administration is or includes a site proximate to the tumor resection site (e.g., within 4 inches, within 3.5 inches, within 3 inches, within 2.5 inches, within 2 inches, within 1.5 inches, within 1 inch, within 0.5 inches, within 0.4 inches, within 0.3 inches, within 0.2 inches, within 0.1 inches, or less; e.g., within 10 centimeters, within 9 centimeters, within 8 centimeters, within 7 centimeters, within 6 centimeters, within 5 centimeters, within 4 centimeters, within 3 centimeters, within 2 centimeters, within 1 centimeter, within 0.5 centimeters, or less). In some embodiments, the target site of administration is or includes a sentinel lymph node. In some embodiments, the target site of administration is or includes a draining lymph node.

[0291] As will be appreciated by those skilled in the art, compositions useful according to the present disclosure may be administered to a target site of a subject in need of administration using suitable delivery techniques known in the art. For example, in some embodiments, the provided techniques may be applicable for administration by injection. In some embodiments, the provided techniques may be suitable for administration, for example, by minimally invasive surgery (MIS), which typically involves one or more small incisions, such as robotic-assisted MIS, robotic surgery, and / or laparoscopic surgery. In some embodiments, the provided techniques may be suitable for administration in the context of accessible excision and / or skin resection. In some embodiments, the provided techniques may be suitable for administration (e.g., by injection) during surgery as part of a minimally invasive procedure, such as minimally invasive surgery (MIS), such as robotic-assisted MIS, robotic surgery, and / or laparoscopic surgery, and / or a procedure involving one or more accessible excision and / or skin resection. In some embodiments, the provided techniques may be suitable for administration (e.g., by injection), which, for example, in some embodiments, requires a robotic surgery system (e.g., da Vinci System) for minimally invasive administration. For example, in some embodiments, compositions that may be useful for injection and / or in the context of minimally invasive procedures, e.g., minimally invasive surgery (MIS), e.g., robotic-assisted MIS, robotic surgery, and / or laparoscopic surgery, and / or procedures involving one or more accessible resections and / or skin resections, are liquids, and the polymer combination preparations provided in such compositions transition from a liquid solution state to a polymer network state (e.g., hydrogel) upon injection into a target site (e.g., tumor resection site) in a subject, and in some embodiments, such transition is caused by exposure to the subject's body temperature (e.g., a viscous polymer solution) of the polymer combination preparation. In some embodiments, the polymer combination preparations in preformed polymer network biomaterials that are compressible without adversely affecting structural integrity may be injected, for example, via minimally invasive procedures, e.g., minimally invasive surgery (MIS), e.g., robotic-assisted MIS, robotic surgery, and / or laparoscopic surgery and / or laparoscopic procedures.

[0292] In some embodiments, the techniques provided herein may be applicable to administration by implantation. For example, in some embodiments, the polymer combination preparation provided in the composition according to the present disclosure is a preformed polymer network biomaterial. An exemplary polymer network biomaterial is or includes a hydrogel. For example, in some embodiments, the compositions provided may be administered to a tumor resection site (e.g., a cavity volume created by tumor resection) by surgical implantation. In some embodiments, the compositions provided may be administered to a tumor resection site by surgical implantation and may be secured by a bioadhesive. In some embodiments, administration may be performed intraoperatively (i.e., immediately after tumor resection).

[0293] In some embodiments, the amount of the polymer combination preparation and / or therapeutic agent incorporated therein to achieve a desired therapeutic effect(s), such as, for example, anti-tumor immunity, may vary from subject to subject depending, for example, on the subject's sex, age, and general condition, the type and / or severity of the cancer, the efficacy of the polymer biomaterial agonist of innate immunity, etc.

[0294] In some embodiments, the present disclosure provides techniques where administration of a composition comprising a polymer combination preparation (e.g., as described herein) is sufficient to provide anti-tumor immunity and thus does not necessarily require administration of a tumor antigen and / or adoptive transfer of immune cells (e.g., T cells) to a subject in need thereof (e.g., as described herein). Thus, in some embodiments, the techniques provided herein do not include administering a tumor antigen to the subject within, for example, one month or less (including, for example, within 3 weeks, within 2 weeks, within 1 week, within 5 days, within 3 days, within 1 day, within 12 hours, within 6 hours) after the subject is administered a composition described and / or utilized herein. In certain embodiments, the techniques provided herein do not include adoptive transfer of immune cells (e.g., T cells) into a subject, e.g., within one month or less (including, e.g., within three weeks, within two weeks, within one week, within five days, within three days, within one day, within 12 hours, within 6 hours) after the subject has been administered a composition described and / or utilized herein.

[0295] In certain embodiments, the present disclosure provides techniques such that administration of polymer combination preparations is particularly effective, for example, when administered as a combination therapy with tumor antigens and / or adoptive transfer of immune cells (e.g., T cells, NK cells, etc.). In certain embodiments, the techniques provided herein include adoptive transfer of immune cells (e.g., T cells, NK cells, etc.) to a subject, for example, within one month or less (including, for example, within three weeks, within two weeks, within one week, within five days, within three days, within one day, within twelve hours, within six hours) after the subject has been administered a composition described and / or utilized herein.

[0296] In some embodiments, the techniques provided herein are useful for treating cancer in a subject. In some embodiments, the techniques provided herein are used to treat resectable tumors. In some embodiments, the techniques provided herein are used to treat solid tumors (such as, but not limited to, blastomas, carcinomas, germ cell tumors, and / or sarcomas). In some embodiments, the techniques provided herein are used to treat lymphomas present in tissues other than the spleen or lymphatic system, such as the thyroid or stomach.

[0297] In some embodiments, the technology provided herein is directed to treating cancers including, but not limited to, acoustic neuroma; adenocarcinoma; adrenal carcinoma; anal carcinoma; angiosarcoma (e.g., lymphangiosarcoma, lymphangioendothelial sarcoma, angiosarcoma); appendix cancer; benign monoclonal gammopathy; biliary tract cancer (e.g., cholangiocarcinoma); bile duct carcinoma; bladder cancer; bone cancer; breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, breast cancer, medullary carcinoma of the breast); brain cancer (e.g., meningioma, glioblastoma, glioma (e.g., astrocytoma, oligodendroglioma), medulloblastoma); bronchial carcinoma; carcinoid tumor; cardiac tumor; cervical cancer (e.g., cervical adenocarcinoma); choriocarcinoma; notochord cancer. tumors;craniopharyngioma;colorectal cancer (e.g., colon carcinoma, rectal carcinoma, colorectal adenocarcinoma);connective tissue cancer;epithelial carcinoma;ductal intraepithelial carcinoma;ependymoma;endothelial sarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma);endometrial cancer (e.g., uterine carcinoma, uterine sarcoma);esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma);Ewing's sarcoma;eye cancer (e.g., intraocular melanoma, retinoblastoma);familial hypereosinophilia;gallbladder cancer;gastric cancer (e.g., gastric adenocarcinoma);gastrointestinal stromal tumor (GIST);germ cell carcinoma;head and neck cancer (e.g., head and neck squamous cell carcinoma), oral cancer (e.g., oral squamous cell carcinoma), laryngeal cancer (e.g., pharyngeal ( laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer); hematopoietic cancer (e.g., lymphoma, primary pulmonary lymphoma, bronchus-associated lymphoid tissue lymphoma, splenic lymphoma, nodal marginal zone lymphoma, childhood B-cell non-Hodgkin's lymphoma); hemangioblastoma; histiocytosis; hypopharyngeal carcinoma; inflammatory myofibroblastic tumor; immune cell amyloidosis; kidney cancer (e.g., nephroblastoma (also known as Wilms' tumor), renal cell carcinoma); liver cancer (e.g., hepatocellular carcinoma (HCC), malignant hepatoma); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), pulmonary adenocarcinoma);leiomyosarcoma (LMS);melanoma;midline carcinoma;multiple endocrine neoplasia syndrome;muscle carcinoma;mesothelioma;nasopharyngeal carcinoma;neuroblastoma;neurofibroma (e.g., type 1 or type 2 neurofibromatosis (NF), schwannoma);neuroendocrine carcinoma (e.g., gastrointestinal pancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor);osteosarcoma (e.g., bone cancer);ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma);papillary adenocarcinoma;pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumor);parathyroid carcinoma;papillary adenocarcinoma;penile cancer (e.g., Paget's disease of the penis and scrotum);Pharyngeal cancer;Pinealoma;Pituitary carcinoma;Pleuropulmonary blastoma;Primitive neuroectodermal tumor (PNT);Plasma cell neoplasms;Paraneoplastic syndromes;Intraepithelial neoplasia;Prostate cancer (e.g., prostatic adenocarcinoma);Rectal cancer;Rhabdomyosarcoma;Retinoblastoma;Salivary gland cancer;Skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC));Small intestinal cancer (e.g., appendix cancer);Soft tissue sarcomas (e.g., malignant fibrous histiocytoma (MFH), lipoma) The compounds are useful for treating cancers including liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma; sebaceous gland carcinoma; gastric cancer; small intestine cancer; sweat gland carcinoma; synovium; testicular cancer (e.g., seminoma, testicular embryonal carcinoma); thymic carcinoma; thyroid cancer (e.g., papillary thyroid carcinoma, papillary thyroid carcinoma (PTC), medullary thyroid carcinoma); urethral cancer; uterine cancer; vaginal cancer; vulvar cancer (e.g., Paget's disease of the vulva), or any combination thereof;

[0298] In certain embodiments, the cancer is breast cancer. In certain embodiments, the cancer is skin cancer. In certain embodiments, the cancer is melanoma. In certain embodiments, the cancer is lung cancer. In certain embodiments, the cancer is kidney cancer. In certain embodiments, the cancer is liver cancer. In certain embodiments, the cancer is pancreatic cancer. In certain embodiments, the cancer is colorectal cancer. In certain embodiments, the cancer is bladder cancer. In certain embodiments, the cancer is lymphoma. In certain embodiments, the cancer is prostate cancer. In certain embodiments, the cancer is thyroid cancer. In certain embodiments, the cancer is brain cancer. In certain embodiments, the cancer is gastric cancer. In certain embodiments, the cancer is esophageal cancer.

[0299] In some embodiments, the technology provided herein is directed to the treatment of adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma, appendix cancer, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, bronchial cancer, carcinoid tumor, cardiac tumor, cervical cancer, choriocarcinoma, chordoma, colorectal cancer, connective tissue cancer, craniopharyngioma, ductal carcinoma in situ, endothelial sarcoma, endometrial cancer, ependymoma, epithelial cancer, esophageal cancer, Ewing's sarcoma, eye cancer, familial hypereosinophilia, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell cancer, head and neck cancer, hemangioblastoma, histiocytosis, Hodgkin's lymphoma, hypopharyngeal cancer, inflammatory myofibroblastic tumor, supraspinal tumor, sarcoma ... Intradermal neoplasms, immune cell amyloidosis, Kaposi's sarcoma, kidney cancer, liver cancer, lung cancer, leiomyosarcoma (LMS), melanoma, midline carcinoma, multiple endocrine neoplasia syndrome, muscle cancer, mesothelioma, myeloproliferative disorders (MPD), nasopharyngeal carcinoma, neuroblastoma, neurofibroma, neuroendocrine carcinoma, non-Hodgkin's lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer, paraneoplastic syndromes, parathyroid carcinoma, papillary adenocarcinoma, penile cancer, pharyngeal cancer, pheochromocytoma, pinealoma, pituitary cancer, pleuropulmonary blastoma, primitive neuroectodermal tumor (PNT), plasma cell neoplasms, prostate cancer, rectal cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sebaceous gland cancer, skin cancer, small intestine cancer bowel cancer, small intestine cancer, soft tissue sarcoma, gastric cancer, sweat gland cancer, synovial tumor, testicular cancer, thymic cancer, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vascular cancer, vulvar cancer, or a combination thereof.

[0300] In some embodiments, the methods provided herein may include administering a provided composition (e.g., as described herein) to a target site of a tumor resection in a subject, and optionally monitoring the tumor resection site or a distant site for risk or occurrence of tumor regrowth or tumor outgrowth in the subject after administration, for example, every 3 months or longer (including, for example, every 6 months, every 9 months, every year, or longer) after administration. If the subject is determined to have a risk or occurrence of tumor recurrence based on the monitoring report, in some embodiments, the subject may be administered a second composition (e.g., as described herein) and / or undergo a different treatment regimen (e.g., chemotherapy).

[0301] In some embodiments, the techniques provided herein may be useful for treating a subject suffering from metastatic cancer. For example, in some embodiments, the methods provided herein may include administering to a target site (e.g., as described herein) of a subject suffering from one or more metastases who has undergone tumor resection (e.g., surgical resection of a primary tumor), and optionally monitoring at least one metastatic site in the subject after administration, for example, every 3 months or longer (e.g., including every 6 months, every 9 months, every year, or longer) after administration. Based on the results of the monitoring report, in some embodiments, the subject may be administered a second composition (e.g., as described herein) and / or undergo a different treatment regimen (e.g., chemotherapy).

[0302] In certain embodiments, the methods described herein do not include administering the provided compositions before tumor resection. In certain embodiments, the methods described herein include administering the provided compositions before tumor resection. In certain embodiments, the techniques provided herein include administering the provided compositions to the tumor resection site at the same time as tumor resection. In certain embodiments, the techniques provided herein include administering the provided compositions to the tumor resection site after tumor resection.

[0303] It will also be understood that the compositions described herein may be administered in combination with one or more additional pharmaceutical agents and / or treatment regimens. For example, in some embodiments, the compositions described herein may be administered as part of a combination therapy. For example, the compositions may be administered in combination with additional pharmaceutical agents that reduce and / or modify their metabolism, inhibit their excretion, and / or modify their distribution within the body. In some embodiments, the compositions described herein are administered in combination with a systemic treatment, e.g., chemotherapy, radiation therapy, and / or immunomodulatory therapy. In some embodiments, the immunomodulatory therapy may include systemic and / or local administration of agents, e.g., small molecules, peptides, proteins, saccharides, steroids, antibodies, fusion proteins, nucleic acid agents (e.g., but not limited to, antisense polynucleotides, ribozymes, and small interfering RNA), peptidomimetics, and the like. For example, in some embodiments, the combination therapy may include a composition described herein and an immune checkpoint inhibitor therapy (e.g., by inhibition of the PD-1 / PD-L1 pathway). In some embodiments, the combination therapy may include a composition described herein and a chemotherapeutic agent. Suitable chemotherapeutic agents may be found in any of a variety of anti-cancer drug classes, including, but not limited to, alkylating agents, antimetabolites, topoisomerase inhibitors, and / or mitotic inhibitors. In some embodiments, the compositions described herein are administered before, during, and / or after at least one or more additional therapies as part of a combination therapy. It will also be understood that the additional therapies used may achieve a desired effect for the same disorder and / or may achieve a different effect. In certain embodiments, the additional pharmaceutical agent is not an adoptively transferred cell. In certain embodiments, the additional pharmaceutical agent is not a T cell. In certain embodiments, the additional pharmaceutical agent is administered multiple days or multiple weeks after administration of the compositions described herein.

[0304] In some embodiments, the polymer preparations provided herein can be useful for providing sustained release of a payload (eg, resiquimod) incorporated therein.

[0305] In certain embodiments, the technology provided herein may be useful for treating subjects suffering from a wide range of diseases for which localized drug release may be advantageous. In certain embodiments, the technology provided herein may be used in regenerative medicine. In certain embodiments, the technology provided herein may be used in tissue engineering. In certain embodiments, the technology provided herein may be used to assist in medical imaging (e.g., x-rays, CT scans, and / or radioisotope imaging). In certain embodiments, the technology provided herein may be used in dentistry (e.g., tooth restoration). In certain embodiments, the technology provided herein may be used in dermatology applications (e.g., injections to treat facial wrinkles and / or folds). In certain embodiments, the technology provided herein may be used in cosmetics and / or plastic surgery. In certain embodiments, the technology provided herein may be used in orthopedic applications (e.g., bone wound healing, osteoarthritis, spinal fusion, and / or intervertebral discs). In certain embodiments, the technology provided herein may be used to treat incontinence and other urological indications (e.g., urinary and / or anal). In certain embodiments, the technology provided herein may be used to treat heart failure. In certain embodiments, the technology provided herein may be used to treat hearing loss. In certain embodiments, the technology provided herein may be used for epidermal wounds and / or internal wound dressings. In certain embodiments, the technology provided herein may be used to prevent post-surgical adhesions. In certain embodiments, the technology provided herein may be used for cancer immunotherapy, including localized sustained delivery of immunomodulatory molecules. In certain embodiments, the technology provided herein may be used to treat autoimmune and / or rheumatic diseases (e.g., by localized and / or sustained delivery of immunomodulatory molecules).In certain embodiments, the technology provided herein may be used to treat fibrosis and / or scarring (e.g., by localized and / or sustained delivery of antifibrotic molecules for the prevention or healing of fibrosis and / or scarring). In certain embodiments, the technology provided herein may be used to treat infection (e.g., by localized and / or sustained delivery of anti-infective molecules, such as azithromycin, remdesivir, and / or any suitable antibiotic and / or antiviral agent known in the art, for the prevention and / or treatment of infection). In certain embodiments, the technology provided herein may be used to relieve pain (e.g., by localized and / or sustained delivery of analgesic molecules, such as ketorolac, bupivacaine, and / or any suitable analgesic agent known in the art, for the relief of pain).

[0306] In certain embodiments, the technology provided herein may be particularly useful for sustained release of molecules for the treatment of ocular pathology. In certain embodiments, the technology provided may be particularly suitable for intravitreal injection. In certain embodiments, the technology provided may be particularly suitable for local administration. In certain embodiments, the technology provided may be used to treat glaucoma and / or high intraocular pressure (e.g., by local and / or sustained release of beta (adrenergic) blockers, prostaglandin analogs, carbonic anhydrase inhibitors, parasympathetic analogs, alpha 2 adrenergic agonists, Rho kinase inhibitors, and / or docosanoids). In certain embodiments, the technology provided may be used to treat age-related macular degeneration (e.g., by local and / or sustained release of any anti-VEGF drug, VEGF inhibitor, anti-VEGFR drug, and / or VEGFR inhibitor known in the art). In certain embodiments, the provided technology may be used to treat symptomatic vitreomacular adhesions (e.g., by local and / or sustained release of ocriplasmin and / or any α2 antiplasmin degrading agent known in the art). In certain embodiments, the provided technology may be used to treat post-operative inflammation after any ophthalmic surgery (e.g., by local and / or sustained release of ketorolac, loteprednol, dexamethasone, corticosteroids, and / or any suitable anti-inflammatory agent known in the art). In certain embodiments, the provided technology may be used to deliver anesthetics for ophthalmic procedures (e.g., local and / or sustained delivery of lidocaine and / or any suitable anesthetic agent known in the art). In certain embodiments, the provided technology may be used to treat allergic conjunctivitis (e.g., by local and / or sustained delivery of histamine H1 receptor antagonists and / or dexamethasone) by either topical or intracanalicular administration. In certain embodiments, the provided technology may be used to treat bacterial conjunctivitis and / or corneal ulcers (e.g., localized and / or sustained delivery of fluoroquinolones and / or other suitable antimicrobial agents known in the art).In certain embodiments, the provided technology may be used to treat cystinosis (e.g., topical and / or sustained delivery of cysteamine hydrochloride and / or other suitable cysteine ​​removing agents and / or somatostatin inhibitors known in the art). In certain embodiments, the provided technology may be used to treat neurotrophic keratitis (e.g., topical and / or sustained delivery of nerve growth factor and / or other suitable anti-neurotrophic keratitis agents known in the art). In certain embodiments, the provided technology may be used to treat macular edema following branch retinal vein occlusion or central retinal vein occlusion (e.g., topical and / or sustained delivery of dexamethasone and / or other suitable corticosteroid agents known in the art). In certain embodiments, the provided technology may be used to treat dry eye (e.g., topical and / or sustained delivery of cyclosporine and / or other suitable immunomodulatory agents). In certain embodiments, the provided technology may be used to treat HSV-mediated keratitis (e.g., localized and / or sustained delivery of trifluridine and / or other suitable antiviral agents known in the art).

[0307] In certain embodiments, the subject being treated is a mammal. In certain embodiments, the subject is a human. In certain embodiments, the subject is a human subject undergoing tumor resection. In certain embodiments, the subject is a human subject not suitable for tumor resection surgery. In certain embodiments, the subject is a human patient undergoing (pre-operative) neoadjuvant therapy. In certain embodiments, the subject is a human patient not undergoing neoadjuvant therapy. In certain embodiments, the subject is a human patient undergoing (pre-operative) neoadjuvant chemotherapy. In certain embodiments, the subject is a human patient not undergoing (pre-operative) neoadjuvant chemotherapy. In certain embodiments, the subject is a human patient undergoing neoadjuvant radiation therapy. In certain embodiments, the subject is a human patient not undergoing neoadjuvant radiation therapy. In certain embodiments, the subject is a human patient undergoing neoadjuvant chemotherapy and radiation therapy. In certain embodiments, the subject is a human patient undergoing neoadjuvant molecular targeted therapy. In certain embodiments, the subject is a human patient not undergoing neoadjuvant molecular targeted therapy. In certain embodiments, the subject is a human patient who has not received neoadjuvant chemotherapy. In some embodiments, the subject is receiving, has received, or will receive immune checkpoint blockade therapy. In certain embodiments, the subject is receiving immune checkpoint blockade therapy. In certain embodiments, the subject is a human patient who has received and / or is receiving molecular targeted therapy (e.g., a therapy such as those described as neoadjuvant and / or adjuvant therapy) as the only therapeutic intervention (e.g., a subject for whom surgical resection is not a viable option). In some embodiments, the subject is receiving, has received, or will receive certain other cancer therapies (e.g., including but not limited to, costimulation, oncolytic viruses, CAR T cells, transgenic TCRs, TILs, vaccines, BiTEs, ADCs, cytokines, modulators of natural immunity, or any combination thereof).In certain embodiments, the subject is a human patient who has received neoadjuvant immunotherapy including immune checkpoint blockade (e.g., anti-CTLA-4, anti-PD-1, and / or anti-PD-L1). In certain embodiments, the subject is a human patient who has not received and / or will not receive neoadjuvant immunotherapy including immune checkpoint blockade (e.g., anti-CTLA-4, anti-PD-1, and / or anti-PD-L1). In certain embodiments, the subject is a human patient whose tumor has not responded objectively to neoadjuvant therapy (as defined by Response Evaluation Criteria in Solid Tumors (RECIST) or Immune Related Response Criteria (irRC)) and / or is objectively unresponsive (e.g., stable disease, progression). In certain embodiments, the subject is a human patient whose target lesions have responded objectively to neoadjuvant therapy and / or are objectively responding (e.g., partial response, complete response). Non-target lesions may show incomplete response, stable disease, or progression. In certain embodiments, the subject is a human patient suitable for receiving immunotherapy as a (postoperative) adjuvant therapy. In certain embodiments, the subject is a domesticated animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a companion animal, such as a dog or cat. In certain embodiments, the subject is a livestock animal, such as a cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a zoo animal. In another embodiment, the subject is a research animal, such as a rodent, pig, dog, or non-human primate. In certain embodiments, the subject is a non-human transgenic animal, such as a transgenic mouse or a transgenic pig.

[0308] kit The present disclosure also provides kits that are useful for carrying out the techniques provided herein. In some embodiments, the kits include the compositions or pharmaceutical compositions described herein and containers (e.g., vials, ampoules, bottles, syringes, and / or dispenser containers, or other suitable containers). In some embodiments, the kits include delivery technologies, such as syringes, bags, etc., or components thereof, that can be provided as single-use and / or multi-use items. In some embodiments, one or more component(s) of the compositions or pharmaceutical compositions described herein are provided separately in one or more containers. For example, the individual components of the polymer combination preparation (e.g., those described herein, such as, but not limited to, poloxamer and a second polymer, such as hyaluronic acid and / or chitosan or variants thereof) can be provided in separate containers in some embodiments. In some such embodiments, the individual components of the biomaterial (e.g., as described herein, including but not limited to, a poloxamer and a second polymer, such as hyaluronic acid and / or chitosan or variants thereof) may each be provided independently as a dry lyophilized powder, dry particles, or liquid. In some embodiments, the individual components of the polymer combination preparation (e.g., as described herein, including but not limited to, a poloxamer and a second polymer, such as hyaluronic acid and / or chitosan or variants thereof) may be provided as a single mixture in a container. In some such embodiments, the single mixture may be provided as a dry lyophilized powder, dry particles, or liquid (e.g., a homogenous liquid).

[0309] In some embodiments, the polymer combination preparation (e.g., those described herein) may be provided as a preformed polymer network biomaterial in a container. In some embodiments, such a preformed polymer network biomaterial (e.g., a hydrogel) may be provided in a dry state. In some embodiments, a preformed polymer network biomaterial (in the form of a viscous polymer solution) may be provided in a container.

[0310] In some embodiments, the provided kits may optionally include a container containing a pharmaceutical excipient for diluting or suspending the compositions or pharmaceutical compositions described herein. In some embodiments, the provided kits may include a container containing an aqueous solution. In some embodiments, the provided kits may include a container containing a buffer solution.

[0311] In some embodiments, the kit provided may include resiquimod (e.g., one or more solid forms described herein). For example, in some embodiments, resiquimod may be provided in a separate container so that it can be added to a liquid mixture of a polymer combination preparation (e.g., as described herein) prior to administration to a subject. In some embodiments, resiquimod may be incorporated into a polymer combination preparation described herein.

[0312] In certain embodiments, the kits described herein further comprise instructions for carrying out the methods described herein. The kits described herein may also include information from regulatory agencies, such as the U.S. Food and Drug Administration (FDA), if necessary. In certain embodiments, the information included in the kits provided herein is, for example, prescription information for the treatment of cancer. Instructions may be present in the kit in various forms, one or more of which may be present in the kit. One form in which these instructions may be present is as information printed on a suitable medium or substrate, such as a paper or multiple papers on which the kit packaging, insert, and other information is printed. Yet another means may be a computer readable medium on which the instruction information is recorded, such as a diskette, CD, USB drive, and the like. Yet another means that may be present is a website address that can be used via the Internet to access the instruction information. Any convenient means may be present in the kit.

[0313] Other features of the present invention will become apparent through the following description of exemplary embodiments, which are provided for the purpose of illustrating the present invention and are not intended to limit the present invention. EXAMPLES

[0314] The following examples are not intended to limit the scope of any claims. The following non-limiting examples are presented to further illustrate the teachings of the present invention. Those skilled in the art will recognize that in light of this specification, many changes can be made to the specific embodiments shown herein and still obtain similar or equivalent results without departing from the spirit and scope of the teachings of the present invention.

[0315] Example 1. Preparation and characterization of solid forms of resiquimod Materials and Methods X-ray powder diffraction (XRPD): XRPD patterns were collected on a PANalytical DYY884-AERIS-300 diffractometer using the following parameters: [Table 20]

[0316] Differential Scanning Calorimetry (DSC): DSC thermograms were collected on a TA Instruments DSC250 instrument using the following parameters: [Table 21]

[0317] Thermogravimetric Analysis (TGA): TGA thermograms were collected on a TA Instruments TGA550 instrument using the following parameters: [Table 22]

[0318] Resiquimod morphology I A representative procedure for preparing Form I of resiquimod is given below. Resiquimod (10 mg) obtained by anti-solvent recrystallization with DCM and heptane (described below to obtain Form I) was added to a clear glass vial with a screw thermoset PTFE liner cap and diethyl ether (5 mL) was added. The mixture was vortexed for 1 min and kept in a ThermoMixer (Eppendorf) for heating and cooling temperature cycling (50-5 °C). Heating was fixed at 50 °C for 6 h and cooling at 5 °C for 6 h in the ThermoMixer. The heating rate was maintained at 3 °C / min and the cooling rate at 1 °C / min. After completion of five heating-cooling cycles at 1000 RPM, the suspension was centrifuged and the residue was dried under vacuum (-700 mmHg) at room temperature.

[0319] The XRPD pattern of resiquimod Form I is shown in FIG. 1 and the corresponding data are summarized below. [Table 23]

[0320] As shown in the DSC curve in Figure 2, the sample exhibited an endotherm with an onset temperature of 193.6°C and a peak temperature of 195.1°C. Figure 3 shows the TGA curve of the sample, which showed a weight loss of 1.985% up to 200°C.

[0321] Form I of resiquimod was also prepared by adding resiquimod (10 mg) obtained by anti-solvent recrystallization with DCM and heptane (described below to obtain Form I) to a clear glass vial followed by the addition of acetone (0.5 mL). The mixture was vortexed for 1 min and kept in a ThermoMixer (Eppendorf) for heating and cooling temperature cycling (50-5°C). Heating was fixed at 50°C for 6 h and cooling at 5°C for 6 h in the ThermoMixer. The heating rate was maintained at 3°C / min and the cooling rate at 1°C / min. After completion of five heating / cooling cycles at 1000 RPM, the suspension was centrifuged and the residue was dried under vacuum (-700 mmHg) at room temperature.

[0322] Form I of resiquimod was also prepared according to the following procedure. A mixture of 2-(ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]quinoline 5-oxide (34.0 kg) in DCM was added to the reactor and the mixture was cooled to 0-10 °C. Trichloroacetyl isocyanate (1.00 kg) was added slowly to the reaction mass under argon atmosphere while maintaining the temperature at 0-10 °C. The reaction mixture was stirred at 0-10 °C for 20-30 minutes and then warmed to 25-35 °C and then to 40-45 °C. The reaction mixture was stirred at 40-45 °C for 1-2 hours. After that, about 3-4 volumes of solvent were removed under vacuum. Methanol (2.4 L) was added and then 3-4 volumes of solvent were distilled off under vacuum. Methanol (2.4 L) was added again and then 3-4 volumes of solvent were distilled off under vacuum. Methanol (16.0 L) was then added followed by a solution of sodium methoxide in methanol (5.2 kg). The reaction mixture was then heated to 50-55°C for 1-2 hours. Approximately 3-4 volumes of solvent were then distilled off under vacuum and the mixture was cooled to 35°C. DCM (4.0 L) was added followed by 3-4 volumes of solvent distilled off under vacuum and this process was repeated twice. DCM (16 L) and water (16 L) were added, the mixture was stirred and the organic layer was collected. The aqueous layer was extracted twice more with DCM (16 L). The combined organic layers were dried over sodium sulfate and filtered. Approximately 3-4 volumes of the filtrate were removed under vacuum and ethyl acetate (2.4 L) was added. This was repeated twice. The mixture was then warmed to 50-55°C before being cooled to 25-35°C and stirred for 1-2 hours. A solid formed which was collected by filtration and dried under vacuum to obtain crude resiquimod. Crude resiquimod (375 g) was added to dichloromethane (37.5 L) at 25-35°C. The mixture was refluxed for 1-2 h, then cooled to 25-35°C and filtered. The filtrate was concentrated to a volume of about 15 L and then heated to 35-40°C. Heptane (15 L) was then added slowly to the mixture at 35-40°C. The mixture was stirred at 35-40°C for 1-2 h, then cooled to 25-35°C and stirred for 1-2 h. The resulting solid was collected by filtration and dried under reduced pressure at 50-55°C to obtain resiquimod form I.

[0323] Morphology of resiquimod II A representative procedure for preparing resiquimod form II is given below. Resiquimod (10 mg) was added to a clear glass vial and methyl isopropyl ketone (0.5 mL) was added. The mixture was vortexed for 1 min and kept in a ThermoMixer (Eppendorf) for heating and cooling temperature cycling (50-5 °C). Heating was fixed at 50 °C for 6 h and cooling at 5 °C for 6 h in the ThermoMixer. The heating rate was maintained at 3 °C / min and the cooling rate at 1 °C / min. After completion of five heating / cooling cycles at 1000 RPM, the suspension was centrifuged and the residue was dried under vacuum (-700 mmHg) at room temperature.

[0324] The XRPD pattern of resiquimod Form II is shown in FIG. 4 and the corresponding data are summarized below. [Table 24]

[0325] As shown in the DSC curve in Figure 5, the sample exhibited a first endotherm with an onset temperature of 140.7°C and a peak temperature of 145.4°C, and a second endotherm with an onset temperature of 156.7°C and a peak temperature of 159.7°C. Figure 6 shows the TGA curve of the sample, which exhibited a weight loss of 15.941% up to 170°C.

[0326] Resiquimod morphology III A representative procedure for preparing resiquimod form III is shown below. Resiquimod (10 mg) was added to a clear glass vial and 1,4-dioxane (0.5 mL) was added. The mixture was vortexed for 1 min and stirred at 50° C. for 7 days. After 7 days, the suspension was centrifuged and the residue was dried under vacuum (−700 mmHg) at room temperature.

[0327] The XRPD pattern of resiquimod Form III is shown in FIG. 7 (top spectrum) and the corresponding data are summarized below. [Table 25]

[0328] Upon drying, resiquimod Form III was converted to resiquimod Form I. The dried sample exhibited the DSC curve shown in Figure 8, which showed an endotherm with an onset temperature of 193.6°C and a peak temperature of 195.3°C, consistent with conversion to Form I. Figure 9 shows the TGA curve of the dried sample, which showed virtually no weight loss below 200°C, also consistent with conversion to Form I.

[0329] Resiquimod morphology IV A representative procedure for preparing resiquimod form IV is shown below. Resiquimod (15 mg) was added to a clear glass vial and tetrahydrofuran (0.5-1 mL) was added. The mixture was stirred at 40° C. for 4 hours. The mixture was then filtered through a 0.45 μm PVDF filter and the filtrate was kept at 2-8° C. overnight and then at −20° C. for 1 day. The solution was allowed to evaporate at ambient conditions for 1 day and then evaporated under vacuum at room temperature for 2 days before the solid was collected.

[0330] The XRPD pattern of resiquimod form IV is shown in FIG. 10 and the corresponding data are summarized below. [Table 26]

[0331] As shown in the DSC curve in Figure 11, the sample exhibited an endotherm with an onset temperature of 193.1° C. and a peak temperature of 194.7° C. Figure 12 shows the TGA curve of the sample, which showed a weight loss of 6.965% up to 200° C.

[0332] Resiquimod morphology V A representative procedure for preparing resiquimod form V is shown below. Resiquimod (15 mg) was added to a clear glass vial and methyl ethyl ketone (0.5-1 mL) was added. The mixture was stirred at 40°C for 4 hours. The mixture was then filtered through a 0.45 μm PVDF filter and the filtrate was kept at 2-8°C overnight and then at -20°C for 1 day. The crystallized sample was isolated and dried prior to XRPD.

[0333] The XRPD pattern of resiquimod form V is shown in FIG. 13 (top spectrum) and the corresponding data are summarized below. [Table 27]

[0334] It was observed that samples of resiquimod Form V converted to Form I upon drying and / or grinding. See Figure 13 (bottom three spectra).

[0335] As shown in the DSC curve in Figure 14, the sample exhibited a first endotherm with an onset temperature of 55.2°C and a peak temperature of 62.2°C, and a second endotherm with an onset temperature of 194.5°C and a peak temperature of 195.4°C. Figure 15 shows the TGA curve of the sample, which exhibited a weight loss of 3.836% up to 110°C.

[0336] Morphology of resiquimod VI A representative procedure for preparing resiquimod Form VI is shown below. Resiquimod (15 mg) was added to a clear glass vial and anisole (0.5-1 mL) was added. The mixture was stirred at 40°C for 4 hours. The mixture was then filtered through a 0.45 μm PVDF filter and the filtrate was kept at 2-8°C overnight and then at -20°C for 1 day. The crystallized sample was isolated and dried prior to XRPD.

[0337] The XRPD pattern of resiquimod Form VI is shown in FIG. 16 and the corresponding data are summarized below. [Table 28]

[0338] As shown in the DSC curve in Figure 17, the sample exhibited a first endotherm with an onset temperature of 72.7° C. and a peak temperature of 78.2° C., and a second endotherm with an onset temperature of 193.7° C. and a peak temperature of 195.1° C. Figure 18 shows the TGA curve of the sample, which exhibited a weight loss of 7.508% up to 110° C.

[0339] Morphology of resiquimod VII A representative procedure for preparing resiquimod form VII is shown below. Resiquimod (10 mg) was added to a clear glass vial and dissolved in a minimal amount (0.25-1 mL) of 2-methyltetrahydrofuran with stirring and occasional sonication. The solution was filtered through a 0.45 μm filter and methyl t-butyl ether (3-5 mL) was added slowly under stirring. The mixture was stirred at 2-8 °C for 16 h. The suspension was centrifuged and the residue was dried under vacuum (-700 mmHg) at room temperature.

[0340] The XRPD pattern of resiquimod Form VI is shown in FIG. 19 and the corresponding data are summarized below. [Table 29]

[0341] As shown in the DSC curve in Figure 20, the sample exhibited a first endotherm with an onset temperature of 95.0° C. and a peak temperature of 108.9° C., and a second endotherm with an onset temperature of 160.9° C. and a peak temperature of 171.8° C. Figure 21 shows the TGA curve of the sample.

[0342] Example 2. Screening for polymorphs of resiquimod Screening for polymorphs by heating-cooling temperature cycling Resiquimod form I (10 mg) was weighed into a clear glass vial and 0.5 mL of solvent was added. The mixture was vortexed for 1 min and kept in a ThermoMixer (Eppendorf) for heating and cooling temperature cycling (50-5 °C). Heating was fixed at 50 °C for 6 h and cooling at 5 °C for 6 h in the ThermoMixer. The heating rate was maintained at 3 °C / min and the cooling rate at 1 °C / min. After completion of five heating / cooling cycles at 1000 RPM, the suspension was centrifuged and the residue was dried under vacuum (-700 mmHg) at room temperature. The solution was evaporated under vacuum (-700 mmHg) at room temperature. All solid samples were analyzed by XRPD for the assessment of new polymorphic forms. The results are summarized in Table 1. [Table 1]

[0343] Slurry polymorph screening Resiquimod form I (10 mg) was weighed into a clear glass vial and 0.5 mL of solvent was added. The mixture was vortexed for 1 min and placed under stirring at 50° C. After 7 days the suspension was centrifuged and the residue was dried under vacuum (−700 mmHg) at room temperature. The solution was evaporated under vacuum (−700 mmHg) at room temperature. The results are summarized in Table 2. [Table 2]

[0344] Screening for polymorphs by cooling crystallization Resiquimod form I (15 mg) was weighed into a clear glass vial and 0.5-1 mL of solvent was added. The mixture was stirred at 40 °C for 4 h. The mixture was then filtered through a 0.45 μm PVDF filter and the filtrate was kept at 2-8 °C overnight and then at -20 °C for 1 day. The crystallized sample was isolated and dried prior to XRPD. The solution was allowed to evaporate at ambient conditions for 1 day and then vacuum evaporated at room temperature for 2 days. The results are summarized in Table 3. [Table 3]

[0345] Screening for polymorphs by antisolvent crystallization Resiquimod form I (10 mg) was weighed into a clear glass vial and dissolved in a minimum amount of solvent (0.25-1 mL) by stirring and occasional sonication. These solutions were filtered through a 0.45 μm filter and 3-5 mL of antisolvent was added slowly under stirring. The mixtures were left under stirring at 2-8°C for 16 h. The suspensions were centrifuged and the residues were dried under vacuum (-700 mmHg) at room temperature. The solutions were evaporated under vacuum (-700 mmHg) at room temperature for 2 days. The results are summarized in Table 4. [Table 4]

[0346] Overview of resiquimod solid forms Seven solid forms of resiquimod were identified from the polymorph screening described herein, and are summarized in Tables 5 and 6. [Table 5] [Table 6]

[0347] Example 3. Visual solubility assessment of resiquimod form I The solubility was assessed by adding solvent to a sample of resiquimod Form I (10 mg) in two steps (0.25 mL per step) and stirring at room temperature, and the results are summarized in Table 7. [Table 7-1] [Table 7-2]

[0348] Example 4. Exemplary Materials and Methods of Preparation and Characterization of Exemplary Polymer Combination Preparations Described herein and Reference Polymer Biomaterials This example relates to the preparation and characterization of the exemplary polymer combinations described herein.In some embodiments, a generality can be observed that as the concentration of one biomaterial (e.g., poloxamer) increases, the concentration of at least one additional biomaterial (e.g., hyaluronic acid and / or chitosan / modified chitosan) required to create a suitable polymer network tends to decrease.In some embodiments, this generality applies in the opposite direction (e.g., a suitable polymer network formed using a lower concentration of poloxamer may use a higher concentration of at least one additional biomaterial).

[0349] An exemplary polymer combination formulation including poloxamer and hyaluronic acid is shown below:

[0350] A preparation containing 13.5% (w / w) poloxamer 407 and 0.65% (w / w) 1.5 MDa hyaluronic acid in 0.1 M NaHCO3, 0.9% saline (pH 8.1) or 25 mM phosphate buffer (pH 7.4 or pH 8).

[0351] Preparations containing 10-12.5% ​​(w / w) poloxamer 407 and 0.65-1% (w / w) 1.5 MDa hyaluronic acid in 0.1 M NaHCO3, 0.9% saline (pH 8.1) or 25 mM phosphate buffer (pH 7.4 or pH 8).

[0352] A preparation comprising 9-10% (w / w) poloxamer 407 and 1-1.2% (e.g., 1.1%) (w / w) 1.5 MDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0353] A preparation containing 8-9% (w / w) poloxamer 407 and 1.65-1.75% (w / w) 1.32 MDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0354] A preparation containing 10% (w / w) poloxamer 407 and 1-1.5% (e.g., 1.3%) (w / w) 773 kDa hyaluronic acid in 10 mM PBS (pH 7.4) or 25 mM phosphate buffer (pH 7.4 or pH 8).

[0355] A preparation containing 9-10% (w / w) poloxamer 407 and 1.2-2.5% (w / w) 730 kDa hyaluronic acid in 10 mM PBS (pH 7.4) or 25 mM phosphate buffer (pH 7.4 or pH 8).

[0356] A preparation containing 9-10% (w / w) poloxamer 407 and 1.2-2.5% (w / w) 730 kDa hyaluronic acid in 10 mM PBS (pH 8) or 25 mM phosphate buffer (pH 8).

[0357] Preparations containing 9-11.5% (w / w) poloxamer 407 and 2-2.75% (w / w) 730 kDa hyaluronic acid in 10 mM PBS (pH 7.4) or 25 mM phosphate buffer (pH 7.4 or pH 8).

[0358] A preparation containing 12.3% (w / w) poloxamer 407 and 1.625% (w / w) 730 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0359] A preparation containing 8% (w / w) poloxamer 407 and 1.75% to 2.25% (w / w) 337 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0360] A preparation containing 10% (w / w) poloxamer 407 and 2-6% (w / w) 309 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0361] A preparation containing 10% (w / w) poloxamer 407 and 2-6% (w / w) hyaluronic acid of 264-310 kDa in 22.5 mM phosphate buffer (pH 7.4 or pH 8).

[0362] A preparation containing 8-12.5% ​​(w / w) poloxamer 407 and 1-4% (w / w) hyaluronic acid of 264-310 kDa in 22.5 mM phosphate buffer (pH 7.4 or pH 8).

[0363] A preparation containing 8-12.5% ​​(w / w) poloxamer 407 and 1-4% (w / w) 119 or 120 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0364] A preparation containing 10% (w / w) poloxamer 407 and 2-6% (w / w) 119 or 120 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0365] A preparation containing 8-12.5% ​​(w / w) poloxamer 407 and 1-4% (w / w) 187 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0366] A preparation containing 10% (w / w) poloxamer 407 and 2-6% (w / w) 187 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0367] A preparation containing 8-10% (w / w) poloxamer 338 and 1-1.5% (w / w) 1.32 MDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0368] A preparation containing 8-10% (w / w) poloxamer 338 and 1.4-2% (w / w) 730 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0369] A preparation containing 8-10% (w / w) poloxamer 338 and 1.75-2.5% (w / w) 119 kDa hyaluronic acid in 25 mM phosphate buffer (pH 7.4 or pH 8).

[0370] Exemplary polymer combination formulations including poloxamer and chitosan or modified chitosan are shown below.

[0371] Preparations containing 13.5% (w / w) poloxamer 407 and 0.65–1.3% (w / w) carboxymethyl chitosan in 10 mM PBS, 33 mM NaHCO3, 0.45% saline (pH 8.1) or 25 mM phosphate buffer (pH 7.4).

[0372] Preparations containing 8-12.5% ​​(w / w) poloxamer 407 and 2.5-5% (w / w) carboxymethyl chitosan in 10 mM PBS, 33 mM NaHCO3, 0.45% saline (pH 8.1) or 25 mM phosphate buffer (pH 7.4).

[0373] An exemplary polymer combination formulation including poloxamer, hyaluronic acid, and chitosan or modified chitosan is shown below.

[0374] A preparation containing 8-12.5% ​​(w / w) poloxamer 407, 2-6% (w / w) 119 kDa hyaluronic acid, and 0.2-5% (w / w) carboxymethyl chitosan in 25 mM phosphate buffer (pH 7.4).

[0375] A preparation containing 8-12.5% ​​(w / w) poloxamer 407, 2-6% (w / w) 187 kDa hyaluronic acid, and 0.2-5% (w / w) carboxymethyl chitosan in 25 mM phosphate buffer (pH 7.4).

[0376] A preparatio...

Claims

1. A method for preparing a composition, the composition comprising: Resiquimod and a polymer combination preparation comprising at least a first and a second polymer component, wherein the first polymer component is or comprises a poloxamer and the second polymer component is not a poloxamer, and wherein the polymer combination preparation transitions from a precursor state to a polymer network state in response to a gelation trigger; the polymer network state has a viscosity that is significantly higher than the viscosity of the precursor state; the gelation trigger is or comprises a temperature equal to or greater than a critical gelation temperature (CGT) of the polymer combination formulation, a polymer component ratio equal to or greater than a critical gelation weight ratio of the at least first and second polymer components, a molecular weight of the at least first and / or second polymer components, or a combination thereof; the polymer network state includes crosslinks not present in the precursor state; the crosslinks are or include intramolecular crosslinks, intermolecular crosslinks, or both; the first polymer component is present in the polymer combination preparation at a concentration of 12.5% ​​(w / w) or less; The method comprises: providing at least one solid form of resiquimod selected from the group consisting of Form I, Form II, Form III, Form IV, Form V, Form VI, and Form VII; combining the solid form of resiquimod with the first polymer component and the second polymer component; The method of claim 1, wherein the composition is obtained by comprising:

2. at least one solid form of resiquimod (i) Form I, wherein Form I is characterized in an XRPD pattern having peaks at about 8.72, about 12.24, about 16.29, about 17.56, about 19.51, about 21.31, and about 29.15 degrees 2θ; (ii) Form II, wherein said Form II is characterized in an XRPD pattern by peaks at about 7.75, about 9.65, about 11.23, about 14.38, about 19.90, about 20.80, and about 22.65 degrees 2θ; (iii) Form III, wherein said Form III has an XRPD pattern characterized by peaks at about 8.69, about 9.18, about 9.48, about 11.97, about 14.41, about 18.53, and about 19.70 degrees 2θ; (iv) Form IV, characterized in that the peaks in its XRPD pattern are at about 6.01, about 12.00, about 12.15, about 16.14, about 19.24, about 20.21, about 21.19, about 22.12, and about 24.50 degrees 2θ; (v) Form V, wherein Form V is characterized in an XRPD pattern by peaks at about 8.13, about 10.20, about 10.44, about 16.29, and about 24.56 degrees 2θ; (vi) Form VI, characterized in that its XRPD pattern has peaks at about 9.40, about 13.02, about 18.13, about 18.93, about 20.38, about 23.16, and about 27.78 degrees 2θ; or (vii) Form VII, characterized in that its XRPD pattern has peaks at about 6.25, about 9.92, about 10.96, about 16.51, about 18.99, about 23.75, and about 24.24 degrees 2θ; The method of claim 1.

3. The method of claim 1 or 2, wherein the polymer combination preparation does not contain covalent crosslinks.

4. 2. The method of claim 1, wherein the CGT of the polymer combination preparation is 30-39°C or 20-30°C.

5. 10. The method of claim 1, wherein the polymer combination preparation comprises a total polymer content of at least 5% (w / w) or at least 10% (w / w).

6. 2. The method of claim 1, wherein the critical gelation weight ratio of the first polymer component to the second polymer component is from 1:1 to 22:1, from 1:1 to 18:1, from 1:1 to 14:1, or from 1:1 to 10:

1.

7. The method of claim 1 , wherein the polymer network state is a viscous solution or a colloid.

8. The method of claim 1 , wherein the polymer network state is a hydrogel.

9. The method of claim 1 , wherein the second polymer component is or comprises a carbohydrate polymer.

10. 10. The method of claim 9, wherein the carbohydrate polymer in the polymer combination preparation is present at a concentration of about 10% (w / w) or less, or about 5% (w / w) or less.

11. 11. The method of claim 9 or 10, wherein the carbohydrate polymer is or comprises hyaluronic acid.

12. 11. The method of claim 9 or 10, wherein the carbohydrate polymer is or comprises chitosan or modified chitosan.

13. The method of claim 12, wherein the modified chitosan is or comprises carboxymethylchitosan.

14. The polymer network state is a. a storage modulus in the range of 100 Pa to about 10,000 Pa measured at 37° C. and pH 5-8; b. a storage modulus that is at least 40% lower than the storage modulus of a hydrogel formed from a solution having the poloxamer at a solution concentration of 18% (w / w), and c. A storage modulus measured at 37°C that remains substantially the same after its precursor state is stored at 2-8°C for one month (or no more than 20% of the polymer combination preparation degrades in one month when measured at 37°C). The method of claim 1 characterized by one or more material properties selected from:

15. 2. The method of claim 1, wherein the poloxamer is or comprises poloxamer 407.

16. 10. The method of claim 1, wherein the first polymer component is present in the polymer combination preparation at a concentration of 11% (w / w) or less.

17. Resiquimod and 8-12.5% ​​(w / w) poloxamer 407; 1-4% (w / w) hyaluronic acid having a molecular weight of about 100 kDa to about 500 kDa; A composition comprising:

18. 18. The composition of claim 17, comprising 0.005 mg / mL to 1.00 mg / mL of resiquimod.

19. 19. The composition of claim 17 or 18, wherein the composition is administered to a subject in need thereof.

20. 20. The composition of claim 19, wherein the subject in need thereof is a tumor resection subject.

21. (i) at least one solid form of resiquimod selected from the group consisting of Form I, Form II, Form III, Form IV, Form V, Form VI, and Form VII; (ii) a first polymer component; and (iii) a second polymer component; and A kit comprising: When combined, a polymer combination preparation is provided, wherein the first polymer component is or comprises a poloxamer and the second polymer component is not a poloxamer, and wherein the polymer combination preparation transitions from a precursor state to a polymer network state in response to a gelation trigger; the polymer network state has a viscosity that is significantly higher than the viscosity of the precursor state; the gelation trigger is or comprises a temperature equal to or greater than a critical gelation temperature (CGT) of the polymer combination formulation, a polymer component ratio equal to or greater than a critical gelation weight ratio of the at least first and second polymer components, a molecular weight of the at least first and / or second polymer components, or a combination thereof; the polymer network state includes crosslinks not present in the precursor state; the crosslinks are or include intramolecular crosslinks, intermolecular crosslinks, or both; The kit, wherein the first polymer component is present in the polymer combination preparation at a concentration of 12.5% ​​(w / w) or less.

22. A composition comprising resiquimod Form I and at least one solid form selected from the group consisting of Form II, Form III, Form IV, Form V, Form VI, and Form VII.

23. 1. A process for preparing resiquimod Form I, comprising: (i) providing resiquimod; (ii) dissolving resiquimod in a suitable solvent (e.g., dichloromethane); (iii) adding a suitable anti-solvent (e.g., heptane); to obtain Form I of resiquimod.

24. 1. A solid form of resiquimod, said solid form comprising: (a) Form II, characterized in an XRPD pattern having peaks at about 7.75, about 9.65, about 11.23, about 14.38, about 19.90, about 20.80, and about 22.65 degrees 2θ; (b) Form V, characterized in an XRPD pattern by peaks at about 8.13, about 10.20, about 10.44, about 16.29, and about 24.56 degrees 2θ; or (c) Form VII, characterized in that the XRPD pattern has peaks at about 6.25, about 9.92, about 10.96, about 16.51, about 18.99, about 23.75, and about 24.24 degrees 2θ; A solid form of resiquimod.

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