Compositions and methods for localized therapeutic delivery to brain tumors

Abstract

A composition for localized delivery of a drug to an intracranial region, such as brain tissue or a brain tumor. A method for localized delivery of a drug or therapy to an intracranial region. The composition may comprise a hydrogel in which a chemotherapy or immunotherapy drug is dispersed. A kit comprising a composition or solutions that may be combined to form a composition.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 334,643, filed April 25, 2022, which is incorporated herein by reference. [Background technology]

[0002] Current therapies for patients with brain cancer, such as those with glioblastoma (GBM), are limited for a variety of reasons, which typically result in poor patient outcomes. From 1971 to 2011, brain cancer survival rates improved from approximately 8% to approximately 13%, while average survival rates from all cancers increased from approximately 25% to 50% during the same period.

[0003] Glioblastoma is the most common and aggressive primary central nervous system tumor, constituting approximately 48% of all malignant brain tumors, with a median age at diagnosis of 65 years. Standard treatment for GBM includes surgical resection followed by combination chemotherapy and radiation. Surgery provides clinical relief, allows for the collection of diagnostic tissue samples, and improves survival. However, GBM has a diffuse infiltration pattern, and it is not possible to remove all cells, as resection must be balanced with the preservation of healthy tissue. As a result, radiation therapy and chemotherapy are used to eliminate residual tumor cells after surgery, but these techniques are limited by their inability to reach all tumor cells, lack of specificity for tumor cells, and the ability of tumor cells to develop resistance to these therapies.

[0004] Approximately 95% of patients are diagnosed after the age of 40 (median age = 65 years). Primary GBM tumors arise de novo and account for 90% of cases, while secondary tumors arise from low-grade gliomas and account for 10% of all tumors. Patients with both primary and secondary tumors typically present with symptoms of increased intracranial pressure, such as headaches, neurological deficits, seizures, etc. The diagnosis of GBM is based on the presence of several histologic features, including anaplasia, mitotic activity, microvascular proliferation, and necrosis.

[0005] Current GBM therapies typically include surgical resection, radiation therapy, and oral temozolomide chemotherapy; this therapeutic regimen results in a median overall survival of approximately 15 months.

[0006] The ability to achieve more effective therapy is limited by several factors, including drug toxicity, drug delivery, and tumor biology. The toxicity of certain drugs can cause hypophysitis, uveitis, and orbital inflammation, pneumonia, adrenal insufficiency, enteritis, arthralgia, pancreatitis, and autoimmune diabetes, rashes and vitiligo, hepatitis, hypothyroidism, xerostomia, or a combination thereof.

[0007] Although GLIADEL® wafers are the only FDA-approved product that directly addresses the challenges associated with therapeutic delivery to the brain, the product is believed to have several drawbacks, including its use of less potent chemotherapeutics that may be susceptible to wafer migration, mechanical mismatch with soft tissue, rapid drug release, slow material degradation, and / or resistance. For comparison purposes, a plot of the survival percentage of 9L glioma-bearing rats when treated with doxorubicin-loaded polyanhydride wafers (i.e., GLIADEL® delivery technology) is provided in Figure 1.

[0008] There remains a need for improved compositions and methods for treating brain cancers, such as GBM, including compositions and methods for the localized delivery of various therapies to brain tumors that overcome one or more of the aforementioned disadvantages. Summary of the Invention

[0009] Methods and compositions, such as hydrogels (e.g., adhesive hydrogels), that can effectively deliver therapeutic agents to brain tumors or brain tissue are provided herein. In some embodiments, the compositions include spray-dispersed drug-loaded hydrogels. The compositions described herein can be injected during biopsy or via intraventricular shunts. The compositions and methods described herein can achieve increased drug diffusion from the resection cavity to disseminate to tumor nests, provide an alternative therapy to temozolomide, have a safe product profile after treatment with radiation therapy, promote wound healing, and / or reduce or eliminate scar formation.

[0010] Embodiments of the compositions provided herein include hydrogels, which may be soft and / or adhesive, and may provide a competitive advantage over other products, such as GLIADEL® wafers (Azurity Pharmaceuticals, USA). For example, the compositions provided herein may provide a higher hydrogel:tissue interface, improved stability at the resection site due to adhesive properties, mechanical stiffness equivalent to soft tissue, slower and / or more controlled drug delivery than wafers, and / or the potential for combination delivery. The compositions provided herein may also deliver doxorubicin, which may be more potent than current therapies, less susceptible to resistance, and / or have multiple mechanisms of action, including the ability to induce immunogenic cell death.

[0011] In one aspect, a drug delivery composition is provided. In some embodiments, the drug delivery composition comprises a hydrogel and a drug dispersed in the hydrogel. The drug may comprise a chemotherapy drug, an immunotherapy drug, or a combination thereof. In some embodiments, the hydrogel comprises a polymer component, the polymer component comprising a polymer having three or more aldehyde groups, and a dendrimer component, the dendrimer component comprising a dendrimer having at least two branches comprising one or more surface groups. The one or more surface groups may react reversibly or irreversibly with the three or more aldehyde groups.

[0012] In another aspect, a method of treating a patient is provided. In some embodiments, the method comprises locally delivering a drug delivery composition as described herein to an intracranial region of a patient. Localized delivery of the drug delivery composition may comprise injecting or spraying the drug delivery composition in an intracranial region, which may include brain tissue, a brain tumor, a site of a previously resected tumor, or a combination thereof.

[0013] In yet another aspect, a kit is provided. In some embodiments, the kit includes a first part including a first solution containing a polymer component, the polymer component comprising a polymer, and a second part including a second solution containing a dendrimer component, the dendrimer component comprising a dendrimer having at least two branches comprising one or more surface groups, and at least one drug is disposed in the first solution, the second solution, or both the first solution and the second solution, and the drug comprises a chemotherapy drug, an immunotherapy drug, or a combination thereof.

[0014] Additional aspects will be set forth in part in the description which follows and in part will be obvious from the description or may be learned by practice of the aspects described herein. The advantages described herein may be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive. [Brief description of the drawings]

[0015] [Figure 1] The survival percentages of the various treatments are shown. [Diagram 2] 1 illustrates one embodiment of a multi-compartment syringe. [Diagram 3] 1 shows plots of cell viability versus concentration for various treatments. [Figure 4] 1 shows the degradation rate of an embodiment of a hydrogel. [Figure 5A] 1 shows the release profile of an embodiment of the composition. [Figure 5B] 1 shows the release profile of an embodiment of the composition. [Figure 6A] 1 shows the release profile of an embodiment of the composition. [Figure 6B] 1 shows the release profile of an embodiment of the composition. [Figure 7A] 1 shows the release profile of an embodiment of the composition. [Figure 7B] 1 shows the release profile of an embodiment of the composition. [Figure 8A] 1 shows the release profile of an embodiment of the composition. [Figure 8B] 1 shows the release profile of an embodiment of the composition. [Figure 9] 1 shows the cumulative release of doxorubicin from an embodiment of the composition. [Figure 10] 1 shows the cumulative release of capped doxorubicin from one embodiment of the composition. [Figure 11] 1 shows the cumulative release of doxorubicin from one embodiment of a composition and one embodiment of encapsulated doxorubicin. [Figure 12A] 1 shows the drug release profile from one embodiment of the composition having different drug loadings. [Figure 12B] 1 shows the drug release profile from one embodiment of the composition having different drug loadings. [Figure 13] 1 shows plots of average radiant efficiency for embodiments of the composition. [Figure 14] 1 shows plots of survival probability for embodiments of the composition. [Figure 15] 1 shows a plot of survival probability for one embodiment of the composition and one embodiment of an empty hydrogel. [Figure 16] 1 shows plots of survival probability for embodiments of the composition. [Figure 17] 1 shows plots of total flux for embodiments of the composition. [Figure 18] 1 shows plots of total flux for embodiments of the composition. [Figure 19]1 shows plots of survival percentages for embodiments of the composition. [Figure 20A] 1 shows plots of cell viability for embodiments of the composition. [Figure 20B] 1 shows plots of cell viability for embodiments of the composition. [Figure 21] 1 shows a plot of survival probability following administration of an embodiment of the composition. [Figure 22] 1 shows a plot of survival probability following administration of an embodiment of the composition. [Figure 23] 1 shows a plot of survival probability following administration of an embodiment of the composition. [Figure 24] 1 shows a plot of survival probability following administration of an embodiment of the composition. [Figure 25A] 1 shows a plot of survival probability following administration of an embodiment of a composition containing 7.5 micrograms of an embodiment of the drug. [Figure 25B] 1 shows a plot of survival probability following administration of an embodiment of a composition containing 15 micrograms of an embodiment of the drug. [Figure 25C] 1 shows a plot of survival probability following administration of an embodiment of a composition containing 7.5 micrograms of an embodiment of the drug. [Figure 26] 1 shows plots of survival percentages following administration of embodiments of the composition. [Figure 27] 1 shows a plot of survival probability following administration of an embodiment of the composition. [Figure 28] 1 shows a plot of survival probability following administration of an embodiment of the composition. [Figure 29] 1 shows plots of total flux following injection of hydrogel embodiments. [Diagram 30] 1 shows data collected from studies to determine the effect of embodiments of the composition on T cells in lymph nodes. [Diagram 31] 1 shows data collected from a study to determine the effect of embodiments of the composition on central memory CD8 T cells in the spleen. [Diagram 32]1 shows data collected from a study to determine whether macrophages contributed to an increase in CD45+ cells in tumors upon treatment with one embodiment of the composition. [Diagram 33] 1 shows the results of testing M1 polarization markers in CD80+ / Macs 206 tumors treated with an embodiment of the composition. [Diagram 34] 1 shows the results of a test designed to determine a shift in dendritic cell population. [Diagram 35] FIG. 1 is a schematic diagram of one embodiment of anti-cancer immune activity. [Diagram 36] FIG. 1 is a schematic diagram of the possible behavior of nanoparticles in the cGAS-STING pathway. [Figure 37] FIG. 1 is a schematic diagram of a possible mechanism of action. [Figure 38] 1 shows one embodiment of a nanoparticle-based STING agonist formulation. [Figure 39] 1 presents data showing stimulation of dendritic cells and macrophages by one embodiment of a cyclic dinucleotide nanoparticle. [Figure 40A] 1 shows the total release and cumulative release profiles, respectively, of embodiments of the composition. [Figure 40B] 1 shows the total release and cumulative release profiles, respectively, of embodiments of the composition. [Diagram 41] Results of a local immunotherapy pilot study are presented. [Figure 42-1] FIG. 1 is a schematic diagram of treatment mechanisms and outcomes. [Figure 42-2] FIG. 1 is a schematic diagram of treatment mechanisms and outcomes. [Figure 43A] 1 shows the results regarding cell viability upon treatment with one embodiment of the composition. [Figure 43B] 1 shows the results regarding CRT expression upon treatment with one embodiment of the composition. [Diagram 44] FIG. 1 is a schematic diagram of the test described herein. [Figure 45-1] 1 shows the results of an examination of BMDM CD86 activation marker expression. [Figure 45-2]1 shows the results of an examination of BMDM CD86 activation marker expression. [Diagram 46] 13 shows results demonstrating that combination therapy increased the ratio of pro-inflammatory to anti-inflammatory BMDMs. [Figure 47] FIG. 1 is a schematic diagram of one embodiment of a treatment plan. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] The drug delivery compositions described herein may include a hydrogel and a drug. The drug may be selected from a chemotherapy drug, an immunotherapy drug, or a combination thereof. In some embodiments, the drug includes a nucleic acid, a nanoparticle, an antibody, a small molecule, an immunomodulator, or a combination thereof. The hydrogel may include those known in the art. Non-limiting examples of hydrogels are described, for example, in U.S. Pat. No. 8,802,072 and U.S. Pat. No. 10,736,914, which are incorporated herein by reference.

[0017] The drug delivery compositions described herein may include a hydrogel and a drug dispersed in the hydrogel. The drug may be substantially uniformly dispersed in the hydrogel. In some embodiments, the drug is dispersed in the hydrogel in a manner that forms a concentration gradient of the drug.

[0018] The hydrogels described herein may include a polymeric component, where the polymeric component comprises a polymer having three or more aldehyde groups, and a dendrimer component, where the dendrimer component comprises a dendrimer having at least two branches that include one or more surface groups.

[0019] Drugs The drug of the drug delivery compositions described herein can include a chemotherapy drug, an immunotherapy drug, or a combination thereof.

[0020] The chemotherapy drug may include any known chemotherapy drug, including, but not limited to, bleomycin, busulfan, carboplatin, carmustine, cisplatin, cladobruvin, dactinomycin, daunorubicin, doxorubicin, estramustine, interferon, irinotecan, levamisole, methotrexate, mitomycin, paclitaxel, pentostatin, plicamycin, tamoxifen, temozolomide, vinblastine, vindesine, etc. Additionally or alternatively, the chemotherapy drug may include one or more radiosensitizers, including 5-halo-uracil, antiangiogenic compounds, including thalidomide and tranilast, natural or synthetic peptide hormones, including octreotide, and apoptosis-inducing compounds, including butyrate. In some embodiments, the chemotherapy drug includes doxorubicin.

[0021] The immunotherapy drug may include any known immunotherapy drug. As used herein, the phrase "immunotherapeutic drug" refers to any drug that can induce, enhance, suppress, or otherwise modify immune response. In some embodiments, the immunotherapy drug is selected from (a) those that target CTLA-4, such as ipilimumab or tremelimumab, (b) those that target PD-1, such as nivolumab, pidilizumab, or pembrolizumab, AMP-224, (c) those that target PD-L1, such as MPDL-3280A, MSB0010718C, or MEDI4736, or (d) those that target GITR, such as TRX518 or MK4166. Non-limiting examples of immunotherapy drugs are disclosed by U.S. Pat. No. 11,186,640.

[0022] In some embodiments, the immunotherapy drug comprises a cyclic dinucleotide. "Cyclic dinucleotide" or "CDN" may include a class of molecules that contain 2'-5' and / or 3'-5' phosphodiester bonds between two purine nucleotides. This includes 2'-5'-2',5', 2'-5'-3'5', and 3',5'-3',5' linkages. CDNs may activate the cytoplasmic surveillance pathway through direct binding of two cytoplasmic pattern recognition receptors (PRRs), DEAD (aspartic acid-glutamic acid-alanine-aspartic acid)-box helicase 41 (DDX41) and stimulator of interferon genes (STING). Type I interferon responses to infection with intracellular bacteria may result from the secretion of cyclic di-adenosine monophosphate (cdAMP) or its related cyclic dinucleotide (CDN), cyclic di-guanine monophosphate (cdGMP). CDNs can bind with high affinity to DDX41 and complex with STING adaptor proteins, resulting in activation of the TBK1 / IRF3 signaling pathway and induction of IFN-β and other IRF-3-dependent gene products that strongly activate innate immunity. CDNs can include second messengers expressed by most bacteria and can regulate diverse processes, including motility and biofilm formation. Endogenous CDNs can also be produced in response to cytoplasmic DNA by the host enzyme cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS) in tumors or during infection. In some embodiments, the CDN is the canonical bacterial CDN, cyclic di-guanine monophosphate (cdGMP). In some embodiments, the CDN is an endogenous product of cGAS. In some embodiments, the CDN is an agonist of STING.

[0023] Cyclic dinucleotides that can be used include those known in the art, such as those disclosed in U.S. Patent Nos. 7,709,458 and 7,592,326, WO2007 / 054279, U.S. Patent Application Publication No. 2014 / 0205653, and Yan et al. Bioorg.Med.Chem Lett.18:5631 (2008). Non-limiting examples of cyclic dinucleotides include cdAMP, cdGMP, cdIMP, c-AMP-GMP, c-AMP-IMP, and c-GMP-IMP, and analogs thereof, including but not limited to phosphorothioate analogs.

[0024] In some embodiments, the cyclic dinucleotide is an agonist of STING (stimulator of interferon genes). As described herein, the STING signaling pathway in immune cells can be a central mediator of the innate immune response and, when stimulated, can induce the expression of various interferons, cytokines, and T cell recruiting factors that amplify and enhance immune activity. Recent studies have shown that STING agonists, described for example in Dubensky, T., et al., Therapeutic Advances in Vaccines, Vol. 1(4): 131-143 (2013), and Hanson, M., et al., The Journal of Clinical Investigation, Vol. 125(6): 2532-2546 (2015), can be effective adjuvants and can efficiently induce immune responses. In some embodiments, the STING agonist is chemically synthesized. In some embodiments, the STING agonist is an analog of a naturally occurring cyclic dinucleotide. STING agonists, including cyclic dinucleotide analogues, suitable for use in the present disclosure are provided in U.S. Patent Nos. 7,709,458 and 7,592,326, and U.S. Patent Application Publication No. 2014 / 0205653.

[0025] Drugs, such as chemotherapy drugs and / or immunotherapy drugs (e.g., cyclic dinucleotides), can be encapsulated in the liquid. The encapsulation can be performed before the drugs are dispersed in the hydrogel. For example, chemotherapy drugs can be encapsulated in the liquid, immunotherapy drugs (e.g., cyclic dinucleotides) can be encapsulated in the liquid, or chemotherapy drugs and immunotherapy drugs (e.g., cyclic dinucleotides) can be encapsulated together in the same liquid or separately in different liquids. The encapsulated drug can be in the form of liquid nanoparticles (e.g., nanospheres), such as any liquid nanoparticles known in the art. A non-limiting example of encapsulating liquid nanoparticles is disclosed by U.S. Pat. No. 11,207,418, which is incorporated by reference. In some embodiments, the chemotherapy drug is doxorubicin, and the doxorubicin can be encapsulated in the liquid particles (e.g., liquid spheres). The encapsulated doxorubicin can be DOXIL® anthracycline topoisomerase inhibitor (Baxter, USA). The nanoparticles can have any suitable average diameter, such as from about 1 nm to about 100 nm, from about 10 nm to about 90 nm, from about 20 nm to about 80 nm, from about 30 nm to about 70 nm, or from about 40 nm to about 60 nm.

[0026] The drugs described herein may be present in any amount in the drug delivery composition. In some embodiments, the drugs are present in the drug delivery composition in a total amount of about 1 μg to about 5000 μg, about 1 μg to about 1000 μg, about 1 μg to about 500 μg, about 50 μg to about 500 μg, or about 50 μg to about 500 μg. For example, a composition containing a "total amount" of 500 μg of a drug may contain 500 μg of a chemotherapy drug, 500 μg of an immunotherapy drug, 400 μg of a chemotherapy drug, and 100 μg of an immunotherapy drug, etc. The drugs described herein may be present in the drug delivery composition in any concentration. In some embodiments, the drug is present in the drug delivery composition at a total concentration of about 0.1 μg / μL to about 100 μg / μL, about 1 μg / μL to about 80 μg / μL, about 1 μg / μL to about 60 μg / μL, about 1 μg / μL to about 40 μg / μL, about 1 μg / μL to about 30 μg / μL, about 1 μg / μL to about 20 μg / μL, about 1 μg / μL to about 10 μg / μL, about 2 μg / μL to about 8 μg / μL, or about 2 μg / μL to about 5 μg / μL.

[0027] Treatment methods Provided herein is a method for treating a patient, including a patient with a brain tumor. The patient may be any mammal, such as a human. In some embodiments, the method for treating a patient includes locally delivering a drug delivery composition described herein to the intracranial region of the patient.

[0028] The localized delivery of the drug delivery composition can be accomplished in any manner. In some embodiments, the localized delivery of the drug delivery composition includes injecting or spraying the drug delivery composition. The injection of the drug delivery composition can be performed during a biopsy, via an intraventricular shunt, or a combination thereof.

[0029] The intracranial region may include brain tissue, a brain tumor, the site of a previously resected tumor, or a combination thereof.

[0030] In some embodiments, the method also includes administering a second drug or therapy to the patient before, during, and / or after the local delivery of the drug delivery composition. The administration of the second drug or therapy can be accomplished in any manner, such as by orally administering the second drug or therapy. The second drug can include a second chemotherapy drug or therapy, such as temozolomide or a PD1 checkpoint blockade therapy.

[0031] The hydrogels described herein can provide controlled drug release, such as sustained drug release. The components of the hydrogel can be adjusted to achieve a desired release profile. The drug can be released continuously or intermittently. In some embodiments, the drug is released from the drug delivery composition continuously for at least 12 hours, at least 24 hours, at least 36 hours, at least 48 hours, at least 72 hours, at least 100 hours, at least 200 hours, or at least 300 hours.

[0032] In some embodiments, the cumulative percentage of drug released from the drug delivery composition is at least 80%, 85%, or 90% by weight of the drug within 10 days or less, 12 days or less, 14 days or less, 16 days or less, 18 days or less, 20 days or less, 22 days or less, 24 days or less, 26 days or less, 28 days or less, 30 days or less, 32 days or less, 34 days or less, 36 days or less, 38 days or less, 40 days or less, 50 days or less, 60 days or less, 80 days or less, or 100 days or less after topical delivery of the drug delivery composition.

[0033] Dendrimer Components In some embodiments, the dendrimer component includes a dendrimer having amines on at least a portion of its surface groups, commonly referred to as "terminal groups" or "end groups". The dendrimer may have amines on 20% to 100% of its surface groups. In some embodiments, the dendrimer has amines on 100% of its surface groups. In some embodiments, the dendrimer component includes a dendrimer having amines on less than 75% of its surface groups. As used herein, the term "dendrimer" refers to any compound having a multivalent core covalently attached to two or more dendritic branches. In some embodiments, the multivalent core is covalently attached to three or more dendritic branches. In some embodiments, the amines are primary amines. In some embodiments, the amines are secondary amines. In some embodiments, one or more surface groups have at least one primary amine and at least one secondary amine.

[0034] In some embodiments, the dendrimer extends through at least two generations. In some embodiments, the dendrimer extends through at least three generations. In some embodiments, the dendrimer extends through at least four generations. In some embodiments, the dendrimer extends through at least five generations. In some embodiments, the dendrimer extends through at least six generations. In some embodiments, the dendrimer extends through at least seven generations.

[0035] In some embodiments, the dendrimer has a molecular weight of about 1,000 to about 1,000,000 daltons. In some embodiments, the dendrimer has a molecular weight of about 3,000 to about 120,000 daltons. In some embodiments, the dendrimer has a molecular weight of about 10,000 to about 100,000 daltons. In some embodiments, the dendrimer has a molecular weight of about 20,000 to about 40,000 daltons. Unless otherwise specified, the "molecular weight" of a dendrimer refers to the weight average molecular weight.

[0036] Generally, dendrimers can be made using any known method. In some embodiments, dendrimers are made by oxidizing a starting dendrimer having surface groups that include at least one hydroxyl group, such that at least a portion of the surface groups include at least one amine. In some embodiments, dendrimers are made by oxidizing a starting fifth generation (G5) dendrimer having surface groups that include at least one hydroxyl group, such that at least a portion of the surface groups include at least one amine. In some embodiments, dendrimers are made by oxidizing a starting G5 dendrimer having surface groups that include at least one hydroxyl group, such that about 25% of the surface groups include at least one amine. In some embodiments, the dendrimer is a G5 dendrimer having primary amines on about 25% of the surface groups of the dendrimer.

[0037] In some embodiments, the dendrimer is a poly(amidoamine) derived (PAMAM) dendrimer. In some embodiments, the dendrimer is a G5 PAMAM derived dendrimer. In some embodiments, the dendrimer is a G5 PAMAM derived dendrimer having primary amines on about 25% of the surface groups of the dendrimer.

[0038] In some embodiments, the dendrimer is a poly(propyleneimine) derived dendrimer. In some embodiments, the dendrimer component is combined with a liquid to form a dendrimer component solution. In some embodiments, the dendrimer component solution is an aqueous solution. In some embodiments, the solution comprises water, phosphate buffered saline (PBS), Dulbecco's modified Eagle's medium (DMEM), or any combination thereof. In some embodiments, the dendrimer component concentration in the dendrimer component solution is about 5% to about 25% by weight. In some embodiments, the dendrimer component concentration in the dendrimer component solution is about 10% to about 20% by weight. In some embodiments, the dendrimer component concentration in the dendrimer component solution is about 11% to about 15% by weight.

[0039] In some embodiments, the dendrimer component or dendrimer component solution further comprises one or more additives. Generally, the amount of additive may vary depending on the application, tissue type, concentration of the dendrimer component solution, type of dendrimer component, concentration of the polymer component solution, and / or type of polymer component. Examples of suitable additives include, but are not limited to, pH adjusters, thickeners, antimicrobial agents, colorants, surfactants, and radiopaque compounds. Specific examples of these types of additives are described herein. In one embodiment, the dendrimer component solution comprises a foaming additive.

[0040] Polymer Component Generally, the polymeric component comprises a polymer and / or oligomer having one or more functional groups capable of reacting with one or more functional groups on the biological tissue and / or one or more functional groups on the dendrimer component.

[0041] In some embodiments, the polymer is at least one polysaccharide. In some embodiments, the at least one polysaccharide may be linear, branched, or have both linear and branched sections in its structure. In general, the at least one polysaccharide may be natural, synthetic, or modified, for example, by crosslinking, altering the polysaccharide's substituents, or both. In one embodiment, the at least one polysaccharide is plant-based. In some embodiments, the at least one polysaccharide is animal-based. In some embodiments, the at least one polysaccharide is a combination of plant-based and animal-based polysaccharides. Non-limiting examples of polysaccharides include, but are not limited to, dextran, chitin, starch, agar, cellulose, hyaluronic acid, or combinations thereof.

[0042] In some embodiments, at least one polymer has a molecular weight of about 1,000 to about 1,000,000 daltons. In some embodiments, at least one polymer has a molecular weight of about 5,000 to about 15,000 daltons. Unless otherwise specified, the "molecular weight" of a polymer refers to the weight average molecular weight.

[0043] In some embodiments, the polymer is functionalized such that its structure contains one or more functional groups that react with one or more functional groups on the biological tissue and / or one or more functional groups on the dendrimer component. In some embodiments, the polymer is functionalized such that its structure contains three or more functional groups that react with one or more functional groups on the biological tissue and / or one or more functional groups on the dendrimer component. In some embodiments, the functional group incorporated into the structure of the polymer is an aldehyde.

[0044] In some embodiments, the degree of functionalization of the polymer is adjustable. "Degree of functionalization" generally refers to the number or percentage of groups on the polymer that are substituted or converted to one or more desired functional groups. The one or more functional groups, in certain embodiments, include aldehydes, substituents capable of photoreversible dimerization, or combinations thereof. In some embodiments, the degree of functionalization is adjusted based on the type of tissue to which the adhesive is applied, the concentration of the components, and / or the type of polymer or dendrimer used in the adhesive. In some embodiments, the degree of functionalization is about 10% to about 75%. In some embodiments, the degree of functionalization is about 15% to about 50%. In some embodiments, the degree of functionalization is about 20% to about 30%.

[0045] In some embodiments, the polymer is a dextran having a molecular weight of about 10 kDa. In some embodiments, the polymer is a dextran having about 50% of its hydroxyl groups converted to aldehydes. In some embodiments, the polymer is a dextran having a molecular weight of about 10 kDa and about 50% of its hydroxyl groups converted to aldehydes.

[0046] In some embodiments, the polysaccharide is oxidized to contain a desired percentage of one or more aldehyde functional groups. The polysaccharide may be oxidized, for example, to convert about 10% to about 100%, about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, about 40% to about 60%, or about 50% (mol %) of its hydroxyl groups to aldehydes. Generally, the oxidation may be carried out using any known means. For example, suitable oxidizing agents include, but are not limited to, periodates, hypochlorites, ozone, peroxides, hydroperoxides, persulfates, and percarbonates. In one embodiment, the oxidation is carried out using sodium periodate. Typically, different amounts of oxidizing agents may be used to modify the degree of functionalization.

[0047] In some embodiments, the polymer component is combined with a liquid to form a polymer component solution. In some embodiments, the polymer component solution is an aqueous solution. In some embodiments, the solution comprises water, PBS, DMEM, or any combination thereof.

[0048] Generally, the polymer component solution can have any suitable concentration of the polymer components. In some embodiments, the polymer component concentration in the polymer component solution is from about 5% to about 40% by weight. In some embodiments, the polymer component concentration in the polymer component solution is from about 5% to about 30% by weight. In some embodiments, the polymer component concentration in the polymer component solution is from about 5% to about 25% by weight. Typically, the concentration can be adapted and / or adjusted based on the particular application, tissue type, and / or type and concentration of dendrimer components used.

[0049] The polymer component or polymer component solution may also include one or more additives. In some embodiments, the additive is compatible with the polymer component. In some embodiments, the additive does not contain a primary or secondary amine. Generally, the amount of additive varies depending on the application, tissue type, concentration of the polymer component solution, and type of polymer component and / or dendrimer component. Examples of suitable additives include, but are not limited to, pH adjusters, thickeners, antimicrobial agents, colorants, surfactants, radiopaque compounds, and other additives described herein. In other embodiments, the polymer component solution includes a foaming agent.

[0050] In some embodiments, the pH adjuster is an acidic compound. Examples of acidic pH adjusters include, but are not limited to, carboxylic acids, inorganic acids, and sulfonic acids. In some embodiments, the pH adjuster is a basic compound. Examples of basic pH adjusters include, but are not limited to, hydroxides, alkoxides, nitrogen-containing compounds other than primary and secondary amines, basic carbonates, and basic phosphates.

[0051] Generally, the thickening agent may be selected from any known viscosity modifying compound, including, but not limited to, polysaccharides and their derivatives, such as starch or hydroxyethyl cellulose.

[0052] Generally speaking, a surfactant can be any compound that reduces the surface tension of water.In one embodiment, the surfactant is an ionic surfactant, such as sodium lauryl sulfate.In another embodiment, the surfactant is a neutral surfactant.Examples of neutral surfactants include, but are not limited to, polyoxyethylene ethers, polyoxyethylene esters, and polyoxyethylene sorbitan.

[0053] In some embodiments, the radiopaque compound is barium sulfate, gold particles, or a combination thereof.

[0054] Hydrogels In general, the hydrogels described herein can be formed by combining the polymer component or polymer component solution and the dendrimer component or dendrimer component solution in any manner. In some embodiments, the polymer component or polymer component solution and the dendrimer component or dendrimer component solution are combined prior to localized delivery of the drug delivery composition. In some embodiments, the polymer component or polymer component solution and the dendrimer component or dendrimer component solution are combined in any order in the intracranial region. In some embodiments, the localized delivery of the composition includes locally delivering the polymer component or polymer component solution to the intracranial region and locally delivering the dendrimer component or dendrimer component solution to the intracranial region, where the polymer component and the dendrimer component contact each other in the intracranial region.

[0055] The hydrogel may be locally delivered using any suitable tool and method: double barrel syringes with rigid or flexible ejection tips and optional extension tubes, needles, stents, catheters, and other devices known in the art are envisioned.

[0056] As used herein, a hydrogel is a "therapeutic" if it stops, reverses, or reduces the rate of tumor growth, improves survival odds, extends the lifespan of a patient, or a combination thereof.

[0057] After contacting one or more biological tissues in the intracranial region, the hydrogel may be given sufficient time to harden or gel. When a hydrogel "hardens" or "gels" as those terms are used herein, it means that reactive groups on the polymer component, the dendrimer component, and one or more biological tissues have undergone one or more reactions. Without wishing to be bound by any particular theory, it is believed that the hydrogels described herein are effective because the polymer component reacts with both the dendrimer component and the surface of the biological tissue. In some embodiments, the aldehyde functional group of the polymer component reacts with one or more amines of (i) a drug such as doxorubicin, (ii) a dendrimer component, and / or (iii) one or more biological tissues to form an imine bond. The formation of the imine bond may be reversible, and the formation and hydrolysis of the imine bond may control or contribute to the release kinetics of the drug from the hydrogel, and therefore the number / ratio of functional groups involved in imine bond formation may be configured to achieve the desired release kinetics. In some embodiments, it is believed that the amines on the dendrimer component react with a high percentage of the aldehydes on the polymer component, thereby reducing the toxicity of the hydrogel and increasing its biocompatibility. Typically, the time required to cure or gel the hydrogel will vary based on many factors, including, but not limited to, the characteristics of the polymer and / or dendrimer components, the concentration of the polymer and / or dendrimer component solutions, and the characteristics of one or more biological tissues. In some embodiments, the hydrogel will cure sufficiently to provide the desired bond or seal immediately after the components are combined. The gelation or cure time should provide that the mixture of components can be delivered to the target area in a fluid form before becoming too viscous or solid, and then cure rapidly thereafter once applied to the target area. In one embodiment, the gelation or cure time is less than 120 seconds. In some embodiments, the gelation or cure time is between 3 and 60 seconds. In some embodiments, the gelation or cure time is between 5 and 30 seconds. Before or after the hydrogel is cured, the substituents capable of photoreversible dimerization can be activated or deactivated as desired.

[0058] In certain embodiments, one or more effervescent agents are added to the polymer component solution and / or the dendrimer component solution before the solutions are combined. In one embodiment, the effervescent agent comprises a two-part liquid system comprising part 1 and part 2, where part 1 comprises a bicarbonate salt and part 2 comprises an aqueous solution of a di- or polyaldehyde and a titrant. A wide range of di- or polyaldehydes exists, their usefulness being limited primarily by their availability and their solubility in water. For example, aqueous glyoxal (ethanedial) is useful, as is aqueous glutaraldehyde (pentadial). Water-soluble mixtures of di- and polyaldehydes prepared by oxidative cleavage of appropriate carbohydrates with periodate, ozone, etc. may also be useful.

[0059] A titrant is most preferably used in the liquid solution of Part 2. More specifically, the titrant is an organic or inorganic acid, buffer, salt, or salt solution that can react with the bicarbonate component of Part 1 to produce carbon dioxide and water as reaction by-products. The carbon dioxide gas produced forms a foam-like structure in the hydrogel and expands the volume of the hydrogel.

[0060] Most preferably, the titrant is an inorganic or organic acid present in an amount to impart an acidic pH to the resulting mixture of Part 1 and Part 2 components. Preferred acids that can be used in the practice of the invention include phosphoric acid, sulfuric acid, hydrochloric acid, acetic acid, and citric acid.

[0061] Hydrogel Kit In another embodiment, a kit is provided that includes a first part comprising a polymer component or a polymer component solution and a second part comprising a dendrimer component or a dendrimer component solution. The kit may further include an applicator or other device means, such as a multi-compartment syringe, for storing, combining and delivering the two parts and / or the resulting hydrogel to the intracranial region.

[0062] At least one drug as described herein may be disposed in the polymer component solution, the dendrimer component solution, or in both the polymer component solution and the dendrimer component solution. The drugs as described herein may include chemotherapy drugs, immunotherapy drugs, or combinations thereof.

[0063] In some embodiments, the kit includes separate reservoirs for the polymer component solution and the dendrimer component solution. In some embodiments, the kit includes reservoirs for the polymer component solutions of different concentrations. In some embodiments, the kit includes reservoirs for the dendrimer component solutions of different concentrations.

[0064] In some embodiments, the kit includes instructions for selecting an appropriate concentration or amount of at least one of the polymer component, polymer component solution, dendrimer component, dendrimer component solution, or drug to compensate for or take into account at least one characteristic of one or more biological tissues, treatment regimens, or the like.

[0065] In some embodiments, the kit includes at least one syringe. In some embodiments, the syringe includes separate reservoirs for the polymer component solution and the dendrimer component solution. The syringe may also include a mixing tip that combines the two solutions when the plunger is depressed. The mixing tip may be releasably securable to the syringe (to allow for mixing tip exchange), and the mixing tip may include a static mixer. In some embodiments, the reservoirs in the syringe may have different sizes or may contain different volumes of solutions. In some embodiments, the reservoirs in the syringe may be the same size or may contain the same volumes of solutions. In some embodiments, one reservoir may contain part 1 of the foam composition described herein above, and a second reservoir may contain part 2 of the foam composition.

[0066] FIG. 2 shows one embodiment of a syringe 100. The syringe 100 includes a body 110 having two reservoirs (130, 140). The dendrimer component solution is disposed in the first reservoir 130 and the polymer component solution is disposed in the second reservoir 140. The two reservoirs (130, 140) are emptied by depressing the plunger 120, which forces the contents of the two reservoirs (130, 140) into the mixing tip 150 and out of the syringe 100. In a further embodiment, one or more of the syringe's reservoirs may be removable. In this embodiment, the removable reservoir may be replaced with a reservoir containing a desired concentration of the polymer component solution or the dendrimer component solution. In a preferred embodiment, the kit is sterile. For example, the kit components may be packaged together, for example, in a tray, pouch, and / or box. The packaged kits may be sterilized using known techniques at suitable wavelengths (where applicable), such as electron beam irradiation, gamma irradiation, ethylene oxide sterilization, or other suitable techniques.

[0067] All referenced publications are incorporated herein by reference in their entirety. Furthermore, to the extent that a definition or use of a term in a reference incorporated herein by reference is inconsistent with or contradicts the definition of that term provided herein, the definition of that term provided herein shall apply and the definition of that term in the reference shall not apply.

[0068] Although certain aspects of the prior art have been discussed to facilitate disclosure of various embodiments, applicants do not in any way disclaim these technical aspects, and it is contemplated that the present disclosure may include one or more of the prior art technical aspects discussed herein.

[0069] The present disclosure may address one or more of the problems and deficiencies of known methods and processes. However, it is contemplated that the various embodiments may prove useful in addressing other problems and deficiencies in certain technical fields. Thus, the present disclosure should not be construed as being limited to addressing any of the specific problems or deficiencies discussed herein.

[0070] Where a document, act, or article of knowledge is referenced or discussed in this specification, such reference or discussion is not an admission that that document, act, or article of knowledge, or any combination thereof, was publicly available, known to the public, part of common general knowledge, or otherwise constitutes prior art under applicable statutory provisions or was known to be relevant to any attempt to solve any problem to which this specification pertains.

[0071] In the description provided herein, the terms "includes," "is," "containing," "having," and "comprises" are used in an open-ended manner and should thus be interpreted as "including, but not limited to." Unless otherwise specified, when a composition, kit, or method is claimed or described in terms of "comprising" various steps or components, the composition, kit, or method can also "consist essentially of" or "consist of" the various steps or components.

[0072] The terms "a," "an," and "the" are intended to include multiple alternatives, e.g., at least one. For example, disclosure of a "polymer," a "drug," etc., is meant to encompass one or mixtures or combinations of two or more polymers, drugs, etc., unless otherwise specified.

[0073] Various numerical ranges may be disclosed herein. When applicants disclose or claim any type of range, applicants' intention is to disclose or claim individually each possible number that such ranges can reasonably encompass, including the endpoints of the ranges, and any subranges and combinations of subranges encompassed therein, unless otherwise specified. Moreover, all numerical endpoints of ranges disclosed herein are approximations. As a representative example, applicants disclose that in some embodiments, the drug is present in the drug delivery composition at a total concentration of about 1 μg / μL to about 10 μg / μL. This range should be interpreted to encompass about 1 μg / μL and about 10 μg / μL, and further encompasses each of "about" 2 μg / μL, 3 μg / μL, 4 μg / μL, 5 μg / μL, 6 μg / μL, 7 μg / μL, 8 μg / μL, and 9 μg / μL, including any ranges and subranges between any of these values.

[0074] As used herein, the term "about" means plus or minus 10% of the numerical value of the number with which it is used.

[0075] List of embodiments The following is a non-limiting list of embodiments of the present disclosure. Embodiment 1. A drug delivery composition comprising a hydrogel, e.g., an adhesive hydrogel, and a drug dispersed in the hydrogel, wherein the drug comprises a chemotherapy drug, an immunotherapy drug, or a combination thereof.

[0076] Embodiment 2. The composition of embodiment 1, wherein the hydrogel comprises a polymer component, for example a polymer component comprising a polymer having three or more aldehyde groups.

[0077] Embodiment 3. The composition of any of the preceding embodiments, wherein the hydrogel comprises a dendrimer component, e.g., a dendrimer component comprising a dendrimer having at least two branches comprising one or more surface groups.

[0078] Embodiment 4. The composition of any of the preceding embodiments, wherein 100% of the one or more surface groups comprise at least one primary or secondary amine.

[0079] Embodiment 5. The composition of any of the preceding embodiments, wherein less than 95%, less than 90%, less than 85%, less than 80%, or less than 75% of the one or more surface groups comprise at least one primary or secondary amine.

[0080] Embodiment 6. The composition of any of the preceding embodiments, wherein the dendrimer is a fifth generation polyamidoamine (G5 PAMAM) dendrimer.

[0081] Embodiment 7. The composition of any of the preceding embodiments, wherein the polymer having three or more aldehyde groups comprises a polysaccharide, e.g., an oxidized polysaccharide.

[0082] Embodiment 8. The composition of any of the preceding embodiments, wherein about 10% to about 100%, about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, about 40% to about 60%, or about 50% (mol %) of the hydroxyl groups of the polysaccharide are oxidized to an aldehyde.

[0083] Embodiment 9. The composition of any of the preceding embodiments, wherein the polymer comprises dextran.

[0084] Embodiment 10. The composition of any of the preceding embodiments, wherein the drug comprises or consists of a chemotherapy drug.

[0085] Embodiment 11. The composition of any of the preceding embodiments, wherein the chemotherapy drug comprises bleomycin, busulfan, carboplatin, carmustine, cisplatin, cladobruvin, dactinomycin, daunorubicin, doxorubicin, estramustine, interferon, irinotecan, levamisole, methotrexate, mitomycin, paclitaxel, pentostatin, plicamycin, tamoxifen, temozolomide, vinblastine, vindesine, or a combination thereof.

[0086] Embodiment 12. The composition of any of the preceding embodiments, wherein the chemotherapy drug comprises one or more radiosensitizers, e.g., 5-halo-uracil, anti-angiogenic compounds (e.g., natural or synthetic peptide hormones, including thalidomide, tranilast, octreotide, and / or compounds that induce apoptosis, including butyrate).

[0087] Embodiment 13. The composition of any of the preceding embodiments, wherein the chemotherapy drug comprises or consists of doxorubicin.

[0088] Embodiment 14. The composition of any of the preceding embodiments, wherein the immunotherapy drug comprises: (a) one or more agents that target CTLA-4, e.g., ipilimumab or tremelimumab; (b) one or more agents that target PD-1, e.g., nivolumab, pidilizumab, or pembrolizumab, AMP-224; (c) one or more agents that target PD-L1, e.g., MPDL-3280A, MSB0010718C, or MEDI4736; (d) one or more agents that target GITR, e.g., TRX518 or MK4166; or (e) a combination thereof.

[0089] Embodiment 15. A composition according to any of the preceding embodiments, wherein the immunotherapy drug comprises or consists of a cyclic dinucleotide.

[0090] Embodiment 16. The composition of any of the preceding embodiments, wherein the chemotherapy drug, immunotherapy drug, or chemotherapy drug and immunotherapy drug are encapsulated in a liquid, and the encapsulation may form liquid particles (e.g., liquid nanoparticles), e.g., liquid spheres (e.g., liquid nanospheres).

[0091] Embodiment 17. The composition of any of the preceding embodiments, wherein the chemotherapy drug and the immunotherapy drug are encapsulated together in the same liquid particle, or the chemotherapy drug and the immunotherapy drug are encapsulated separately in different liquid particles.

[0092] Embodiment 18. A composition according to any of the preceding embodiments, wherein the liquid particles, e.g., liquid nanoparticles, comprise a poly-beta-amino-ester.

[0093] Embodiment 19. The composition of embodiment 18, wherein the cyclic dinucleotide is conjugated to a poly-beta-amino-ester via a cathepsin-sensitive bond, and the poly-beta-amino-ester is optionally modified with arginine.

[0094] Embodiment 20. The composition of any of the preceding embodiments, wherein the liquid particles are liquid nanoparticles (e.g., liquid nanospheres) having an average diameter of about 1 nm to about 100 nm, about 10 nm to about 90 nm, about 20 nm to about 80 nm, about 30 nm to about 70 nm, or about 40 nm to about 60 nm.

[0095] Embodiment 21. The composition of any of the preceding embodiments, wherein the hydrogel comprises phosphate buffered saline (PBS).

[0096] Embodiment 22. The composition of any of the preceding embodiments, wherein the hydrogel has a solids content of about 5% to about 40%, about 5% to about 35%, about 5% to about 30%, about 5% to about 25%, about 8% to about 25%, about 10% to about 20%, or about 12% to about 18% by weight.

[0097] Embodiment 23. The composition of any of the preceding embodiments, wherein the drug is present in the drug delivery composition in a total amount of about 1 μg to about 5000 μg, about 1 μg to about 1000 μg, about 1 μg to about 500 μg, about 50 μg to about 500 μg, or about 50 μg to about 500 μg.

[0098] Embodiment 24. The composition of any of the preceding embodiments, wherein the drug is present in the drug delivery composition at a total concentration of from about 0.1 μg / μL to about 100 μg / μL, from about 1 μg / μL to about 80 μg / μL, from about 1 μg / μL to about 60 μg / μL, from about 1 μg / μL to about 40 μg / μL, from about 1 μg / μL to about 30 μg / μL, from about 1 μg / μL to about 20 μg / μL, from about 1 μg / μL to about 10 μg / μL, from about 2 μg / μL to about 8 μg / μL, or from about 2 μg / μL to about 5 μg / μL.

[0099] Embodiment 25. A method of treating a patient, the method comprising locally delivering a drug delivery composition of any of the preceding embodiments to the patient, e.g., to the intracranial region of the patient, the patient optionally being a human.

[0100] 26. (A) localized delivery of the drug delivery composition comprises injecting or spraying the drug delivery composition; (B) the polymer component / solution and the dendrimer component / solution are contacted prior to localized delivery of the composition to the intracranial region; and / or (C) The method of any of the preceding embodiments, wherein the localized delivery of the drug delivery composition comprises localized delivery of a polymer component / solution and localized delivery of a dendrimer component / solution to the intracranial region in any order, sequentially, at least partially simultaneously, or a combination thereof.

[0101] Embodiment 27 The method of any of the preceding embodiments, wherein the injection of the drug delivery composition is performed during a biopsy, via an intraventricular shunt, or a combination thereof.

[0102] Embodiment 28 The method of any of the preceding embodiments, wherein the intracranial region comprises brain tissue, a brain tumor, the site of a previously resected tumor, or a combination thereof.

[0103] Embodiment 29. The method of any of the preceding embodiments, further comprising administering to the patient a second drug or therapy before, during, and / or after the local delivery of the drug delivery composition.

[0104] Embodiment 30 The method of any of the preceding embodiments, wherein administering the second drug or therapy comprises orally administering the second drug or therapy.

[0105] Embodiment 31. The method of any of the preceding embodiments, wherein the second drug or therapy comprises a second chemotherapy drug or therapy, including, but not limited to, any of those disclosed herein, such as any of those of embodiment 11.

[0106] Embodiment 32 The method of any of the preceding embodiments, wherein the second chemotherapy drug or therapy is not present in the drug delivery composition.

[0107] Embodiment 33 The method of any of the preceding embodiments, wherein the second chemotherapy drug or therapy comprises or consists of temozolomide.

[0108] Embodiment 34. The method of any of the preceding embodiments, wherein the second drug or therapy comprises an immunotherapy drug, such as any of those disclosed herein, e.g., any of those of embodiment 14 or 15.

[0109] Embodiment 35. The method of any of the preceding embodiments, wherein the second drug or therapy comprises aPD1 checkpoint blockade therapy.

[0110] Embodiment 36. The method of any of the preceding embodiments, wherein the drug is released from the drug delivery composition continuously for at least 12 hours, at least 24 hours, at least 36 hours, at least 48 hours, at least 72 hours, at least 100 hours, at least 200 hours, or at least 300 hours.

[0111] Embodiment 37. The method of any of the preceding embodiments, wherein the cumulative percentage of drug released from the drug delivery composition is at least 80% by weight within 10 days or less, 12 days or less, 14 days or less, 16 days or less, 18 days or less, 20 days or less, 22 days or less, 24 days or less, 26 days or less, 28 days or less, 30 days or less, 32 days or less, 34 days or less, 36 days or less, 38 days or less, 40 days or less, 50 days or less, 60 days or less, 80 days or less, or 100 days or less after local delivery of the drug delivery composition.

[0112] Embodiment 38. The method of any of the preceding embodiments, wherein the cumulative percentage of drug released from the drug delivery composition is at least 90% by weight within 10 days or less, 12 days or less, 14 days or less, 16 days or less, 18 days or less, 20 days or less, 22 days or less, 24 days or less, 26 days or less, 28 days or less, 30 days or less, 32 days or less, 34 days or less, 36 days or less, 38 days or less, 40 days or less, 50 days or less, 60 days or less, 80 days or less, or 100 days or less after local delivery of the drug delivery composition.

[0113] Embodiment 39. A kit for making a drug delivery composition, comprising: a first part comprising a first solution comprising the polymer component of any of the preceding embodiments; and a second part comprising a second solution comprising the dendrimer component of any of the preceding embodiments.

[0114] Embodiment 40. A kit according to any of the preceding embodiments, wherein at least one drug is disposed in the first solution, the second solution, or both the first solution and the second solution.

[0115] Embodiment 41. The kit of embodiment 40, wherein (i) the drug comprises or consists of a chemotherapy drug disposed in the first solution, the second solution, or both the first solution and the second solution; (ii) the drug comprises or consists of an immunotherapy drug disposed in the first solution, the second solution, or both the first solution and the second solution; or (iii) the drug comprises or consists of a chemotherapy drug and an immunotherapy drug both disposed in the first solution, both disposed in the second solution, both disposed in the first and second solutions, or the chemotherapy drug is disposed in one of the first and second solutions and the immunotherapy drug is disposed in the other.

[0116] Embodiment 42. A kit according to any of the preceding embodiments, wherein the kit comprises a syringe.

[0117] Embodiment 43 A kit according to any of the preceding embodiments, wherein the syringe includes a mixing tip, such as the mixing tip shown in FIG.

[0118] Embodiment 44. A kit according to any of the preceding embodiments, wherein the syringe comprises a first reservoir and a second reservoir, and the first solution and the second solution are disposed in the first reservoir and the second reservoir, respectively.

[0119] Embodiment 45. A composition, kit, or method of any of the preceding embodiments, wherein an aldehyde group of the polymer component reacts, e.g., reacts reversibly, with (i) a drug, e.g., doxorubicin, (ii) a dendrimer component, and / or (iii) one or more amines of one or more biological tissues to form an imine bond.

[0120] Embodiment 46 The composition, kit, or method of embodiment 45, wherein imine bond formation and imine bond hydrolysis controls or contributes to the release kinetics of the drug from the hydrogel.

[0121] Embodiment 47. The composition, kit or method of embodiment 45, wherein the formation of imine bonds and hydrolysis of the imine bonds achieves sustained delivery of the drug. EXAMPLES

[0122] The present invention is further illustrated by the following examples, which are not to be construed as imposing limitations on its scope in any way. On the contrary, it is clearly understood that after reading the description herein, resort may be made to various other aspects, embodiments, modifications, and equivalents thereof, which may suggest themselves to those skilled in the art, without departing from the spirit of the invention or the scope of the appended claims. Thus, other aspects of the present invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein.

[0123] Example 1 - Testing of Various Therapies The efficacy of several therapies against GBM cell lines was measured. Doxorubicin showed increased efficacy against GBM cell lines compared to standard of care chemotherapy. [Table 1]

[0124] FIG. 3 shows a plot of cell viability versus concentration for each of the therapies listed in the table above.

[0125] Studies have demonstrated that doxorubicin more potently eliminated GBM spheroids in vitro than other tested chemotherapies, however, other tested chemotherapies may be used with the methods and compositions described herein.

[0126] However, based on these studies, an initial screening of dextran-dendrimer hydrogels was performed. [Table 2]

[0127] Testing of the compositions in the preceding table revealed that Sample 5 had a faster degradation time than Sample 1, as shown in FIG.

[0128] Based on the results of the initial screening, several hydrogel formulations were manufactured and tested, including those listed in the table below. [Table 3]

[0129] The doxorubin release profile of the above compositions was examined, and as shown in Figures 5A and 5B, increasing the w / v % of dextran did not substantially affect the doxorubin release profile.

[0130] The studies in this example also revealed that altering the solids content of the compositions tested did not substantially affect the doxorubicin release profile, as shown in Figures 6A and 6B.

[0131] Further testing of this example also demonstrated that loading doxorubicin with dextran or dendrimer did not substantially alter the doxorubicin release profile, as shown in Figures 7A and 7B.

[0132] However, varying the amount of doxorubicin loaded into the above compositions slightly altered the doxorubicin release profile, as shown in Figures 8A and 8B.

[0133] However, each of the hydrogels tested in this example provided sustained release of doxorubicin, as shown in Figure 9. After 19 days, the "12% dendrimer, 10% dextran" hydrogel had a cumulative release percentage just below 90%, the "15% dendrimer, 12.5% ​​dextran" hydrogel had a cumulative release percentage above 90%, and the "18% dendrimer, 15% dextran" had a cumulative release percentage of 100%.

[0134] The release of primary amine capped doxorubicin was also tested. The tests showed that the capped doxorubicin achieved a higher cumulative release percentage than doxorubicin, as shown in Figure 10. The cumulative release percentages of Doxil and Doxorubicin were also compared. A greater cumulative percentage of doxorubicin was released over 200 hours, as shown in Figure 11.

[0135] The in vitro release of acriflavine was also tested. Injectable hydrogel formulations (75% oxidized dextran, 12.5% ​​w / v, PAMAM dendrimer, 15% w / v, pH 9.2) were prepared at four different loading concentrations (0.625 mg, 1.25 mg, 2.5 mg, and 5 mg, each of which had previously been injected locally in the brain). The cumulative release profiles were similar for the different loading amounts, as shown in Figures 12A and 12B. Acriflavine retained its activity upon release, as expected.

[0136] After injection of the example hydrogel into the brains of mice, magnetic resonance imaging (MRI) was used to assess the persistence of the hydrogel. The hydrogel persisted for at least 20 days, according to MRI scans collected at the time of injection and 2, 12, and 20 days thereafter.

[0137] In addition to their durability, the degradation of the hydrogels was also examined, specifically, in vivo hydrogel degradation by IVIS for formulations with different solids content. A plot of the mean radiant efficiency versus days from hydrogel injection is shown in FIG.

[0138] A therapy retention and tissue penetration study was designed and performed. The purpose of this study was to evaluate the retention, cellular uptake, and tissue penetration of doxorubicin upon release from the hydrogel and compare these to intratumoral injection of the therapy. The following table describes the study. [Table 4]

[0139] The results of this study showed that hydrogel delivery of doxorubicin improved retention of doxorubicin compared to intratumoral injection. Increased doxorubicin tissue penetration over time was also observed as doxorubicin was released from the hydrogel.

[0140] Further testing demonstrated that the solids content of the hydrogel formulation did not significantly affect the outcome of topical dox therapy, as shown in FIG.

[0141] Another study was designed and performed to evaluate whether sustained local delivery of doxorubicin using the hydrogel of this example could extend survival in orthotopic GL261 tumor-bearing mice. The table below describes the study, and the steps were applied to four groups (untreated control, empty hydrogel, intratumoral injection of doxorubicin, and hydrogel delivery of doxorubicin (N=6-10 per group, C57BL / 6 mice)). [Table 5]

[0142] This study demonstrated that local hydrogel delivery of doxorubicin significantly enhanced survival in orthotopic tumor-bearing mice compared to intratumoral (IT) injection, as shown in Figure 15. The results also demonstrated that the survival efficacy of hydrogel delivery of doxorubicin was dose-dependent, as shown in Figure 16. Figure 16 shows that survival probability was similar for control and empty hydrogels, and that hydrogels containing 15 μg of doxorubicin provided a greater survival probability than hydrogels containing 7.5 μg of doxorubicin.

[0143] Another study was designed and tested to determine whether local delivery of doxorubicin could enhance the efficacy of current oral temozolomide chemotherapy. The table below describes the study, which was applied to six groups (untreated control, empty hydrogel, oral temozolomide (TMZ), hydrogel delivery of doxorubicin, hydrogel delivery of doxorubicin + oral TMZ) (N=6-10 per group, C57BL / 6 mice). [Table 6]

[0144] The results demonstrated that local delivery of doxorubicin reduced tumor burden in mice bearing intracranial tumors, as shown in Figure 17. Tumor burden bioluminescence analysis was performed throughout the study and the data is presented in Figure 18.

[0145] For this example study, there was no significant benefit in median OS for mice receiving combined oral TMZ and hydrogen doxorubicin therapy, a conclusion based, at least in part, on the plot in FIG.

[0146] Comparative studies have shown that Doxil has comparable efficacy to doxorubicin in clearing glioblastoma cells in vitro, as evidenced by Figures 20A and 20B, which show cell viability following therapy. [Table 7]

[0147] Further testing showed that clinically available nanoparticle DOX (DOXIL® chemotherapy) resulted in increased survival when delivered to tumor-bearing mice using the hydrogel of this example. This conclusion was based, at least in part, on the data shown in Figure 21. As shown in Figure 22, clinically available nanoparticle DOX (DOXIL® chemotherapy) was as effective as hydrogel-free doxorubicin at half the concentration (7.5 μg).

[0148] The study was also designed to test whether local doxorubicin delivered via hydrogel could eradicate patient-derived xenograft orthotopic tumors in nude mice. The study, described in the table below, was applied to six groups (untreated control, empty hydrogel, oral TMZ, hydrogel delivery of doxorubicin (7.5 μg), hydrogel delivery of doxorubicin (15 μg), and hydrogel delivery of DOXIL® chemotherapy (7.5 μg) (N=6-10 per group, nude mice) (PDX characteristics: MGMT methylated, EGFR non-amplified, TP53 non-mutated). [Table 8]

[0149] The results of the study, shown in Figure 23, demonstrated that, as seen in the syngenic model, survival was significantly extended in orthotopic PDX tumor-bearing mice treated with hydrogel doxorubicin therapy. Sixty days after tumor inoculation, hydrogel delivery of DOXIL chemotherapy (7.5 μg) resulted in a survival probability of over 50%, while hydrogel delivery of doxorubicin (15 μg) had a survival probability just below 50%.

[0150] In this example, orthotopic PDX tumor-bearing NSG mice treated with various hydrogel doxorubicin regimens had significantly extended survival. These results are shown in FIG.

[0151] This example study also showed that there was no significant difference between PDX nude mice and GL261 C57BL6 mice survival (see FIG. 25A (DOXIL® chemotherapy hydrogel) (7.5 μg), FIG. 25B (doxorubicin hydrogel) (15 μg), FIG. 25C (doxorubicin hydrogel) (7.5 μg)). It should be noted that insignificant differences were obtained between nude and C57BL6 mice survival curves for all treatments, but different cells were used. These results suggested that adaptive immune responses may not play a significant role in eliminating tumors against the primary challenge.

[0152] Another study was performed to evaluate whether topical doxorubicin therapy conferred protection to long-term surviving mice against tumor rechallenge. This study followed the following parameters: [Table 9]

[0153] All long-term surviving syngenic mice treated with topical hydrogel therapy rejected tumors upon contralateral hemisphere rechallenge. This conclusion was based on the data presented in FIG.

[0154] Furthermore, long-term surviving PDX nude mice did not reject tumor upon contralateral hemisphere re-challenge, as shown in FIG. 27.

[0155] Another study was also performed to determine whether topical hydrogel doxorubicin chemotherapy could synergize with systemic aPD1 checkpoint blockade therapy. The dose in this study was 200 μg aPD1 administered intraperitoneally, and four groups were tested (untreated, systemic aPD1, intracranial doxorubicin hydrogel, intracranial doxorubicin hydrogel + systemic aPD1) (C57BL / 6 mice, N = 6–10 per group). [Table 10]

[0156] As shown in Figure 28, the combination of local doxorubicin therapy with systemic aPD1 did not enhance the survival of glioblastoma GL261-bearing mice. Without wishing to be bound by any particular theory, it was believed that one or more factors may be driving the lack of synergy between local doxorubicin chemotherapy and aPD1. For example, with regard to the drug release and biodistribution profile of the therapy, doxorubicin may reduce infiltrating T cells and / or systemic aPD1 may not reach the target tissue with the required pharmacokinetic profile. With regard to the immunosuppressive tumor microenvironment (TME), factors may include limited T cell infiltration and penetration into the TME with chemotherapy and / or chemotherapy cannot eliminate or restore immunosuppressive macrophages.

[0157] Primary tumor immunophenotyping - Five mice were studied for each treatment group (untreated, doxorubicin hydrogel) and mice were sacrificed 14 days after tumor inoculation (4 days after therapy). Tissues studied included tumor, spleen, bone marrow, blood, and tumor-draining lymph nodes. The panel for this study included memory T cells, macrophages, and dendritic cells / MDSC. [Table 11]

[0158] The majority of untreated tumors did not have significant numbers of immune cells for the T cell panel. CD45+ and CD3+ subsets of live cells were <1000 events in 3 / 5 untreated tumors. Different enrichment steps (magnetic beads, different Percoll gradients) were tested to increase the number of immune cells in the samples. A plot of total flux (photons / sec) versus days after tumor implantation is shown in Figure 29.

[0159] The increase in naive T cells in lymph nodes 4 days after therapy compared to untreated mice is shown in Figure 30. No other notable differences in T cells were observed in the tumor-draining lymph nodes. Only a slight decrease in central memory CD8 T cells was observed in the spleen, as shown in Figure 31. The only other difference in splenic cell immune infiltration was a slight decrease in CD45+ cells in the treatment group.

[0160] Macrophages contributed to the increase in CD45+ cells in tumors upon doxorubicin hydrogel treatment, as shown in Figure 32. A slight increase in the M1 polarization marker CD80+ in doxorubicin-treated tumors was observed, as shown in Figure 33.

[0161] Preliminary analysis suggested a significant shift in dendritic cell populations, as shown in Figure 34. However, it should be noted that the low numbers of DCs in untreated tumors (approximately 1000s) complicated the analysis of cDC / pDC populations.

[0162] Example 2 - Further testing and analysis STING (cGAS / stimulator of interferon genes agonist) is understood to drive anti-cancer immune activity through the production of type I interferon and other pro-inflammatory cytokines (Figure 35). Nanoparticles can be used in the cGAS-STING pathway. Cyclic dinucleotides (CDNs) can be potential cancer immunotherapy drugs, and nanoparticle-mediated delivery can be used (Figure 36).

[0163] It is hypothesized that local hydrogel delivery of combination chemoimmunotherapy may (1) modulate key components of the tumor microenvironment, (2) enhance antitumor immune priming, and / or (3) increase the efficacy of checkpoint blockade therapy (Figure 37).

[0164] Nanoparticle-based STING agonist formulations may have or facilitate high biodegradability, low toxicity, nucleic acid encapsulation (amine groups), high endosomal escape (good buffering capacity), and easy synthesis (high versatility) (Figure 38).

[0165] CDN nanoparticles stimulated dendritic cells and macrophages, as shown in Figure 39. Example hydrogel compositions mediated the release of CDN nanoparticles in vitro, as shown in Figures 40A and 40B.

[0166] The high hydrophilicity and negative charge of CDNs may hinder their delivery to cells and thus their clinical potential, limiting their delivery to tumors to intratumoral injection.CDNs are currently undergoing clinical trials, but there are concerns that simple intratumoral CDN injection is a suboptimal means to stimulate the cytoplasmic STING signaling pathway.However, this example describes a polymer-based CDN-conjugated nanoparticle that allows for systemic delivery of CDNs that would otherwise be cleared from circulation within minutes and programmed to be released in the cytoplasm, and its safety, efficacy, and mechanism of action have been tested in multiple mouse tumor models.

[0167] These NPs were made with PBAE modified with arginine residues that enhanced the biocompatibility and endosomal escape ability of the NPs and complexed with CDN-conjugated PBAE chains bearing cathepsin-sensitive bonds that allowed the release of the CDN in the cytoplasm.

[0168] A local immunotherapy MTD pilot study was designed and performed. The study parameters are displayed in the table below. GL261-luc tumor-bearing mice were studied, with each group containing 3 mice. Mice received 10 μL hydrogel injections and IVIS imaging was used to assess luciferase-tumor burden, AF568-hydrogel degradation, and AF647-therapy release. [Table 12] [Table 13]

[0169] The results of the localized immunotherapy pilot study are shown in Figure 41. IVIS bioluminescence-based tumor burden imaging (day 18) was collected for blank hydrogel, aPD-1 hydrogel (150 micrograms), aPD-L1 hydrogel (150 micrograms), and CDN dendrimer NP hydrogel (40 micrograms CDN).

[0170] It was determined that doxorubicin treatment increased surface expression of calreticulin on GBM cells but did not affect HMGB1 excretion at relevant concentrations. The results are shown in Figure 42. As shown in Figure 43, doxorubicin induced calreticulin (CRT) exposure, which indicates immunogenic cell death.

[0171] We also tested whether chemoimmunotherapy could synergize to enhance bone marrow-derived macrophage activation (Figure 44). As shown in Figure 45, BMDM CD 86 activation marker expression was enhanced by chemoimmunotherapy treatment. As shown in Figure 46, the combination therapy increased the ratio of pro-inflammatory to anti-inflammatory BMDM.

[0172] It was determined that the hydrogels of the aforementioned examples, including adhesive hydrogels, effectively delivered therapy locally to treat malignant brain tumors in mice. For example, hydrogel delivery of free or nanoparticle doxorubicin extended survival of mice bearing orthotopic syngenic tumors of PDX tumors. Also, 100% of long-term surviving mice treated with local hydrogel doxorubicin therapy rejected tumors upon rechallenge. Hydrogel delivery of CDN nanoparticles is another strategy for treating GBM alone and in combination with doxorubicin chemotherapy.

[0173] The compositions and methods described herein can allow for immune manipulation with materials that enhance therapeutic efficacy (Figure 47).

Claims

1. A hydrogel, wherein the hydrogel is an adhesive hydrogel, A drug dispersed in the hydrogel, The drug delivery composition comprises a drug, the drug being a chemotherapy drug, an immunotherapy drug, or a combination thereof. The drug delivery composition is configured to deliver the drug locally. The drug delivery composition.

2. The drug comprises the chemotherapy drug, and the chemotherapy drug comprises doxorubicin. The drug delivery composition according to claim 1, wherein the hydrogel comprises (i) a polymer component, the polymer component comprising a polymer having three or more aldehyde groups, and (ii) a dendrimer component, the dendrimer component comprising a dendrimer having at least two branches comprising one or more surface groups.

3. The aforementioned hydrogel A polymer component, wherein the polymer component includes a polymer having three or more aldehyde groups, The drug delivery composition according to claim 1, comprising a dendrimer component, wherein the dendrimer component includes a dendrimer having at least two branches containing one or more surface groups.

4. The drug delivery composition according to claim 3, wherein 100% of the one or more surface groups comprises at least one primary or secondary amine.

5. The drug delivery composition according to claim 3, wherein less than 75% of the one or more surface groups comprises at least one primary or secondary amine.

6. The drug delivery composition according to claim 1, wherein the dendrimer is a fifth-generation polyamidoamine (G5 PAMAM) dendrimer.

7. The drug delivery composition according to claim 1, wherein the polymer comprises dextran.

8. The drug delivery composition according to claim 1, wherein the chemotherapy drug comprises doxorubicin, and the doxorubicin is encapsulated in a liquid sphere.

9. The drug delivery composition according to claim 8, wherein the liquid sphere is a liquid nanosphere.

10. The drug delivery composition according to claim 1, wherein the immunotherapy drug comprises a cyclic dinucleotide.

11. The cyclic dinucleotide is encapsulated in nanoparticles, and the nanoparticles are (i) comprising a poly-beta-aminoester in which the cyclic dinucleotide is conjugated via a cathepsin-sensitive bond, wherein the poly-beta-aminoester is optionally modified with arginine, (ii) Having an average diameter of approximately 30 nm to approximately 70 nm, or (iii) A combination thereof, the drug delivery composition according to claim 10.

12. The drug delivery composition according to claim 1, wherein the drug delivery composition has a solid content of about 8% to about 25% by weight.

13. The drug delivery composition according to claim 1, wherein the hydrogel comprises phosphate-buffered saline (PBS).

14. The drug delivery composition according to claim 1, wherein the drug is present in the drug delivery composition in (i) a total amount of about 50 μg to about 500 μg, (ii) a total concentration of about 1 μg / μL to about 10 μg / μL, or (iii) a combination thereof.

15. The drug delivery composition according to claim 3, wherein the three or more aldehyde groups of the polymer reversibly react with (i) the drug, (ii) the dendrimer, and / or (iii) one or more amines of one or more biological tissues to form imine bonds, and the formation of the imine bonds and the hydrolysis of the imine bonds control or contribute to the release kinetics of the drug from the hydrogel.

16. A kit for preparing a drug delivery composition, wherein the kit is A first part comprising a first solution containing a polymer component, wherein the polymer component comprises a polymer, A second part comprising a second solution containing a dendrimer component, wherein the dendrimer component includes a dendrimer having at least two branches containing one or more surface groups, At least one drug is present in the first solution, the second solution, or both the first and second solutions. A kit comprising the drug a chemotherapy drug, an immunotherapy drug, or a combination thereof.

17. The kit according to claim 16, wherein the chemotherapy drug comprises doxorubicin.

18. The kit according to claim 17, wherein the doxorubicin is enclosed in a liquid sphere.

19. The kit according to claim 18, wherein the liquid sphere is a liquid nanosphere.

20. The kit according to any one of claims 16 to 19, wherein the immunotherapy drug comprises a cyclic dinucleotide.

21. The kit according to claim 20, wherein the cyclic dinucleotide is encapsulated in nanoparticles having an average diameter of about 30 nm to about 70 nm.

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