Compositions and methods for targeting tumor-associated macrophages

Abstract

Provided herein are methods for treating a disease or disorder, comprising administering to a subject a composition comprising a compound, the compound comprising a targeting moiety, a glucan backbone, and an active ingredient. Also provided herein are methods for targeting monocytes, macrophages, dendritic cells, and other cells that recruit to disease sites, comprising administering to a subject a composition comprising a compound, the compound comprising a targeting moiety, a glucan backbone, and an active ingredient.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority from U.S. Provisional Patent Application No. 63 / 447,628, entitled "COMPOSITIONS AND METHODS FOR TARGETING TUMOR-ASSOCIATED MACROPHAGES," filed February 22, 2023, the contents of which are incorporated by reference in their entirety.

[0002] The present disclosure provides a method for treating a disease or disorder, comprising administering to a subject a composition comprising a compound, the compound comprising a targeting moiety, a glucan backbone, and an active ingredient. The present disclosure also provides a method for targeting monocytes, macrophages, dendritic cells, and other cells that recruit to disease sites, comprising administering to a subject a composition comprising a compound, the compound comprising a targeting moiety, a glucan backbone, and an active ingredient. [Background technology]

[0003] Treatment of solid tumors has improved in recent years with the advent of a wide range of targeted therapies, including small molecules and biologics. While small molecules have excellent solid tumor penetration, many of them act by inhibiting signaling pathways, resulting in growth inhibition, rather than direct tumor cytotoxicity. If the entire tumor is not killed, cancer cells may develop resistance to small molecules. Biologics, such as antibodies and antibody-drug conjugates (ADCs), can have significant efficacy against certain malignancies and favorable blood stability. However, antibodies have limited solid tumor penetration, and tumor cells may develop resistance to ADCs through various cellular modifications (Collins, et al., Acquired Resistance to Antibody-Drug Conjugates. Cancers (Basel) 11, (2019)). There is a need for improved methods for targeting, penetrating, and killing cancer cells in solid tumors. Embodiments are provided to meet this need. Summary of the Invention

[0004] Provided herein is a method for treating cancer in a subject, comprising administering to the subject having cancer a composition comprising a compound, the compound comprising: i) a tumor-associated macrophage (TAM) targeting moiety; ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient, the active ingredient being attached to the glucan backbone; and iv) a targeting linker connecting the targeting moiety to the glucan backbone, the targeting linker comprising a carbamate group and a chain moiety, the carbamate group being connected to the backbone monomer and the chain moiety connecting the carbamate group to the tumor-associated macrophage targeting moiety.

[0005] In some embodiments, the plurality of backbone monomers comprises a plurality of D-glucose monomers linked by α-1,6 glycosidic bonds or β-1,4 glycosidic bonds. In some embodiments, the plurality of D-glucose monomers is n, where n=5 to 167. In some of the embodiments, the plurality of D-glucose monomers is n, where n=50 to 65. In some of the embodiments, the glucan backbone is a linear dextran molecule. In some of the embodiments, the glucan backbone is a cyclodextrin molecule, where n=6 to 16.

[0006] In some of any of the embodiments, the tumor-associated macrophage targeting moiety comprises mannose, galactose, collagen, fucose, sulfated N-acetylgalactosamine, N-acetylglucosamine, luteinizing hormone, thyroid-stimulating hormone, phospholipase A2 or a fragment thereof, or chondroitin sulfate. In some of any of the embodiments, the targeting moiety is mannose. In some embodiments, the ratio of mannose to backbone monomer is from about 1:5 to about 1:33. In some of any of the embodiments, the ratio of mannose to backbone monomer is from about 1:6 to about 1:19. In some of any of the embodiments, the degree of substitution of mannose on the cyclodextrin ranges from about 0.1 to about 7. In some of any of the embodiments, the degree of substitution of mannose on the cyclodextrin ranges from about 0.5 to about 5.

[0007] In some of the embodiments, the targeting linker is connected to the glucan backbone via the oxygen atom of the carbamate group. In some of the embodiments, the chain portion of the targeting linker comprises a C3-C7 alkylene chain. In some of the embodiments, the chain portion of the targeting linker comprises a C6-alkylene moiety. In some of the embodiments, the chain portion of the targeting linker is an unsubstituted C6-alkylene moiety. In some of the embodiments, the carbon atom of the carbamate group of the targeting linker is the only sp2 hybridized carbon when the linker is attached to mannose. In some of the embodiments, the tumor-associated macrophage targeting moiety is a moiety that targets CD205 (DEC205), CD206, CD207 (Langerin), CD209 (DC-SIGN), CD280 (ENDO180), or CD301 (MGL). In some embodiments, the tumor-associated macrophage targeting moiety is a CD206 targeting moiety.

[0008] In some of the embodiments, the compounds have a molar ratio of TAM targeting moiety to active ingredient of about 1:1 to about 1:10. In some of the embodiments, the molar ratio of TAM targeting moiety to active ingredient is about 1:1, about 1:2, or about 1:3. In some of the embodiments, the active ingredient is attached to the glucan backbone via a payload linker. In some of the embodiments, the active ingredient is a cytotoxic drug. In some embodiments, the cytotoxic agent is selected from the group consisting of auristatin, dolastatin, auristatin E, monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), dimethylvaline-valine-dolaisoloiin-dolaproine-phenylalanine-p-phenylenediamine (AFP), 5-benzoylvaleric acid-auristatin E ester (AEVB), auristatin EB (AEB), ansamitocin, ivlertansine / emtansine (DMI), ravtansine / soravtansine (DM4), duocarmycin, calicheamicin, and pyrrolobenzodiazepine. In some embodiments, the cytotoxic agent is MMAE.

[0009] In some embodiments, the payload linker is a non-cleavable linker. In some embodiments, the payload linker comprises a carbamate group and a chain moiety, wherein the carbamate group is attached to the backbone monomer and the chain moiety connects the carbamate group to the active ingredient. In some embodiments, the payload linker is a cleavable linker. In some embodiments, the cleavable linker is cleavable by a protease. In some embodiments, the protease is a lysosomal protease or an endosomal protease. In some embodiments, the cleavable linker is cleavable by a pH change. In some embodiments, the payload linker comprises a Val-Cit moiety.

[0010] In some of any of the embodiments, the cancer is a solid tumor selected from the group consisting of carcinoma, lymphoma, blastoma, sarcoma, glioma, leukemia, lymphoid malignancy, squamous cell carcinoma, squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, peritoneal cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, gallbladder cancer, hepatoma, breast cancer, colon, rectal cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, renal cancer, prostate cancer, vulvar cancer, thyroid cancer, hepatic carcinoma, anal cancer, penile cancer, and head and neck cancer. In some embodiments, the solid tumor is a sarcoma or glioblastoma. In some embodiments, the cancer is a soft tissue sarcoma. In some embodiments, the cancer is undifferentiated pleomorphic sarcoma (UPS).

[0011] In some of any of the embodiments, the subject and / or cancer comprises a cell population (a) resistant to temozolomide (TMZ) and / or doxorubicin, and / or (b) resistant to inhibition by TMZ and / or doxorubicin. In some of any of the embodiments, the composition is administered to the subject 6 times a day, 5 times a day, 4 times a day, 3 times a day, 2 times a day, once a day, every other day, 3 times a week, twice a week, at least once a week, once a week, every two weeks, once every three weeks, once a month, once every two months, or once a month. In some embodiments, the composition is administered to the subject every two weeks, once every three weeks, or once a month.

[0012] In some of any of the embodiments, the method further comprises administering to the subject one or more additional therapeutic agent(s), adjuvant therapy, and / or radiation therapy. In some embodiments, the one or more additional active agents are selected from the group consisting of chemotherapeutic agents, DNA hypomethylating agents, alkylating agents, topoisomerase inhibitors, therapeutic antibodies that specifically bind to cancer antigens, hematopoietic growth factors, cytokines, antibiotics, COX-2 inhibitors, CDK inhibitors, immunomodulators, antithymocyte globulin, immunosuppressants, and corticosteroids or pharmacological derivatives thereof. In some embodiments, the one or more additional active agent(s) is a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is TMZ, doxorubicin, and / or paclitaxel. In some of any of the embodiments, the chemotherapeutic agent is administered prior to, concurrently with, and / or after initiation of administration of the composition to the subject. In some embodiments, the adjuvant therapy and / or radiation therapy is administered prior to, concurrently with, and / or after initiation of administration of the composition to the subject.

[0013] In some of the embodiments, the composition is administered to the subject intravenously, intraperitoneally, subcutaneously, intramuscularly, intracranially, or by pump infusion. In some embodiments, the composition is administered to the subject intravenously or by pump infusion.

[0014] Provided herein is a method for treating glioblastoma in a subject, comprising administering a composition to the subject having glioblastoma, the composition comprising: i) a tumor-associated macrophage (TAM) targeting moiety; ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient comprising MMAE, the active ingredient being attached to the glucan backbone; and iv) a targeting linker connecting the targeting moiety to the glucan backbone, the targeting linker comprising a carbamate group and a chain moiety, the carbamate group being connected to the backbone monomer and the chain moiety connecting the carbamate group to the tumor-associated macrophage targeting moiety; and the subject and / or glioblastoma comprises a cell population that is (a) resistant to TMZ and / or (b) resistant to inhibition by TMZ.

[0015] Provided herein is a method for treating brain metastases in a subject, comprising administering a composition to the subject having brain metastases, the composition comprising: i) a tumor-associated macrophage (TAM) targeting moiety; ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient comprising MMAE, the active ingredient being attached to the glucan backbone; and iv) a targeting linker connecting the targeting moiety to the glucan backbone, the targeting linker comprising a carbamate group and a chain moiety, the carbamate group being connected to the backbone monomer and the chain moiety connecting the carbamate group to the tumor-associated macrophage targeting moiety; and the subject and / or brain metastases comprising: (a) TMZ-resistant and / or (b) a cell population resistant to inhibition by TMZ.

[0016] Provided herein are methods for treating UPS in a subject, comprising administering a composition to the subject having UPS, the composition comprising: i) a tumor-associated macrophage (TAM) targeting moiety; ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient comprising MMAE, the active ingredient being attached to the glucan backbone; and iv) a targeting linker connecting the targeting moiety to the glucan backbone, the targeting linker comprising a carbamate group and a chain moiety, the carbamate group being connected to the backbone monomer and the chain moiety connecting the carbamate group to the tumor-associated macrophage targeting moiety; and wherein the subject and / or UPS is (a) resistant to doxorubicin and / or (b) comprises a cell population resistant to doxorubicin.

[0017] In some of any of the embodiments, the plurality of backbone monomers comprises a plurality of D-glucose monomers linked by α-1,6 glycosidic bonds or β-1,4 glycosidic bonds. In some of any of the embodiments, the plurality of D-glucose monomers is n, where n=5 to 167. In some of any of the embodiments, the plurality of D-glucose monomers is n, where n=50 to 65. In some of any of the embodiments, the glucan backbone is a linear dextran molecule. In some of any of the embodiments, the glucan backbone is a cyclodextrin molecule, where n=6 to 16.

[0018] In some of the embodiments, the tumor-associated macrophage targeting moiety comprises mannose, galactose, collagen, fucose, sulfated N-acetylgalactosamine, N-acetylglucosamine, luteinizing hormone, thyroid-stimulating hormone, phospholipase A2 or a fragment thereof, or chondroitin sulfate. In some of the embodiments, the targeting moiety is mannose. In some embodiments, the ratio of mannose to backbone monomer is about 1:5 to about 1:33. In some of the embodiments, the ratio of mannose to backbone monomer is about 1:6 to about 1:33. In some of the embodiments, the degree of substitution of mannose on the cyclodextrin ranges from about 0.1 to about 7. In some of the embodiments, the degree of substitution of mannose on the cyclodextrin ranges from about 0.5 to 5. In some of the embodiments, the targeting linker is attached to the glucan backbone via the oxygen atom of the carbamate group. In some of any of the embodiments, the chain portion of the targeting linker comprises a C3-C7 alkylene chain. In some of the embodiments, the chain portion of the targeting linker comprises a C6-alkylene moiety. In some of the embodiments, the chain portion of the targeting linker is an unsubstituted C6-alkylene moiety.

[0019] In some of the embodiments, the carbon atom of the carbamate group of the targeting linker is the only sp2 hybridized carbon when the linker is attached to mannose. In some of the embodiments, the tumor-associated macrophage targeting moiety is a moiety that targets CD205 (DEC205), CD206, CD207 (Langerin), CD209 (DC-SIGN), CD280 (ENDO180), or CD301 (MGL). In some embodiments, the tumor-associated macrophage targeting moiety is a CD206 targeting moiety. In some of the embodiments, the compound has a molar ratio of TAM targeting moiety to active ingredient of about 1:1 to about 1:10. In some of the embodiments, the molar ratio of TAM targeting moiety to active ingredient is about 1:1, about 1:2, or about 1:3.

[0020] In some of these embodiments, the active ingredient is connected to the glucan backbone via a payload linker. In some embodiments, the payload linker is a non-cleavable linker. In some embodiments, the payload linker comprises a carbamate group and a chain moiety, wherein the carbamate group is connected to a backbone monomer and the chain moiety connects the carbamate group to the active ingredient. In some embodiments, the payload linker is a cleavable linker. In some embodiments, the cleavable linker is cleavable by a protease. In some embodiments, the protease is a lysosomal protease or an endosomal protease. In some embodiments, the cleavable linker is cleavable by a pH change. In some embodiments, the payload linker comprises a Val-Cit moiety.

[0021] In some embodiments, the method further comprises administering radiation therapy to the subject. In some embodiments, the method further comprises administering TMZ and / or doxorubicin to the subject. In some embodiments, the method attenuates tumor growth in the subject. In some embodiments, the subject is immunocompetent or immunocompromised. In some embodiments, the method includes: i) the tumor-associated macrophage (TAM) targeting moiety is a CD206 targeting moiety; and ii) the active ingredient is attached to the glucan backbone via a payload linker, wherein the active ingredient is MMAE and the payload linker is a Val-Cit linker. In some embodiments, the method includes: i) the tumor-associated macrophage (TAM) targeting moiety is a CD206 targeting moiety; and ii) the active ingredient is attached to the glucan backbone via a payload linker, wherein the active ingredient is MMAE and the payload linker is a Val-Cit linker. In some of any of the embodiments, the methods include administering Compound A to the subject.

[0022] Provided herein is a kit comprising a composition comprising a compound and instructions for administering the compound to a subject to treat cancer, the compound comprising: i) a tumor-associated macrophage (TAM) targeting moiety; ii) a glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient, the active ingredient being attached to the glucan backbone; and iv) a targeting linker connecting the targeting moiety to the glucan backbone, the targeting linker comprising a carbamate group and a chain moiety, the carbamate group being connected to the backbone monomer and the chain moiety connecting the carbamate group to the tumor-associated macrophage targeting moiety. [Brief explanation of the drawings]

[0023] [Figure 1] 1 is a diagrammatic representation of a candidate compound in which the tumor-associated macrophage (TAM) targeting moiety is labeled with MMAE. [Figure 2] A and B show the therapeutic effect of Target 5 on tumor volume (A) and body weight (B) in a subcutaneous HT1080 fibrosarcoma model. [Figure 3] 1 shows the therapeutic effect of Target 5 on tumor size (measured by bioluminescence) upon intracranial challenge with HT1080 cells in the STS brain metastasis model. [Figure 4A] Figure 1 shows the efficacy of Target 5 in doxorubicin-resistant UPS PDX models. Figure 2 shows the effect of Target-5 on tumor volume. [Figure 4B] Figure 1 shows the efficacy of Target 5 in doxorubicin-resistant UPS PDX models. Figure 2 shows the effect of Target-5 on tumor volume. [Figure 4C] Figure 1 shows the efficacy of Target 5 in a doxorubicin-resistant UPS PDX model. Figure 2 shows the effect of Target-5 on tumor volume compared to doxorubicin. [Figure 4D] Shows the efficacy of Target 5 in doxorubicin-resistant UPS PDX models. Representative tumors were excised from mice in each treatment group. [Figure 5]A, B show the efficacy of Target 5 on tumor volume (A) and body weight (B) in a subcutaneous glioma model (U87MG cell line). [Figure 6-1] The efficacy of Target 5 on tumor volume (A) and body weight (B) in immune-competent mice subcutaneously implanted with GL261 tumor cells (moderately resistant to temozolomide) is shown. [Figure 6-2] 1 shows the efficacy of Target 5 on tumor volume in immune-competent mice subcutaneously implanted with GL261 tumor cells (moderately resistant to temozolomide). [Figure 7] 1 shows plasma concentrations of free MMAE in mice after administration of Target-5. [Figure 8] A, B. Target-5 exhibits anti-cancer efficacy in a treatment-naive PDX model of undifferentiated pleomorphic sarcoma. Mean tumor volume (A) and body weight (B) are shown. [Figure 9] A (tumor volume) and B (histology of resected tumor) show that Target-5 exhibits anti-cancer efficacy, whereas compound B (Target-5 without MMAE) has no anti-cancer activity. [Figure 10] Demonstrates the anti-cancer efficacy of Target-5 compared to doxorubicin in a myxofibrosarcoma PDX model. DETAILED DESCRIPTION OF THE INVENTION

[0024] Provided herein are methods for treating a disease or disorder, the methods comprising administering to a subject a composition comprising a compound, the compound comprising a targeting moiety, a glucan backbone, and an active ingredient. In some embodiments, the targeting moiety is a tumor-associated macrophage targeting moiety. In some embodiments, the methods comprise administering a compound that targets monocytes, macrophages, and other cells (such as dendritic cells), particularly cells that recruit to disease sites. In some embodiments, the targeting moiety (e.g., tumor-associated macrophage targeting moiety) is attached to the glucan backbone. In certain embodiments, the methods comprise administering a composition comprising a compound, the compound comprising a glucan backbone, a targeting moiety, a targeting moiety linker, a payload, and optionally a payload linker.

[0025] CD206 + Cells, particularly macrophages, have been targeted with a variety of molecules with the aim of delivering diagnostic and therapeutic agents to sites where such cells congregate. One example of such a molecule can be found in US2017 / 0209584, entitled "Compositions for Targeting Macrophages and Other CD206 High Expressing Cells and Methods of Treating and Diagnosis." The molecules disclosed in this reference and others target the CD206 of interest. + Although capable of targeting cells, these molecules have a number of drawbacks.

[0026] Soft tissue sarcoma (STS) is a rare but deadly cancer in children and adults. The American Cancer Society estimates that there are approximately 13,000 new cases of STS annually in the United States, with approximately 5,130 expected deaths and a 5-year survival rate of only 16% for metastatic disease. Traditional treatments, including surgery, chemotherapy, and radiation therapy, have limited success in treating STS. For example, undifferentiated pleomorphic sarcoma (UPS), a highly aggressive adult sarcoma, has a median overall survival of only 15.5 months for metastatic UPS. Ewing sarcoma / primitive neuroectodermal tumor (PNET), an aggressive sarcoma associated with adolescents, has a 5-year survival rate of only 15% for metastatic cases.

[0027] Doxorubicin (DOX) is one of the most active drugs available for the treatment of sarcomas and is often the first-line treatment for undifferentiated / unclassified STS, but doxorubicin's cardiotoxicity can be dose-limiting. Response rates to a single doxorubicin treatment are significantly reduced after drug exposure, indicating that increased resistance to doxorubicin can lead to treatment failure (Das, B. et al. Commun Biol 4, 1312 (2021)).

[0028] Glioblastoma multiforme (GBM) is a severe and almost always fatal cancer. Treatment includes surgery, radiation, and temozolomide (TMZ), followed by 6–12 months of maintenance TMZ. Despite maximal treatment, 50% of patients with a confirmed GBM diagnosis die within 12–15 months of diagnosis. Treatment is labor-intensive and often carries the additional burden of worsening neurological deficits. These symptoms can impose a significant burden on patients and their families. Since TMZ was approved in 2005, no new pharmacologic therapies have been approved to extend survival for GBM. Even with the addition of the alternating current electric field electrode device, Optune (Novocure, St. Helier, Jersey), the 5-year survival rate for GBM is only 13% (Stupp et al., JAMA. 2017;318(23):2306–2316).

[0029] Although TMZ is a first-line chemotherapy drug for the treatment of GBM, tumor resistance and recurrence are nearly inevitable. Due to widespread exposure to TMZ and the heterogeneous and mutation-prone nature of GBM, it is very common for GBM tumors to develop resistance to TMZ. Currently, more than 50% of GBM patients treated with TMZ do not respond to this therapy (Singh et al. Cancer Drug Resist. 2021;4(1):17-43). There is a great need for new treatments with limited toxicity to treat drug-resistant cancers (e.g., soft tissue sarcomas and glioblastomas).

[0030] New tumor-agnostic approaches are needed to overcome the limitations of current state-of-the-art technologies and further improve treatment outcomes. For example, STS includes more than 50 different histological and molecular subtypes, each of which exhibits diverse clinical behaviors (Katz et al., "More Than 50 Subtypes of Soft Tissue Sarcoma: Paving the Path for Histology-Driven Treatments," Am Soc Clin Oncol Educ Book, 2018, 38, 925-938). Partly due to this diversity, the efficacy of current treatment options is limited, and no single-agent or combination treatments consistently and effectively treat all STS subtypes.

[0031] Tumors are often characterized by a high proportion of tumor-associated macrophages (TAMs), often representing 30–50% or more of the total cells within a tumor (Vinogradov et al., Nanomedicine (Lond). 2014 Apr;9(5):695-707). Targeting TAMs as receiver cells for delivery of tumor-killing payloads has the advantage of enabling localized delivery of anticancer therapies without adversely affecting TAMs. This approach not only avoids the difficulty of targeting heterogeneous cancer cells, but also circumvents the evolved resistance of most types of cancer cells. Provided herein is a treatment method that does not require consideration of tumor heterogeneity and the evolved drug resistance associated with current therapeutics (e.g., doxorubicin, temozolomide). Provided herein is a treatment method that does not require consideration of the specificities of each cancer type.

[0032] All publications, including patent documents, scientific articles, and databases, referenced in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. To the extent that a definition set forth herein conflicts or contradicts a definition set forth in a patent, application, publication, or other publication incorporated herein by reference, the definition set forth herein shall control over the definition incorporated herein by reference.

[0033] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0034] I. Compound Provided herein are methods for treating a disease or disorder (e.g., cancer) in a subject, the methods comprising administering a composition comprising a compound. In some embodiments, the compound comprises a glucan backbone, a targeting moiety, a targeting moiety linker, a payload, and an optional payload linker. In some embodiments, the arrangement of these components results in a compound that targets tumor-associated macrophages (TAMs). In some embodiments, the methods disclosed herein utilize TAMs as receiver cells for picking up, processing, and delivering the payload to the tumor environment. For example, sarcomas are characterized by abundant tumor-associated macrophages (TAMs) (Fujiwara et al., "Role of tumor-associated macrophages in sarcomas," Cancers (Basel), 2021, 13(5):1086). Anti-CD206 immunohistochemistry on a human sarcoma tissue array containing 59 specimens encompassing 19 sarcoma subtypes has shown high and relatively uniform expression of this TAM receptor in essentially all specimens.

[0035] In some embodiments, the method involves detecting a cell present in a tumor-associated macrophage (e.g., CD206 + This includes internalization of administered compounds by tumor-associated macrophage (TCM) cells. By being able to internalize into cells present in tumor-associated macrophages, administered compounds and compositions can deliver payloads to disease sites where such cells are concentrated, such as solid tumor cancers and granulomatous diseases.

[0036] In some embodiments, the methods disclosed herein involve administering a composition comprising a compound that is larger than typical small molecules but smaller than antibody-drug conjugates, allowing for excellent penetration to the targeted site with minimal leakage into normal tissues, thereby limiting potential toxicity. Furthermore, while rapidly dividing and mutating heterogeneous cancer cells often develop resistance to conventional therapies, terminally differentiated macrophages are not subject to the selective pressure to develop resistance to the methods of treatment disclosed herein.

[0037] A. Glucan backbone In certain embodiments, the methods described herein involve administering a compound comprising a glucan backbone, which is a linear, branched, or cyclic oligosaccharide or polysaccharide comprising multiple glucose monomers linked primarily by C-1→C-6 glycosidic bonds. In certain embodiments, other glycosidic linkages, such as α-1,3 or α-1,4 linkages, may also be present. In some embodiments, the multiple glucose monomers are linked by α-1,6 and α-1,3 glycosidic linkages. In some embodiments, the multiple glucose monomers are linked by a combination of α-1,6 and α-1,4 glycosidic linkages. A glucan backbone can also be defined as a polymer of glucose with different glycosidic linkage positions.

[0038] In some embodiments, a glucan backbone can be defined as a polymer of glucose with different glycosidic bond positions. In some embodiments, a glucan backbone can include alpha or beta isomers of glucose. Glucan backbones include, but are not limited to, dextrans, which are linear or branched compounds, and cyclodextrins, which are cyclic glucans.

[0039] The mass and molecular weight of the glucan backbone can vary, depending in part on the number of glucose monomers. In some embodiments, the glucan backbone can have a molecular weight ranging from 1 to 30 kilodaltons (kDa). Preferred embodiments include glucan backbones of approximately 1 kDa, 3 kDa, 6 kDa, 10 kDa, 20 kDa, or 30 kDa. In some embodiments, the glucan backbone can have a molecular mass ranging from 1,000 to 30,000 grams per mole (g / mol). In some embodiments, the glucan backbone can contain a number of glucose monomers ranging from 5 to 167. The glucan backbone can be linear, branched, cyclic, or a combination thereof. For example, dextran is an example of a linear or branched glucan backbone. Cyclodextrin is another example of a glucan backbone. The backbones described herein can be substituted or unsubstituted. For example, a substituted cyclodextrin is a cyclodextrin derivative that is hydrophobic, hydrophilic, ionic, non-ionic, or any other variation thereof.

[0040] In some embodiments, the glucan backbone comprises a plurality of backbone monomers, the plurality of backbone monomers comprising a plurality of D-glucose monomers linked via α-1,6 glycosidic bonds or β-1,4 glycosidic bonds. In certain embodiments, the plurality of backbone monomers comprises a plurality of D-glucose monomers linked via β-1,4 glycosidic bonds. In some embodiments, the plurality of D-glucose monomers is n, where n=16-111. In some embodiments, n=50-65. In some embodiments, the plurality of D-glucose monomers is n, where n=5-167. In some embodiments, the glucan backbone is a linear dextran molecule. In some embodiments, the glucan backbone is a cyclodextrin molecule comprising 6-16 D-glucose monomers.

[0041] B. Targeting part In certain embodiments, the methods described herein involve administering a compound comprising a targeting moiety attached to a glucan backbone. In some embodiments, the targeting moiety is a tumor-associated macrophage targeting moiety. In some embodiments, the targeting moiety binds to a receptor present on or near a tumor, including, but not limited to, CD205 (DEC205), CD206, CD207 (Langerin), CD209 (DC-SIGN), CD280 (ENDO180), or CD301 (MGL), allowing targeted delivery of a payload to the tumor. In some embodiments, the targeting moiety is a CD205 targeting moiety. In some embodiments, the targeting moiety is a CD206 targeting moiety. In some embodiments, the targeting moiety is a CD207 targeting moiety. In some embodiments, the targeting moiety is a CD209 targeting moiety. In some embodiments, the targeting moiety is a CD280 targeting moiety. In some embodiments, the targeting moiety is a CD301 targeting moiety. In some embodiments, the targeting moiety is a CD206 ligand.

[0042] In some embodiments, the target receptor is located in tumor-associated macrophages. In some embodiments, the target receptor is located in cancer or tumor cells. The targeting moiety can be a molecule, compound, structure, or any combination thereof that targets one or more pattern recognition receptors in tumor-associated macrophages or cancer or tumor cells. The targeting moiety can target pattern recognition receptors also characterized as C-type lectin receptors. In certain embodiments, the targeting moiety targets CD206, the mannose receptor.

[0043] In some embodiments, the targeting moiety comprises mannose, galactose, collagen, fucose, sulfated N-acetylgalactosamine, N-acetylglucosamine, luteinizing hormone, thyroid-stimulating hormone, phospholipase A2 or a fragment thereof, or chondroitin sulfate. In some embodiments, the targeting moiety comprises mannose (including its D- and L-isomers). In some embodiments, the targeting moiety comprises a furanose. In some embodiments, the targeting moiety comprises a pyranose. In some embodiments, the targeting moiety is D-mannose. It should be understood that a tumor-associated macrophage targeting moiety may also be referred to as a targeting moiety.

[0044] In some embodiments, the targeting moieties are attached to about 10% to about 50% of the glucose residues in the glucan backbone, or about 20% to about 45% of the glucose residues, or about 25% to about 40% of the glucose residues.

[0045] C. Targeting Linker to Scaffold Ratio The density of targeting moieties per backbone subunit is expressed using the ratio of targeting moieties to backbone subunits for linear, branched, or cyclic polysaccharide backbones. For example, the degree of substitution (ds) is used to express the density of targeting moieties on a glucan backbone. The ratio of targeting moieties to glucan backbone refers to the number of targeting moieties substituting one or more backbone subunits. For example, a ratio of 1:7 means that there is one targeting moiety for every seven glucose subunits in the glucan backbone. ds represents the average number of substitutions or substitution positions per base unit. For example, a ds of 0.9 means that one backbone subunit is substituted with an average of 0.9 targeting moieties. In some embodiments, the ratio of targeting moieties to backbone subunits is about 1:5 to about 1:25. In some embodiments, the ratio of targeting moiety to backbone subunit is at least 1:50 (e.g., at least 1:33, at least 1:35, at least 1:40, or at least 1:45) to about 1:5. In some embodiments, the ratio of targeting moiety to backbone subunit is about 1:6 to about 1:19. In some embodiments, the ratio of targeting moiety to backbone subunit is 1:1 or about 1:1. In some embodiments, the ratio of targeting moiety to backbone subunit is 2:1 or about 2:1. In some embodiments, the ratio of targeting moiety to backbone subunit is 3:1 or about 3:1. In some embodiments, ds is about 0.1 to about 7. In some embodiments, ds is about 0.5 to 5. In some embodiments, in conjunction with the above or below embodiments, the targeting moiety comprises mannose.

[0046] D. Targeting Linkers In certain embodiments, the methods described herein include administering a compound comprising a targeting moiety attached to a glucan backbone by a targeting linker. In some embodiments, the targeting linker is a cleavable or non-cleavable linker. In some embodiments, the cleavable linker can be cleaved by an enzyme (e.g., a protease), a change in temperature, a change in pH, a chemical stimulus, or any combination thereof. In some embodiments, the cleavable linker can include a protease cleavage site. In some embodiments, the cleavable linker can be cleaved by a lysosomal protease or an endosomal protease.

[0047] In some embodiments, the targeting linker can include a carbamate group. In some embodiments, the targeting linker includes a carbamate group and a chain moiety, where the carbamate group is connected to the backbone monomer and the chain moiety connects the carbamate group to the targeting moiety. As defined herein, the carbamate functional group has its plain and ordinary meaning derived from the field of organic chemistry. In some embodiments, the chain moiety of the targeting linker includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) units selected from the group consisting of an optionally substituted alkylene chain, an optionally substituted CO-alkylene chain, a peptide chain, a polymer chain, and a heteroatom selected from the group consisting of an O atom, an S atom, and an optionally substituted N atom. In some embodiments, the chain moiety is a C1-C 12 In some embodiments, the chain portion comprises a C3-C7 alkylene chain. In some embodiments, the chain portion comprises a C6 alkylene chain. In some embodiments, the chain portion is a C6 alkylene chain. In some embodiments, the alkylene chain is an oxo, OH, NH2, SH, C1-C 12 Alkyl, C1-C 12 Haloalkyl, O(C1-C 12 alkyl), O(C1-C 12 haloalkyl), NH(C1-C 12 alkyl), NH(C1-C12 Haloalkyl), N(C1-C 12 alkyl)2, N(C1-C 12 haloalkyl)2, S(C1-C 12 alkyl), S(C1-C 12 haloalkyl), C(O)OH, C(O)O(C1-C 12 alkyl), C(O)O(C1-C 12 haloalkyl), C(O)NH(C1-C 12 alkyl), C(O)NH(C1-C 12 haloalkyl), C(O)N(C1-C 12 alkyl), C(O)N(C1-C 12 haloalkyl)2, C(O)S(C1-C 12 alkyl), and C(O)S(C1-C 12 haloalkyl). In some embodiments, the alkylene chain is unsubstituted.

[0048] In some embodiments, one or more targeting moieties are attached to the glucan backbone via a linker. The linker may be attached to about 1 to about 50% of the backbone moieties. In some embodiments, the targeting linker is 1-12 comprising an alkylene chain and a carbamate group, the carbamate group being connected to a backbone monomer; 1-12 An alkylene chain connects the carbamate group and the targeting moiety.

[0049] E. Active Ingredients In certain embodiments, the methods described herein include administering a compound comprising an active ingredient. In some embodiments, the active ingredient is a molecule or compound that can be used for diagnostic purposes, therapeutic purposes, or a combination thereof. The active ingredient may also be referred to as a payload. In some embodiments, the active ingredient may be or include a cytotoxic agent, an imaging agent, or a combination thereof. In some embodiments, the payload may facilitate targeted delivery of the compound described herein to tumor-associated macrophages or cancer cells. In some embodiments, the payload is a hydrophobic payload (e.g., topoisomerase inhibitor I, topoisomerase inhibitor II, or temozolomide). In some embodiments, the payload is a hydrophilic payload.

[0050] In some embodiments, the active ingredient is a therapeutic agent. The therapeutic agent can be any compound known to be useful in treating macrophage-mediated diseases. Therapeutic agents include, but are not limited to, chemotherapeutic agents such as doxorubicin; alkylating agents such as temozolomide; anti-infective agents such as antibiotics (e.g., tetracycline, streptomycin, rifampin, and isoniazid), antiviral agents, antifungal agents, and antiparasitic agents; immunoadjuvants; steroids; nucleotides such as DNA, RNA, RNAi, siRNA, CpG, or poly(I:C); peptides; proteins; antibody-drug conjugates (e.g., trastuzumab deruxtecan), or metals such as silver, gallium, or gadolinium.

[0051] In some embodiments, the therapeutic agent is selected from the group including, but not limited to, cytostatic agents, alkylating agents, antimetabolites, antiproliferative agents, tubulin binding agents, hormones and hormone antagonists, anthracyclines, vinca drugs, mitomycins, bleomycins, cytotoxic nucleosides, pteridine drugs, diynenes, podophyllotoxins, toxic enzymes, and radiosensitizing agents.

[0052] In some embodiments, the therapeutic agent is lomustine, epirubicin, topotecan, irinotecan, pemetrexed, docetaxel, oxaliplatin, altretamine, valrubicin, sarcin, temozolomide, mechlorethamine, triethylenephosphoramide, cyclophosphamide, ifosfamide, chlorambucil, busulfan, melphalan, triaziconazole, nitrosourea compounds, adriamycin, carminomycin, daunorubicin (daunomycin), doxorubicin, isoniazid, rifampin, indomethacin, gallium(III), 68 gallium(III), aminopterin, methotrexate, methopterin, mithramycin, streptonigrin, dichloromethotrexate, mitomycin C, actinomycin-D, porfiromycin, 5-fluorouracil, floxuridine, ftorafur, 6-mercaptopropyl benzoate, benzocaine, benzodiazepine, benzocaine, benzophenone, benzocaine ... Ibuprofen, cytarabine, cytosine arabinoside, podophyllotoxin, etoposide, etoposide phosphate, melphalan, vinblastine, vincristine, leurocidin, vindesine, leurocidin, taxol, taxane, cytochalasin B, gramicidin D, ethidium bromide, emetine, tenoposide, colchicine, dihydroxyanthracin dione, mitoxantrone, procaine, tetracaine, lidocaine, propranolol The compound is selected from the group consisting of lanolol, puromycin, ricin subunit A, abrin, diphtheria toxin, botulinum toxin, cyanguinosin, saxitoxin, shiga toxin, tetanus toxin, tetrodotoxin, trichothecenes, verrucologens, corticosteroids, progestins, estrogens, antiestrogens, androgens, aromatase inhibitors, calicheamicin, esperamicin, deruxtecan, and dynemicin.

[0053] In some embodiments, the active ingredient is or comprises a cytotoxic agent. In some embodiments, the cytotoxic agent is a chemotherapeutic agent, an antitubulin agent, a DNA modifying agent, or a small interfering ribonucleic acid. In some embodiments, the cytotoxic agent is selected from the group consisting of auristatin, dolastatin, auristatin E, monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), dimethylvaline-valine-dolaisoloiin-dolaproine-phenylalanine-p-phenylenediamine (AFP), 5-benzoylvaleric acid-auristatin E ester (AEVB), auristatin EB (AEB), ansamitocin, ibreltansine / emtansine (DMI), ravtansine / soravtansine (DM4), duocarmycin, calicheamicin, and pyrrolobenzodiazepine. In certain embodiments, the active ingredient is MMAE.

[0054] F. Payload Linker In certain embodiments, the active ingredient or payload is directly linked to the glucan backbone. In some embodiments, the active ingredient is connected to the glucan backbone via a linker. The linker may be cleavable or non-cleavable. In some embodiments, one or more therapeutic agents are linked via a biodegradable linker. In some embodiments, the biodegradable linker is acid-sensitive, for example, a hydrazone linker. The use of an acid-sensitive linker allows for the transport of the drug into cells and the release of the drug substantially inside the cells. In some embodiments, the payload linker is a Val-Cit linker.

[0055] In some embodiments, the payload linker can include a carbamate group. In some embodiments, the payload linker includes a carbamate group and a chain moiety, where the carbamate group is connected to the backbone monomer and the chain moiety connects the carbamate group to the active ingredient. As used herein, the carbamate functional group has its plain and ordinary meaning derived from the field of organic chemistry. In some embodiments, the chain moiety of the payload linker includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) units selected from the group consisting of an optionally substituted alkylene chain, an optionally substituted CO-alkylene chain, a peptide chain, a polymer chain, and a heteroatom selected from the group consisting of an O atom, an S atom, and an optionally substituted N atom. In some embodiments, the chain moiety is a C1-C 12 In some embodiments, the chain portion comprises a C3-C7 alkylene chain. In some embodiments, the chain portion comprises a C6 alkylene chain. In some embodiments, the chain portion is a C6 alkylene chain. In some embodiments, the alkylene chain is an oxo, OH, NH2, SH, C1-C 12 Alkyl, C1-C 12 Haloalkyl, O(C1-C 12 alkyl), O(C1-C 12 haloalkyl), NH(C1-C 12 alkyl), NH(C1-C 12 Haloalkyl), N(C1-C 12 alkyl)2, N(C1-C 12 haloalkyl)2, S(C1-C 12 alkyl), S(C1-C 12 haloalkyl), C(O)OH, C(O)O(C1-C 12 alkyl), C(O)O(C1-C 12 haloalkyl), C(O)NH(C1-C 12 alkyl), C(O)NH(C1-C 12 haloalkyl), C(O)N(C1-C 12 alkyl), C(O)N(C1-C 12 haloalkyl)2, C(O)S(C1-C 12alkyl), and C(O)S(C1-C 12 haloalkyl). In some embodiments, the alkylene chain is unsubstituted.

[0056] In some embodiments, the molar ratio of targeting moiety (e.g., mannose) to payload is about 1:10 to about 10:1. In some embodiments, the molar ratio of targeting moiety to payload is at least about 1:10 (e.g., about 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, and 4:1). In some embodiments, the molar ratio of targeting moiety to payload is about 1:1. In some embodiments, the molar ratio of targeting moiety to payload is about 1:2. In some embodiments, the molar ratio of targeting moiety to payload is about 1:3. In some embodiments, the molar ratio of targeting moiety to payload is about 1:4.

[0057] In some embodiments, the payload linker is -C(O)-C 1-12 The carbamate group is connected to the backbone monomer and includes an alkylene chain and a carbamate group, and the carbamate group is connected to the backbone monomer and includes a —C(O)—C 1-12 An alkylene chain connects the carbamate group to the payload.

[0058] G. Secondary Payloads and Linkers In addition to the targeting, diagnostic, and therapeutic payloads, the compounds used in the methods disclosed herein may include secondary agents that can be attached to the glucan backbone to add additional functional capabilities. Typically, the secondary payload is attached to a linker in a manner similar to that used to attach a targeting moiety to a targeting linker. In some embodiments, the secondary payload can facilitate targeted delivery of the compositions described herein to macrophages or cancer cells. In some embodiments, the secondary payload is a hydrophobic payload (e.g., topoisomerase inhibitor I, topoisomerase inhibitor II, or temozolomide). In some embodiments, the secondary payload is a hydrophilic payload.

[0059] In some embodiments, the secondary payload can be a radiosensitizer. See, e.g., Zhang et al., "Application of Radiosensitizers in Cancer Radiotherapy," Int J Nanomedicine, 2021, 16:1083-1102. In some embodiments, the radiosensitizer is a small molecule radiosensitizer, including, but not limited to, monomethyl auristatin E (MMAE), mitomycin C, misonidazole, curcumin, or paclitaxel. In some embodiments, the secondary payload can be an anti-tuberculosis drug (e.g., rifampin or isoniazid).

[0060] The secondary payload can include additional agents, for example, for diagnostic imaging, therapy, or other purposes. Specifically, in some embodiments, a combination of therapeutic and imaging agents can be linked to the glucan backbone to combine diagnostic and therapeutic functions. In other embodiments, various amino acids, such as cysteine ​​or lysine, can be attached to the linker to crosslink the molecule to its target.

[0061] The secondary payload linker is a cleavable or non-cleavable linker that connects the glucan backbone to the secondary payload moiety. The cleavable linker can be cleaved by an enzyme (e.g., a protease), a change in temperature, a change in pH, a chemical stimulus, or any combination thereof. The cleavable linker can include a protease cleavage site. In some embodiments, the cleavable linker can be cleaved by a lysosomal or endosomal protease.

[0062] The secondary payload linker can include a carbamate group. In some embodiments, the secondary payload linker includes a carbamate group and a chain moiety, where the carbamate group is connected to the backbone monomer and the chain moiety connects the carbamate group to the secondary agent. As used herein, the carbamate functional group has its plain and ordinary meaning from the field of organic chemistry. In some embodiments, the chain moiety of the secondary payload linker includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) units selected from the group consisting of an optionally substituted alkylene chain, an optionally substituted CO-alkylene chain, a peptide chain, a polymer chain, and a heteroatom selected from the group consisting of an O atom, an S atom, and an optionally substituted N atom. In some embodiments, the chain moiety is a C1-C 12 In some embodiments, the chain portion comprises a C3-C7 alkylene chain. In some embodiments, the chain portion comprises a C6 alkylene chain. In some embodiments, the chain portion is a C6 alkylene chain. In some embodiments, the alkylene chain is an oxo, OH, NH2, SH, C1-C 12 Alkyl, C1-C 12 Haloalkyl, O(C1-C 12 alkyl), O(C1-C 12 haloalkyl), NH(C1-C 12 alkyl), NH(C1-C 12 Haloalkyl), N(C1-C 12 alkyl)2, N(C1-C 12 haloalkyl)2, S(C1-C 12 alkyl), S(C1-C12 haloalkyl), C(O)OH, C(O)O(C1-C 12 alkyl), C(O)O(C1-C 12 haloalkyl), C(O)NH(C1-C 12 alkyl), C(O)NH(C1-C 12 haloalkyl), C(O)N(C1-C 12 alkyl), C(O)N(C1-C 12 haloalkyl)2, C(O)S(C1-C 12 alkyl), and C(O)S(C1-C 12 haloalkyl). In some embodiments, the alkylene chain is unsubstituted.

[0063] In some embodiments, one or more secondary payload moieties are attached to the glucan backbone via linkers, which may be attached to about 1 to about 50% of the backbone moieties.

[0064] II. Exemplary Compounds In some embodiments, the compositions described herein comprise Compound A or a pharmaceutically acceptable salt thereof. [ka]

[0065] In some embodiments, unless otherwise specified, the monomers of the type designated a, c, or d in Compound A may be in a block copolymer configuration, randomly arranged within the polymer, or any combination thereof. In some embodiments, a, c, and d in Compound A may each independently represent an integer of 0, at least 1, about 1 to about 165, about 16 to about 111, about 50 to about 65, about 5 to about 167, or about 6 to about 16. In some embodiments, the glucan backbone of Compound A is linear, branched, cyclic, or a combination thereof. In some embodiments, the terminal group of the glucan backbone of Compound A may be a hydroxyl terminal group of the monomer. In some embodiments, the terminal group of the glucan backbone of Compound A may be any terminal group recognizable by one of ordinary skill in the art.

[0066] In some embodiments, the glucan backbone of Compound A is linear, branched, cyclic, or a combination thereof, and a, c, and d are each independently selected from the group consisting of 0, an integer of at least 1, an integer from about 1 to about 165, an integer from about 16 to about 111, an integer from about 5 to about 167, an integer from about 50 to about 65, and an integer from about 6 to about 16, and the terminal groups of the glucan backbone are hydroxy terminal groups of the monomers. In some embodiments, the glucan backbone of Compound A is cyclic, and a, c, and d are each independently selected from the group consisting of 0, an integer of at least 1, an integer from about 1 to about 165, an integer from about 16 to about 111, an integer from about 5 to about 167, an integer from about 50 to about 65, and an integer from about 6 to about 16. In some embodiments, the glucan backbone of Compound A is linear, and a, c, and d are each independently selected from the group consisting of 0, an integer of at least 1, an integer from about 1 to about 165, an integer from about 16 to about 111, an integer from about 5 to about 167, an integer from about 50 to about 65, and an integer from about 6 to about 16, and the terminal groups of the glucan backbone are hydroxyl end groups of the monomers. In some embodiments, the glucan backbone of Compound A is branched, and a, c, and d are each independently selected from the group consisting of 0, an integer of at least 1, an integer from about 1 to about 165, an integer from about 16 to about 111, an integer from about 5 to about 167, an integer from about 50 to about 65, and an integer from about 6 to about 16, and the terminal groups of the glucan backbone are hydroxyl end groups of the monomers.

[0067] In some embodiments in combination with the above or below embodiments, the glucan backbone of Compound A is about 6 kDa, and the glucan backbone is dextran. In some embodiments in combination with the above or below embodiments, the a, c, and d groups of Compound A are interspersed. In some embodiments in combination with the above or below embodiments, the terminal groups of the glucan backbone are natural terminal groups of glucose, such as hydroxyl terminal groups. In some embodiments in combination with the above or below embodiments, the ratio of targeting moiety to backbone monomer of Compound A is about 1:30 to 1:40 (e.g., 1:33). In some embodiments in combination with the above or below embodiments, the ratio of MMAE to mannose of Compound A is about 1:1 to 1:3 (e.g., 1:1) or about 1:3 to 1:5 (e.g., 1:4).

[0068] In some embodiments, provided herein is Compound A, wherein the glucan backbone is dextran, the molecular weight of the glucan backbone is about 6 kDa, the a, c, and d groups are interspersed, the terminal groups of the glucan backbone are natural terminal groups of glucose, such as hydroxyl terminal groups, the ratio of targeting moiety to backbone monomer is about 1:30 to 1:40 (e.g., 1:33), and the ratio of MMAE to mannose is about 1:1 to 1:3 (e.g., 1:1).

[0069] In some embodiments, provided herein is Compound A, wherein the glucan backbone is dextran, the glucan backbone has a molecular weight of about 6 kDa and is interspersed with a, c, and d groups, the terminal groups of the glucan backbone are natural terminal groups of glucose, such as hydroxyl terminal groups, the ratio of targeting moiety to backbone monomer is about 1:33 to 1:40 (e.g., 1:37), and the ratio of MMAE to mannose is about 1:3 to 1:5 (e.g., 1:4).

[0070] III. Pharmaceutical Compositions Compositions comprising the compounds, such as pharmaceutical compositions and formulations, are also provided. In some embodiments, the methods described herein include administering the compositions for the treatment of a disease. In some embodiments, the disease is cancer. Pharmaceutical formulations are provided that include the compounds and additional agents for combination treatment or therapy. Pharmaceutical compositions and formulations typically include, optionally, one or more pharmaceutically acceptable carriers or excipients. In some embodiments, the composition includes at least one additional therapeutic agent.

[0071] The term "pharmaceutical formulation" refers to a preparation that is in a form such that the biological activity of the active ingredient contained therein is effective and that does not contain additional ingredients that are unacceptably toxic to a subject to which the formulation will be administered. A "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation, other than the active ingredient, that is non-toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0072] In some embodiments, the choice of carrier will be determined in part by the method of administration. Accordingly, there are a variety of suitable formulations. For example, the pharmaceutical composition may contain a preservative. Suitable preservatives may include, for example, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. In some embodiments, a mixture of two or more preservatives is used. The preservative or mixtures thereof are typically present in an amount of about 0.0001% to about 2% by weight of the total composition. Carriers are described, for example, in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). Pharmaceutically acceptable carriers are generally non-toxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citric acid, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol alcohol, butyl alcohol, or benzyl alcohol; alkyl parabens such as methyl paraben or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polyisoprene; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as polyethylene glycol (PEG).

[0073] In some embodiments, a buffering agent is included in the composition. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some embodiments, a mixture of two or more buffering agents is used. The buffering agent or mixture thereof is typically present in an amount of about 0.001% to about 4% by weight of the total composition. Methods for preparing administrable pharmaceutical compositions are known. Exemplary methods are described in more detail, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0074] A formulation or composition may contain multiple active ingredients useful for the particular indication, disease, or condition being treated by the compound or composition. Such active ingredients are preferably present in a combination in amounts effective for the intended purpose. Thus, in some embodiments, a pharmaceutical composition further comprises another pharmaceutically active agent or drug, such as a chemotherapeutic agent, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, temozolomide, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc. In some embodiments, the compound or composition is administered in the form of a salt, e.g., a pharmaceutically acceptable salt. Suitable pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid, and sulfuric acid, and organic acids such as tartaric acid, acetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid, succinic acid, and arylsulfonic acids, e.g., p-toluenesulfonic acid, etc. In certain embodiments, the formulation or composition comprises doxorubicin, temozolomide, and / or paclitaxel.

[0075] The active ingredient may be incorporated into microcapsules, colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or macroemulsions. In certain embodiments, the pharmaceutical composition is formulated as an inclusion complex, such as a cyclodextrin inclusion complex, or as a liposome. Liposomes can function to target host cells (e.g., T cells or NK cells) to specific tissues. Many methods are available for preparing liposomes, such as those described in Szoka et al., Ann. Rev. Biophys. Bioeng., 9:467 (1980), and U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

[0076] In some embodiments, the pharmaceutical composition may use a sustained-release, delayed-release, or extended-release delivery system so that the composition is delivered before and long enough to cause sensitization of the treated area. Many types of release delivery systems are available and known. Such systems can increase convenience for patients and physicians by avoiding repeated administration of the composition.

[0077] In some embodiments, the pharmaceutical composition contains a compound in an amount effective to treat or prevent a disease or condition, for example, an amount therapeutically or prophylactically effective to treat a disease or disorder. The therapeutic or prophylactic efficacy of some embodiments is monitored by periodic evaluation of the treated subject. In the case of repeated administration over several days or longer, depending on the condition, treatment is repeated until a desired suppression of disease symptoms occurs. However, other administration regimens may be useful and can be determined. The desired dosage can be delivered by a single bolus administration of the composition, multiple bolus administrations of the composition, or continuous infusion administration of the composition. In some embodiments, administration by continuous infusion can be achieved through the use of a pump.

[0078] The compositions can be administered using standard administration techniques, formulations, and / or devices. Formulations and devices such as syringes and vials for storing and administering the compositions are provided. Formulations include those for oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. In some embodiments, the compositions are administered parenterally. As used herein, the term "parenteral" includes intravenous, intramuscular, subcutaneous, rectal, vaginal, intracranial, intrathoracic, and intraperitoneal administration. In some embodiments, the compositions are administered to a subject using peripheral systemic delivery via intravenous, intraperitoneal, or subcutaneous injection.

[0079] Compositions in some embodiments are provided as sterile liquid preparations, such as isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which may, in some aspects, be buffered to a selected pH. Liquid preparations are generally easier to prepare than gels, other viscous compositions, and solid compositions. Furthermore, liquid compositions offer somewhat greater convenience for administration, especially by injection. Viscous compositions, on the other hand, can be formulated within an appropriate viscosity range to prolong contact with specific tissues. Liquid or viscous compositions can include a carrier, which can be a solvent or dispersion medium containing, for example, water, saline, phosphate-buffered saline, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol), and suitable mixtures thereof.

[0080] Sterile injectable solutions can be prepared by incorporating the binding molecule in a solvent, for example, by mixing with a suitable carrier, diluent, or excipient, such as sterile water, saline, glucose, dextrose, etc. The composition can also be lyophilized. Depending on the desired route of administration and preparation, the composition can contain auxiliary substances such as wetting agents, dispersing or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or viscosity-enhancing additives, preservatives, flavoring agents, coloring agents, etc. In some embodiments, suitable formulations may be prepared with reference to standard textbooks.

[0081] Various additives may be added, including antimicrobial preservatives, antioxidants, chelating agents, and buffers, which enhance the stability and sterility of the compositions. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, etc. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0082] Sustained-release preparations may also be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules.

[0083] The formulations to be used for in vivo administration are generally sterile. Sterility may be readily accomplished, for example, by filtration through sterile filtration membranes.

[0084] Pharmaceutical compositions for combination therapy are also provided. Any additional agent for combination therapy described herein can be prepared and administered as one or more pharmaceutical compositions containing the compound. Combination therapy can be administered in one or more pharmaceutical compositions.

[0085] IV. Methods and Uses Provided herein are methods, such as methods of treatment, methods of use, and uses of compounds and / or pharmaceutical compositions and formulations thereof, e.g., in the treatment or prevention of diseases or disorders. Also provided are methods of combination therapy including the compounds and / or pharmaceutical compositions for the treatment or prevention of diseases or disorders. Also provided are methods of targeting tumor-associated macrophages (TAMs). In some embodiments, the methods of use may be for targeting macrophages for the treatment of intracellular pathogens (e.g., M. tuberculosis, F. tularensis, S. typhi). In some embodiments, the methods disclosed herein may be used to target tumor-associated macrophages. In some embodiments, the methods disclosed herein may be used to treat cancer. In some embodiments, the methods disclosed herein may be used to treat drug-resistant cancers, cancer cells, and / or tumors (e.g., doxorubicin-resistant cancers, temolodide-resistant cancers). In some embodiments, the treatment method comprises administering a composition comprising a compound comprising a mannosylated dextran backbone connected to monomethyl auristatin E (MMAE) by a valine-citrulline linker.

[0086] Provided herein are methods for treating or preventing macrophage-associated and other diseases or disorders associated with cells highly expressing CD205 (DEC205), CD206, CD207 (Langerin), CD209 (DC-SIGN), CD280 (ENDO180), or CD301 (MGL). These diseases or disorders may include, but are not limited to, cancer (e.g., soft tissue sarcoma or glioblastoma) or non-malignant tumors (e.g., meningioma hemangioblastoma or giant cell tumor), chronic infectious diseases (e.g., tuberculosis), or granulomatous diseases (e.g., sarcoidosis). In some embodiments, non-malignant tumors include, but are not limited to, all grades of meningioma (e.g., grade 1 meningioma, grade 2 meningioma, or grade 3 meningioma), schwannoma, schwannomatosis, neurofibroma, neurofibromatosis type 1 (NF1), or neurofibromatosis type 2 (NF2).

[0087] In some embodiments, the cancer may be any cell in a subject undergoing unregulated proliferation. The cancer may be any cancer cell capable of metastasizing. For example, the cancer may be a sarcoma, glioma, lymphoma, leukemia, carcinoma, blastoma, or germ cell tumor. In some embodiments, the cancer is selected from the group consisting of carcinoma, lymphoma, blastoma, sarcoma, glioma, leukemia, lymphoid malignancies, squamous cell carcinoma, squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, peritoneal cancer, hepatocellular carcinoma, gastric cancer, stomach cancer, gastrointestinal cancer, esophageal squamous cell, hepatocellular carcinoma, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, gallbladder cancer, liver cancer, breast cancer (e.g., HR+ / HER2- breast cancer, HR- / HER2- breast cancer, HR+ / HER2+ breast cancer, or HR- / HER2+ breast cancer), colon cancer, rectal cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, hepatic carcinoma, anal cancer, penile cancer, and head and neck cancer. In some embodiments, the non-malignant tumor is a meningioma hemangioblastoma or giant cell tumor. In certain embodiments, the cancer is a brain metastasis from a cancer. In certain embodiments, the cancer is a solid tumor.

[0088] In certain embodiments, the cancer is a soft tissue sarcoma. In certain embodiments, the soft tissue sarcoma may be, but is not limited to, angiosarcoma, dermatofibrosarcoma protuberans, epithelioid sarcoma, gastrointestinal stromal tumor (GIST), Kaposi's sarcoma, leiomyosarcoma, liposarcoma, malignant peripheral nerve sheath tumor, myxofibrosarcoma, rhabdomyosarcoma, solitary fibrous tumor, synovial sarcoma, undifferentiated pleomorphic sarcoma (UPS), desmoid, hemangiopericytoma, fibrosarcoma, angiosarcoma, alveolar soft part sarcoma (ASPS), clear cell sarcoma and melanoma of the soft tissue, extraskeletal myxoid chondrosarcoma (EMC), Ewing's sarcoma, or desmoplastic round cell tumor. In certain embodiments, the cancer is undifferentiated pleomorphic sarcoma (UPS).

[0089] In certain embodiments, the cancer is a glioma. In certain embodiments, the glioma can be an astrocytoma, an ependymoma, or an oligodendroglioma. In certain embodiments, the cancer is an astrocytoma. In certain embodiments, the cancer is a glioblastoma (glioblastoma multiforme).

[0090] In certain embodiments, the methods described herein comprise administering to a subject in need thereof a composition comprising a compound comprising a mannosylated dextran backbone connected to monomethyl auristatin E (MMAE) by a valine-citrulline linker for the treatment of soft tissue sarcoma. In certain embodiments, the methods described herein comprise administering to a subject in need thereof a composition comprising a compound comprising a mannosylated dextran backbone connected to monomethyl auristatin E (MMAE) by a valine-citrulline linker for the treatment of undifferentiated pleomorphic sarcoma (UPS). In certain embodiments, the methods described herein comprise administering to a subject in need thereof a composition comprising a compound comprising a mannosylated dextran backbone connected to monomethyl auristatin E (MMAE) by a valine-citrulline linker for the treatment of glioblastoma multiforme.

[0091] In some embodiments, the cancer, cancer cells, and / or tumor are resistant to commonly administered therapies. In some embodiments, the cancer, cancer cells, or tumor are resistant to commonly administered chemotherapeutic agents. In some embodiments, the cancer, cancer cells, and / or tumor are resistant to inhibition by doxorubicin or temozolomide. In certain embodiments, the methods described herein comprise administering to a subject in need thereof compounds and / or pharmaceutical compositions for the treatment of drug-resistant cancers, cancer cells, and / or tumors. In certain embodiments, the methods described herein comprise administering to a subject in need thereof compounds and / or pharmaceutical compositions for the treatment of doxorubicin-resistant cancers, cancer cells, and / or tumors. In certain embodiments, the methods described herein comprise administering to a subject in need thereof compounds and / or pharmaceutical compositions for the treatment of temozolomide-resistant cancers, cancer cells, and / or tumors.

[0092] In certain embodiments, the methods described herein comprise administering to a subject in need thereof a composition comprising a compound comprising a mannosylated dextran backbone connected to monomethyl auristatin E (MMAE) by a valine-citrulline linker for the treatment of doxorubicin-resistant cancers, cancer cells, and / or tumors. In certain embodiments, the methods described herein comprise administering to a subject in need thereof a composition comprising a compound comprising a mannosylated dextran backbone connected to monomethyl auristatin E (MMAE) by a valine-citrulline linker for the treatment of temozolomide-resistant cancers, cancer cells, and / or tumors.

[0093] In certain embodiments, methods of treatment include administering an effective amount of a compound and / or pharmaceutical composition for the treatment of an autoimmune disease, such as rheumatoid arthritis, lupus (SLE), or vasculitis. In certain embodiments, methods of treatment include administering an effective amount of a compound and / or pharmaceutical composition for the treatment of an inflammatory disease, such as Crohn's disease, inflammatory bowel disease, or collagen vascular disease. In certain embodiments, methods of treatment include administering a compound and / or pharmaceutical composition for the treatment of a macrophage-mediated disorder.

[0094] The compositions disclosed herein can also be used to treat lysosomal storage diseases, including, but not limited to, cholesteryl ester storage disease, Wolman disease, Hunter syndrome, Hurler disease, Fabry disease, Gaucher disease, Krabbe disease (globoid cell leukodystrophy), metachromatic leukodystrophy, Niemann-Pick disease, Sandhoff disease, Tay-Sachs disease, Batten disease, cystinosis, Danon disease, and Pompe disease.

[0095] A. Medication and Administration In some embodiments, the methods disclosed herein include administering a compound or pharmaceutical composition to a subject via any suitable method, such as by injection, e.g., intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjunctival injection, sub-Tenon injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, the compound or composition is administered parenterally, intrapulmonary, and intranasally, and, if desired for localized treatment, by intralesional administration. Parenteral injections include intramuscular, intravenous, intraarterial, intraperitoneal, intracranial, intrathoracic, or subcutaneous administration. In some embodiments, the methods include parenterally administering a compound or pharmaceutical composition intraparenchyma or into the circulation so that the disclosed compound reaches the target tissue (e.g., where cancer cells may be present). In some embodiments, the methods involve administering the compound or pharmaceutical composition directly into or adjacent to the tumor mass.

[0096] Parenteral administration of compound, when used, is generally characterized by injection.Injection can be prepared in conventional form, either as solution or suspension, solid form suitable for dissolving suspension before injection, or emulsion.The revised method for parenteral administration involves the use of slow-release or sustained-release system to maintain a constant dosage.

[0097] Dosage and administration can depend in part on whether the administration is short-term or long-term. Various dosing schedules include, but are not limited to, single or multiple doses at various times, bolus doses, and pulse infusions. In some embodiments, a pump can be used for continuous administration.

[0098] For the prevention or treatment of disease, the appropriate dosage of the compound or pharmaceutical composition will depend on the type of disease being treated, the severity and course of the disease, whether the compound or pharmaceutical composition is being administered for prophylactic or therapeutic purposes, previous therapy, the clinical history of the subject, and the discretion of the attending physician. The compound or pharmaceutical composition, in some embodiments, is suitably administered to the patient at one time or over a series of treatments.

[0099] In some embodiments, a dose of a compound or composition is administered to a subject as a single dose, or administered only once within a period of two weeks, one month, three months, six months, one year, or longer. In some embodiments, a dose of a compound or composition is administered to a subject in multiple doses. In some embodiments, the dose is administered to a subject once daily. In some embodiments, the dose is administered to a subject multiple times daily. In some embodiments, the dose is administered to a subject 6 times a day, 5 times a day, 4 times a day, 3 times a day, 2 times a day, once a day, every other day, 3 times a week, twice a week, at least once a week, once a week, once every two weeks, once every three weeks, once a month, once every two months, or once a month. In certain embodiments, the dose is administered to a subject once every two weeks, once every three weeks, or once a month.

[0100] B. Combination Therapy Also provided herein are methods for treating a disease or disorder (e.g., cancer) comprising administering to a subject in need thereof a combination therapy comprising an effective amount of a compound and / or pharmaceutical composition and an additional therapeutic agent or intervention. In some embodiments, the compound and / or pharmaceutical composition is administered simultaneously with the additional therapeutic agent or intervention. In some embodiments, the compound and / or pharmaceutical composition is administered sequentially, in any order, with the additional therapeutic agent or intervention. In some embodiments, the additional therapeutic agent or intervention may include, but is not limited to, a chemotherapeutic agent, a DNA hypomethylating agent, an alkylating agent, a topoisomerase inhibitor, a therapeutic antibody that specifically binds to a cancer antigen, a hematopoietic growth factor, a cytokine, an antibiotic, a COX-2 inhibitor, a CDK inhibitor, an immunomodulatory agent, an antithymocyte globulin, an immunosuppressant, a corticosteroid or a pharmacological derivative thereof, radiation, surgery, and adjuvant therapy. In some embodiments, the compound and / or pharmaceutical composition is administered in combination with a cytotoxic or therapeutic agent. In some embodiments, the compound and / or pharmaceutical composition is administered in combination with a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent comprises temozolomide (TMZ), doxorubicin, and / or paclitaxel. In some embodiments, the compound and / or pharmaceutical composition is administered with adjuvant therapy. In some embodiments, the compound and / or pharmaceutical composition is administered with radiation therapy. In some embodiments, the combination therapy comprises the compound and / or pharmaceutical composition and one additional therapeutic agent or intervention. In some embodiments, the combination therapy comprises the compound and / or pharmaceutical composition and multiple additional therapeutic agents or interventions.

[0101] In certain embodiments, the methods disclosed herein comprise administering a compound and / or pharmaceutical composition in combination with an effective amount of doxorubicin to a subject in need thereof. In certain embodiments, the methods disclosed herein comprise administering a compound and / or pharmaceutical composition in combination with an effective amount of temozolomide to a subject in need thereof. In certain embodiments, the methods disclosed herein comprise administering a compound and / or pharmaceutical composition in combination with paclitaxel to a subject in need thereof. In certain embodiments, the methods described herein comprise administering a composition comprising a compound comprising a mannosylated dextran backbone connected to MMAE by a valine-citrulline linker in combination with an effective amount of doxorubicin to a subject in need thereof. In certain embodiments, the methods described herein comprise administering a composition comprising a compound comprising a mannosylated dextran backbone connected to MMAE by a valine-citrulline linker in combination with an effective amount of temozolomide to a subject in need thereof. In certain embodiments, the methods described herein include administering to a subject in need thereof a composition comprising a compound comprising a mannosylated dextran backbone connected to MMAE by a valine-citrulline linker in combination with an effective amount of paclitaxel.

[0102] In some embodiments, the compound and / or pharmaceutical composition is co-administered with one or more additional therapeutic agents or in connection with another therapeutic intervention, either simultaneously or sequentially in any order. In some circumstances, the compound and / or pharmaceutical composition is co-administered sufficiently closely with another therapy so that its administration can enhance the effect of the one or more additional therapeutic agents, or vice versa. In some embodiments, the compound and / or pharmaceutical composition is administered before the one or more additional therapeutic agents. In some embodiments, the compound and / or pharmaceutical composition is administered after the one or more additional therapeutic agents.

[0103] V. Manufactured Articles or Kits Also provided are articles of manufacture or kits containing the compounds and / or compositions comprising the same. The articles of manufacture may include a container and a label or package insert associated with or on the container. Suitable containers include, for example, bottles, vials, syringes, test tubes, IV infusion bags, etc. The containers may be formed from a variety of materials, such as glass or plastic. In some embodiments, the container has a sterile access port. Exemplary containers include intravenous infusion bags and vials (including those with needle-pierceable stoppers). The articles of manufacture or kits may further include a package insert indicating that the composition can be used to treat a particular condition, such as a condition described herein (e.g., cancer). Alternatively, or in addition, the articles of manufacture or kits may further include a separate container or the same container containing a pharmaceutically acceptable buffer. Other materials, such as other buffers, diluents, filters, needles, and / or syringes, may also be included.

[0104] The label or package insert may indicate that the composition is used to treat a disease, disorder, or condition (e.g., cancer) in an individual. A label or package insert on or associated with the container may indicate instructions for reconstituting and / or using the formulation. The label or package insert may further indicate that the formulation is useful or intended for subcutaneous administration, intravenous administration, or other modes of administration to treat or prevent an autoimmune disease, disorder, or condition in an individual. In some embodiments, the label or package insert may include instructions for use, e.g., instructions for administering the compound or composition according to any of the methods or uses described herein in some embodiments.

[0105] In some embodiments, the container holds the composition alone or in combination with another composition effective for treating, preventing, and / or diagnosing a condition. An article of manufacture or kit can include (a) a first container containing a composition comprising the compound (i.e., a first medicament) and (b) a second container containing a composition comprising an additional agent, such as a cytotoxic agent or other therapeutic agent (i.e., a second medicament), wherein the article or kit further comprises instructions on a label or package insert for treating a subject with an effective amount of the second medicament.

[0106] VI. General Synthetic Methods The compositions of the present disclosure will now be described with reference to the following exemplary synthetic schemes for their general preparation, followed by specific examples. To obtain the various compositions herein, those skilled in the art will recognize that starting materials can be appropriately selected so that the ultimately desired substituents are retained throughout the reaction scheme, with or without protection as necessary, to yield the desired products. Alternatively, it may be necessary or desirable to use, in place of the ultimately desired substituent, a suitable group that is retained throughout the reaction scheme and can be appropriately replaced with the desired substituent. Furthermore, those skilled in the art will recognize that protecting groups can be used to protect certain functional groups (amino, carboxy, or side chain groups) from reaction conditions, and that such groups are appropriately removed under standard conditions.

[0107] Where it is desired to obtain a particular isomer of a compound or to otherwise purify a reaction product, chromatography, recrystallization, and other conventional separation techniques may be used on intermediates or final products.

[0108] General methods for preparing the compositions described herein are illustrated in the following exemplary methods.

[0109] In some embodiments, the compositions described herein can be synthesized according to the procedure shown in Scheme A1.

[0110] Scheme A1 [ka]

[0111] Scheme A2 [ka]

[0112] As seen in the above scheme, a glucan compound (e.g., dextran or cyclodextrin) is reacted with an activating agent. The resulting activated glucan derivative can then be reacted with an appropriate reagent to introduce a targeting moiety attached to the glucan backbone via a targeting linker and an active ingredient linked to the glucan backbone via a payload linker. Those skilled in the art will recognize that the above scheme is illustrative and that the order of the various reagents and synthetic steps can be modified as necessary to achieve the intended final product. For example, a, b, and c can each independently refer to an integer of 0, at least 1, at least 1, about 1 to about 165, about 16 to about 111, about 5 to about 167, about 50 to about 65, or about 6 to about 16. It should be understood that the types of monomers designated a, b, or c may be in a block copolymer configuration, randomly arranged within the polymer, or any combination thereof, unless otherwise specified. It should also be understood that the glucan backbone in the above scheme can be linear, branched, cyclic, or a combination thereof. Groups not specified in the above scheme, such as any terminal group of the glucan backbone, can be any terminal group recognizable by those skilled in the art. For example, the terminal group of the glucan backbone can be a hydroxyl end group of a monomer.

[0113] VII. Definition Unless otherwise defined, all technical terms, notations, and other technical and scientific terms or terminology used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some instances, terms having a commonly understood meaning are defined herein for clarity and / or ready reference, and the inclusion of such definitions herein should not necessarily be construed as representing a substantial difference from what is commonly understood in the art.

[0114] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, "a" or "an" means "at least one" or "one or more." Aspects, embodiments, and variations described herein should be understood to include "comprising," "consisting of," and / or "consisting essentially of" aspects, embodiments, and variations.

[0115] Throughout this disclosure, various aspects of the claimed subject matter are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the claimed subject matter. Accordingly, the description of a range should be considered to have specifically disclosed all possible subranges and individual numerical values ​​within that range. For example, when a range of values ​​is presented, it should be understood that each intervening value between the upper and lower limits of that range, and every other stated or intervening value within that stated range, is encompassed by the claimed subject matter. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed by the claimed subject matter, subject to any specifically excluded limit in the stated range. When a stated range includes one or both limits, ranges excluding either or both of those included limits are also encompassed by the claimed subject matter. This applies regardless of the width of the range.

[0116] The term "about" as used herein refers to a normal error range for each value, which is readily apparent to one skilled in the art. As used herein, reference to a value or parameter with "about" includes (and represents) an embodiment directed to that value or parameter itself. For example, a statement referring to "about X" includes the statement of "X."

[0117] As used herein, a "composition" refers to a mixture of two or more products, substances, or compounds, which may be a solution, suspension, liquid, powder, paste, aqueous, non-aqueous, or any combination thereof.

[0118] As used herein, unless otherwise specified, "alkyl" refers to an alkyl group having a specified number of carbon atoms (i.e., C-C 10 "C" refers to a monovalent saturated hydrocarbon chain, which may be linear (i.e., unbranched) or branched, or a combination thereof, having from 1 to 20 carbon atoms ("C-C" means 1 to 10 carbon atoms). Particular alkyl groups are those having from 1 to 20 carbon atoms ("C-C" means 1 to 20 carbon atoms). 20 alkyl"), those having 1 to 10 carbon atoms ("C1-C 10 alkyl"), those with 6 to 10 carbon atoms ("C6-C 10 alkyl"), those having 1 to 6 carbon atoms ("C1-C6 alkyl"), those having 2 to 6 carbon atoms ("C2-C6 alkyl"), or those having 1 to 4 carbon atoms ("C1-C4 alkyl"). Examples of alkyl groups include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, and the like.

[0119] As used herein, "alkylene" refers to the same residue as alkyl, but with divalency. Particular alkylene groups are those having 1 to 20 carbon atoms ("C-C 20 alkylene), those having 1 to 10 carbon atoms ("C1-C10 alkylene), those with 6 to 10 carbon atoms ("C6-C 10 alkylene"), those having 1 to 6 carbon atoms ("C1-C6 alkylene"), those having 1 to 5 carbon atoms ("C1-C5 alkylene"), those having 1 to 4 carbon atoms ("C1-C4 alkylene"), or those having 1 to 3 carbon atoms ("C1-C3 alkylene"). Examples of alkylene include, but are not limited to, groups such as methylene (-CH-), ethylene (-CHCH-), propylene (-CHCHCH-), isopropylene (-CHCH(CH)-), butylene (-CH(CH)CH-), isobutylene (-CHCH(CH)CH-), pentylene (-CH(CH)CH-), hexylene (-CH(CH)CH-), heptylene (-CH(CH)CH-), octylene (-CH(CH)CH-), and the like.

[0120] As used herein, "halo" or "halogen" refers to Group 17 elements having atomic numbers 9-85. Preferred halo groups include fluorine, chlorine, bromine, and iodine radicals. When a residue is substituted with more than one halogen, it may be designated by using a prefix corresponding to the number of halogen moieties attached; for example, dihaloaryl, dihaloalkyl, trihaloaryl, etc., refer to aryl and alkyl substituted with two ("di") or three ("tri") halo groups, which may, but are not necessarily, the same halogen; thus, 4-chloro-3-fluorophenyl is within the scope of dihaloaryl. An alkyl group in which each hydrogen is replaced with a halo group is referred to as a "perhaloalkyl." A preferred perhaloalkyl group is trifluoromethyl (-CF3). Similarly, "perhaloalkoxy" refers to an alkoxy group in which a halogen replaces each H in the hydrocarbon comprising the alkyl portion of the alkoxy group. One example of a perhaloalkoxy group is trifluoromethoxy (-OCF3).

[0121] As used herein, "carbamate" refers to the group -OC(=O)-NH-. Unless otherwise specified, the nitrogen atom of the carbamate group is understood to be unsubstituted (i.e., bearing a hydrogen atom).

[0122] As used herein, "oxo" refers to the moiety =0.

[0123] As used herein, "optionally substituted" means, unless otherwise specified, that a group can be unsubstituted or substituted with one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the substituents listed for that group, which may be the same or different. In some embodiments, an optionally substituted group has one substituent. In another embodiment, an optionally substituted group has two substituents. In another embodiment, an optionally substituted group has three substituents. In another embodiment, an optionally substituted group has four substituents. In some embodiments, an optionally substituted group has 1 to 2, 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4, or 2 to 5 substituents. In some embodiments, an optionally substituted group is unsubstituted. [Example]

[0124] VIII. Working Examples The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention.

[0125] Example 1: Structure of Target 5, a targeted chemotherapeutic drug composed of a mannosylated dextran ligand linked to the toxin monomethylauristatin E The compound described herein was synthesized. This compound (Target 5) consists of a mannosylated dextran backbone (to form the mannose-binding site targeting moiety) connected to monomethyl auristatin E (MMAE) via a valine-citrulline linker. The structure of Target 5 is shown in Figure 1. The compound in Figure 1 is sometimes referred to as Compound A.

[0126] Synthesis of Compound A I. Synthesis of Val-Cit-PAB-MMAE [ka] To Fmoc-Val-Cit-PAB-PNP (470 mg, 0.613 mmol) in DMF (12 mL) was added pyridine (5 mL), a DMF solution of HOBt (80 mg, 0.593 mmol) (5 mL), a DMF solution of DIPEA (86 mg, 0.667 mmol) (5 mL), and a DMF solution of MMAE (400 mg, 0.557 mmol) (12 mL). The mixture was stirred at room temperature for 2 days. It was diluted with EtOAc (500 mL), washed with H2O (300 mL x 5) and brine (200 mL), and dried over Na2SO4. The solution was loaded onto a silica column and developed with a MeOH / DCM gradient (0% to 10%) to give Fmoc-Val-Cit-PAB-MMAE (570 mg, 0.423 mmol, 76%) as a white solid.

[0127] [ka] To a solution of Fmoc-Val-Cit-PAB-MMAE (560 mg, 0.416 mmol) in DMF (9 mL) was added piperidine (2.3 mL). The mixture was stirred at room temperature for 1 hour. It was concentrated to near dryness under high vacuum (water bath temperature 20-28 °C). EtO (30 mL) was added. The supernatant was decanted. The residue was triturated with EtO (20 mL x 4), and the EtO layer was discarded. The residue was then filtered, washed with EtO (10 mL), and dried under high vacuum to give Val-Cit-PAB-MMAE (409 mg, 0.364 mmol, 88%) as a white solid.

[0128] II. Synthesis of Mannosamine (Compound 7) [ka] To a mixture of amine 1 (9.00 g, 76.9 mmol) in 1 M NaOH (83.8 mL, 83.8 mmol) in an ice-water bath, CbzCl (14.04 g, 82.30 mmol) was added dropwise over 10 minutes (min). The ice-water bath was removed, and the mixture was stirred for 1 h. DCM (60 mL) was added. The mixture was stirred at room temperature for 1 day. It was diluted with DCM (100 mL). The organic layer was separated, washed with brine (100 mL), dried over Na2SO4, and filtered through a short silica plug using DCM followed by 1:1 DCM / EtOAc as eluents. The product-containing fractions were combined and concentrated. The residue was washed with hexane (400 mL) to give the protected amine 2 (13.50 g, 53.78 mmol, 70%) as a white solid.

[0129] [ka] To a solution of mannose 3 (10.00 g, 55.56 mmol) in pyridine (70 mL) was added AcO (30.24 g, 296.5 mmol). The mixture was stirred at room temperature for 2 days. It was added to 1 N HCl (1.0 L). The mixture was extracted with EtOAc (300 mL). The organic layer was washed with HO (200 mL) and brine (200 mL), dried over NaSO, and concentrated to give mannose pentaacetate 4 (21.65 g, 55.51 mmol, quantitative yield) as a pale yellow gel.

[0130] [ka] To a solution of compound 4 (21.65 g, 55.51 mmol) in DCM (500 mL) under N2, compound 2 (9.50 g, 37.8 mmol) and SnCl4 (15.77 g, 60.54 mmol) were added. The mixture was stirred at room temperature for 1 day. The mixture was poured into ice-cold saturated NaHCO3 (1.2 L). It was stirred for 15 minutes, filtered through a plug of Celite, and washed with DCM (300 mL). The organic layer of the filtrate was separated. The aqueous layer was extracted with DCM (300 mL x 2). The combined organic phases were washed with brine (500 mL), dried over Na2SO4, and concentrated. The residue was purified by column chromatography using an EtOAc / DCM gradient (0% to 10%) to give compound 5 (6.94 g, 11.9 mmol, 32%) as a pale yellow oil.

[0131] [ka] To a solution of compound 5 (6.94 g, 11.9 mmol) in MeOH (150 mL) was added 25 wt% NaOMe in MeOH (0.85 g, 3.94 mmol). The mixture was stirred at room temperature for 4 hours. Amberlite IR 120(H) resin (5.50 g) was added to adjust the pH to approximately 6. The mixture was filtered. The filtrate was concentrated to give compound 6 (4.80 g, 11.6 mmol, 98%) as a colorless oil.

[0132] [ka] A solution of compound 6 (4.80 g, 11.6 mmol) in MeOH (200 ml) was purged with N for 15 min. Pd / C (10 wt%, 1.30 g) was added. The mixture was degassed and refilled with H. A H balloon was attached and the mixture was stirred at room temperature for 5 h. It was filtered and washed with MeOH (100 ml). The filtrate was concentrated and dried under high vacuum to give mannose amine 7 (3.24 g, 11.6 mmol, quantitative yield) as a yellow oil.

[0133] III. Synthesis of Compound A [ka] To a DMSO solution (60 ml) of dextran 8 (MW approximately 6000, 2.00 g, 12.3 mmol of glucose units) was added pyridine (60 ml). The mixture was cooled in an ice-water bath. Compound 9 (2.48 g, 12.3 mmol) and DMAP (250 mg, 2.05 mmol) were added. The mixture was stirred at 4°C for 4 hours. EtOH (600 ml) was added. The mixture was allowed to stand at room temperature for 10 minutes. It was filtered, washed with EtOH (100 ml), EtO (100 ml), and dried under high vacuum to give compound 10 (2.20 g, 1 H NMR revealed approximately 22 mol% carbonate per mole of glucose unit. The average molecular weight of the repeating unit is approximately 198 g / mol.

[0134] [ka] To a solution of compound 10 (930 mg, 1.03 mmol of carbonate salt), a DMSO solution (5.0 mL) of HOBt (148 mg, 1.10 mmol), a pyridine solution (8.4 mL) of DIPEA (148 mg, 1.15 mmol), and a DMSO solution (12.0 mL) of Val-Cit-PAB-MMAE (232 mg, 0.207 mmol) were added. The mixture was stirred at room temperature for 90 min. Then, a DMSO solution (5.0 mL) of mannose amine 7 (232 mg, 0.832 mmol) was added. The mixture was stirred at room temperature for 20 h. EtOH (500 mL) was added. The mixture was allowed to stand at room temperature for 2 h. It was filtered, washed with EtOH (200 mL), and dried under high vacuum to give target 5-6k (900 mg) as a white solid. 1 According to H NMR, per mole of glucose unit, this compound contains 3.1 mol% MMAE and 11.6 mol% mannose (and the polymer has an average of 37 glucose units, so 1 MMAE unit and 4 mannose units). The average molecular weight of the repeating units is about 234 g / mol, with a Mw of about 8600.

[0135] Example 2: Administration of Target 5 shows potent anti-cancer efficacy in multiple murine soft tissue sarcoma (STS) tumor models Subcutaneous model The anti-cancer efficacy of target 5 was determined in a subcutaneous HT1080 mouse fibrosarcoma model. Briefly, HT1080 cells (1 x 10 cells) were injected into the right flank of 7-8 week-old female athymic nude mice (strain code 490, Charles River Laboratories). 6 The tumors were subcutaneously inoculated with 150 mm (150 mm / animal) before the start of treatment with target 5. 3 Tumors were allowed to grow until tumor volume reached 1000 μg / kg. Tumors were measured by length and width in millimeters three times a week. Tumor volume was calculated using the formula V = L x W x W / 2. If a second tumor developed in any animal, the volumes of both tumors were measured and combined. Target 5 was administered twice a week at 5 mg / kg or 10 mg / kg via the lateral tail vein. Saline was administered via the tail vein, and doxorubicin (4 mg / kg) was injected intraperitoneally.

[0136] As can be seen from Figure 2, target 5 was able to reduce tumor volume in a dose-dependent manner. Mice administered 10 mg / kg of target 5 had reduced tumor volume comparable to mice treated with doxorubicin (Figure 2A). Based on the stable weight gain of the mice, there was no significant toxicity from target 5, whereas administration of doxorubicin caused significant toxicity, as indicated by a decrease in body weight (Figure 2B).

[0137] Intracranial model The therapeutic efficacy of Target 5 was analyzed in a highly invasive STS brain metastasis model. Luciferase-expressing HT1080 cells (HT1080-Luc) were implanted intracranially on day 0. On day 4, mice were assigned to treatment groups with equal mean tumor size, as determined by mean bioluminescence intensity. Mice were administered saline or 10 mg / kg Target 5 on days 4 and 7, and imaging was performed on day 10. As shown in Figure 3, mice treated with Target 5 (right panel) showed a significant reduction in tumor volume (determined by a visible reduction in bioluminescence) after two doses compared with saline-treated mice (left panel), whose tumors increased in size. The reduction in tumor volume in Target 5-treated mice was statistically significantly different from saline-treated mice (p<0.0001), as determined by two-way ANOVA. Although STS rarely metastasizes to the brain, these data demonstrate the efficacy of Target 5 treatment in such cases and suggest that Target 5 treatment may have broad applicability in primary CNS tumors and CNS metastases from other cancers.

[0138] Doxorubicin-resistant undifferentiated pleomorphic sarcoma (UPS) PDX model The efficacy of target 5 was determined in the UPS PDX model (Certis Oncology Solutions PDX model CRT00001.001 (doxorubicin-resistant undifferentiated pleomorphic sarcoma)). UPS is a doxorubicin-resistant subtype of STS. Tumor fragments (CRT0000.001) were implanted subcutaneously into the right hind flank of male and female nude mice (n = 8 per group). Tumors ranged from 72 to 294 mm. 3 Range (average 150mm3 ), mice were randomized to their respective treatment groups and dosed within 24 hours. Target 5 was administered intravenously using 2 mg / kg, 4 mg / kg, 8 mg / kg, or 10 mg / kg (all dosed twice weekly). Control mice received intravenous saline twice weekly. Tumor volume and body weight were measured twice weekly. Mice were observed and dosed once the animals reached a tumor volume of 1,500 mm 3 The study was continued for up to 14 days until the time point at which the tumor size reached 100% or the humane endpoint, whichever occurred first. As can be seen in Figures 4A and 4B, Target-5 exhibited dose-dependent tumor suppression effects in the doxorubicin-resistant sarcoma PDX model. Mice treated with 4 mg / kg, 8 mg / kg, and 10 mg / kg of Target-5 showed similar reductions in tumor volume, demonstrating the large therapeutic index of Target-5. The mean mouse body weights were comparable across treatment groups.

[0139] Complete blood counts (CBC) and blood chemistry values ​​were determined for PDX model CRT0000.001 at the end of the study. Values ​​represent the average of four mice (randomly selected) from the saline control group, the 4 mg / kg Target-5 treatment group, and the 10 mg / kg Target-5 treatment group. Comparison of analyte means between treatment groups demonstrates the broad tolerability of Target-5, even at the highest dose. [Table 1]

[0140] Tumor fragments were implanted subcutaneously into the right hind flank of 45 female nude mice. Tumors ranged from 72 to 294 mm 3 Range (average 150mm 3 When tumor volume reached 1,500 mm, 30 mice were randomized to each treatment group and dosed within 24 hours. Target 5 was administered intravenously at 10 mg / kg (dosed twice weekly). Control mice received intravenous saline twice weekly. Tumor volume and body weight were measured twice weekly. Mice were observed and dosed when the animals reached a tumor volume of 1,500 mm. 3The study was continued for up to 26 days until the tumor reached a critical mass or humane endpoint, whichever occurred first. At the end of the study, or when the animals reached the humane endpoint, three to four tumors per group were excised and processed as FFPE specimens. Figure 4C shows that target 5 administered at 10 mg / kg per dose demonstrated potent anticancer efficacy, controlling tumor growth more effectively than doxorubicin in this doxorubicin-resistant model. A representative tumor dissection at the end of the study is shown in Figure 4D. These data demonstrate that target 5 is therapeutically effective in treating doxorubicin-resistant cancers.

[0141] Example 3: Administration of Target 5 in a mouse glioma model shows potent anti-cancer efficacy Subcutaneous glioma model To determine the anti-cancer efficacy of target 5 in a glioma model, eight groups of athymic mice (n = 8 per group) were implanted with a human glioma cell line (U87MG) on day 0. By day 14 (when tumor volume was approximately 100 mm), 3 Starting with the initial dose of 100 mg / kg, mice were treated twice weekly with various doses of target 5, saline, or temozolomide (12.5 mg / kg). Currently, temozolomide is the standard of care. Target 5 was administered intravenously to mice, and temozolomide was administered by oral gavage. Figure 4 shows that target 5 was able to reduce tumor volume in a dose-dependent manner (Figure 5A), while there was no significant toxicity as judged by the effect on mouse body weight (Figure 5B).

[0142] Temozolomide-resistant glioma model The therapeutic efficacy of Target 5 in moderately temozolomide-resistant tumors was analyzed. Five groups of immune-competent mice (C57BL / 6; n = 10 per group; Jackson Laboratories) were treated with 5x10 6GL261 tumor cells were implanted subcutaneously on day 0. GL261 tumor cells are moderately resistant to temozolomide chemotherapy. Mice were administered saline, temozolomide (12.5 mg / kg by gavage), or various concentrations of Target 5 (5 mg / kg, 7.5 mg / kg, or 10 mg / kg by tail vein injection). Figure 5 shows the effect of Target 5 on GL261 tumor volume. On day 21, mean tumor volume for vehicle treatment = 1687 mm 3 , SD=928.9mm 3 ; Mean tumor volume for treatment with 5 mg / kg of Target 5 = 440.5 mm 3 , SD=159.2mm 3 ;Mean tumor volume for treatment with 7.5 mg / kg Target 5 = 275.1 mm 3 , SD=120.7mm 3 ; Mean tumor volume for treatment with 10 mg / kg of Target 5 = 117.2 mm 3 , SD=89.6mm 3 ; Mean tumor volume after treatment with temozolomide = 302.6 mm 3 ; SD=99.7. These data indicate that the protection provided by Target-5 is greater than that provided by temozolomide, a standard chemotherapy drug used to treat glioblastoma. Target-5 was able to reduce tumor volume in a dose-dependent manner (Figure 6A) without adversely affecting mouse body weight (Figure 6B). Furthermore, dissection of representative tumors at the end of the study showed that tumor volume was reduced in Target-5-treated mice compared with saline-treated mice (Figure 6C). These data indicate that Target-5 administration can exert potent antitumor efficacy against moderately resistant tumors without appreciable toxicity.

[0143] Example 4: Efficacy of Target 5 in inhibiting the growth of temozolomide-resistant tumors in an immune-competent intracranial glioma model This exemplary method determines whether target 5 can be targeted across the blood-brain barrier and exhibit anti-cancer efficacy in temozolomide-resistant tumors in an immune-competent intracranial glioma model.

[0144] In this exemplary method, 55 C57B / 6 mice (including spares) are purchased (equal numbers of males and females). Mice are 8 weeks old at the time of implantation and weigh >20 gm on the day of implantation to ensure sufficient headroom for the planned intervention. Recognizing that the GL261 luciferase line may be immunogenic (Sanchez et al., 2020), a non-transgenic GL261 model is used to assess anti-cancer efficacy by Kaplan-Meier (KM) survival assay. Low-passage GL261 cells (ATCC) are grown and cultured at 5x10 in 2 μl. 4 Cells are stereotactically implanted into the brain. Mice with obvious cell leakage from the burr hole are excluded. Mice are assigned based on body weight into four test groups (n=10 / group): saline twice weekly control, target 5 at 10 mg / kg twice weekly and 6.75 mg / kg three times weekly, and temozolomide at 10 mg / kg twice weekly). All treatments are administered via intravenous tail vein injection, with the first administration occurring six days after implantation. Body weights are collected three times weekly for four weeks. Mice are closely observed and euthanized when euthanasia criteria are met (see the Vertebrate section).

[0145] At study completion, brains were collected from healthy and moribund mice (excluding dead mice) to calculate KM survival data and assess differences in tumor size, followed by sectioning, hematoxylin staining, and IHC for CD206 at a 1:500 dilution (Abcam ab64693, Cambridge, UK) to assess CD206 expression in this intracranial model. Blood was collected for analysis, including CBC, reticulocyte count, and chemistry values, to identify signs of liver, kidney, or hematologic toxicity. Quantitative IHC for P-glycoprotein (Pgp), the gene product of MDR1, was performed.

[0146] All in vivo studies are performed with 10 mice per group and are conducted in a blinded manner. Data (weight and survival) are collected for each individual mouse in each group. Statistical significance is determined by a two-tailed t-test for equal / unequal variances comparing the control group with the test substance-treated group to determine p-values.

[0147] Example 5: Efficacy of Target 5 in inhibiting temozolomide-resistant tumor growth in PDX models of glioma In this exemplary method, the efficacy of target 5 in glioma PDX models is determined. PDX models are selected based on high temozolomide IC50 and MGMT methylation status. Selection of models derived from pretreated and untreated patients is important. Pilot studies are conducted using both subcutaneous and intracranial implants of selected PDX lines in athymic mice to generate predictive growth curves for subsequent studies. Tumor size is measured twice weekly by caliper for subcutaneous tumors and by MRI for intracranial tumors.

[0148] Once growth curves of temozolomide-resistant PDX lines in athymic mice are established, subcutaneous anti-cancer efficacy studies using target 5 will be performed on both PDX models. Tumors in over 40 animals per PDX model will be measured at 120-200 mm by caliper. 3 When it is determined that the animals are randomized and assigned to test groups.

[0149] Target 5 is administered intravenously at 5 mg / kg twice weekly, 10 mg / kg twice weekly, or 6.7 mg / kg three times weekly. Mice treated with temozolomide receive temozolomide at a dose of 10 mg / kg twice weekly, while control mice receive saline intravenously twice weekly. Animals are evaluated for moribundity, death, or tumor volume. 3 1500mm 3 The study will continue until it reaches 100%. The antitumor efficacy of Target 5 in temozolomide-resistant tumors in PDX models will be evaluated.

[0150] For intracranial testing using PDX models, a dosing regimen will be developed that will be selected based on the efficacy of target 5 in subcutaneous models. Forty-five athymic mice (1.5x spares, equal numbers of males and females) per model will be weighed three times per week and tumor volume will be measured weekly by MRI. Volumetric tumor size measurements will be determined by MRI rather than relying on the more commonly used 2D measurements. Tumors in more than 30 animals will exceed 5 mm. 3When the tumor is measured as >100%, the animal is randomized and placed on study. Mice continue to be studied using moribundity or death as the endpoint. Data include KM survival curves and tumor volume measurements. For each group, data are analyzed for mean, median (with standard deviation and error), and for individual mice in each group. Statistical significance is determined by a two-tailed t-test for equal / unequal variances across the entire data set to determine p-values.

[0151] Data from these studies will be used to evaluate the anti-cancer efficacy of Target 5 as an effective single-agent treatment in patients with glioblastoma who have or develop resistance to temozolomide.

[0152] Example 6: Efficacy of Target 5 in inhibiting growth of doxorubicin-resistant UPS PDX models In this exemplary method, the anti-cancer efficacy of Target 5 is determined in a second UPS PDS model. In this study, Target 5 is administered intravenously at 10 mg / kg and doxorubicin at 2 mg / kg twice weekly. A saline control and Target 5 without MMAE (10 mg / kg twice weekly) are also administered. The dose of doxorubicin administered is one that provides anti-cancer efficacy without dose-limiting toxicity. Cryopreserved tumor fragments are implanted into the right flank as described above (n=10 / group, 60 mice total, 1.5x spares, equal numbers of males and females). Tumors are measured as described above to be between 120-200 mm by caliper measurement. 3 Once this is reached, the animals are randomized, assigned to test groups, and observed.

[0153] All in vivo studies are performed blinded, with enough mice per group to ensure statistical significance. For each group, data are analyzed for the mean, median (including standard deviation and error), and for individual mice within each group. Statistical significance using a two-tailed t-test with equal / unequal variances is applied to data comparing the vehicle control group with the test substance-treated group to determine p-values.

[0154] Example 7: Efficacy of Target 5 in inhibiting synovial sarcoma, leiomyosarcoma, and Ewing's / PNET This exemplary method evaluates the efficacy of Target 5 in a PDX model of the STS subtype, which strongly expresses CD206. Anticancer efficacy studies are conducted as described above, using equal numbers of male and female mice, n=10 per group (saline, Target 5, doxorubicin). At study termination, a subset of mice in each PDX study undergoes hematology and chemistry panels and necropsy. At least one study will include satellite PK animals for assessment of Target 5 and MMAE exposure in plasma and tissues (tumor, brain, heart, liver, kidney, and gastrointestinal tract).

[0155] Example 8: Quantification of MMAE in the systemic circulation in mice after Target 5 administration The amount of free MMAE in the systemic circulation of normal, non-tumor-bearing mice was determined after administration of Target 5. Seven groups of female CD-1 mice (n=3 per time point) received a single intravenous dose of 10 mg / kg of Target 5 at time point 0. Mice were sacrificed at the indicated time points (0, 0.5, 1, 2, 4, 8, and 24 hours), and plasma concentrations of free MMAE were determined. [Table 2] As shown in Figure 7, the pharmacokinetics of target 5 was consistent with high exposure (C0 and AUC inf ), very low clearance (Cl), moderate volume of distribution (V ss ), long half-life (t 1 / 2 ) and low variability. These data demonstrate that very little free MMAE (0.2%) is present in the circulation of mice. Example 9: Anti-cancer efficacy of target 5 in a mouse PDX sarcoma model (CRT0028.001)

[0156] The anti-cancer efficacy of target 5 was investigated in a second PDX model (CRT0028.001, treatment-naive undifferentiated pleomorphic sarcoma). Seventeen athymic mice (male and female) were subcutaneously implanted with CRT00288.001 (Certis Oncology Solutions, San Diego), an undifferentiated pleomorphic sarcoma. Tumors were grown in a sequential fashion until they reached 120–150 mm. 3When t1 was measured, mice were randomized into three groups and dosed twice weekly with either saline as a vehicle control, 10 mg / kg Target-5, or 1 mg / kg doxorubicin.

[0157] Figure 8A shows the mean tumor volume for each group according to the day of the study. This figure shows that 10 mg / kg of Target-5 exhibits greater antitumor efficacy than 1 mg / kg of doxorubicin. Figure 8B shows that mice treated with 10 mg / kg of Target-5 exhibited stable body weights, which is a measure of the broad health and tolerability of Target-5.

[0158] Example 10: CD206 involvement in the absence of MMAE The effect of Target-5 without MMAE was evaluated. Figure 9 shows the effect of saline, Compound B (Target-5 without MMAE), and Target-5 on tumor volume in GL261-bearing mice. Wild-type C57BL / 6 mice (12 males and 12 females) were injected with 5x10 cells of the glioma cell line GL261. 6 The tumors were subcutaneously transplanted. The average size was 102 mm. 3 When the tumor growth factor (TGF-β) reached 1.0, mice were randomized into three treatment groups: saline control, 10 mg / kg Compound B, and 9 mg / kg Target-5 (equimolar concentrations in each test group) and administered twice weekly for 18 days. Two-way ANOVA was performed using GraphPad Prism 9.0 and fitted to the principal column of column effects. *P<0.05, **P<0.001. TGI=complete growth inhibition. As can be seen in Figure 9A, Compound B does not exhibit anticancer efficacy, while Target-5 exhibits potent antitumor activity.

[0159] Figure 9B shows representative examples of GL261 tumors excised at the end of the study from saline-treated, Compound B-treated (10 mg / kg), and Target-5-treated (9 mg / kg) mice. Compound B and Target-5 doses are molar equivalents. Tumors were immunostained with anti-CD206 (Cell cat#24595S, dilution: 1:1600). The upper left box in each panel shows tumors dissected at the end of the study. As can be seen in Figure 9B, Target-5-treated mice had a significant reduction in tumor size compared to Compound B-treated mice and strong CD206 expression, indicating no downregulation of CD206 expression by Target-5 treatment.

[0160] Example 11: Anti-cancer efficacy of target 5 in a mouse PDX myxofibrosarcoma model The anti-cancer efficacy of Target-5 was investigated in a sarcoma PDX model (myxofibrosarcoma). Tumor fragments were subcutaneously implanted into the right hind flank of 21 male athymic mice. Mice were randomized into three groups and treated twice weekly with either saline as a vehicle control, 10 mg / kg Target-5, or 1 mg / kg doxorubicin. As shown in Figure 10, in this model, mice treated with Target-5 had reduced tumor volume compared with mice treated with saline or doxorubicin. These data demonstrate that Target-5 exhibits anti-cancer efficacy in the myxofibrosarcoma PDX model.

Claims

1. 1. A method of treating cancer in a subject having cancer, comprising administering to said subject a composition comprising a compound: i) a tumor-associated macrophage (TAM) targeting moiety; and ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient, said active ingredient being attached to the glucan backbone; iv) a targeting linker that connects the targeting moiety to the glucan backbone; wherein the targeting linker comprises a carbamate group and a chain moiety, the carbamate group being attached to a backbone monomer, and the chain moiety connecting the carbamate group and the tumor-associated macrophage targeting moiety.

2. 2. The method of claim 1, wherein the plurality of backbone monomers comprises a plurality of D-glucose monomers linked together in α-1,6 glycosidic or β-1,4 glycosidic bonds.

3. 3. The method of claim 1, wherein the number of D-glucose monomers is n, and n=5 to 167.

4. The method of any one of claims 1 to 3, wherein the number of D-glucose monomers is n, and n = 50 to 65.

5. The method according to any one of claims 1 to 4, wherein the glucan backbone is a linear dextran molecule.

6. The method according to any one of claims 1 to 4, wherein the glucan backbone is a cyclodextrin molecule and n = 6 to 16.

7. 7. The method of any of claims 1 to 6, wherein the tumor-associated macrophage targeting moiety comprises mannose, galactose, collagen, fucose, sulfated N-acetylgalactosamine, N-acetylglucosamine, luteinizing hormone, thyroid-stimulating hormone, phospholipase A2 or a fragment thereof, or chondroitin sulfate.

8. The method of any of claims 1 to 7, wherein the targeting moiety is mannose.

9. 9. The method of claim 8, wherein the ratio of mannose to backbone monomer is from about 1:5 to about 1:

33.

10. 10. The method of claim 8 or 9, wherein the ratio of mannose to backbone monomer is from about 1:6 to about 1:

19.

11. The method according to any one of claims 1 to 7, wherein the degree of substitution of mannose in the cyclodextrin ranges from about 0.1 to about 7.

12. 12. The method according to claim 1, wherein the degree of substitution of mannose in the cyclodextrin is in the range of about 0.5 to 5.

13. The method of any one of claims 1 to 12, wherein the targeting linker is attached to the glucan backbone via the oxygen atom of the carbamate group.

14. The chain portion of the targeting linker is C ３ -C ７ The method of any one of claims 1 to 13, comprising an alkylene chain.

15. The chain portion of the targeting linker is C ６ The method of any one of claims 1 to 14, wherein the alkylene moiety is -

16. The chain portion of the targeting linker is an unsubstituted C ６ The method according to any one of claims 1 to 15, wherein the alkylene moiety is -

17. 17. The method of any one of claims 1 to 16, wherein the carbon atom of the carbamate group of the targeting linker is the only sp2 hybridized carbon when the linker is attached to mannose.

18. 18. The method of any of claims 1 to 17, wherein the tumor-associated macrophage targeting moiety is a moiety that targets CD205 (DEC205), CD206, CD207 (Langerin), CD209 (DC-SIGN), CD280 (ENDO180), or CD301 (MGL).

19. 19. The method of claim 18, wherein the tumor-associated macrophage targeting moiety is a CD206 targeting moiety.

20. 20. The method of any of claims 1-19, wherein the compound has a molar ratio of the TAM targeting moiety to the active ingredient of about 1:1 to about 1:

10.

21. 21. The method of any of claims 1-20, wherein the molar ratio of the TAM targeting moiety to the active ingredient is about 1:1, about 1:2, or about 1:

3.

22. The method of any one of claims 1 to 21, wherein the active ingredient is attached to the glucan backbone via a payload linker.

23. The method of any one of claims 1 to 22, wherein the active ingredient is a cytotoxic drug.

24. 24. The method of claim 23, wherein the cytotoxic agent is selected from the group consisting of an auristatin, a dolastatin, auristatin E, monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), dimethylvaline-valine-dolaisoloiin-dolaproine-phenylalanine-p-phenylenediamine (AFP), 5-benzoylvaleric acid-auristatin E ester (AEVB), auristatin EB (AEB), ansamitocin, ibreltansine / emtansine (DMI), ravtansine / sorabutansine (DM4), duocarmycin, a calicheamicin, and a pyrrolobenzodiazepine.

25. 25. The method of claim 24, wherein the cytotoxic agent is MMAE.

26. 23. The method of claim 22, wherein the payload linker is a non-cleavable linker.

27. 27. The method of claim 26, wherein the payload linker comprises a carbamate group and a chain moiety, the carbamate group being connected to the backbone monomer and the chain moiety connecting the carbamate group and the active ingredient.

28. 23. The method of claim 22, wherein the payload linker is a cleavable linker.

29. 29. The method of claim 28, wherein the cleavable linker is cleavable by a protease.

30. 30. The method of claim 29, wherein the protease is a lysosomal or endosomal protease.

31. 29. The method of claim 28, wherein the cleavable linker is cleavable by a change in pH.

32. 29. The method of claim 28, wherein the payload linker comprises a Val-Cit moiety.

33. 33. The method of any of claims 1 to 32, wherein the cancer is a solid tumor selected from the group consisting of carcinoma, lymphoma, blastoma, sarcoma, glioma, leukemia, lymphoid malignancy, squamous cell carcinoma, squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, peritoneal cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, gallbladder cancer, liver cancer, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, hepatic carcinoma, anal cancer, penile cancer, and head and neck cancer.

34. 34. The method of claim 33, wherein the solid tumor is a sarcoma or a glioblastoma.

35. 35. The method of claim 34, wherein the cancer is a soft tissue sarcoma.

36. 36. The method of claim 35, wherein the cancer is undifferentiated pleomorphic sarcoma (UPS).

37. 37. The method of any of claims 1-36, wherein the subject and / or the cancer comprises a cell population that is (a) resistant to temozolomide (TMZ) and / or doxorubicin, and / or (b) resistant to inhibition by TMZ and / or doxorubicin.

38. 38. The method of any of claims 1-37, wherein the composition is administered to the subject 6 times per day, 5 times per day, 4 times per day, 3 times per day, 2 times per day, once per day, every other day, 3 times per week, twice per week, at least once per week, once per week, once every two weeks, once per three weeks, once per month, once every two months, or once per three months.

39. 39. The method of claim 38, wherein the composition is administered to the subject every two weeks, every three weeks, or once a month.

40. 40. The method of any of claims 1-39, wherein the method further comprises administering to the subject one or more additional therapeutic agent(s), adjuvant therapy, and / or radiation therapy.

41. 41. The method of claim 40, wherein the one or more additional active agents are selected from the group consisting of chemotherapeutic agents, DNA hypomethylating agents, alkylating agents, topoisomerase inhibitors, therapeutic antibodies that specifically bind to cancer antigens, hematopoietic growth factors, cytokines, antibiotics, cox-2 inhibitors, CDK inhibitors, immunomodulatory agents, antithymocyte globulin, immunosuppressants, and corticosteroids or pharmacological derivatives thereof.

42. 42. The method of claim 41, wherein the one or more additional active agent(s) is a chemotherapeutic agent.

43. 43. The method of claim 42, wherein the chemotherapeutic agent is TMZ, doxorubicin, and / or paclitaxel.

44. 44. The method of any of claims 40-43, wherein the chemotherapeutic agent is administered prior to, simultaneously with, and / or after the start of administration of the composition to the subject.

45. 41. The method of claim 40, wherein the adjuvant therapy and / or radiation therapy is administered prior to, concurrently with, and / or after the initiation of administration of the composition to the subject.

46. 46. ​​The method of any of claims 1-45, wherein the composition is administered to the subject intravenously, intraperitoneally, subcutaneously, intramuscularly, intracranially, or by pump infusion.

47. 47. The method of claim 46, wherein the composition is administered to the subject by intravenous administration or pump infusion.

48. 1. A method of treating glioblastoma in a subject having glioblastoma, comprising administering to said subject a composition comprising: i) a tumor-associated macrophage (TAM) targeting moiety; and ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient comprising MMAE, said active ingredient being attached to the glucan backbone; iv) a targeting linker that connects the targeting moiety to the glucan backbone; the targeting linker comprises a carbamate group and a chain moiety, the carbamate group being connected to a backbone monomer, and the chain moiety connecting the carbamate group and the tumor-associated macrophage targeting moiety; The method, wherein the subject and / or the glioblastoma comprises a cell population that is (a) resistant to TMZ and / or (b) resistant to inhibition by TMZ.

49. 1. A method of treating brain metastases in a subject having brain metastases, comprising administering to said subject a composition comprising: i) a tumor-associated macrophage (TAM) targeting moiety; and ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient comprising MMAE, said active ingredient being attached to the glucan backbone; iv) a targeting linker that connects the targeting moiety to the glucan backbone; the targeting linker comprises a carbamate group and a chain moiety, the carbamate group being connected to a backbone monomer, and the chain moiety connecting the carbamate group and the tumor-associated macrophage targeting moiety; The method, wherein the subject and / or the brain metastases comprise a cell population that is (a) resistant to TMZ and / or (b) resistant to inhibition by TMZ.

50. 1. A method of treating UPS in a subject comprising administering to the subject a composition comprising: i) a tumor-associated macrophage (TAM) targeting moiety; and ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient comprising MMAE, said active ingredient being attached to the glucan backbone; iv) a targeting linker that connects the targeting moiety to the glucan backbone; the targeting linker comprises a carbamate group and a chain moiety, the carbamate group being connected to a backbone monomer, and the chain moiety connecting the carbamate group and the tumor-associated macrophage targeting moiety; The method, wherein the subject and / or the UPS (a) is resistant to doxorubicin and / or (b) comprises a cell population that is resistant to doxorubicin.

51. 51. The method of any one of claims 48 to 50, wherein the plurality of backbone monomers comprises a plurality of D-glucose monomers linked together via α-1,6 glycosidic or β-1,4 glycosidic bonds.

52. 52. The method of any one of claims 48 to 51, wherein the number of D-glucose monomers is n, and n=5 to 167.

53. 53. The method of any one of claims 48 to 52, wherein the number of D-glucose monomers is n, and n=50 to 65.

54. 54. The method of any one of claims 48 to 53, wherein the glucan backbone is a linear dextran molecule.

55. 54. The method of any one of claims 48 to 53, wherein the glucan backbone is a cyclodextrin molecule and n=6 to 16.

56. 56. The method of any of claims 48-55, wherein the tumor-associated macrophage targeting moiety comprises mannose, galactose, collagen, fucose, sulfated N-acetylgalactosamine, N-acetylglucosamine, luteinizing hormone, thyroid-stimulating hormone, phospholipase A2 or a fragment thereof, or chondroitin sulfate.

57. 57. The method of any of claims 48 to 56, wherein the targeting moiety is mannose.

58. 58. The method of claim 57, wherein the ratio of mannose to backbone monomer is from about 1:5 to about 1:

33.

59. 59. The method of claim 57 or 58, wherein the ratio of mannose to backbone monomer is from about 1:6 to about 1:

33.

60. 57. The method of any one of claims 48 to 56, wherein the degree of substitution of mannose in the cyclodextrin ranges from about 0.1 to about 7.

61. 61. The method of any one of claims 48 to 57 or 60, wherein the degree of substitution of mannose in the cyclodextrin ranges from about 0.5 to 5.

62. 62. The method of any one of claims 1 to 61, wherein the targeting linker is attached to the glucan backbone via the oxygen atom of the carbamate group.

63. The chain portion of the targeting linker is C ３ -C ７ 63. The method of any one of claims 1 to 62, comprising an alkylene chain.

64. The chain portion of the targeting linker is C ６ - alkylene moiety.

65. The chain portion of the targeting linker is an unsubstituted C ６ - alkylene moiety.

66. 66. The method of any one of claims 1 to 65, wherein the carbon atom of the carbamate group of the targeting linker is the only sp2 hybridized carbon when the linker is attached to mannose.

67. 67. The method of any of claims 1-66, wherein the tumor-associated macrophage targeting moiety is a moiety that targets CD205 (DEC205), CD206, CD207 (Langerin), CD209 (DC-SIGN), CD280 (ENDO180), or CD301 (MGL).

68. 68. The method of claim 67, wherein the tumor-associated macrophage targeting moiety is a CD206 targeting moiety.

69. 69. The method of any of claims 48-68, wherein the compound has a molar ratio of the TAM targeting moiety to the active ingredient of about 1:1 to about 1:

10.

70. 70. The method of any of claims 48-69, wherein the molar ratio of the TAM targeting moiety to the active ingredient is about 1:1, about 1:2, or about 1:

3.

71. The method of any one of claims 48 to 70, wherein the active ingredient is attached to the glucan backbone via a payload linker.

72. 72. The method of claim 71, wherein the payload linker is a non-cleavable linker.

73. 73. The method of claim 72, wherein the payload linker comprises a carbamate group and a chain moiety, the carbamate group being connected to the backbone monomer and the chain moiety connecting the carbamate group and the active ingredient.

74. 72. The method of claim 71, wherein the payload linker is a cleavable linker.

75. 75. The method of claim 74, wherein the cleavable linker is cleavable by a protease.

76. 76. The method of claim 75, wherein the protease is a lysosomal or endosomal protease.

77. 75. The method of claim 74, wherein the cleavable linker is cleavable by a change in pH.

78. 75. The method of claim 74, wherein the payload linker comprises a Val-Cit moiety.

79. The method of any of claims 48 to 78, wherein the method further comprises administering radiation therapy to the subject.

80. 80. The method of any of claims 48 to 79, wherein the method further comprises administering to the subject TMZ and / or doxorubicin.

81. The method of any of claims 1-80, wherein the method attenuates tumor growth in the subject.

82. 82. The method of any of claims 1-81, wherein the subject is immunocompetent or immunocompromised.

83. 10. The method of claim 1, i) the tumor-associated macrophage (TAM) targeting moiety is a CD206 targeting moiety; ii) the active ingredient is linked to the glucan backbone via a payload linker, wherein the active ingredient is MMAE and the payload linker is a Val-Cit linker; The method.

84. 51. The method according to any one of claims 48 to 50, i) the tumor-associated macrophage (TAM) targeting moiety is a CD206 targeting moiety; ii) the active ingredient is linked to the glucan backbone via a payload linker, wherein the active ingredient is MMAE and the payload linker is a Val-Cit linker; The method.

85. 2. The method of claim 1, wherein the method comprises administering Compound A to the subject.

86. 51. The method of any of claims 48 to 50, wherein the method comprises administering Compound A to the subject.

87. 1. A kit comprising a composition comprising a compound and instructions for administration to a subject to treat cancer, wherein the compound is i) a tumor-associated macrophage (TAM) targeting moiety; and ii) a glucan backbone, the glucan backbone comprising a plurality of backbone monomers; iii) an active ingredient, said active ingredient being attached to the glucan backbone; iv) a targeting linker that connects the targeting moiety to the glucan backbone; wherein the targeting linker comprises a carbamate group and a chain moiety, the carbamate group being attached to a backbone monomer, and the chain moiety connecting the carbamate group and the tumor-associated macrophage targeting moiety.

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