Treatment of solid tumors with implantable polymers having small molecules

Abstract

Implantable polymers that release a therapeutic agent near a tumor in a patient are disclosed. In some embodiments, a polyethylene glycol hydrogel can release a small molecule, such as a STING agonist, near a phosphatase and tensin homolog (PTEN)-null triple negative breast cancer (TNBC) in the patient.

Description

[0001] This application claims the benefit and priority of U.S. Provisional Application No. 63 / 596,834, filed November 7, 2023, the entire contents of which are incorporated herein by reference.

[0002] All patents, patent applications, and publications cited herein are hereby incorporated in their entirety by reference. The disclosures of these publications are also incorporated herein by reference in their entirety to provide a more comprehensive description of the state of the prior art known to a person skilled in the art as of the date of the invention described and claimed herein.

[0003] This patent disclosure contains copyrighted material. The copyright holder does not object to any reproduction or copying of this patent document or patent disclosure as it appears in the patent documents and records of the United States Patent and Trademark Office, but otherwise reserves all and all copyrights. Technical Field

[0004] An implantable biocompatible polymer is disclosed that continuously releases therapeutic agents (e.g., small molecules containing STING agonists) in vivo to treat cancer. Background Technology

[0005] Injectable delivery of certain therapeutic agents to tumors in a subject may be toxic to the immune cells (e.g., T cells) that attack the tumor. In these cases, alternative delivery modalities are sought. Summary of the Invention

[0006] This document discloses implantable biocompatible polymers (e.g., polyethylene glycol-based hydrogels) that can release therapeutic agents, including STING agonists, into a subject for the treatment of cancer. In some embodiments, the biocompatible polymer containing the STING agonist can be implanted near a solid tumor in a subject. In some embodiments, the cancer may be breast cancer. In some embodiments, the cancer may be triple-negative breast cancer (TBNC) that is ineffective against phosphatase and tensin homolog (PTEN).

[0007] A method for delivering a therapeutic agent to a subject for treating a solid tumor is disclosed by implanting a biocompatible polymer containing a therapeutic agent into the subject. The biocompatible polymer continuously releases the therapeutic agent into the subject over time. In some embodiments, the cancer treated may be breast cancer, including TNBC, or PTEN-ineffective TBNC. In some embodiments, the implanted biocompatible polymer may contain a STING agonist.

[0008] In some embodiments, the implantable biocompatible polymer may be a biopsy marker. In some embodiments, the biopsy marker may have a core of poly(p-dioxanone) (PDO), polylactide (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), or copolymers thereof; and a sealant surrounding the core, the sealant comprising a polyethylene glycol (PEG)-based hydrogel. In some embodiments, the biopsy marker may be HydroMARK. ™ Biopsy markers.

[0009] In some embodiments, this document discloses a method for treating a patient's breast cancer by hydrating a dehydrated biopsy marker with a STING agonist and implanting the hydrated biopsy marker into the patient at a location close to the breast cancer cells. The dehydrated biopsy marker may comprise a polyethylene glycol (PEG)-based hydrogel.

[0010] In some embodiments, this document discloses a method for implanting a hydrated HydroMARK containing a STING agonist near the breast cancer without removing the breast cancer. ™ A method for treating PTEN-ineffective TNBC in subjects using biopsy markers.

[0011] This article discloses HydroMARK with small molecules. ™ Biopsy markers containing STING agonists. Attached Figure Description

[0012] This patent or application document contains at least one drawing in color. A copy of this patent or application disclosure with a color drawing will be provided by the Patent Office upon request and payment of the necessary fees.

[0013] Figure 1 shows that defective STING transport makes TNBC cells highly sensitive to STING agonism. Figure 1A Images of STING staining depicting TNBC tissue microarrays (TMA) with scoring criteria are shown (n=63). 0 = no staining in tumor cells; 1+ = weak staining in >10% of tumor cells; 2+ = moderate staining in >10% of tumor cells; 3+ = strong staining in >10% of tumor cells. Figure 1B Images from two cases are depicted, showing differences in staining intensity between the core and the center. Case 24, score 3+ and 0; Case 89, score 2 and 0. Figure 1C A representative image of PTEN staining of TNBC with STING 3+ is shown. Figure 1D An immunoblot of a plate of TNBC cell line against PTEN and STING is shown. Figure 1EImmunoblots against PTEN and STING were shown for unaltered MDAMB231 (parental) cells, cells containing CRISPR control vectors (randomized), or cells containing PTEN knockout vectors (Sg1, Sg2, Sg3). Cells were treated with PBS as a control or with 10 µM or 50 µM ADU for 48 hours. CXCL10 expression in the supernatant was then analyzed by ELISA (n=3). Two-dimensional ANOVA using Tukey's MCT was performed. *, P<0.05; **, P<0.005; ***, P<0.0002; ****, P<0.0001. Figure 1F Immunoblots against Rab7 and STING were shown for MDA-MB231 parental cells or those containing CRISPR control vectors (randomized) or Rab7 knockout vectors (Sg1, Sg2). Cells were treated with 50 µM ADU for 48 hours, and CXCL10 expression in the supernatant was analyzed by ELISA (n=2). ANOVA by Tukey's MCT showed *, P<0.05; ***, P<0.0002; ****, P<0.0001. Figure 1G Immunofluorescence staining for STING (green) and lysosome (red) colocalization was shown after MDAMB231 cells were treated with 50 µM ADU or 100 nM CID1067700 or a combination thereof for 1 h and 6 h (representing n=3 independent experiments). Colocalization was quantified using ImageJ (n=4). Two-dimensional ANOVA with Tukey's MCT showed **, P<0.005; ***, P<0.0002. Figure 1H The growth inhibition assays of PTEN-ineffective and PTEN WT cell lines over 6 weeks are shown. MDA-MB468 and HCC70 were treated twice weekly with 10 μM or 50 µM ADU. MDAMB231 and HCC1806 were treated twice weekly with 10 µM or 50 µM ADU + / - 100 nM CID1067700. Cell growth was visualized by crystal violet staining and then quantified using absorbance at 590 nm (n=3). Figure 1I ). Figure 1J The growth inhibition assays are shown, in which MDA-MB231 disordered and PTEN KO cell lines (Sg1, Sg3) are compared with... Figure 1H Similarly, the procedure was performed, with crystal violet staining for visualization, followed by quantification using absorbance at 590 nm (n=3). ANOVA using MCT with Tukey's method showed *, P<0.05; **, P<0.005; ***, P<0.0002; ****, P<0.0001. Figure 1K ). Figure 1LImmunoblots of MDMAB468 or Rab7 knockout vectors (Sg1, Sg2) containing CRISPR control vectors (randomized) against Rab7 and STING are shown. Figure 1M Immunofluorescence staining for STING (green) and lysosome (red) colocalization was shown after MDAMB231 cells were treated with 50 µM ADU, 100 nMCID1067700, or a combination thereof for 1 h and 6 h. Colocalization was quantified using ImageJ (n=4). Two-dimensional ANOVA with Tukey's MCT was performed, **, P<0.005; ***, P<0.0002. Figure 1N The growth inhibition assay is shown. Figure 10 The effects of various treatments on CXCL10 are shown. Figure 1P The H-score analysis of STING levels in the PTEN-negative (n=38) and PTEN-positive (n=2) subsets is shown. Six PTEN-low positive TNBCs were included in the PTEN-negative subset. Figure 1Q The quantitative analysis and frequency distribution of H-scores for STING expression in the PTEN-negative subset are shown (n=38). An H-score ≤ 30 indicates low to negative expression. An H-score > 75 indicates high expression.

[0014] Figure 2. Impaired STING turnover enhances STING agonist-induced T cell migration. Figure 2A Luminex cytokine profiling analysis of disordered and Rab7 knockout cells using PTENWT cell lines MDAMB231 and HCC1806, and the PTEN null cell line MDAMB468, is shown. Log-2 fold changes (n=2–4) of cytokine / chemokine differences are shown relative to untreated disordered cells, Rab7 knockout, ADU-treated disordered cells, and ADU-treated Rab7 knockout cells. Figure 2B The results measured by ELISA are shown. Figure 1A The expression of CXCL10, IL-8, and IL-6 in the culture medium (CM) of the samples described in the experiment was evaluated. ANOVA using MCT with Tukey showed *, P < 0.05; **, P < 0.005; ***, P < 0.0002; ****, P < 0.0001. Figure 2C A schematic diagram depicts the assay of CD8+ T cell migration using a 3D microfluidic device. Figure 2DImages of T cells (yellow) migrating to MDAMB231 spheroids (DAPI) are shown. MDAMB231 spheroids were pretreated with 50 µM ADU for 18 hours before loading the 3D microfluidic device, followed by Rab7 knockout. Migrating CD8+ T cells (n=8–9) were quantified after 48 hours. ANOVA by MCT with Tukey showed ***, P<0.0002; ****, P<0.0001. Figure 2E It shows T cells turning towards Figure 1D Images of HCC1806 spheroids migrating in similarly treated cells. Quantification of migrating CD8+ T cells (n=7–10) after 48 hours. ANOVA by MCT with Tukey, ***, P<0.05; ****, P<0.005. Figure 2F CXCL10 quantification in MDAMB231 and HCC1806 cells treated with 50 µM or 10 µM ADU for 48 h in the presence or absence of escalating doses of CID1067700 is shown. CXCL10 in the supernatant was analyzed by ELISA (n=3–5). ANOVA by Tukey's MCT: *, P<0.05; ***, P<0.0002; ****, P<0.0001. Figure 2G A schematic diagram of Jurkat cell migration assays using a 3D microfluidic device is depicted. Representative images of Jurkat cells (pink) migrating to MDA-MB231 spheroids (DAPI) after treatment with 100 nM CID1067700 + / - 50 μM ADU-S100 are shown. Migrating Jurkat cells (n=4) were quantified after 48 hours. ANOVA by MCT with Tukey is shown in the following figures: *, P < 0.05; ***, P < 0.0002; ****, P < 0.0001. Figure 2H It shows in Figure 1A CCL5 expression in HCC1806 was assessed by ELISA in the culture medium (CM) used. ANOVA with Tukey's MCT showed ***, P < 0.0002; ****, P < 0.0001. Figure 2I Representative images of Jurkat cells (pink) migrating to MDAMB231 spheroids (DAPI) after treatment with 100 nM CID1067700 + / - 50 µM ADU. Quantification of migrating Jurkat cells (n=4) after 48 hours. ANOVA by MCT with Tukey, **, P < 0.005; ***, P < 0.0002; ****, P < 0.0001. Figure 2JMultiplex cytokine profiles of the control PTEN null cell lines MDAMB468 and PTEN WT cell lines MDAMB231 and HCC1806 are shown compared to Rab7 KO - / + ADU treatment. The heatmap illustrates the control cell normalized log⁻² fold change (L2FC) of key STING-IRF3 and STING-NFκB cytokines / chemokines. Figure 2K It is an immunoblot of HCC1806 parent or containing a CRISPR control vector (Acramble) or a Rab7 knockout vector (Sg1, Sg2).

[0015] Figure 3 shows that impregnation of STING agonist in PEG hydrogel biopsy markers leads to their local delivery to TNBC. Figure 3A Images of dried radiographic biopsy markers and their corresponding images after 2 hours of hydration are shown. Figure 3B Images of PEG markers implanted in mice on days 0 and 4 are shown, along with magnified views of the markers on days 8 and 25. Figure 3C A schematic diagram of the THP-1 assay is shown. A known amount of ADU-S100 was added to THP1 cells, followed by ELISA to determine the corresponding CXCL10 level. Figure 3D Showing from Figure 3C The average of three repetitions was used to plot the average to produce a standard curve. Figure 3E A schematic diagram is shown for determining the amount of ADU released from a PEG-labeled compound containing 100 µg of ADU over time. Figure 3F It shows the relationship with Figure 3E The corresponding CXCL10 ELISA value is used to utilize Figure 3D The standard curve generated in the process determines the amount of ADU released at each time point. Figure 3G The amount of ADU-S100 released from each biopsy marker over time after immersion in 100 µg of ADU is shown. The average of three replicates is shown. Figure 3H The size of tumors that developed in BALB / c mice after day 4, when 4T1 cells (10kJ) were seeded into the mammary fat pads and then implanted with a PEGylated marker containing either PBS or 100µg ADU, is shown. Individual tumor volume measurements are shown (n=5 mice / group). Figure 3I The size of tumors that developed in BALB / c mice after inoculation of 4T1 cells (100kJ) into the mammary fat pads is shown. Mice were implanted with PEG-labeled material soaked in PBS or 100µg ADU when the tumor size reached 150mm³ on day 6. Individual tumor volume measurements are shown (n=7 mice / group). Figure 3JThe image shows the result of treatment with PEG+PBS on day 25. Figure 3I Images of 4T1 mice. Anatomy of the tumor reveals a complete PEG marker with visible metal clamps. Figure 3K (Left) Shows the volume of PEG markers at each time point after tumor implantation in mice. (Right) Shows the amount of ADU released from various biopsy markers embedded with 100 µg of ADU.

[0016] Figure 4 shows that STING agonist monotherapy is toxic to T cells and does not induce T cell clonal activity. Figure 4A The percentages of CD45+ leukocytes, lymphocytes, and T cells (CD3+, CD4+, CD8+) in primary human TNBC explants treated with 50 µM ADU or PBS for 24 hours, then dissociated into single-cell suspensions and analyzed by flow cytometry are shown. Figure 4B The percentage of cell viability in human CD8+ T cells treated with different doses of ADU for 24 hours is shown; cell viability was determined by CellTiter-Glo (CTG) assay at 24 hours (n=3). ANOVA by MCT with Tukey, ****, P<0.0001. Figure 4C Percentage of CD3+, CD4+, and CD8+ T cell viability. Spleen cells were harvested from C57BL / 6 mice and treated with different doses of ADU for 24 hours. CD3+, CD4+, and CD8+ T cell viability (n=2) was determined by flow cytometry. ANOVA by Tukey's MCT: *, P<0.05; **, P<0.005; ***, P<0.0002. Figure 4D (Left) Shows human CD8+ T cells treated with conditioned medium and observed at different time points. Figure 3E The percentage of viable cells after PEG collection, which is soaked in 100 µg of ADU. Figure 4D (Right) Shows CD8+ T cells treated with direct addition of PBS, direct addition of 100 µg ADU, PEG+PBS, or PEG+ADU containing 100 µg ADU. Cell viability was measured by CTG at 24 hours (n=3). The first plate shows cells treated for 5 days without culture medium replacement, with cell viability measured by CTG (N=3). The second plate shows cells treated in the same manner, with surviving ADU-treated cells replated on day 5, stimulated with IL-2, IL-7, and IL-15, and then CTG measured on day 10 (n=3). Figure 4EThe tumor size is shown after 4T1 cells (10kJ) were seeded into the mammary fat pads of BALB / c mice and implanted with a PEGylated marker containing PBS or 100µg ADU on day 4. On day 30, another PEGylated marker containing PBS or 100µg ADU was implanted. Individual tumor volume measurements are shown (PBS n=6, ADU n=10). * indicates mice used for single-cell RNA sequencing (scRNA-seq) and T-cell receptor sequencing (TCR-seq). Figure 4F Depicting Figure 4E A schematic diagram of the experimental design used. Figure 4G A UMAP plot is shown, displaying unsupervised clusters of scRNA-seq data from 4T1 tumors from E-cells treated with PEG+PBS and PEG+ADU. Cell allocation was based on comparison with cluster gene expression tags from established literature. Activated T cells were the dominant cluster exhibiting any relevant clonoid. Figure 4H A UMAP diagram is shown, which displays clusters with any associated TCR clones. Figure 4I The pie chart shows the percentage of cells with unique TCR clones at frequencies of 1, 2, and 5 in the PBS and ADU treatment groups. Figure 4J Primary human TNBC explants were shown after treatment with 50 µm ADU or PBS for 24 hours and then dissociation into single-cell suspensions. Flow cytometry was performed to analyze the percentages of viable CD45+ leukocytes, CD3+ lymphocytes, and T cells (CD4+, CD8+), as shown in the pie chart.

[0017] Figure 5A Additional studies on the use of STING agonists in PEG hydrogel biopsy markers. Figure 5A Studies without PEG hydrogels are shown. In established 4T1 tumors, either PBS or 100 µg ADU was injected intratumorally on day 6. Mean tumor volume measurements are shown (n = 4–5 mice / group). Figure 5B Representative IHC images of pSTAT1 and pTBK1 staining in 4T1 tumors treated with PEG+PBS or PEG+100µg ADU for 4 days are shown. Scale bar, 100µm. Figure 5C The diagram illustrates the in situ implantation of 100K 4T1 cells, followed by implantation with PEG+PBS or PEG+100µg ADU. Fourteen days later, scRNA-seq and TCR-seq were performed on the four pooled tumors / groups. Figure 5D (Left) UMAP plot, showing from Figure 5C Collected CD45 +Unsupervised clusters of scRNA-seq data from cells. The volcano plot (right) shows an overview of differentially expressed genes (DEGs) in the APC+ macrophage population (K) treated with PEG+ADU versus PEG+PBS. Red dots indicate significantly upregulated genes. Blue dots indicate significantly downregulated genes. Figure 5E and Figure 5F Gene set enrichment analysis (GSEA) based on fold change values ​​of ADU versus PBS-treated tumors in APC+ macrophage clusters is shown. GSEA plots of IFNγ and IFNα responses are shown in (E). Bar plots represent the normalized enrichment scores (NES) of the top upregulated marker gene set (F). Quantitative data are presented as mean ± SEM; p-values ​​were calculated by two-way ANOVA followed by Sidak's post-hoc test (A), ns, not significant; ****, p < 0.0001. Detailed Implementation

[0018] This article discloses a method for administering a dose of a small molecule (containing a STING agonist) to a solid tumor (e.g., immune-mediated clearance of STING-positive TNBC) to avoid negative effects on T cells while still effectively treating the tumor. First, this article discloses certain breast cancers, including triple-negative breast cancer (TNBC) containing phosphatase-free and tensin homologous (PTEN), which, in some embodiments, may be highly sensitive to STING agonistic effects due to defective Rab7 transport, and / or may lead to enhanced T cell migration to the cancer. Second, we have found that certain biocompatible polymers (e.g., small molecules, including STING agonists) implanted near a subject's solid tumor can continuously release the therapeutic agent over time. In the case of STING agonists, release from the biocompatible polymer avoids the severe effects on T cells observed when STING agonists are injected intratumorally.

[0019] In some embodiments, the biocompatible polymer can be an expandable solid form of a polyethylene glycol (PEG)-based hydrogel. PEG can be dry / dessicated and can expand upon fluid contact. In some embodiments, the polymer can be part of a biopsy site marker or biopsy clip. The biopsy site marker may include a metallic marker component. In some embodiments, the biopsy site marker may include HydroMARK. ™ Device.

[0020] In some implementations, this platform provides local delivery of STING agonists and other small molecules into the tumor microenvironment of solid cancer.

[0021] In some implementations, the methods disclosed herein can be used to treat human tumors.

[0022] definition

[0023] This document provides specific embodiments of one or more implementations. However, it should be understood that the invention may be embodied in various forms. Therefore, the specific details disclosed herein should not be construed as limiting, but rather serve as the basis for the claims and as a representative basis for teaching those skilled in the art to employ the invention in any appropriate manner.

[0024] Unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “the / said” include plural indicators. In the claims and / or description, the use of the words “a” or “an” in conjunction with the term “comprising” may refer to “an,” but is also consistent with “one or more,” “at least one,” and “one or more.”

[0025] Wherever phrases such as “for example,” “such as,” or “contains” are used in this document, unless otherwise expressly stated, they should be understood as followed by the phrase “and but not limited to.” Similarly, “example,” “exemplary,” etc., should be understood as non-restrictive.

[0026] The term “substantially” allows for deviations from descriptors that do not negatively impact the intended purpose. Even if the word “substantially” is not explicitly listed, descriptive terms should be understood as being modified by the term “substantially.”

[0027] The terms “comprising,” “including,” “having,” and “involving” are used interchangeably and have the same meaning. Specifically, each of these terms is defined in accordance with the general U.S. patent law definition of “comprising” and is therefore to be interpreted as an open term meaning “at least the following,” and also to not exclude additional features, limitations, aspects, etc. Thus, for example, “a process involving steps a, b, and c” means a process comprising at least steps a, b, and c. Wherever the terms “a” or “an” are used, they should be understood as “one or more” unless such interpretation is meaningless in the context.

[0028] As used herein, the term “about” means approximately, roughly, around, or within a range. When used in conjunction with a numerical range, the term modifies the range by extending the boundaries above and below the stated value. Typically, the term “about” is used herein to modify values ​​above and below the stated value by a variation of, for example, 20% upwards or downwards (higher or lower).

[0029] Biocompatible polymers

[0030] In some embodiments, the polymers disclosed herein are biocompatible. In some embodiments, the polymers are implantable. The polymer can retain a therapeutic agent, which can be released from the polymer (e.g., diffuse out of the polymer) after implantation into a subject. In some embodiments, the therapeutic agent can be continuously released from the polymer in the subject over a period of time (e.g., sustained release).

[0031] In some embodiments, the therapeutic agent may be adsorbed onto the polymer. Upon implantation, the therapeutic agent may be desorbed from the polymer. In some embodiments, the therapeutic agent may be absorbed by the polymer. For example, the therapeutic agent may be dissolved in a solvent (e.g., water) that can be used to hydrate and dehydrate the polymer (e.g., an aqueous swellable polymer) (absorption). Upon implantation, as body fluids replace the solvent, the therapeutic agent may be removed from the hydrated polymer along with the solvent.

[0032] In some embodiments, the biocompatible polymer can be a biodegradable polymer, a non-biodegradable polymer, or a natural polymer. In some embodiments, the biodegradable polymer can be poly(p-dioxanone) (PDO), polyethylene glycol (PEG), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), poly(lactic acid-co-glycolytic acid) (PLGA), poly(caprolactone) (PCL), or combinations thereof.

[0033] In some implementations, the non-biodegradable polymer may be poly(siloxane) (silicone) or poly(ethylene-vinyl acetate).

[0034] In some implementations, the natural polymer can be albumin, cellulose, chitosan, collagen, elastin, gelatin, milk protein, silk, etc.

[0035] In some embodiments, the biocompatible polymer may be hydrophilic. In some embodiments, the biocompatible polymer may be a hydrogel. In some embodiments, the biocompatible polymer may be a PEG-containing hydrogel. The hydrogel may be chemically linked (e.g., covalently cross-linked), physically linked (e.g., non-covalently linked), or a combination thereof.

[0036] In some embodiments, the biocompatible polymer may be poly(p-dioxanone) (PDO), polylactide (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), copolymers thereof, etc.

[0037] In some embodiments, the biocompatible polymer may have a core that can be surrounded by a layer. In some embodiments, the layer surrounding the core may be a sealant. In some embodiments, the core may have poly(p-dioxanone) (PDO), polylactide (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), and / or copolymers thereof. In some embodiments, the core may be surrounded by a sealant layer. The sealant layer may be a polyethylene glycol-based hydrogel.

[0038] In some embodiments, the biocompatible polymer (e.g., the core) may have a metallic marker. In some embodiments, the marker may be titanium or stainless steel.

[0039] In some embodiments, the biocompatible polymer may have pores ranging in size from 10 µm to 500 µm. The spacing between the pores in the polymer may be from about 5 nm to 100 nm.

[0040] biopsy markers

[0041] In some embodiments, the biocompatible polymer described herein may be part of a biopsy marker or biopsy clip. The biopsy marker may have a metallic marker, such as a titanium or stainless steel marker. The biopsy site marker can be used to mark the body area where a biopsy is performed. The implanted biopsy marker may be retained indefinitely and used for preoperative localization when a biopsy identifies cells that must be removed, or for future site marking when pathology indicates that the biopsy cells are benign.

[0042] In this document, any biopsy marker can be used to deliver therapeutic agents. In some embodiments, the biopsy marker may be a polyethylene glycol-based substance, collagen, beta-glucan, or polylactic-co-glycolic acid. In some embodiments, the biopsy marker may be HydroMARK. ™ Biopsy markers, MammoMARK ® Biopsy markers, MammoSTAR ® Biopsy markers or SecurMark ® Biopsy markers. In some implementations, HydroMARK ™ The biopsy markers are marketed by Mammotome, Devdor Medical Products, Inc., which is part of Leica Biosystems in the United States.

[0043] HydroMARK ™The device can be made from an expandable solid cylinder of polymerized and dried polyethylene glycol (PEG)-based gel. The device expands upon fluid contact, and the implanted device remains visible under ultrasound for 6 weeks to 12 months. The device may also contain embedded metallic components, such as titanium or stainless steel, which are permanently visible via X-ray and MR imaging.

[0044] In some implementations, the biopsy markers may be dehydrated or dried. Figure 3A In some implementations, the biopsy marker may be hydrated. The length of the hydrated marker that is ultimately implanted in the subject may be approximately 7 mm, 9 mm, 11 mm, 13 mm, 15 mm, 17 mm, or 19 mm.

[0045] In some embodiments, this document discloses biopsy markers that have been "loaded" with one or more therapeutic agents. In some embodiments, this document discloses HydroMARKs that have been loaded with a STING agonist. ™ Biopsy markers.

[0046] cancer

[0047] The compositions and methods disclosed herein can be used to treat a variety of cancers. In some embodiments, the cancer may be a solid tumor. In some embodiments, the cancer may be a primary solid tumor or a metastatic solid tumor. In some embodiments, the cancer may be carcinoma, lymphoma, melanoma, or sarcoma. In some embodiments, the cancer may be bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colorectal cancer, gastrointestinal cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, or thyroid cancer. In some embodiments, the cancer cells may lack or have reduced amounts of functional phosphatases and tensin homologs (PTEN). In some embodiments, PTEN may be inactivated or ineffective.

[0048] In some implementations, the cancer being treated may be breast cancer. Breast cancer may include ductal carcinoma in situ (DCIS), HER2-positive breast cancer, invasive ductal carcinoma (IDC), invasive lobular carcinoma (ILC), mixed ductal and lobular carcinoma, inflammatory breast cancer, or triple-negative breast cancer (TNBC). In some implementations, breast cancer cells may lack or have reduced amounts of functional phosphatases and tensin homologs (PTEN). In some implementations, PTEN may be inactivated or nullified.

[0049] In some implementations, the cancer can be a cancer that overexpresses STING. STING (interferon gene stimulator) is a transmembrane protein encoded by TMEM173. STING is a regulator of the innate immune response to cytoplasmic double-stranded DNA (dsDNA) in cancer. Upon activation, STING undergoes lysosomal turnover mediated by the Ras-associated protein Rab-7a (Rab7). In some cells, including triple-negative breast cancer (TNBC) cells, this physiological STING degradation can be disrupted by maintaining Rab7 in a hyperphosphorylated, inactive state.

[0050] We recently reported that STING expression in tumor cells is elevated in TNBC, while luminal subtypes A and B are predominantly STING negative. Furthermore, when examining STING expression in a tumor cell microarray (TMA) of 64 individuals with TNBC, there was a clear trend towards STING-high and STING-low TNBC, with approximately 61% of the TNBC expressing STING.

[0051] In TNBC, cells with inactivated or nullified phosphatase and tensin homolog (PTEN) maintain Rab7 in an inactive state. STING transport to lysosomes is impaired in these cells, and the cells are highly sensitive to STING. Rab7 inhibitors exist and can be used to increase the sensitivity of non-PTEN-deficient cells to STING.

[0052] Therapeutic agents

[0053] The therapeutic agent released from the implanted biocompatible polymer can be of various types. In some embodiments, the therapeutic agent can be a small molecule.

[0054] In some implementations, the therapeutic agent may be a STING agonist. STING agonists can cause an increase in STING activity. In some examples, STING agonists can cause an increase in inflammatory cytokines, which can remodel the tumor microenvironment and enhance anti-tumor T cell responses.

[0055] In this document, any STING agonist may be used. In some embodiments, the STING agonist may include ADU-S100, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING, E7766, TAK-676, SNX281, SYNB1891, etc., or combinations thereof.

[0056] In some implementations, the therapeutic agent may be a small molecule serving as an estrogen receptor-targeted therapy. Estrogen receptor-targeted therapy may be a selective estrogen receptor modulator (SERM). In some implementations, the SERM may be tamoxifen, raloxifene, etc.

[0057] In some implementations, the therapeutic agent may be a Rab7 inhibitor.

[0058] In some implementations, the biocompatible polymer may release two or more small molecules and / or STING agonists.

[0059] method

[0060] In some implementations, the biocompatible polymer may be "loaded" with a therapeutic agent. The therapeutic agent may be loaded into the biocompatible polymer before and / or after implantation of the polymer into a subject.

[0061] In some embodiments, the biocompatible polymer can be dehydrated prior to loading the therapeutic agent. The therapeutic agent can be loaded into the polymer by rehydrating it with a solution containing the therapeutic agent. Different volumes of solutions containing the therapeutic agent can be used to hydrate the polymer. In some embodiments, the biocompatible polymer can be hydrated in a volume of less than about 20 µl.

[0062] In some implementations, the biocompatible polymer can be implanted into the body of a subject. The biocompatible polymer can be implanted near the tumor or cancer requiring treatment. The biocompatible polymer can be implanted within 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, or 10 mm of a solid tumor. The biocompatible polymer can be implanted within the solid tumor. The biocompatible polymer can be implanted around the periphery of the solid tumor. The biocompatible polymer can remain in the subject's body for different time periods. In some implementations, the polymer can remain in the subject's body for days, weeks, or months.

[0063] In some embodiments, the tumor is not removed before / during the implantation of the biocompatible polymer. In other embodiments, the tumor may be removed before / during the implantation of the biocompatible polymer.

[0064] Implantation of biocompatible polymers may include surgical implantation or implantation using a large-hole needle. In some embodiments, a variety of biocompatible polymers (i.e., emboli) may be implanted.

[0065] In some embodiments, after implantation, the biocompatible polymer may be loaded with a therapeutic agent. This can be done using a syringe containing the therapeutic agent to be loaded. The needle of the syringe is inserted through the subject's skin and into the implanted polymer. The syringe can deliver the therapeutic agent (e.g., a STING agonist) into the implanted polymer. In some embodiments, other substances may be injected into the implanted biocompatible polymer. In some embodiments, a dehydrated biocompatible polymer (e.g., a biopsy marker) may be loaded with a therapeutic agent (e.g., a STING agonist) and implanted into the subject. Subsequently, the same or another therapeutic agent and / or substance may be injected into the implanted biocompatible polymer.

[0066] In some embodiments, a large-hole needle can be used to insert a dehydrated biocompatible polymer (e.g., a biopsy marker) through the skin and place it near a tumor in a subject. In some embodiments, another syringe containing a therapeutic agent can then be inserted through the skin and into the placed biocompatible polymer. The therapeutic agent can be delivered to the placed biocompatible polymer using the syringe.

[0067] As disclosed herein, the implantable biocompatible polymer that delivers a therapeutic agent to a solid tumor can be used alone to treat a subject. In some embodiments, this treatment can be used in combination with other treatments. In some embodiments, the treatment disclosed herein can be used in combination with conventional therapies. In some embodiments, conventional therapies may include chemotherapy, endocrine therapy, etc.

[0068] Composition

[0069] This document discloses implantable biocompatible polymers comprising therapeutic agents. Embodiments of these biocompatible polymers, combined with therapeutic agents that can be combined with the biocompatible polymers, are described in full throughout this application.

[0070] In some embodiments, compositions of biocompatible polymers and therapeutic agents that may include metal markers or clamps are disclosed. The biocompatible polymer having the therapeutic agent may be hydrated.

[0071] In some embodiments, the biocompatible polymer may be a polymer core comprising a metal marker, surrounded by a polyethylene glycol (PEG)-based hydrogel. In some embodiments, the polymer core may be poly(p-dioxanone) (PDO), polyethylene glycol (PEG), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), poly(lactic acid-co-glycolytic acid) (PLGA), poly(caprolactone) (PCL), or a combination of these polymers. The PEG-based hydrogel surrounding the core may be referred to as a "sealing agent." In some embodiments, the metal marker or clamp may be titanium, stainless steel, etc.

[0072] In some embodiments, the biocompatible polymer may be a biopsy marker. In some embodiments, the biopsy marker may be a combination of biodegradable polymers surrounded by a hydrogel. In various embodiments, the biopsy marker may be of many different types, including HydroMARK. ™ MammoMARK ® MammoSTAR ® SecurMark ® wait.

[0073] Therapeutic agents bound to biocompatible polymers can be absorbed into and / or adsorbed onto the biocompatible polymer. In some embodiments, the dehydrated biocompatible polymer can be hydrated in a solution or suspension containing the therapeutic agent. The biocompatible polymer may have one or more associated therapeutic agents.

[0074] In some embodiments, the therapeutic agent may be a small molecule. Exemplary small molecules may be STING agonists as described herein. Exemplary STING agonists may be ADU-S100, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING, E7766, TAK-676, SNX281, SYNB1891, and other molecules with the same or similar functions.

[0075] Example

[0076] The following examples are provided to facilitate a more complete understanding of the invention. The examples illustrate exemplary modes of preparing and practicing the invention. However, the scope of the invention is not limited to the specific embodiments disclosed in these examples for illustrative purposes only, as similar results can be obtained using alternative methods.

[0077] Example 1

[0078] Defective STING transport makes TNBC cells highly sensitive to STING agonism. .

[0079] STING immunohistochemical studies were performed on TNBC tissue microarrays (TMA) derived from untreated patients (n=63) with clinicopathological features as described in Table 1. Breast pathologists visually scored the samples using an intensity scale: 0 = no staining in tumor cells with intact expression as an internal control using endothelial and inflammatory cells; 1+ = weak staining in >10% of tumor cells; 2+ = moderate staining in >10% of tumor cells; 3+ = strong staining in >10% of tumor cells. Figure 1AMore than half of the TNBCs showed STING expression, and four samples showed high variability in staining from 0 to 2+ / 3+, with similar histology in both high and low STING expression areas. Figure 1B In STING 3+TNBC, 5 out of 6 samples were available for PTEN staining and were PTEN IHC negative or low. No association was observed between heterogeneity and age, lymph node status, tumor size, or grade (Table 1).

[0080] Table 1. Clinicopathological features of triple-negative breast cancer patients. 1、2

[0081]

[0082] 1. American Joint Committee on Cancer (8th Edition) pT and pN Categories

[0083] 2N / A is not applicable.

[0084] Previous studies have shown that PTEN (phosphatase and tensin homolog) is present in TNBC without tumor suppressor factors, which maintains the lysosomal protein Rab7 in an inactive state. Therefore, the transport of STING to lysosomes may be impaired, leading to baseline upregulation of STING protein and increased sensitivity to STING agonism (12). Therefore, PTEN IHC was performed on TMA, and this inverse relationship between PTEN and STING was confirmed. Figure 1C Analysis of larger TMAs showed that the PTEN ineffective state tended to have elevated STING levels in tumor cells. Figure 1P and 1Q Similarly, using a set of TNBC cell lines, the PTEN blank cell lines (MDAMB468, HCC1937, HCC70) showed higher STING protein expression than the PTEN WT cell lines (HCC1143, HCC1806, MDAMB231). Figure 1D Furthermore, when PTEN is knocked out in MDAMB231, the STING protein is upregulated and becomes sensitive to ADU-S100, resulting in higher levels of the interferon regulator factor 3 (IRF3)-regulated chemokine CXCL10. Figure 1E ).

[0085] To broaden the use of STING agonists for PTEN WT TNBC, we directly targeted Rab7 via both protein knockout and the use of a Rab7 inhibitor (CID1067700), which acts as a competitor to the GTP and GDP binding sites in Rab7, thereby preventing nucleotide exchange and activation (24, 25). Rab7 knockout in MDAMAB231 resulted in a significant increase in STING protein and a sensitization response to ADU-S100, with a significant increase in CXCL10 ( Figure 1F As a control, Rab7 knockout in PTEN WTMDAMB468 cells did not alter STING protein levels. Figure 1L ).

[0086] To investigate the immediate effects of Rab7 inhibition, MDAMB231 cells were treated with ADUS100, the Rab7 inhibitor CID1067700, and combinations thereof. Immunofluorescence staining of STING and lysosomes was then performed to monitor co-localization. Following ADU-S100 stimulation, significant co-localization of STING with lysosomes for degradation was observed, which was disrupted in the presence of Rab7 inhibition. Figure 1G ; Figure 1M In previous growth assays, chronic ADU-S100 treatment of PTEN-ineffective cell lines MDAMB468 and HCC70 resulted in significant growth inhibition at a low dose of 10 µM ADU-S100, which was not observed in MDAMB231 and HCC1806. The addition of a Rab7 inhibitor sensitized MDAMB231 and HCC1806 to ADU-S100, with a more pronounced effect in the latter. Figure 1H ), which is quantified by the absorbance of crystal violet ( Figure 1I ).

[0087] Because MDAMB231 exhibits a relatively weak response to Rab7 inhibitors, particularly ADUS100 at 10 µM, treatment of MDAMB231 PTEN knockout cells with ADU-S100+ / - Rab7 inhibitors revealed that PTEN loss sensitized cells to lower concentrations of ADUS100 (10 µM), and the addition of Rab7 inhibitors further enhanced this effect. Figure 1J and Figure 1K ; Figure 1N Furthermore, combination therapy increased CXCL10 production in MDMAMB231 and HCC1806, but not in MDMAB468 where Rab7 was inactivated, and a robust increase in CXCL10 was achieved with ADU-S100 alone. Figure 10 ).

[0088] These data indicate that PTEN-ineffective TNBC has a regulatory aberration of STING, which prevents its transport to lysosomes for degradation, and in PTEN WT TNBC, this can be summarized by Rab7 knockout or the addition of a Rab7 inhibitor.

[0089] Example 2

[0090] Impaired STING turnover enhances STING agonist-induced T cell migration .

[0091] Previous data showed that Rab7 knockout upregulated CXCL10. To better analyze these resulting cytokines, MDAMB231 and HCC1806 were measured using Luminex assays with MDAMB468 as a control. Additionally, disordered control cells and RAB7 knockout cells from each cell line were treated with 50 µM ADU-S100. Figure 2K The aim was to assess whether cytokine levels could be further increased. Key induced cytokines were TBK1-IRF3-regulated CXCL10 and CCL5, and NF-κB-regulated cytokines IL-8 and IL-6. Figure 2A and Figure 2J CXCL10 and CCL5 are both involved in T cell recruitment, while IL-6 and IL-8 are associated with T cell recruitment and immunosuppression (26). ELISA was then performed to confirm the upregulation of CXCL-10, IL-8, and IL-6. Figure 2B The MDMAMB468 control did not show any further increase in cytokine production from Rab7 knockout because Rab7 was already inactivated in PTEN-deficient cells, and adding ADUS100 to both confounding and Rab7 knockout cells did not show any additive effect. However, MDA-MB231 cells with Rab7 knockout not only resulted in increased CXCL10 production but also increased IL-8 and IL-6 at baseline and in response to ADU-S100 stimulation. Similarly, HCC1806 Rab7 knockout cells showed increased CXCL10 levels, which were further enhanced with ADU-S100. Compared to disordered production, stimulation of Rab7 knockout HCC1806 cells with ADU-S100 resulted in increased CCL5 ( Figure 2H IL-1, IL-8, and IL-6 also increased significantly.

[0092] Given that activation of cytotoxic T lymphocytes depends on successful transport and infiltration into tumor tissue (27), and that cytokines upregulated by Rab7 knockout are primarily involved in T cell transport, the potential in vivo effects of immune cell recruitment were modeled using the previously described 3D microfluidic T cell migration assay (27). Figure 2C(28). MDA-MB231 scrambled and Rab7 knockout spheroids were pretreated with 50 µM ADU-S100 and loaded into the migration zone of a 3D microfluidic device. Primary human CD8+ T cells were then introduced into the lateral channels, and T cell transport to the migration zone was assessed after 48 hours. Figure 2D As shown, even with ADU-S100 treatment, minimal T cell migration to MDAMB231 disordered globules was observed. Significant T cell migration to MDAMB231 Rab7 knockout globules was observed due to increased CXCL10 levels from Rab7 knockout in PTEN WT TNBC, and ADU-S100 treatment further increased T cell transport to the migration zone. HCC 1806 showed a similar T cell migration pattern. Figure 2E ).

[0093] To assess whether these findings could be replicated using the Rab7 inhibitor CID1067700, it was first confirmed that the addition of the Rab7 inhibitor increased CXCL10 levels in both untreated and ADU-S100-treated MDAMB231 and HCC1806 cells. Next, the dosage of the Rab7 inhibitor containing ADU-S100 was optimized to achieve the highest possible increase in CXCL10 levels in both MDAMB231 and HCC1806 cells. Figure 2F Using MDAMB231, spheroids were treated with 100 nM Rab7 inhibitor in a 3D microfluidic device with or without 50 µM ADU-S100, followed by introduction of CXCR3-expressing Jurkat cells through a lateral channel. Pretreatment with ADU-S100 resulted in significant migration of Jurkat cells, which was further enhanced by the Rab7 inhibitor. Figure 2G ; Figure 2I ).

[0094] In summary, these findings identify PTEN-deficient or Rab7-knockout / repressed PTEN WTs as a genomic background in TNBC, for which STING agonists can be used not only to induce cellular intrinsic toxicity but also to recruit T cells to the TME.

[0095] Example 3

[0096] Impregnation of the STING agonist in the PEG hydrogel biopsy marker results in its localized delivery to the TNBC. .

[0097] In human clinical trials, intratumoral administration has been considered an obstacle to therapeutic efficacy (14). In current clinical practice, metallic markers are deployed to mark the biopsy area after a breast biopsy. Recently, metallic markers have been embedded in hydrogels that swell to optimize their visualization on ultrasound. These markers are FDA-approved and can be made from polyethylene glycol, collagen, and beta-glucan. Once hydrated, these markers can fully swell within 6 hours, with the size of the PEG hydrogel increasing proportionally to its maximum ( ). Figure 3A In the example, the dehydrated PEG measures 5×1×1 mm and, at maximum expansion, 14×4×4 mm, and can absorb at least 350 µl of ddH2O, which can be used to deliver a certain amount of the desired drug.

[0098] We embedded ADU-S100 within a PEG hydrogel marker in an attempt to retain it within the TME. To ensure that the PEG would swell as expected, it was implanted subcutaneously into the tissue of Balb / c mice. Significant swelling of the PEG was observed by day 4, with maximum expansion between days 11 and 15, and some reduction in PEG size by day 25. Figure 3B , Figure 3K ).

[0099] In human breast tissue, PEG was observed to remain visible even after 6 to 12 months. To assess whether PEG hydrogel could release ADU-S100 in a controlled manner, a reporter gene assay based on THP-1 cells was developed. THP-1 cells were first treated with phorbol 12-myristate 13-acetate to differentiate into macrophages sensitive to STING agonist treatment and secreting CXCL10. We first established a standard curve from which the amount of ADU-S100 secreted from PEG could be derived from the corresponding amount of CXCL10 released. THP-1 cells were treated with a known amount of ADU-S100, and after 24 hours, the supernatant was harvested for CXCL10 ELISA analysis. Figure 3C ), to generate a standard curve ( Figure 3D Next, 100 µg of ADU-S100 was reconstituted into 20 µl of ddH2O, and the drug was absorbed by the PEG hydrogel. Then, PEG+ADU-S100 was coated with collagen to simulate an in vivo tissue barrier, with 1 ml of culture medium placed on top, and the medium was replaced with fresh medium every 24 hours for 5 consecutive days. The conditioned medium harvested at the 5 time points was then added to differentiated THP-1 cells for CXCL10 analysis. Figure 3E ).

[0100] By using the previous Figure 3DThe standard curve generated in the figure was used to derive the amount of ADU-S100 released at each time point. Figure 3F As expected, the PEG hydrogel was able to release ADU-S100 in a slow and sustained manner, with 70 µg of the embedded 100 µg released by day 5. Unlike the PEG hydrogel, collagen and β-glucan breast markers did not release ADU-S100. Figure 3G ).

[0101] Next, the TNBC 4T1 model was treated with PEG hydrogel containing 100 µg of ADU-S100 relative to PBS. Given the invasiveness of the 4T1 mouse model, the PEG hydrogel was first implanted into a low tumor burden model on day 4 after 10K 4T1 cells were injected into the fourth mammary fat pad. PEG+ADU-S100 prevented 4T1 cells from establishing tumors, while PEG+PBS showed robust tumor growth. Figure 3H However, when PEG+ADU-S100 was implanted near established 4T1 tumors, growth was initially suppressed, and exponential growth was observed by day 20. Figure 3I On day 25, PEG markers adjacent to the tumor could be easily identified, and resection showed that the PEG markers remained intact. Figure 3J ).

[0102] These data demonstrate that PEG hydrogel markers are a novel drug delivery method for ADU-S100.

[0103] In another experiment, we will Figure 3H and Figure 3I The results shown were compared with those of direct intratumoral injection of ADU-S 100, and minimal response to tumor growth was found compared with PEG delivery. Figure 5A ).

[0104] Example 4

[0105] STING agonist monotherapy is toxic to T cells and does not induce T cell clonal expansion. .

[0106] T cells can become hyperresponsive to STING agonists, activating different groups of genes that lead to apoptosis (34, 35). To investigate this potential toxicity, primary human TNBC explants and human CD8+ T cells were used. As previously described (35), explants were prepared from patient TNBC and treated with PBS or 50 µM ADU-S100 for 24 hours. TNBCs were examined by flow cytometry and showed a slight increase in CD45+ leukocytes and a small decrease in lymphocytes with ADU-S100. A significant decrease in CD3+, CD4+, and CD8+ T cells was observed in the case of ADU-S100 treatment ( Figure 4A and Figure 4J Treatment of primary human CD8+ T cells with increased concentrations of ADU-S100 showed a corresponding increase in T cell death. Figure 4B ).

[0107] Next, spleen cells were obtained from wild-type mice, and the T population was sorted into CD3+, CD4+, and CD8+ subsets by flow cytometry. Each subset was then treated with escalating concentrations of ADU-S100 for 24 hours, resulting in a corresponding increase in cell death, with CD8+ T cells being the most sensitive to treatment. Figure 4C ).

[0108] Given that PEG hydrogels can deliver STING agonists in a controlled and sustained manner ( Figure 3F It was hypothesized that embedding ADU-S100 in a hydrogel could limit T-cell toxicity. For example, if 100 µg of ADU-S100 is injected, T cells are immediately exposed to the full dose of the drug, which could lead to immediate toxicity, while if 100 µg is delivered over time in a controlled manner, toxicity could be minimized. Therefore, human CD8+ T cells were seeded in triplicate and then treated with direct PBS, direct ADU-S100, PEG+PBS, or PEG+ADUS100. Direct ADU-S100 treatment resulted in significantly higher T-cell toxicity compared to embedding the drug in PEG. Figure 4D ).

[0109] Example 5

[0110] Additional studies on the delivery of STING agonists using PEG hydrogel biopsy markers .

[0111] Based on the data shown in Example 3, we continued to investigate the potential of this PEG marker for in vivo delivery of ADU-S100 using the established triple-negative breast cancer 4T1 syngeneic model (Aslakson, Cheryl J. and Fred R. Miller., “Selective events in the metastatic process defined by analysis of the sequential dissemination of subpopulations of a mouse mammary tumor.” Cancer Research 52.6, 1992, 1399-1405).

[0112] To determine whether the reduction in tumor size was consistent with the intrinsic effect of ADU-S100 delivered by PEG in the model, pTBK1 and pSTAT1 staining was performed on treated 4T1 mouse tumors on day 4, revealing evidence of activation of the intermediate-target STING. Figure 5B Next, to better characterize the TME effect of ADU-S100 delivered by PEG, we analyzed CD45 levels from 4T1 tumors 14 days post-treatment. + Single-cell RNA sequencing (scRNA-seq) of immune cells Figure 5C Uniform manifold approximation and projection (UMAP) clusters revealed five distinct immune cell types. Figure 5D Furthermore, we found that antigen-presenting cells (APCs) and macrophage clusters exhibited robust upregulation of ADU-S100-induced IFN-mediated genes, including lfitm3, B2m, Isg15, Irf7, and Cxcl10. Figure 5D ), and significantly enriched marker IFN gene features ( Figure 5E and Figure 5F In summary, these findings confirm the effective delivery of PEG markers to the TME by ADU-S100.

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[0161] *****

[0162] equivalent

[0163] Using only conventional experiments, those skilled in the art will recognize or be able to identify many equivalents of the particular substances and procedures described herein. Such equivalents are considered to be within the scope of the invention and covered by the appended claims.

Claims

1. A method for delivering a therapeutic agent to a subject to treat a solid tumor, the method comprising implanting a biocompatible polymer containing the therapeutic agent into the subject, wherein the biocompatible polymer (embolus) continuously releases the therapeutic agent into the subject over time.

2. The method according to claim 1, wherein the solid tumor is located inside a human body.

3. The method of claim 2, wherein the solid tumor includes cancer.

4. The method of claim 3, wherein the cancer includes carcinoma, lymphoma, melanoma, or sarcoma.

5. The method according to claim 3, wherein the cancer includes bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colorectal cancer, gastrointestinal cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, or thyroid cancer.

6. The method of claim 5, wherein the breast cancer includes ductal carcinoma in situ (DCIS), HER2-positive breast cancer, invasive ductal carcinoma (IDC), invasive lobular carcinoma (ILC), mixed ductal and lobular carcinoma, inflammatory breast cancer, or triple-negative breast cancer (TNBC).

7. The method of claim 5, wherein the TNBC has a mutated or absent phosphatase and tensin homolog (PTEN).

8. The method according to claim 2, wherein the solid tumor is a primary solid tumor or a metastatic solid tumor.

9. The method of claim 2, wherein the solid tumor is not removed prior to the implantation.

10. The method of claim 9, wherein the biocompatible polymer is implanted near the solid tumor.

11. The method of claim 10, wherein the biocompatible polymer is implanted within 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm or 10 mm of the solid tumor.

12. The method of claim 9, wherein the biocompatible polymer is implanted into the solid tumor.

13. The method of claim 9, wherein the biocompatible polymer is implanted around the solid tumor.

14. The method of claim 2, wherein the solid tumor is removed prior to the implantation.

15. The method of claim 1, wherein the biocompatible polymer comprises a biodegradable polymer, a non-biodegradable polymer, or a natural polymer.

16. The method of claim 15, wherein the biodegradable polymer comprises poly(p-dioxanone) (PDO), polyethylene glycol (PEG), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), poly(lactic acid-co-glycolytic acid) (PLGA), poly(caprolactone) (PCL), or combinations thereof.

17. The method of claim 15, wherein the non-biodegradable polymer comprises poly(siloxane) (silicone) or poly(ethylene-vinyl acetate).

18. The method of claim 15, wherein the natural polymer comprises albumin, cellulose, chitosan, collagen, elastin, gelatin, milk protein, or filament.

19. The method of claim 15, wherein the biocompatible polymer comprises a biodegradable polymer that can be retained in the subject for days, weeks or months.

20. The method of claim 1, wherein the biocompatible polymer comprises poly(urethane).

21. The method of claim 1, wherein the biocompatible polymer is hydrophilic.

22. The method of claim 1, wherein the biocompatible polymer comprises a hydrogel.

23. The method of claim 22, wherein the hydrogel comprises pores of 10 µm to 500 µm.

24. The method of claim 23, wherein the spacing between the pores in the hydrogel is about 5 nm to about 100 nm.

25. The method of claim 22, wherein the hydrogel comprises a chemically linked hydrogel or a physically linked hydrogel.

26. The method of claim 22, wherein the biocompatible polymer comprises a PEG-based hydrogel.

27. The method of claim 1, wherein the biocompatible polymer comprises: The core comprises poly(p-dioxanone) (PDO), polylactide (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), or copolymers thereof; and The sealant surrounding the core comprises a polyethylene glycol (PEG)-based hydrogel.

28. The method of claim 27, wherein the core further comprises a titanium or stainless steel marker.

29. The method of claim 1, wherein the biocompatibility marker comprises a biopsy marker.

30. The method of claim 29, wherein the biopsy marker comprises a polyethylene glycol-modified substance, collagen, β-glucan, or polylactic-co-glycolic acid.

31. The method of claim 30, wherein the biopsy marker comprises an aqueous expandable polymer.

32. The method of claim 29, wherein the biopsy marker comprises HydroMARK. ™ Biopsy markers.

33. The method of claim 29, wherein the biopsy marker is selected from the group consisting of: MammoMARK ® MammoSTAR ® and SecurMark ® Biopsy markers.

34. The method of claim 1, wherein the therapeutic agent comprises small molecules, macromolecules, cells, or combinations thereof.

35. The method of claim 34, wherein the therapeutic agent comprises a small molecule.

36. The method of claim 35, wherein the small molecule comprises a STING agonist.

37. The method of claim 36, wherein the STING agonist comprises ADU-S100, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING, E7766, TAK-676, SNX281, SYNB1891 or combinations thereof.

38. The method of claim 35, wherein the small molecule comprises estrogen receptor-targeted therapy.

39. The method of claim 38, wherein the estrogen receptor-targeted therapy comprises a selective estrogen receptor modulator (SERM).

40. The method of claim 39, wherein the SERM comprises tamoxifen or raloxifene.

41. The method of claim 1, wherein the biocompatible polymer is approximately 17 mm, 15 mm, 13 mm, 11 mm, 9 mm, or 7 mm in length when hydrated.

42. The method of claim 1, wherein the implantation comprises surgical implantation or implantation using a large-hole needle.

43. The method of claim 1, wherein a plurality of biocompatible polymer emboli are implanted.

44. The method of claim 2, wherein the method is used in combination with conventional therapy.

45. The method of claim 44, wherein conventional therapy includes chemotherapy or endocrine therapy.

46. ​​A method for treating a solid tumor in a human subject, the method comprising: a. Use therapeutic agents to hydrate and dehydrate biopsy markers; as well as b. Implant the hydrated biopsy marker near the subject's solid tumor.

47. The method of claim 46, wherein the biopsy marker comprises a metal clip embedded in a biocompatible polymer.

48. The method of claim 46, wherein the biopsy marker comprises: a. A core comprising poly(p-dioxanone) (PDO), polylactide (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), or copolymers thereof; and b. A sealant surrounding the core. The sealant comprises a polyethylene glycol (PEG)-based hydrogel.

49. The method of claim 46, wherein the biopsy marker comprises HydroMARK. ™ Biopsy markers.

50. The method of claim 46, wherein the therapeutic agent comprises a small molecule.

51. The method of claim 50, wherein the small molecule comprises a STING agonist.

52. The method of claim 46, further comprising injecting a substance into the implanted biopsy marker after the implantation.

53. A method for treating breast cancer in a human patient, the method comprising: Use STING agonist hydration dehydration biopsy markers; The dehydrated biopsy markers mentioned above include polyethylene glycol (PEG)-based hydrogels; as well as The hydrated biopsy marker is implanted into the patient's body near the location of the breast cancer cells.

54. The method of claim 53, wherein the dehydrated biopsy marker comprises HydroMARK. ™ Biopsy markers.

55. A method for loading a biopsy marker with a therapeutic agent, the method comprising hydrating the biopsy marker with an aqueous solution containing the therapeutic agent to obtain a hydrated combination.

56. The method of claim 55, wherein the biopsy marker is hydrated with an aqueous solution in a volume of less than about 20 µl.

57. The method of claim 55, further comprising implanting the hydration combination into a human subject to continuously release the therapeutic agent.

58. The method of claim 55, wherein the biopsy marker comprises a combination of biodegradable polymers surrounded by a dehydrated hydrogel.

59. The method of claim 55, wherein the biopsy marker comprises HydroMARK. ™ Biopsy markers.

60. The method of claim 55, wherein the therapeutic agent comprises a small molecule.

61. The method of claim 60, wherein the small molecule comprises a STING agonist.

62. A method for treating phosphatase and tensin homolog (PTEN)-ineffective, triple-negative breast cancer (TNBC), the method comprising implanting, without resection of the breast cancer, a hydrated HydroMARK containing a STING agonist near the breast cancer. ™ Biopsy markers.

63. The method of claim 62, wherein the implanted HydroMARK ™ The biopsy marker also contains a Rab7 inhibitor.

64. A HydroMARK ™ Biopsy marker, the HydroMARK ™ Biopsy markers include the STING agonist.

65. A composition comprising: A biocompatible polymer, wherein the biocompatible polymer comprises a metal marker; and A therapeutic agent, wherein the therapeutic agent is absorbed into and / or adsorbed onto the biocompatible polymer.

66. The composition of claim 65, wherein the biocompatible polymer is hydrated.

67. The composition of claim 65, wherein the biocompatible polymer comprises a polymer core containing the metal marker, the polymer core being surrounded by a polyethylene glycol (PEG)-based hydrogel.

68. The composition of claim 67, wherein the polymer core comprises poly(p-dioxanone) (PDO), polyethylene glycol (PEG), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), poly(lactic acid-co-glycolytic acid) (PLGA), poly(caprolactone) (PCL), or combinations thereof.

69. The composition of claim 65, wherein the metal marker comprises a titanium or stainless steel marker.

70. The composition of claim 65, wherein the biocompatible polymer comprises a biopsy marker.

71. The composition of claim 70, wherein the biopsy marker comprises HydroMARK. ™ Biopsy markers, MammoMARK ® Biopsy markers, MammoSTAR ® Biopsy markers or SecurMark ® Biopsy markers.

72. The composition of claim 70, wherein the biopsy marker comprises a composite biodegradable polymer surrounded by a hydrogel.

73. The composition of claim 72, wherein the biopsy marker comprises HydroMARK. ™ Biopsy markers.

74. The composition of claim 65, wherein the therapeutic agent comprises a small molecule.

75. The composition of claim 65, wherein the therapeutic agent comprises a STING agonist.

76. The composition of claim 75, wherein the STING agonist comprises ADU-S100, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING, E7766, TAK-676, SNX281, SYNB1891, or combinations thereof.

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