Method of use of mesoporous silica rods for treating cancer

Abstract

Embodiments of the invention include methods of treating conditions that are ameliorated by stimulating an immune response. In aspects, the methods include subcutaneous injection of mesoporous silica rods into a subject or patient. The mesoporous silica rods can include a cytokine (e.g., IL-2 or IL-12) and / or an adjuvant. The mesoporous silica structures can induce an innate systemic immune response to treat cancer, infection and other ailments. Embodiments also include methods of producing mesoporous silica rods.

Description

METHOD OF USE OF MESOPOROUS SILICA RODS FOR TREATING CANCERFIELD OF THE INVENTION

[0001] The invention relates to cancer therapeutics, and more specifically, to structures and methods of treating acute myeloid leukemia and other cancers using mesoporous silica material to deliver cytokines.BACKGROUND

[0002] The immune system of an organism acts in coordination with myriad biological processes to protect itself from diseases, disorders and ailments. The immune system carries out these processes by detecting and responding to a wide variety of pathogens and foreign objects not native to the body of the organism.

[0003] Dysfunction of the immune system can lead to several diseases, disorders and ailments, including autoimmune diseases, inflammatory diseases and cancer. Other ailments that afflict an immune system are immunodeficiencies. Immunodeficiency occurs when the immune system’s activity and response falls below typically healthy thresholds and can result in recurring and life-threatening infections. Immunodeficiency can manifest as the result of genetic disease, or acquired conditions such as HIV / AIDS, or through the use of immunosuppressive medication.

[0004] Autoimmunity refers to a collection of autoimmune disorders, in which the organism literally attacks itself. Autoimmune disorders result from hyperactivity of the immune system, wherein the immune system attacks normal tissues as if they were pathogens, physical insults, pathogenic insults or other foreign organisms. Examples of autoimmune diseases are diabetes mellitus type 1 , rheumatoid arthritis and systemic lupus erythematosus.

[0005] In the human body, the innate immune system recognizes pathogenic insults as well as dead and defective cells in the body to initiate protective responses. This recognition occurs through a set of germline-encoded pattern recognition receptors (PRRs) that sense pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPS).

[0006] Cytoplasmic PRRs include NOD-like receptors (NLRs) and retinoic acid-inducible gene-l-like receptors (also known as RIG-l-like receptors). The NLRs recognize ligands from various microbial pathogens, host cells and environmental sources. Based on their domain architecture, NLRs are subdivided into NLRPs and NLRCs. Among these, NLRP1 (mouse NLRPI b), NLRP3 and NLR familyapoptosis inhibitory protein / NLCR4 are well established NLRs for their ability to assemble inflammasomes.

[0007] Inflammasomes are multimeric cytosolic protein complexes that assemble in response to DAMPS and PAMPs, leading to the activation of inflammatory responses including enzymatic activation of canonical caspase-1. Inflammasome assembly initiates an inflammatory form of cell death known as pryoptosis, triggering the release of the proinflammatory cytokines interleukin-1 (IL-1 p) and IL-18. The NLRP3 inflammasome responds to cellular perturbations and a wide variety of microbes.Lipopolysaccharides have been found to trigger the activation of a distinct type of inflammasome known as the “non-canonical” inflammasome. Ratherthan activating caspase-1 , the non-canonical inflammasome results in the activation of the related caspase-4 and caspase-5 in humans and caspase- 11 in mice. However, the functional consequences are similar to those in canonical inflammasome responses, which includes pyroptosis as well as IL-1 p and IL-18 release.

[0008] The NLRP3 inflammasome is a multimeric cytosolic protein complex that, like other inflammasomes, assembles in response to cellular perturbations. This assembly leads to the activation of caspase-1 , which promotes maturation and release of the inflammatory cytokines interleukin-1 p (IL-1 p) and IL-18, as well as inflammatory cell death (i.e., pyroptosis). The inflammatory cytokines contribute to the development of systemic low-grade inflammation, and aberrant NLRP3 activation can drive a chronic inflammatory state in the body to modulate the pathogenesis of inflammation-associated diseases.Therefore, targeting NLRP3 or other signaling molecules downstream, such as caspase-1 , IL-1 p or IL-18, has the potential for great therapeutic benefit. However, NLRP3 inflammasome-mediated inflammatory cytokines play dual roles in mediating human disease. While they are detrimental in the pathogenesis of inflammatory and metabolic diseases, they have a beneficial role in numerous infectious diseases and some cancers. Therefore, fine tuning of NLRP3 inflammasome activity is essential for maintaining proper cellular homeostasis and health. The mechanisms of NLRP3 inflammasome activation play divergent roles in the pathogenesis of inflammation-associated diseases such as cancer, atherosclerosis, diabetes and obesity, and therefore offer therapeutic potential when targeted to correct pathways of treatment.

[0009] Foam cells, also known as lipid-laden macrophages, are cholesterol-containing cells that can form a plaque and lead to inflammatory maladies such as atherosclerosis which can lead to heart attack and stroke. Foam cells are lipid-rich cells with an M2 macrophage-like phenotype. However, certain foam cells may be derived from smooth muscle tissue, and these specific foam cells present a limited macrophage-like phenotype. The presence of the NLRP3 inflammasome is known to promote foam cell formation, which has been observed in several histological samples. However, many studies have also reported that NLRP3 inflammasome activation promotes the formation of macrophage foam cells through IL-1 p.

[0010] Aluminum hydroxide, denoted by the chemical formula AI(OH)3 and commonly known in pharmaceutical and related fields as “Alum,” is composed of double layers of hydroxyl groups with aluminum ions occupying two-thirds of the octahedral holes between the double layers. Among several uses, aluminum hydroxide is used as a pharmaceutical adjuvant in some vaccines. Further, aluminum hydroxide is also known to stimulate the immune system by inducing the release of uric acid, which eventually results in stimulation of T cells and B cells. Another commonly used adjuvant also known to be safe in vaccines is cytosine guanosine dinucleotide (CpG). Cytosine guanine dinucleotide (CpG) fragments are potently immunogenic DNA fragments which serve as friend or foe recognition systems between bacterial (hypomethylated) and mammalian (methylated) DNA.

[0011] Recent studies suggest that the nascence and proliferation of cancerous tumors are directly related to inflammatory processes. It has long been known that cancer development and its response to therapy are regulated by inflammation, which either promotes or suppresses tumor progression. Chronic inflammation facilitates tumor progression and treatment resistance, whereas induction of acute inflammatory reactions through the generation of a systemic immune response often stimulates the maturation and differentiation of immune cells and antigen-presentation cells (APCs), leading to antitumor responses.

[0012] Circulating cancer cells can be borne in the circulatory system (blood cancers), lymphatic system (lymphatic cancers), or may result from a tumor cell that has shed into the vasculature or lymphatic system from a primary tumor and is carried around the body by the circulation of lymph or blood. Circulating cancer cells shed from tumors are also known as circulating tumor cells (CTCs) which can extravasate and become seeds for the subsequent growth of additional tumors (metastases) in distant organs or tissues.

[0013] One method of treating circulating cancer cells has been with immunotherapy, including the use of chimeric antigen receptor T cells (CAR-T cells) or immune checkpoint inhibitors. While cancer immunotherapy has shown some success, it is noteworthy that less than 50% of patients treated with single agent immunotherapy actually respond. This even occurs amongst the most immunogenic tumor types. To enhance the efficacy of cancer immunotherapy, physicians often administer combinations of systemic immune checkpoint inhibitors to the patient being treated. Although these combinations can improve treatment outcomes, they frequently result in intolerable or severe adverse events, leading to therapy discontinuation or, in extreme cases, patient mortality. The consensus across immuno-oncology (IO) research is clear: safe combination therapy is essential for developing more effective cancer treatments.

[0014] Next, most blood cancers start in the bone marrow where blood is produced. Stem cells in the bone marrow mature and develop into three types of blood cells: red blood cells, white blood cells, or platelets. In most blood cancers, the normal blood cell development process is interrupted by uncontrolled growth of an abnormal type of blood cell. These abnormal blood cells, or cancerous cells, prevent the blood from performing many of its functions, like fighting off infections or preventing serious bleeding. There are three types of blood cancers: leukemia, lymphoma and myeloma.

[0015] Leukemia is a type of cancer found in the blood and bone marrow, and is caused by the rapid production of abnormal white blood cells. The abnormal white blood cells are not able to fight infection, and impair the ability of the bone marrow to produce red blood cells and platelets.

[0016] Lymphoma is a type of blood cancer that affects the lymphatic system, which removes excess fluids from the body and produces immune cells. Lymphocytes are a type of white blood cell that fight infection. Abnormal lymphocytes become lymphoma cells, which then multiply and collect in the lymph nodes and other tissues. Over time, these cancerous cells impair the immune system.

[0017] Myeloma is a cancer of the plasma cells. Plasma cells are white blood cells that produce disease- and infection-fighting antibodies in the body. Myeloma cells prevent the normal production of antibodies, leaving your body's immune system weakened and susceptible to infection.

[0018] Acute Myeloid Leukemia (AML) is a cancer of the myeloid line of blood cells, characterized by the rapid growth of abnormal cells that build up in the bone marrow and blood and interfere with normal blood cell production. Symptoms may include feeling tired, shortness of breath, easy bruising and bleeding, and increased risk of infection. Occasionally, AML may spread to the brain, skin, or gums.

[0019] Myelodysplastic Syndrome (MDS) is one of a group of cancers in which immature blood cells in the bone marrow do not mature, and as a result do not develop into healthy blood cells. Those afflicted with MDS are usually asymptomatic in the cancer’s early stages, with symptoms later manifesting as fatigue, shortness of breath, bleeding disorders, anemia and increased frequency of infection. Some types of MDS may further develop into AML.

[0020] Clonal Hematopoiesis of Indeterminate Potential (CHIP) is a common aging-related phenomenon in which hematopoietic stem cells (HSCs) or other early blood cell progenitors contribute to the formation of a genetically distinct clonal subpopulation of blood cells. This subpopulation in the blood is characterized by a shared unique mutation in cellular DNA. The establishment of a clonal subpopulation may occur when stem or progenitor cells acquire one or more somatic mutations that giveit a competitive advantage in hematopoiesis over stem and progenitor cells that lack these mutations. CHIP may also arise without a driving mutation, through mechanisms such as neutral drift in the stem cell population. CHIP may occur in subjects who are completely healthy but has also been found in subjects with hematologic diseases. The clonal subpopulation may vary in size depending on the subject, but the incidence of CHIP has been found to rise dramatically with age. Subjects with CHIP have been linked to a more than 10-fold increased risk of developing blood cancer and an increased risk of cardiovascular disease.

[0021] Conventional cancer treatments are directed at removing cancerous tissue and preventing it from spreading. Such treatment options include surgery, chemotherapy, radiation therapy, hormonal therapy, targeted therapy and palliative care. Treatments are usually pursued based on the type, location and grade of the cancer as well as the patient's health and preferences. These options have limitations. They can also be ineffective, particularly when cancer has metastasized and cancer cells are circulating. Moreover, chemotherapy and radiation therapy have a range of side-effects related to cell toxicity.

[0022] Because cancer cells divide faster than most normal cells, they can be sensitive to chemotherapy drugs. However, chemotherapy drugs will also attack other cells in the body, especially fast-dividing cells such as blood cells and the cells lining the mouth, stomach, and intestines.Accordingly, there is a need for improved medications and methods of treating cancer that are more targeted and have less deleterious side effects.

[0023] Cytokine-based therapies, which have shown a potential be be curative in clinical trials, are significantly limited by their toxicity. For instance, Adesleukin, a systemically administered IL-2 cytokine, proved effective in a subset of patients but is was used sparingly due to severe toxicityto the extent it is used, Adesluekin is currently used in combo with the Lovance TIL therapy. Similarly, early clinical trials of systemic IL-12 therapy were halted following treatment-related deaths. As a result, several groups are now developing new IL-12 formulations aimed at reducing toxicity while maximizing curative efficacy.

[0024] The heterogeneity of cancer limits the effectiveness of single-agent therapies, which often only work for a limited period before relapse occurs in late-stage patients. This variability is especially pronounced within the tumor microenvironment (TME), where even patients with the same tumor type can have dramatically different TMEs. This is particularly evident in immunogenic cancers like melanoma and lung cancer.

[0025] The most effective single immuno-oncology agents only benefit 40-50% of patients, with some becoming refractory to treatment. To address this, personalized combination therapies are being explored to increase efficacy and minimize the risk of therapy resistance. By tailoring combinations to thespecific TME of each patient, it is possible to improve outcomes and reduce the likelihood of adverse events, presenting a significant opportunity for innovation, including through the use of the methods and compositions disclosed herein for the treatment of a cancer suffered by a patient.

[0026] Technological improvements in RNA sequencing and multiplex immunohistochemistry (IHC) are revolutionizing our understanding of individual patients' tumors. These advancements allow for unprecedented insights into the tumor microenvironment (TME), paving the way for more targeted and effective therapies, including through the use of the methods and compositions disclosed herein for the treatment of a cancer suffered by a patient.SUMMARY OF THE INVENTION

[0027] The inventions described and claimed herein have many attributes and embodiments including, but not limited to, those set forth or described or referenced in this brief summary. The inventions described and claimed herein are not limited to, or by, the features or embodiments identified in this summary, which is included for purposes of illustration only and not restriction.

[0028] Embodiments of the invention include a biomaterial-based platform technology for immune cell recruitment and modulation to elicit broad immunomodulatory effects for the treatment of cancer and infectious diseases. MSRs are a type of biomaterial that in an embodiment form a scaffold.

[0029] The invention further relates to methods of treating cancers and other conditions that can be ameliorated by initiating a systemic immune response. In aspects, the method includes subcutaneous injection of mesoporous silica rods (MSRs) into a patient or subject. Upon injection, the mesoporous silica rods elicit an innate systemic immune response. The innate systemic immune response can include granulopoiesis - the formulation of a granuloma at the injection site which further stimulates the production of cytokines downstream from the granuloma formed at the injection site. Immune cell stimulation also occurs in the bone marrow, lymph nodes and spleen, which increase production of immune cells. Within the bone marrow, myeloid cells are activated for differentiation into various cells such as neutrophils, macrophages, and other immune cells. Through subcutaneous injection of MSRs, a systemic immune response is catalyzed which acts upon and treats cancerous cells and tissues. Embodiments further include pre-treating a patient or subject with chemotherapy for a predetermined period prior to subcutaneous injection with mesoporous silica rods, concomitantly treating a patient or subject with chemotherapy and subcutaneous injection of mesoporous silica rods, or both.

[0030] In further aspects, the invention relates to methods of treating acute myeloid leukemia (AML) and other circulating cancers by stimulating a systemic immune response. In aspects, the methods include subcutaneous injection of mesoporous silica rods (MSRs) into the patient or subject to stimulate asystemic immune response. The immune response can stimulate differentiation of leukemic or pre- leukemic blasts. The differentiation may occur at the injection site or elsewhere in the body. In yet further aspects, the systemic immune response initiated by the subcutaneous injection of MSRs results in differentiation of myeloid cells, including differentiating cancerous myeloid cells and cancer precursor cells into immune cells. The differentiated leukemic blasts may then be cleared after circulation (e.g., neutrophils live for approximately 24 hours). In aspects, differentiated leukemic blasts and other differentiated immune cells can be cleared within 24 hours, within 48 hours, 72 hours, or within a few weeks to months, depending on the cell type (macrophages can live for months in vivo). In some embodiments, administration of the MSR therapy causes inflammation which causes the differentiated cells to undergo immunogenic cell death. In other embodiments, the “leukemic” neutrophils can result in neutrophil extracellular traps at the injection site. These traps can capture and display AML antigens. In other embodiments the differentiated leukemic blasts can become antigen presenting cells (APCs) and display leukemic antigens. The antigen presentation mechanisms can result in T cell immunity to the tumor. In further aspects, the inflammation at the local injection site can start a cytokine cascade, which includes IL-6, IL-8, IL-12, and IL-1 beta. This cytokine cascade can cause steady state granulopoiesis, emergency granulopoiesis, and / or other systemic effects. The IL-1 beta can also help elicit T cell response, including a memory T cell response. In other embodiments, the injection site inflammation can cause reactive lymph nodes. Embodiments further include pre-treating a patient or subject who has AML or other circulating cancer with chemotherapy for a predetermined period prior to subcutaneous injection with mesoporous silica rods, concomitantly treating a patient or subject who has AML or other circulating cancer with chemotherapy and subcutaneous injection of mesoporous silica rods, or both. The chemotherapy can prime the leukemic blasts for differentiation and / or decrease overall disease burden. In other embodiments, the inflammatory cascade generated at the injection site can recruit in tumor cells. Those tumor cells may be recognized by the immune cells to create a vaccine like response.

[0031] In an embodiment, the rods are maintained in close proximity at the site of administration and remain localized at or near the site of administration. In an embodiment, the rods are maintained in close proximity at the site of administration in order to trigger or induce a localized response.

[0032] In an embodiment, the rods do not spontaneously assemble.

[0033] In embodiments, the invention includes a composition of mesoporous silica rods carrying an immune cell recruitment compound and an immune cell activation compound.

[0034] In embodiments, the mesoporous silica rods carry a cytokine payload for injection into a tumor, into the skin, or into an infected area. In aspects of embodiments, the cytokine is selected from the group of granulocyte-macrophage colony-stimulating factors such as Leukine, Molgramostin andSargramostim. The cytokine payload can be impregnated into the mesoporous silica rod, coated onto the mesoporous silica rod, or coupled to the mesoporous silica rod in any suitable manner known in the art.

[0035] Embodiments also include a method of use for a mesoporous silica rod that has a cytokine payload for injection into a tumor, cancerous lesion, cancerous region, the skin or an infected area. The cytokine payload can be interleukin-12 (IL-12), interleukin-2 (IL-2) or granulocyte-macrophage colonystimulating factor (GM-CSF). The mesoporous silica rod can also have an adjuvant (e.g., aluminum hydroxide, a lipopolysaccharide or a toll like receptor agonist) to stimulate an immune response. The adjuvant can be present in an impregnated payload, a coating, or any other suitable coupling known in the art. The adjuvant and / or cytokine can be physically trapped in the pores of the MSRs. The adjuvant and / or cytokine can be bound to the MSRs through electrostatic interactions.

[0036] Embodiments further include methods of use for subcutaneous injection of mesoporous silica rods containing cytokine payloads and an adjuvant. The methods of use include subcutaneous injection of mesoporous silica rods containing a cytokine payload and an adjuvant into the skin of the patient or subject to stimulate an innate systemic immune response for the treatment of tumor cells associated with acute myeloid leukemia or other cancers.

[0037] Another embodiment is a method of treating an ailment. The method can include the steps of (a) identifying an affected tissue and (b) inserting or injecting a mesoporous silica rod at or near the affected tissue. The mesoporous silica rod can carry a cytokine payload and / or an adjuvant. In aspects, the affected tissue is tumor tissue. In aspects, the affected tissue is infected tissue (e.g., bacterial, fungal or viral infection).

[0038] Another embodiment is a method of potentiating a systemic immune response to act upon an affected tissue. The method can be used, for example, to convert a “cold” tumor to a “hot” tumor. The method can include steps of (a) identifying an affected tissue (e.g., a cold tumor) and (b) inserting or injecting a mesoporous silica rod at or near the affected tissue. The mesoporous silica rod can carry a cytokine payload and / or an adjuvant.

[0039] Yet another embodiment is a method of potentiating a systemic immune response to act upon an affected tissue through subcutaneous injection of MSR that is remote from the affected tissue. The method can be used, for example, to convert a “cold” tumor to a “hot” tumor. The method can include steps of (a) identifying an affected tissue (e.g., a cold tumor) and (b) subcutaneously injecting a mesoporous silica rod into a patient remote from the site of the affected tissue. The mesoporous silica rod can carry a cytokine payload and / or an adjuvant.

[0040] In embodiments the mesoporous silica rod can be cylindrical with a plurality of pores. Each pore can have a diameter of about 2 nanometers. In aspects, the pore diameter is about 3 nanometers, about 4 nanometers, about 5 nanometers, 10 nanometers, 15 nanometers, 20 nanometers, 25 nanometers, 30 nanometers, 35 nanometers, 40 nanometers or larger. In aspects, the diameter of the pore varies (e.g., from about 2 nanometers to about 50 nanometers or larger).

[0041] In another embodiment, a mesoporous silica rod is coated with a cytokine payload and coated with an adjuvant (e.g., aluminum hydroxide, TLR agonist or cytosine guanosine dinucleotide (CpG)) for subcutaneous injection and / or implantation into a tumor, cancerous region, the skin or an inflammatory area of a patient or subject.

[0042] In further embodiments, mesoporous silica rods are coated with granulocyte-macrophage colony-stimulating factor for injection and / or implantation into a tumor, cancerous region, the skin or an inflammatory area of a patient or subject.

[0043] In yet further embodiments, mesoporous silica rods are coated with granulocytemacrophage colony-stimulating factor and / or other therapeutic proteins for subcutaneous injection into the skin of a patient or subject.

[0044] In aspects, the mesoporous silica rod delivers a physical insult to a tumor and creates a localized innate immune response which becomes an adaptive response. This response leads to tumor and cancer cell clearance.

[0045] In aspects, the mesoporous silica rod delivers a physical insult to a tissue of a patient or subject and creates a systemic innate immune response and / or adaptive response which leads to tumor and cancer cell clearance.

[0046] In further aspects, the mesoporous silica rod delivers a physical insult to a tissue of a patient or subject and creates a systemic innate immune response which leads to regression of the signs and symptoms of acute myeloid leukemia (AML).

[0047] In yet other aspects, the mesoporous silica rod can generate a local, controlled immunological microenvironment and serve as a site for regulation of the immune response against acute myeloid leukemia and other cancers.

[0048] In other aspects, the subcutaneous injection of mesoporous silica rods with an adjuvant and / or cytokine payload can generate a localized inflammatory reaction at the site of treatment, therebyeffecting an immune response for treatment of a disease, tumor, cancer, cancerous cell, cancerous lesion or other ailment.

[0049] In further aspects, the subcutaneous injection of mesoporous silica rods with an adjuvant and / or cytokine payload can generate a systemic innate inflammatory response, thereby effecting an immune response for treatment of a disease, tumor, cancer, cancerous cell, cancerous lesion or other ailment.

[0050] Other treatment areas targeted by the embodiments above include, for example, benign cancer tumors, benign cancer cells, non-circulating cancer cells, non-circulating tumor cells, malignant tumors, cancerous lesions, cancerous tissues, sebaceous cysts, acne vulgaris, fatty tumors, inflamed cartilage, inflamed tissue, abscesses or any set of aggregated cells that do not form a healthy tissue structure within a human body, organism, or within the integumentary system of a human body or organism.

[0051] Embodiments also include methods of producing and manufacturing mesoporous silica rods for subcutaneous injection to modulate an immune response.

[0052] Embodiments include a method of producing mesoporous silica rods that includes steps of (a) adding a poloxamer to water to form a solution, (b) mixing the solution, (c) adding an acid, (d) adding a source of silicon dioxide, (e) incubating the solution, (f) sieving and vacuum filtering the solution, and (g) heating the solution to yield the mesoporous silica rods in solution. The final solution can be sterilized. The mesoporous silica rods are generally stable in lyophilized form for storage / shipping.

[0053] Methods of the present invention can also include a step of mixing the mesoporous silica rods with a granulocyte-macrophage colony-stimulating factor (GM-CSF). Methods of the present invention can also include a step of mixing the mesoporous silica rods with an adjuvant. Methods can also include a step of mixing the mesoporous silica rods with a CpG oligodinucleotide. Methods can further include a step of mixing the mesoporous silica rods with a cytokine. Methods can also include a step of lyophilizing the mesoporous silica rods.

[0054] Other features and advantages of aspects of the present invention will become apparent from the following detailed description, taken in conjunction with the accompanying drawings, which illustrate, by way of example, the principles of aspects of the invention.

[0055] In an embodiment, a cancer is a disease of uncontrolled growth and loss of cellular identity. Tumor cancer cells can take on properties of immune cells and use these properties for immune evasion, survival, and metastasis.

[0056] With regard to the present invention, the invention generates a “pull” mechanism of inflammation in the tumor microenvironment, near a tumor draining lymph node. In another embodiment, when administered subcutaneously, the tumor cells change into terminally differentiated, partially differentiated cells that are (1) cells that have shorter lives and will die after a set period of time, (2) able to present their own antigen in an APC-like manner to generate a vaccine-like response, and / or (3) non- pathogenic.

[0057] In an embodiment, an MSR, as well as other biomaterial scaffolds, can generate an immunomodulatory “pull” mechanism, so that a tumor cell transdifferentiates into an immune cell or an immune cell like phenotype. In an embodiment, a biomaterial scaffold is coated or seeded with different immune adjuvants or agonists that direct a response by the tumor cells into the desired cell phenotype. In an embodiment, the response is inflammatory. In a further embodiment, the treatment comprises the addition of chemotherapy to enhance a tumor cells differentiation and the differentiation effect. In an embodiment, IL-12 and other cytokines are used to direct the differentiation of cancer cells, including, but not limited to tumor cells to direct this effect. In another embodiment, a stiff or solid material is used to attract and / or differentiate tumor cells.

[0058] In another embodiment, other materials and / or adjuvants that form a depot can be used with or without a biomaterial scaffold to generate a differentiation form or type of response.

[0059] In a further embodiment, biomaterial scaffolds are seeded with an immune agonist or other therapeutic asset that can attract circulating cancer cells, tumor cells and / or immune cells to generate a vaccine-like response against tumors.

[0060] In an embodiment, an antigen is added to a biomaterial scaffold with immune signals that can influence or enhance an immune response. In another embodiment, this immune response results in tumor cells phagocytosing the MSRs or equivalent biomaterial, thus enhancing the tumor cell interaction with the drug loaded onto the biomaterial scaffold. In one embodiment, a tumor cell following phagocytosis becomes a neutrophil-like cell and undergoes netosis at the site of biomaterial scaffoldgenerated inflammation. In another embodiment, MSR causes netosis, which traps and presents tumor antigens. In another embodiment, the netosis causes local inflammation to generate an anti-tumor response.

[0061] In an embodiment, use of a biomedical scaffold with a therapeutic is used to treat a heme malignancy and / or a solid tumor.

[0062] In another embodiment, a tumor cell can take on a phenotype of a macrophage, a neutrophil, and / or other immune cells. In a further embodiment the phenotype depends on an inflammatory signal that is delivered to the tumor cell.

[0063] In another embodiment, the MSR can serve as a site to generate a vaccine-like response when administered intratumorally with MSR and IL-12, peri lymphatically with MSR and IL-12, and subcutaneously with MSR+IL-12+antigen.BRIEF DESCRIPTION OF THE DRAWINGS

[0064] FIG. 1 depicts the formation of a macro-immune structure induced by the subcutaneous injection of mesoporous silica rods containing therapeutics and adjuvants. The subcutaneous injection of mesoporous silica rods further triggers immune cell trafficking to / from lymph nodes and tumors.

[0065] FIG. 2A is a graph showing survival percentages of mice challenged with 100K c1498-eGFP cells when treated with chemotherapy plus subcutaneous injection of mesoporous silica rods versus mice treated with chemotherapy plus subcutaneous injection of GM-CSF + CpG versus mice treated with chemotherapy alone versus an untreated control group.

[0066] FIG. 2B is a graph showing survival percentages of mice challenged with 250K WEHI-3 cells when treated with chemotherapy plus subcutaneous injection of mesoporous silica rods versus mice treated with chemotherapy plus subcutaneous injection of GM-CSF + CpG versus mice treated with chemotherapy alone versus an untreated control group.

[0067] FIG. 3A is a flow chart depicting the cascading actions of an immune response elicited by subcutaneous injection of mesoporous silica rods into murine subjects at the injection site of the skin.

[0068] FIG. 3B is a flow chart depicting the cascading actions of an immune response elicited by subcutaneous injection of mesoporous silica rods into murine subjects as observed in the blood of the murine subjects.

[0069] FIG. 3C is a flow chart depicting the cascading actions of an immune response elicited by subcutaneous injection of mesoporous silica rods into murine subjects as observed in the lymph nodes of the murine subjects.

[0070] FIG. 3D is a flow chart depicting the cascading actions of an immune response elicited by subcutaneous injection of mesoporous silica rods into murine subjects as observed in the bone marrow of the murine subjects.

[0071] FIG. 3E is a flow chart depicting the cascading actions of an immune response elicited by subcutaneous injection of mesoporous silica rods into murine subjects as observed in the spleen of the murine subjects.

[0072] FIG. 3F is a flow chart depicting the cascading actions of an immune response elicited by subcutaneous injection of mesoporous silica rods into murine subjects as observed in the tumor cells of the murine subjects.

[0073] FIG. 4 depicts an AML blast differentiating into other types of cells such as neutrophils and macrophages as a result of the immune response generated by the subcutaneous injection of mesoporous silica rods.

[0074] FIG. 5A is a graph showing exposure of c1498-eGFP cells stimulated with GM-CSF for 5 days and stained for CD11 b Ly6C, CD11 b Ly6G, CD115 Ly6C, CD45 B220, CD1 15 Ly6G, CD19 B220 or unstimulated cells stained with the same markers. GM-CSF did not differentiate cells. .

[0075] FIG. 5B is a graph showing c1498 GFP-labeled cancer cells (i.e., GFP+ Ly6C+, GFP+ Ly6G+, GFP+ CD11 b+, GFP+ CD115+, GFP+ CD19+, GFP+ B220+, GFP+ CD45.2+) in an unstimulated condition versus appearance of GFP-labeled cancer cells in a stimulated condition following 100ng / mL exposure of GM-CSF for 5 days.

[0076] FIG. 5C is a graph showing the baseline values in c1498 cells and c1498-eGFP cells recorded for presence of B220, CD11 b CD18, CD11 b GR-1 , and CD11 c CD86.

[0077] FIG. 6A is two graphs depicting tumor cell death (c1498-eGFP) in multiple conditions: 100K cells untreated, 100K cells treated with chemotherapy, 100K cells treated with mesoporous silica rods, GM-CSF, and CpG (ATT-01), 100K cells treated with chemotherapy and ATT-01 , 500K cells untreated, 500K cells treated with chemotherapy, 500K cells treated with ATT-01 , and 500K cells treated with chemotherapy and ATT-01 for 24 hours.

[0078] FIG. 6B is two graphs depicting percentage of CD19+ B220+ present in c1498-eGFP in multiple conditions: 100K cells untreated, 100K cells treated with chemotherapy, 100K cells treated with mesoporous silica rods (ATT-01), 100K cells treated with chemotherapy and ATT-01 , 500K cellsuntreated, 500K cells treated with chemotherapy, 500K cells treated with ATT-01 , and 500K cells treated with chemotherapy and ATT-01 for 24 hours.

[0079] FIG. 6C is four graphs depicting percentage of CD11 b+ CD86+ cells present (top row) and percentage of CD11 b+ GR-1 + cells present (bottom row) in c1498-eGFP after treatment with multiple conditions: 100K cells untreated, 100K cells treated with chemotherapy, 100K cells treated with mesoporous silica rods (ATT-01), 100K cells treated with chemotherapy and ATT-01 , 500K cells untreated, 500K cells treated with chemotherapy, 500K cells treated with ATT-01 , and 500K cells treated with chemotherapy and ATT-01 for 24 hours.

[0080] FIG. 7 is a graph illustrating cytokine levels in pig serum up to 21 days post-immunization with 700 pL ATT-01 by subcutaneous injection.

[0081] FIG. 8A shows a histology slide of MSR inside a granuloma of a murine subject.

[0082] FIG. 8B shows a histology slide of an MSR at the injection site.

[0083] FIG. 8C shows a histology slide of a murine injection site with a mature and resolving granuloma at the MSR injection site.

[0084] FIG. 8D shows a histology slide of a murine granuloma at the MSR injection site.

[0085] FIG. 8E shows a histology slide displaying a sheet of lymphoid cells in a control treated murine lymph node.

[0086] FIG. 8F shows a histology slide of sheets of lymphoid cells in a control mouse in lymph node.

[0087] FIG. 8G shows a histology slide displaying follicular hyperplasia of a murine lymph node in an MSR treated mouse.

[0088] FIG. 8H shows a histology slide displaying sinus hyperplasia of a murine lymph node in a control mouse.

[0089] FIG. 8I shows a histology slide of a lymph node from an MSR treated mouse with murine immunoblasts circled in the figure. Mixed reactive hyperplasia was observed.

[0090] FIG. 9 shows graphs depicting murine complete blood cell (CBC) counts over time across three conditions after administration of C1498-eGFP cells: untreated cells, treatment with chemotherapy plus mesoporous silica rods, GM-CSF, and CpG, and treatment with chemotherapy plus GM-CSF plus CpG, as a control.

[0091] FIG. 10A shows an experiment where mice were given 100K c1498-eGFP cells on day 0. Then, mice received chemotherapy (doxorubicin and cytarabine = iCT), iCT plus MSR, GM-CSF, and CpG (Att-01), and iCT, GM-CSF, and CpG. The figure shows graphs depicting percentage of GFP+ positive cells in the bone marrow, at the injection site, in the lymph nodes and in splenocytes of murine subjects 7 days post-immunization.

[0092] FIG. 10B shows an experiment where mice were given 100K c1498-eGFP cells on day 0. Then, mice received iCT, iCT plus MSR, GM-CSF, and CpG (Att-01), and iCT, GM-CSF, and CpG. The figure shows graphs depicting percentage of GFP+ positive cells in the bone marrow, at the implant site, in the lymph nodes and in the spleen of murine subjects 22 days post-immunization.

[0093] FIG. 10C shows an experiment where mice were given 100K c1498-eGFP cells on day 0. Then, mice received iCT, iCT plus MSR, GM-CSF, and CpG (Att-01), and iCT, GM-CSF, and CpG. The figure shows graphs depicting percentage of GFP+ positive cells in the blood, ovaries, and in the liver of murine subjects 22 days post-immunization.

[0094] FIG. 11A shows an experiment where mice were given 100K c1498-eGFP cells on day 0. Then, mice received iCT, iCT plus MSR, GM-CSF, and CpG (Att-01), and iCT, GM-CSF, and CpG. The figure shows graphs depicting immune cell infiltration at the subcutaneous injection site of mesoporous silica rods in murine subjects on day 7 post immunization.

[0095] FIG. 11 B shows an experiment where mice were given 100K c1498-eGFP cells on day 0. Then, mice received iCT, iCT plus MSR, GM-CSF, and CpG (Att-01), and iCT, GM-CSF, and CpG. The figure shows graphs depicting immune cell populations in the bone marrow of murine subjects on day 7 post immunization.

[0096] FIG. 11 C shows graphs depicting immune cell population in the bone marrow of murine subjects on day 22 post immunization.

[0097] FIG. 11 D shows graphs depicting immune cell populations in spleens of murine subjects on day 7 post immunization.

[0098] FIG. 11 E shows graphs depicting immune cell populations in spleens of murine subjects on day 22 post immunization.

[0099] FIG. 11 F shows graphs depicting immune cell populations in draining lymph nodes of murine subjects on day 7 post immunization.

[0100] FIG. 11 G shows graphs depicting immune cell populations in draining lymph nodes of murine subjects on day 22 post immunization.

[0101] FIG. 11 H shows graphs depicting CD86+ cells infiltration in organs of murine subjects on day 7 post immunization.

[0102] FIG. 12 depicts an overview of mesoporous silica rod manufacture. A symmetric triblock copolymer (Pluronic P123) composed of poly (ethylene oxide) and poly (propylene oxide) is employed to form rod-shaped micelles in solution. Tetraethyl Orthosilicate (TEOs) is added to the solution and the silica deposits on the micelles, creating a hexagonal pore structure. The Pluronic P123 is rinsed and calcinated (high temperature treatment) to remove the polymer, leaving a silica mesoporous structure.

[0103] FIG. 13 is a process flowchart for synthesizing mesoporous silica rods (MSRs).

[0104] FIG. 14A is a representative SEM sizing image of standard MSR width.

[0105] FIG. 14B is a representative SEM sizing image of standard MSR length.

[0106] FIG. 15 is a process flowchart of a proposed manufacturing process of mesoporous silica rods (MSRs).

[0107] FIG. 16 shows representative SEM sizing images of MSR length and width. Modified MSRs length (top left) and width (top right). Standard MSRs length (bottom left) and width (bottom right).D[n,0.1](pm), D[n,0.5](pm) D[n,0.9](pm) = biodistribution of MSRs sized at 10% or less, 50% or less, and 90% or less respectively within the total number mean.

[0108] FIG. 17 shows B16F10 tumor cells that were injected in the flank of mice. When tumors averaged 70-90mm3 the mice were treated with ATT-02 which was administered either intratumorally, perimtumorally, or perlymphatic.

[0109] Figure 18 shows the therapeutic effects of single treatment of ATT-02 and ATT-02 OVA in the primary tumor.

[0110] Figure 19 shows the Abscopal effects following a single treatment of ATT-02 and ATT-02 OVA in the secondary tumor.

[0111] Figure 20 shows the detection of IL-12 (20A) and IFNgamma (20B) in the serum of mice that received a single dose of PBS, a control or a therapeutic.

[0112] Figure 21 shows the quantification of a percentage of CD8 positive T cells antigen specific T cells by SIINFEKL tetramer versus the total population in tumors. (A) quantification for Primary Tumors and (B) quantification for Secondary Tumors at 7 days post treatment.

[0113] Fig. 22 shows the analysis of immune cells by quantification of a percentage of myeloid cells after administration of a control or a treatment with a therapeutic versus the total population in tumors. (A) analysis for Primary Tumors and (B) analysis for Secondary Tumors at 7 days post treatment.

[0114] Fig. 23 shows the quantification of a percentage of CD8+ T cells (a, c) and the antigen specific T- cells by SIINFEKL tetramer versus the total population (b, d) staining after PBS, MSR, ATT-02, ATT-02 OVA, IL-12 and IL-12 OVA treatments either i.t. or s.c. in lymph nodes of treated and untreated animals.

[0115] Fig. 24 shows the analysis of immune cells in lymph nodes by quantification of a percentage of macrophages after administration of a control or a therapeutic versus the total population. (A) analysis of macrophages and (B) analysis of monocytes.

[0116] Fig. 25 shows in vitro differentiation of tumor cells with MSR therapeutics. (A) Tumor cells (B16-OVA) were cultured with MSR for 48 hours and showed a phenotypic change. (B) EMT-6, B16-OVA, and CT26 cells were cultured for 24 or 48 hours with MSR treatments: MSR alone, IL-12 + OVA, Att-02 (MSR+ IL-12) and OVA, IL-12 alone, or ATT-02 (MSR+IL-12) and showed a phenotypic change into an immune cell like state. (C). EMT-6, B16-OVA , and CT26 cells were cultured for 24 or 48 hours with MSR treatments: MSR alone, IL-12 + OVA, Att-02 (MSR+ IL-12) and OVA, IL-12 alone, or ATT-02 (MSR+IL- 12) and showed a phenotypic change.

[0117] Fig. 26 shows in vitro differentiation of THP-1 cells (human). (A) Human THP-1 cells (ATCC) were cultured in RPMI culture media + PMA for 24 hours. One hundred thousand cells were exposed to MSR (5mg / dose), and MSR (5mg / dose) containing human GM-CSF (1 ug / dose) and CpG (100ug / dose) for 24 hours. Microscopy of THP-1 cells is performed by brightfield images showing the cellular morphological analysis after exposure to MSR biomaterial and Att-01 . (B) Human THP-1 (ATCC) were differentiated with PMA for 24 hours. One hundred thousand differentiated cells were co-cultured with MSR (5mg / dose), Att-01 (5mg MSR + 1 ug GM-CSF + 100ug CpG), and to 1 ug GM-CSF + 100ug CpG for 24 hours. Untreated cells were used as control. Supernatant was collected and IL-1 b and IL-18 were detected by ELISA (R&D) for inflammasome responses. (C) Undifferentiated human THP-1 cells (ATCC)were cultured in RPMI culture media for one week. 100K human THP-1 cells were treated with iCt (ara-C + Doxorubicin), Att-01 (5mg MSR + 1 ug GM-CSF + 100ug CpG), iCt + Att-01 , GM-CSF (1 ug) + CpG (100ug) and iCt + GM-CSF (1 ug) + CpG (100ug). Untreated cells were used as control. Cells were harvested after 24 hours. Shown are the percentage of T- and B- cells determined by CD3+CD4+, CD3+CD8+, and CD3+B220+ analysis by Flow Cytometry. Active macrophages were determined by the percentage of CD11 c+CD86+, and CD11 c+GR-1 + cells by Flow Cytometry. Active myeloid cells were determined by percentage of CD11 b+CD86+ and CD11 b+ GR-1 + cells by Flow Cytometry. (D) One million human THP-1 cells (ATCC) were exposed to PMA for 24 hours. Cells were then co-cultured with MSR (1 mg), Att-02 (1 mg MSR + 20ug IL-12), Att-02 + CpG (1 mg MSR + 20ug IL-12 + 100ug CpG, CpG (100ug) + IL-12 (20ug), MSR (1 mg) + CpG (100ug), silica NP (1 mg), NP (1 mg) + CpG (100ug), NP (1 mg) + IL-12 (20ug), and NP (1 mg) + IL-12 (20ug) + CpG (100ug) for 24 hours. Untreated cells are used as control. Supernatant was collected. Nets formation was analyzed by Netosis Kit (Abeam) following the manufacturing protocol. Absorbance was read at 405nm.DEFINITIONS

[0118] Reference in this specification to “one embodiment / aspect” or “an embodiment / aspect” means that a particular feature, structure, or characteristic described in connection with the embodiment / aspect is included in at least one embodiment / aspect of the disclosure. The use of the phrase “in one embodiment / aspect” or “in another embodiment / aspect” in various places in the specification are not necessarily all referring to the same embodiment / aspect, nor are separate or alternative embodiments / aspects mutually exclusive of other embodiments / aspects. Moreover, various features are described which may be exhibited by some embodiments / aspects and not by others.Similarly, various requirements are described which may be requirements for some embodiments / aspects but not other embodiments / aspects. Embodiment and aspect can in certain instances be used interchangeably.

[0119] The terms used in this specification generally have their ordinary meanings in the art, within the context of the disclosure, and in the specific context where each term is used. Certain terms that are used to describe the disclosure are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the disclosure. It will be appreciated that the same thing can be said in more than one way.

[0120] Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein. Nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in thisspecification including examples of any terms discussed herein is illustrative only and is not intended to further limit the scope and meaning of the disclosure or of any exemplified term. Likewise, the disclosure is not limited to various embodiments given in this specification.

[0121] Without intent to further limit the scope of the disclosure, examples of instruments, apparatus, methods and their related results according to the embodiments of the present disclosure are given below. Note that titles or subtitles may be used in the examples for convenience of a reader, which in no way should limit the scope of the disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In the case of conflict, the present document, including definitions, will control.

[0122] As applicable, the terms "about" or "generally", as used herein in the specification and appended claims, and unless otherwise indicated, means a margin of + / - 20%. Also, as applicable, the term "substantially" as used herein in the specification and appended claims, unless otherwise indicated, means a margin of + / - 10%. It is to be appreciated that not all uses of the above terms are quantifiable such that the referenced ranges can be applied.

[0123] The term “mesoporous silica” refers to a form of silica that is characterized by its mesoporous structure (i.e., having pores that range from about 2 nm to about 50 nm in diameter). Mesoporous silica is a relatively recent development in nanotechnology. The most common types of mesoporous nanoparticles are MCM-41 and SBA-15. Research continues on the particles, which have applications in catalysis, drug delivery and imaging. Mesoporous ordered silica films have been also obtained with different pore topologies.

[0124] The term “mesoporous silica rod” or “MSR” generally refers to preformed shapes of silica that are characterized by their mesoporous structure (i.e., having pores that range from about 2 nm to about 50 nm in diameter). A mesoporous rod may take the shape of a cylinder, tube, hollow tube, substantially hollow tube, substantially non-hollow tube, prism, sphere, pyramid, cube, bar or any suitable shape known in the art. Length of mesoporous rods can typically range between 50-150 microns, although shorter and longer lengths are also contemplated.

[0125] The term “mesoporous silica rod conjugate” or “MSR conjugate” generally refers to a mesoporous silica rod joined with a therapeutic, an adjuvant, or both. Therapeutics include cytokine payloads, other therapeutic payloads disclosed herein, and any other suitable therapeutic known in the art. Adjuvants include those such as aluminum hydroxide, toll receptor agonists, other adjuvants disclosed herein, and any other suitable adjuvant known in the art. The term “mesoporous silica rodconjugate’’ or “MSR conjugate” is interchangeable with the terms “compound,” “conjugate,” “composition,” “preparation,” “ATT-01 ,” “mesoporous silica rod,” “mesoporous silica material,” “mesoporous silica rod compound,” “mesoporous silica rod preparation,” “MSR,” “MSR material,” “MSR compound,” “MSR composition,” “MSR preparation,” or any suitable equivalent term, phrase or terms known in the art.

[0126] The term “MCM-41 ” or “Mobil Composition of Matter No. 41 ” refers to a mesoporous material with a hierarchical structure from a family of silicate and alumosilicate solids. MCM-41 consists of a regular arrangement of cylindrical mesopores that form a one-dimensional pore system. It is characterized by an independently adjustable pore diameter, a sharp pore distribution, a large surface area and a large pore volume.

[0127] The term “SBA-15” or “Santa Barbara Amorphous-15” refers to a stable mesoporous silica sieve that has high hydrothermal and mechanical stability from a framework of uniform hexagonal pores that feature a narrow pore-size distribution and a tunable pore diameter (i.e., 5 nm to 15 nm) but, most significantly, from its relatively thick walls, which range between 3.1 nm and 6.4 nm. SBA-15 has a high internal surface area, which lends itself to various applications, including environmental adsorption and separation, advanced optics and catalysts.

[0128] The term “active agent” or “active ingredient” refers to a substance, compound, or molecule, which is biologically active or otherwise, induces a biological or physiological effect on a subject to which it is administered to. In other words, “active agent” or “active ingredient” refers to a component or components of a composition to which the whole or part of the effect of the composition is attributed. An active agent can be a primary active agent, or in other words, the component(s) of a composition to which the whole or part of the effect of the composition is attributed. An active agent can be a secondary agent, or in other words, the component(s) of a composition to which an additional part and / or other effect of the composition is attributed.

[0129] A “pharmaceutical composition” can include the combination of an active agent, such as a therapeutic peptide, with a carrier, inert or active, in a sterile composition suitable for diagnostic or therapeutic use in vitro, in vivo or ex vivo.

[0130] As used herein, the term "prevention" means all of the actions by which the occurrence of the disease is restrained or retarded.

[0131] The term “treating” or “treatment” refers to one or more of (1) inhibiting the disease (i.e., arresting further development of the pathology and / or symptomatology); and (2) ameliorating the disease (i.e., reversing the pathology and / or symptomatology) such as decreasing the severity of disease.

[0132] A “subcutaneous injection” generally refers to an injection administered as a bolus into the subcutis - the layer of skin directly below the dermis and the epidermis.

[0133] A “systemic immune response” generally refers to the body’s integrated response to an antigen or other catalyst. As opposed to a local immune response, a systemic immune response recruits immune cells and cytokines from all systems and areas of the body where they are produced and activated.

[0134] The term "administration" refers to the introduction of an amount of a predetermined substance into a patient by a certain suitable method. The composition disclosed herein may be administered via any of the common routes, but is not limited to intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrapulmonary, or intrarectal administration.

[0135] The term “inflammation” refers to part of the complex biological response of body tissues to harmful stimuli, such as pathogens, damaged cells, or irritants, and is a protective response involving immune cells, blood vessels, and molecular mediators. The function of inflammation is to eliminate the initial cause of cell injury, clear out necrotic cells and tissues damaged from the original insult and the inflammatory process, and initiate tissue repair. The five cardinal signs are heat, pain, redness, swelling, and loss of function. Inflammation is a generic response, and therefore it is considered as a mechanism of innate immunity, as compared to adaptive immunity, which is specific for each pathogen. Too little inflammation could lead to progressive tissue destruction by the harmful stimulus (e.g., bacteria) and compromise the survival of the organism. In contrast, too much inflammation, in the form of chronic inflammation, is associated with various diseases, such as hay fever, periodontal disease, atherosclerosis, and osteoarthritis. Inflammation can be classified as either acute or chronic. Acute inflammation is the initial response of the body to harmful stimuli and is achieved by the increased movement of plasma and leukocytes (in particular granulocytes) from the blood into the injured tissues. A series of biochemical events propagates and matures the inflammatory response, involving the local vascular system, the immune system, and various cells within the injured tissue. Prolonged inflammation, known as chronic inflammation, leads to a progressive shift in the type of cells present at the site of inflammation, such as mononuclear cells, and is characterized by simultaneous destruction and healing of the tissue from the inflammatory process.

[0136] The term “inflammatory disorder” or “inflammation condition” refers to a condition in which the immune system mistakenly attacks one’s own cells or tissues. This causes abnormal inflammation that can result in chronic pain, redness, swelling, stiffness and damage to otherwise healthy body tissues.Inflammatory conditions can affect the nervous system (e.g., encephalitis, myelitis, meningitis, arachnoiditis and neuritis). Inflammatory conditions can affect the eyes (e.g., dacryoadenitis, scleritis, episcleritis, keratitis, retinitis, chorioretinitis, blepharitis, conjunctivitis and uveitis). Inflammatory conditions can affect the ears (e.g., Otitis externa, Otitis media, Labyrinthitis and Mastoiditis).Inflammatory conditions can affect the cardiovascular system (e.g., Endocarditis, Myocarditis, Pericarditis, Arteritis, Phlebitis and Capillaritis). Inflammatory conditions can affect the respiratory system (e.g., Sinusitis, Rhinitis Pharyngitis, Laryngitis, Tracheitis, Bronchitis, Bronchiolitis, Pneumonitis, Pleuritis and Mediastinitis. Inflammatory conditions can affect the mouth and digestive system (e.g., Stomatitis, Gingivitis, Gingivostomatitis, Glossitis, Tonsillitis, Sialadenitis / Parotitis, Cheilitis, Pulpitis, Gnathitis, Esophagitis, Gastritis, Gastroenteritis, Enteritis, Colitis, Enterocolitis, Duodenitis, Ileitis, Caecitis, Appendicitis and Proctitis). Inflammatory conditions can affect the accessory digestive organs (e.g., Hepatitis, Ascending cholangitis, Cholecystitis, Pancreatitis and Peritonitis). Inflammatory conditions can affect the integumentary system (e.g., Dermatitis, Folliculitis, Cellulitis and Hidradenitis). Inflammatory conditions can affect the musculoskeletal system (e.g., Arthritis Dermatomyositis, Myositis, Synovitis / Tenosynovitis, Bursitis, Enthesitis, Fasciitis, Capsulitis, Epicondylitis, Tendinitis, Panniculitis, Osteochondritis: Osteitis / Osteomyelitis, Spondylitis, Periostitis and Chondritis). Inflammatory conditions can affect the urinary system (e.g., Nephritis, Glomerulonephritis, Pyelonephritis, Ureteritis, Cystitis and Urethritis). Inflammatory conditions can affect the female reproductive system (e.g., Oophoritis, Salpingitis, Endometritis, Parametritis, Cervicitis, Vaginitis, Vulvitis and Mastitis). Inflammatory conditions can affect the male reproductive system (e.g., Orchitis, Epididymitis, Prostatitis, Seminal vesiculitis, Balanitis, Posthitis and Balanoposthitis. Inflammatory conditions can affect the endocrine system (e.g., Insulitis, Hypophysitis, Thyroiditis, Parathyroiditis and Adrenalitis). Inflammatory conditions can also affect the lymphatic system (e.g., Lymphangitis and Lymphadenitis).

[0137] The term “granulocyte” generally refers to any immune cell that has granules in its cytoplasm.

[0138] The term “granuloma” generally refers to an inflammatory nodule consisting of immune cells attempting to wall off substances perceived as foreign but are unable to eliminate, such as an injected or implanted mesoporous silica rod.

[0139] The term “granulopoiesis” generally refers to the formation and development of granulocytes.

[0140] The term “emergency granulopoiesis” generally refers to a process that occurs in response to an insult to an organism in which immune cells are produced in an accelerated manner.

[0141] The term “autoimmune disease” or “autoimmune disorder” refers to a condition arising from an abnormal immune response to a functioning body part. Common autoimmune diseases include Addison disease, Celiac disease, Dermatomyositis, Graves disease, Hashimoto thyroiditis, Multiple sclerosis, Myasthenia gravis and Pernicious anemia.

[0142] The term "immunotherapy" refers to the treatment of a subject afflicted with, or at risk of contracting or suffering a recurrence of, a disease by a method comprising inducing, enhancing, suppressing or otherwise modifying an immune response. "Treatment" or "therapy" of a subject refers to any type of intervention or process performed on, or the administration of an active agent to, the subject with the objective of reversing, alleviating, ameliorating, inhibiting, slowing down or preventing the onset, progression, development, severity or recurrence of a symptom, complication, condition or biochemical indicia associated with a disease.

[0143] The term "potentiating an endogenous immune response" or “potentiating a systemic immune response” refers to increasing the effectiveness or potency of an existing immune response in a subject. This increase in effectiveness and potency may be achieved, for example, by overcoming mechanisms that suppress the endogenous host immune response, the systemic immune response or by stimulating mechanisms that enhance the endogenous host immune response and the systemic immune response.

[0144] The term “granulocyte-macrophage colony-stimulating factor” (initialism: GM-CSF), also known as colony-stimulating factor 2 (CSF2), refers to a monomeric glycoprotein secreted by macrophages, T cells, mast cells, natural killer cells, endothelial cells and fibroblasts that functions as a cytokine.

[0145] The term “antigen” generally refers to a molecule, moiety, foreign particulate matter, or an allergen, such as pollen, that can bind to a specific antibody or T-cell receptor. Antigens can originate either from within the body ("self-protein" or "self antigens") or from the external environment ("non-self"). The immune system identifies and attacks "non-self1external antigens.

[0146] “Antigen-Presenting Cells” (APCs) generally refer to cells that display antigens bound by major histocompatibility complex (MHC) proteins on their surface. T cells may recognize these complexes using their T cell receptors (TCRs). APCs process antigens and present them to T-cells.

[0147] The term “neoplasia” refers to a disease that is caused by or results in inappropriately high levels of cell division, inappropriately low levels of apoptosis, or both. For example, cancer is an example of a neoplasia. Examples of cancers include, leukemia (e.g., acute leukemia, acute lymphocyticleukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin's disease, non-Hodgkin's disease), Waldenstrom's macroglobulinemia, heavy chain disease, and solid tumors such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodenroglioma, schwannoma, meningioma, melanoma, neuroblastoma, and retinoblastoma). Lymphoproliferative disorders are also considered to be proliferative diseases.

[0148] The term “cancer" refers to human cancers and carcinomas, sarcomas, adenocarcinomas, etc., including solid tumors, kidney, breast, lung, kidney, bladder, urinary tract, urethra, penis, vulva, vagina, cervical, colon, ovarian, prostate, pancreas, stomach, brain, head and neck, skin, uterine, testicular, esophagus, and liver cancer. In any of the embodiments above, one or more cancer therapies, e.g., chemotherapy, radiation therapy, immunotherapy, surgery, or hormone therapy can be coadministered further with the methods described herein.

[0149] The term “circulating cancer” generally refers to any type of cancer that resides in the circulatory system or lymphatic system and afflicts the tissues, cells and functioning of those systems.

[0150] The term “circulating cancer cells” generally refers to cancer cells present in the circulatory system or lymphatic system of a patient or subject.

[0151] The term “circulating tumor cell” or CTC generally refers to cell that has shed into the vasculature or lymphatic system from a primary tumor and is carried around the body in the blood and / or lymphatic circulation.

[0152] The term “acute myeloid leukemia (AML)” refers to a cancer of the myeloid line of blood cells, characterized by the rapid growth of abnormal cells that build up in the bone marrow and blood and interfere with normal blood cell production.

[0153] The term “abscopal effect” refers to a hypothesis in the treatment of metastatic cancer whereby shrinkage of untreated tumors occurs concurrently with shrinkage of tumors within the scope of the localized treatment. It is thought that in the abscopal effect, the immune system is stimulated to fight cancer in the whole body as a result of the local therapy.

[0154] The term “infectious disease” refers to bacterial, protozoan, and viral pathogens that infect humans and cause disease. Viral pathogens include, for example, human immunodeficiency virus, hepatitis B virus, hepatitis C virus, herpes virus. Bacterial and protozoal pathogens can include E. coli, Staphylococcus sp., Streptococcus sp., Mycobacterium tuberculosis, Giardia, Malaria, Leishmania, and Pseudomonas aeruginosa. An infectious pathogen can be capable of establishing chronic infections (e.g., those that are prolonged or persistent).

[0155] The term “skin and soft tissue infections” or “SSTIs” encompass any type of microorganism (i.e., bacterial, viral or fungal) that enters any break in the skin and can invade the subcutaneous tissue (soft tissue under the skin), fascia (connective tissue), and muscles.

[0156] The term “cytokine” or “cytokines” generally refer to any of various small regulatory proteins that regulate the cells of the immune system.

[0157] The term “mesoporous” generally refers to a material having pores of a size between about 2 and about 50 nanometers (nm).

[0158] The term “intratumorally” generally refers to “within a tumor.”

[0159] The term “intertumorally” generally refers to “between tumors.”

[0160] The term “physical insult” generally refers to the action or cause of any kind of injury, disturbance, or disruption to an organism’s body, including but not limited to any action or cause of any kind of injury, disturbance, or disruption to an organism’s tissues.

[0161] The term “inflammasome” generally refers to a multi-protein complex that is responsible for inflammatory rheumatic diseases via activation of capsases.

[0162] The term “pyroptosis” generally refers to a form of programmed cell death associated with antimicrobial responses during inflammation.

[0163] The term “immunogenic cell death” generally refers to any type of cell death eliciting an immune response.

[0164] The term “vaccine” generally refers to a substance given to stimulate the body’s production of antibodies and provide immunity against a disease without causing the disease itself in the treatment.

[0165] The term “inoculation” generally refers to the introduction of an antigenic substance or vaccine into the body to produce immunity to a specific disease.

[0166] The term “neutrophil” refers to a type of white blood cell, also known as neutrocytes or heterophils, that form an essential part of the innate immune system.

[0167] The term “macrophage” refers to a white blood cell that phagocytizes necrotic cell debris and foreign material, including viruses, bacteria and tattoo ink.

[0168] The term “dendritic cells” refers to any cells that have branching processes and which form a part of the mammalian immune system.

[0169] The term “Interleukin / IL” generally refers to any of a group of cytokine proteins important in the regulation of lymphocyte protein.

[0170] The term “lnterleukin-12 / IL-12” generally refers to an effective antitumoral cytokine belonging to the familial group of cytokine proteins.

[0171] The term “bolus injection” generally refers to a single dose of a drug or other medicinal preparation that is administered all at once.

[0172] The term “immunogenicity" generally refers to the ability of a particular substance to provoke an immune response.

[0173] The term “adjuvant” generally refers to a substance that increases or modulates the immune response to a vaccine. An adjuvant can help create a stronger immune response in people receiving a vaccine. Common adjuvants include Aluminum, AS01 B, AS04, CpG 1018, MatrixM™ and MF59.

[0174] The term “adjuvant therapy” or “neoadjuvant therapy” refers to using adjuvants in combination with a primary treatment (e.g., surgery or radiation) to decrease the chance of the cancer returning. It is often used to make the primary treatment more effective.

[0175] The term “NLRP3 inflammasome” refers to a critical component of the innate immune system that mediates caspase-1 activation and the secretion of proinflammatory cytokines IL-1 p / IL-18 in response to microbial infection and cellular damage. However, the aberrant activation of the NLRP3 inflammasome has been linked with several inflammatory disorders, which include cryo pyrin-associated periodic syndromes, Alzheimer’s disease, diabetes, and atherosclerosis. The NLRP3 inflammasome is activated by diverse stimuli, and multiple molecular and cellular events, including ionic flux, mitochondrial dysfunction, and the production of reactive oxygen species. Lysosomal damage has been shown to trigger its activation.

[0176] The term “CCL2,” “chemokine (C-C motif) ligand 2” or “monocyte chemoattractant protein 1 (MCP1)” or “small inducible cytokine A2” refers to a small cytokine that belongs to the CC chemokine family. CCL2 recruits monocytes, memory T cells, and dendritic cells to the sites of inflammation produced by either tissue injury or infection. CCL2 is implicated in pathogeneses of several diseases characterized by monocytic infiltrates, such as psoriasis, rheumatoid arthritis and atherosclerosis.

[0177] The term “immunogenicity” refers to the ability of cells / tissues to provoke an immune response. It is generally considered to be an undesirable physiological response.

[0178] The term “immunogenic tumor” refers to a tumor with sufficient antigens and priming that elicits good T cell responses in the tumor draining lymph node. In contrast, poorly immunogenic tumors fail to generate T cell responses. The ability of tumors to respond to T cell control is not necessarily linked to their ability to prime T cell responses.

[0179] The term “immunogenic cell death,” “ICD” or “immunogenic apoptosis” refers to a form of cell death resulting in a regulated activation of the immune response. This cell death is characterized by apoptotic morphology, maintaining membrane integrity. Endoplasmic reticulum (ER) stress is generally recognized as a causative agent for ICD, with high production of reactive oxygen species (ROS). Two groups of ICD inducers are recognized. Type I inducers cause stress to the ER only as collateral damage, mainly targeting DNA or chromatin maintenance apparatus or membrane components. Type II inducers target the ER specifically. ICD is induced by some cytostatic agents such as anthracyclines, oxaliplatin and bortezomib, or radiotherapy and photodynamic therapy (PDT). Some viruses can be listed among biological causes of ICD. Just as immunogenic death of infected cells induces immune response to the infectious agent, immunogenic death of cancer cells can induce an effective antitumor immuneresponse through activation of dendritic cells (DCs) and consequent activation of specific T cell response. This effect is used in antitumor therapy.

[0180] The term “immune checkpoint’’ or “checkpoint” refers to a regulator of the immune system. Immune checkpoints are crucial for self-tolerance, which prevents the immune system from attacking cells indiscriminately. However, some cancers can protect themselves from attack by stimulating immune checkpoint targets. Checkpoint inhibitor therapy is a form of cancer immunotherapy. The therapy targets immune checkpoints, key regulators of the immune system that when stimulated can dampen the immune response to an immunologic stimulus. Some cancers can protect themselves from attack by stimulating immune checkpoint targets. Checkpoint therapy can block inhibitory checkpoints, restoring immune system function.

[0181] “Tumor cell plasticity” refers to the dynamic ability of cancer cells to change their phenotype in response to various intrinsic and extrinsic signals. This plasticity allows tumor cells to adapt to changing environments, evade therapeutic interventions, and contribute to metastasis, making it a critical factor in cancer progression and treatment resistance.

[0182] “Cancer cell plasticity” is characterized, in an embodiment, by processes such as epithelial- mesenchymal transition (EMT), sternness or stem cell like property acquisition, and metabolic reprogramming, which together endow tumor cells with a high degree of adaptability.

[0183] “Epithelial-mesenchymal transition” is one of the most studied mechanisms of tumor plasticity. During EMT, epithelial tumor cells lose their characteristic cell-cell adhesion properties and acquire mesenchymal traits, allowing them to become more motile and invasive. This transition is often reversible, enabling cells to switch back and forth between epithelial and mesenchymal states depending on the tumor microenvironment. Such plasticity is relevant to the invasion and dissemination of cancer cells but also plays a role in the colonization of metastatic sites.

[0184] Tumor cells can exhibit behaviors that resemble those of macrophages. Some publications describe tumor-macrophage hybrid cells. This plasticity or hybridization event enables tumor cells to adopt characteristics typical of macrophages, such as phagocytosis, expression of macrophage surface markers, and cytokine production. This behavior is observed, for example, in cancers like melanoma and glioblastoma, where tumor cells express macrophage markers such as CD68, CD163, and CD206, allowing them to evade immune surveillance and facilitate tumor progression.

[0185] In addition to aiding immune evasion and metastasis, tumor cells that mimic macrophages can also exhibit phagocytic activity. This ability allows them to engulf dead cells and cellular debris,acquiring additional nutrients and surviving under nutrient-poor conditions. Certain chemotherapies can induce this effect.

[0186] “Differentiation therapy” is a method to treat an advanced cancer in whichare encouraged to differentiate into more mature forms using pharmacological agents. Differentiation therapy is used in APL, a subset of AML, with a very high cure rate. Differentiation therapy has been proposed for solid tumors, and some agents may be working through this mechanism. In addition, studies have shown that biomaterial-induced inflammation can attract tumor cells and influence the microenvironment at the primary tumor site, suggesting that peripheral inflammation can have systemic effects.

[0187] In another embodiment, MSR therapeutics are co-administered in combination with a cell therapy. In an embodiment when an MSR is administered with a cytokine and / or an adjuvant, cells involved in cell therapy are attracted to the site of injection. In a further embodiment, the cells involved in cell therapy are exposed to the silica, cytokines and / or adjuvants and they result in a change in the activation state or result in a proliferative response, depending on the signal. In another embodiment, the MSR plus cytokine, adjuvant, and antigen, generates antigen specific T cells and amplifies / proliferates CAR-T cells, TCR cells, and / or modified NK cells to enhance their effect. In another embodiment, the MSR plus cytokine and / or adjuvant generates a synergistic immune response with a cell therapy. In some embodiments, the MSR therapeutic is ATT-02 (MSR+IL-12) and is combined with a cell therapy.

[0188] Tumor types can be categorized into separate groups based on their response rate to immune checkpoint inhibitors (ICI). In practical terms, which can be understood more clearly in the medical field, tumor types are categorized as either ''hot1' or "cold." Hot tumors have a high mutation load and respond faster to ICIs. They accumulate a lot of mutations, which causes tumor cells to produce specific molecules, neoantigens, on their cell surface. These neoantigens make the tumor more vulnerable to recognition by the immune system and thus more likely to elicit a strong immune response. Malignant tumors considered "hot" include cancer of the bladder, head and neck, kidney cancer, liver cancer, melanoma, and non-small cell lung cancer, as well as tumors of various types with a high rate of microsatellite instability. It is in these types of tumors that inhibitors of immune checkpoints are effective. Cold tumors have a low response rate and are often compared to an impregnable fortress surrounded by a moat. There are few T-cells in their "walls" and it is difficult for them to mobilize an immune response. Common cancers that have "cold" tumors include glioblastoma, ovarian, prostate, and pancreatic cancer.

[0189] The term "administration" refers to the introduction of an amount of a predetermined substance into a patient by a certain suitable method. The composition disclosed herein may be administered via any of the common routes, as long as it is able to reach a desired tissue, for example,but is not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrapulmonary, or intrarectal administration.

[0190] The term “subject” or "patient" refers to any single animal, more preferably a mammal (including such non-human animals as, for example, dogs, cats, horses, rabbits, zoo animals, cows, pigs, sheep, and non-human primates) for which treatment is desired. Most preferably, the patient herein is a human.

[0191] The term “calcination” refers to thermal treatment of a solid chemical compound whereby the compound is raised to high temperature without melting under restricted supply of ambient oxygen, generally for the purpose of removing impurities or volatile substances and / or to incur thermal decomposition.

[0192] The term “poloxamer” refers to nonionic triblock copolymers composed of a central hydrophobic chain of polyoxypropylene (polypropylene oxide)) flanked by two hydrophilic chains of polyoxyethylene (polyethylene oxide)). An important characteristic of poloxamer solutions is their temperature dependent self-assembling and thermo-gelling behavior. Concentrated aqueous solutions of poloxamers are liquid at low temperature and form a gel at higher temperature in a reversible process. The transitions that occur in these systems depend on the polymer composition (molecular weight and hydrophilic / hydrophobic molar ratio). At low temperatures and concentrations (below the critical micelle temperature and critical micelle concentration) individual block copolymers (unimers) are present in solution. Above these values, aggregation of individual unimers occurs in a process called “micellization.” This aggregation is driven by the dehydration of the hydrophobic polyoxypropylene block that becomes progressively less soluble as the polymer concentration or temperature increases. The aggregation of several unimers occurs to minimize the interactions of the PPO blocks with the solvent. Thus, the core of the aggregates is made from the insoluble blocks (polyoxypropylene) while the soluble portion (polyoxyethylene) forms the shell of the micelles.

[0193] The mechanisms on the micellization at equilibrium have shown to depend on two relaxation times: (1) the first and fastest (tens of the microseconds scale) corresponds to the unimers exchange between micelles and the bulk solution and follows the Aniansson-Wall model (step-by-step insertion and expulsion of single polymer chains), and the second and much slower one (in the millisecond range) is attributed to the formation and breakdown of whole micellar units leading to the final micellar size equilibration. Besides spherical micelles, elongated or worm-like micelles can also be formed. The final geometry will depend on the entropy costs of stretching the blocks, which is directly related to their composition (size and polyoxypropylene / polyoxyethylene ratio). The mechanisms involved in the shape transformation are different compared to the dynamics of micellization. Two mechanisms have beenproposed for the sphere-to-rod transitions of block copolymer micelles, in which the micellar growth can occur by (A) fusion / fragmentation of micelles or (B) concomitant fusion / fragmentation of micelles and unimer exchange, followed by smoothing of the rod-like structures.

[0194] With higher increments of the temperature and / or concentration, other phenomena can occur such as the formation of highly ordered mesophases (cubic, hexagonal and lamellar). Eventually, a complete dehydration of the polyoxypropylene blocks and the collapse of the polyoxyethylene chains will lead to clouding and / or macroscopic phase separation. This is due to the fact that hydrogen bonding between the polyoxyethylene and the water molecules breaks down at high temperature and polyoxyethylene becomes also insoluble in water.

[0195] The phase transitions can also be largely influenced by the use of additives such as salts and alcohols. The interactions with salts are related to their ability to act as water structure makers (salting-out) or water structure breakers (salting-in). Salting-out salts increase the self-hydration of water through hydrogen bonding and reduce the hydration of the copolymers, thus reducing the critical micelle temperature and critical micelle concentration. Salting-in electrolytes reduce the water self-hydration and increase the polymer hydration, therefore increasing the critical micelle temperature and critical micelle concentration. The different salts have been categorized by the Hofmeister series according to their ‘salting-out’ power. Different phase diagrams characterizing all these transitions have been constructed for most poloxamers using a great variety of experimental techniques (e.g., SAXS, Differential scanning calorimetry, viscosity measurements, light scattering).

[0196] The term “Pluronic P123” refers to a poloxamer. Because the lengths of the polymer blocks can be customized, many different poloxamers exist that have slightly different properties. Pluronic P123 is a symmetric triblock copolymer comprising polyethylene oxide) (PEO) and polypropylene oxide) (PPO) in an alternating linear fashion, PEO-PPO-PEO. The unique characteristic of PPO block, which is hydrophobic at temperatures above 288 K and is soluble in water at temperatures below 288 K, leads to the formation of micelle consisting of PEO-PPO-PEO triblock copolymers. Some studies report that the hydrophobic core contains PPO block, and a hydrophilic corona consists of PEO block. In 30wt% aqueous solution Pluronic P123 forms a cubic gel phase. The nominal chemical formula of P123 is:HO(CH2CH2O)2Q(CH2CH(CH3)O) 7O(CH2CH20)2OH which corresponds to a molecular weight of around 5800 g / mol. Triblock copolymers based on PEO- PPO-PEO chains are known generically as poloxamer. Poloxamers have behaviors similar to those of hydrocarbon surfactants and will form micelles when placed in a selective solvent such as water. They can form both spherical and cylindrical micelles.

[0197] The term “sargramostim” (tradename Leukine) is a recombinant granulocyte macrophage colony-stimulating factor (GM-CSF) that functions as an immunostimulator.

[0198] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are to be understood as approximations in accordance with common practice in the art. When used herein, the term “about” may connote variation (+) or (-) 1%, 5% or 10% of the stated amount, as appropriate given the context. It is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.

[0199] Many known and useful compounds and the like can be found in Remington’s Pharmaceutical Sciences (13th Ed), Mack Publishing Company, Easton, PA — a standard reference for various types of administration. As used herein, the term “formulation(s)” means a combination of at least one active ingredient with one or more other ingredient, also commonly referred to as excipients, which may be independently active or inactive. The term “formulation” may or may not refer to a pharmaceutically acceptable composition for administration to humans or animals and may include compositions that are useful intermediates for storage or research purposes.

[0200] Other technical terms used herein have their ordinary meaning in the art that they are used, as exemplified by a variety of technical dictionaries. The particular values and configurations discussed in these non-limiting examples can be varied and are cited merely to illustrate at least one embodiment and are not intended to limit the scope thereof.DETAILED DESCRIPTION

[0201] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the subject technology as claimed. Additional features and advantages of the subject technology are set forth in the description below, and in part will be apparent from the description, or may be learned by practice of the subject technology. The advantages of the subject technology will be realized and attained by the structure particularly pointed out in the written description and claims hereof.

[0202] The present invention is thus based on the surprising discovery that subcutaneous injection of mesoporous silica rods (MSRs) stimulates systemic immune activity at high levels that can be directed at targeted tissue. The MSRs can be used as a scaffold to deliver cytokines intratumorally, and to deliver cytokines from areas remote from tumors or cancerous tissue through immune cell production from the bone marrow, blood, lymph nodes and spleen. In various embodiments, the methods described hereinare suitable for treatment of a condition known or expected to be ameliorated by immune stimulation (e.g., cancer or infection). Subcutaneous injection of mesoporous silica rods has been discovered to cause profound changes in bone marrow, blood, lymph nodes and the spleen. The immune response generated by subcutaneous injection of mesoporous silica rods is similar to an acute infection, which provides advantages in treating cancer and cancerous tissues.

[0203] The subcutaneous injection of mesoporous silica rods creates a physical insult that initiates a systemic immune response. A granuloma forms around the injected mesoporous silica rods, developing an induced macro-immune structure composed of immune cells and inflammation that catalyzes downstream production of cytokines and stimulates the systemic production of immune cells. The resulting systemic immune response increases activity in the blood, bone marrow, lymph nodes and spleen by increasing immune cell production from those tissues and organs. Surprisingly, Applicants have discovered that the immune response further allows differentiation of myeloid cells, including cancerous myeloid cells and cancer precursor cells. Additionally, it has been found that this systemic immune response also promotes differentiation of leukemic blasts into cross-presenting antigen-presenting cells (APCs) and other differentiated cells with terminal lifespans that are cleared by the subject or patient over a short time period. The antigen-presenting cells allow the immune system to recognize cancer cells and effectively treat them. Applicants have further discovered that pre-treating a patient or subject with chemotherapy prior to subcutaneous injection of mesoporous silica rods has been shown to prime the leukemic blasts to differentiate into cross-presenting APCs. Methods of the present invention include the step of pre-treating a patient or subject with chemotherapy prior to subcutaneous injection of mesoporous silica rods. Additionally, methods contemplated by the present invention include the step of concomitantly treating a patient or subject with chemotherapy and the subcutaneous injection of mesoporous silica rods.

[0204] The mesoporous silica rods and the methods of using mesoporous silica rods of the invention may include subcutaneous injection of mesoporous silica rods coupled with a cytokine payload and / or an adjuvant and / or an antigen. The cytokine payload, adjuvant and antigen may be an impregnation or coating or may be coupled to the mesoporous silica rods of the present invention by any suitable means known in the art. In aspects the cytokine payload is selected from the group of granulocyte-macrophage colony-stimulating factors such as Leukine, Molgramostin and Sargramostim. In additional aspects, the cytokine payload can be interleukin-12 (IL-12), interleukin-2 (IL-2), or any suitable cytokine payload known in the art. Adjuvants contemplated by the present invention include but are not limited to aluminum hydroxide (alum), a lipopolysaccharide or a toll like receptor agonist (TLR). Antigens may include tumor cell lysates or cancer cell lysates, but myriad antigens are contemplated by the present disclosure and further include any suitable antigen known in the art.Mesoporous Silica Rods (MSRs)

[0205] Mesoporous silica is a form of silica that is characterized by its mesoporous structure, (i.e., having pores that range from about 2 nm to about 50 nm in diameter). Mesoporosity is generally defined between microporous (i.e., < 2 nm) and macroporous (i.e., > 50 nm). Mesoporous silica is a relatively recent development in nanotechnology. Mesoporous ordered silica films have been also obtained with different pore topologies.

[0206] The large surface area of the pores allows the particles to be filled with a drug or a cytotoxin. Like a Trojan Horse, the particles will be taken up by certain biological cells through endocytosis, depending on what chemicals are attached to the outside of the spheres. Some types of cancer cells will take up more of the particles than healthy cells will, suggesting that mesoporous silica rods can be effective to treat certain types of cancer.

[0207] Mesoporous silica conveys several advantages as it is related to interacting with immune cells. The three-dimensional structure created by the MSRs in the subcutaneous space serves as a scaffold that allows the migration and interaction of a population of immune cells. This co-localization allows interactions between immune cells, antigens, and adjuvants which create a granuloma-like structure. For instance, GM-CSF loaded MSR scaffolds recruit not only dendritic cells (DCs) but also a large population of other innate and adaptive immune cells. Histology and cell flow cytometry data have shown that the inflammatory response caused by the MSRs creates an innate and systemic immune response, recruiting diverse populations of immune cells from multiple tissues, organs and systems. Over a period of days, adaptive immune cells are also recruited to the site. When this innate immune response is combined with the delivery of antigen, adjuvants, and cytokines, it further expands to an adaptive response. The benefit of the temporary acute and transient inflammatory response created by the MSRs loaded with immunomodulators help create a robust and durable immune response.

[0208] Mesoporous silica rods also show potential to boost the in vitro and in vivo dissolution of poorly water-soluble drugs. Many drug-candidates coming from drug discovery suffer from poor water solubility. An insufficient dissolution of these hydrophobic drugs in the gastrointestinal fluids strongly limits the oral bioavailability. One example is itraconazole which is an antimycoticum known for its poor aqueous solubility. Upon introduction of itraconazole-on-SBA-15 formulation in simulated gastrointestinal fluids, a supersaturated solution is obtained giving rise to enhanced transepithelial intestinal transport. Also, the efficient uptake into the systemic circulation of SBA-15 formulated itraconazole has been demonstrated in vivo (rabbits and dogs). This approach based on SBA-15 yields stable formulations and can be used for a wide variety of poorly water-soluble compounds.Cytokines

[0209] Cytokines are a broad and loose category of small proteins (e.g., 5 - 25 kDa) that are important in cell signaling. Cytokines are peptides and cannot cross the lipid bilayer of cells to enter the cytoplasm. Cytokines have been shown to be involved in autocrine, paracrine and endocrine signaling as immunomodulating agents.

[0210] Cytokines include chemokines, interferons, interleukins, lymphokines, granulocytemacrophage colony-stimulating factors and tumor necrosis factors, but generally not hormones or growth factors (despite some overlap in the terminology). Cytokines are produced by a broad range of cells, including immune cells like macrophages, B lymphocytes, T lymphocytes and mast cells, as well as endothelial cells, fibroblasts, and various stromal cells; a given cytokine may be produced by more than one type of cell. They act through cell surface receptors and are especially important in the immune system; cytokines modulate the balance between humoral and cell-based immune responses, and they regulate the maturation, growth, and responsiveness of particular cell populations. Some cytokines enhance or inhibit the action of other cytokines in complex ways. Cytokines are important in health and disease, specifically in host immune responses to infection, inflammation, trauma, sepsis, cancer, and reproduction.

[0211] Interleukin-12 (IL-12) is an interleukin that is naturally produced by dendritic cells, macrophages, neutrophils, and human B-lymphoblastoid cells (NC-37) in response to antigenic stimulation. The IL-12 family is unique in comprising the only heterodimeric cytokines, which includes IL- 12, IL-23, IL-27 and IL-35. IL-12 also has anti-angiogenic activity (i.e. , it can block the formation of new blood vessels) thus can be used as an antineoplastic agent. Despite sharing many structural features and molecular partners, they mediate surprisingly diverse functional effects.

[0212] lnterleukin-2 (IL-2) is an interleukin, a type of cytokine signaling molecule in the immune system. It regulates the activities of white blood cells (leukocytes, often lymphocytes) that are responsible for immunity. IL-2 is part of the body's natural response to microbial infection and discriminates between foreign ("non-self") and non-foreign ("self') entities. IL-2 mediates its effects by binding to IL-2 receptors, which are expressed by lymphocytes. The major sources of IL-2 are activated CD4+ T cells and activated CD8+ T cells.

[0213] Applicants have discovered surprising results utilizing the subcutaneous injection of mesoporous silica rods (MSRs) that form induced macro-immune scaffolds that deliver cytokines and promote the generation of immune cells by stimulating a systemic immune response. The combination of the MSRs and cytokines modulate the immune cell population and activation states in a manner that can alter tumor and cancerous-cell microenvironments. This leads to a more effective response by theimmune system and improved survival rates of subjects afflicted with various forms of cancer, inflammatory and infectious disease.

[0214] Without being bound by theory, Applicants propose that the MSRs can trigger multiple pathways of the immune system. It was discovered that macrophage foam cells may be utilized to alter the performance and nature of immune cells, via alternative pathway signaling and temporal control of specific cell recruitment to improve the quality and robustness of the immune response. Through reducing the toxicity of cytokines by controlling and localizing their release, the present invention serves to change the therapeutic window of cytokines such as IL-2 and IL-12, by exposing tumors, cancer cells and areas of inflammation to a locally high concentration in combination with inflammatory signals induced by implanted and / or injected silica materials. In embodiments, the agents (e.g., cytokines) and MSRs work synergistically with one another.

[0215] The treatment of subjects with mesoporous silica rods can trigger a systemic immune response through a physical insult. Through this mechanism, numerous populations of immune cells are recruited to the targeted area of treatment, influencing and activating innate immune pathways. The controlled chemistry and configuration of MSRs and other suitable silica materials activate specific immune pathways in combination with cytokines, IL-12, IL-2, inflammasomes and other regulatory proteins.

[0216] The methods described herein provide several advantages over conventional treatments for cancer, infection and inflammation. The MSR delivery system can be modified or configured to: a) release a variety of cargos (e.g., cytokines) at different rates, b) effectively deliver large cargo utilizing the high surface area of the MSRs, c) increase the half-life of target cytokines, d) control the exposure and concentration of cytokine release, and e) solubilize cargos and safely release them over time (e.g., from days to months).One or more additional agents can be included on the MSRs. In embodiments, adjuvants are included on the MSRs. In embodiments, the MSRs described herein can be targeted to a specific tissue such as a tumor. In other embodiments, the MSRs described herein can be targeted to malignant cancers by generating a systemic immune response. The MSRs can also be used to treat an ailment related to infection and / or inflammation. Further, the MSRs can treat cancerous tissues, cancerous cells, tumors, infections and inflammatory disorders remotely from a subcutaneous injection site of a subject or patient.

[0217] The treatments described herein allow for the creation of an alternative immunological environment within the tumor and / or inflammatory site which creates responses that improve tumor and inflammatory regression. The material composition of the mesoporous silica rods further allows forimmunogenic cell death (ICD) which creates antigen sources for further signaling immune cell proliferation, therefore focusing agents such as neutrophils, macrophages, antigen-presenting cells (APCs), dendritic cells and other immune cells to targeted treatment areas for preferable outcomes such as tumor suppression, tumor regression, tumor-size reduction, tumor elimination, inflammatory reduction, cancer cell elimination, cancer cell reduction, cancer cell suppression, inflammatory suppression, inflammatory regression and elimination of inflammation within targeted areas.Adjuvants

[0218] An adjuvant can be defined as a vaccine component that enhances the magnitude, breadth and / or durability of an immune response. Due to the variety of mechanisms and links between the innate and adaptive immune response, an adjuvant-enhanced innate immune response results in an enhanced adaptive immune response. Specifically, adjuvants may exert their immune-enhancing effects according to five immune-functional activities.

[0219] First, adjuvants may help in the translocation of antigens to the lymph nodes where they can be recognized by T cells. This will ultimately lead to greater T cell activity resulting in a heightened clearance of pathogen throughout the organism. Second, adjuvants may provide physical protection to antigens which grants the antigen a prolonged delivery. This means the organism will be exposed to the antigen for a longer duration, making the immune system more robust as it makes use of the additional time by upregulating the production of B and T cells needed for greater immunological memory in the adaptive immune response. Third, adjuvants may help to increase the capacity to cause local reactions at the injection site (during vaccination), inducing greater release of danger signals by chemokine releasing cells such as helper T cells and mast cells. Fourth, they may induce the release of inflammatory cytokines which helps to not only recruit B and T cells at sites of infection but also to increase transcriptional events leading to a net increase of immune cells as a whole. Finally, adjuvants are believed to increase the innate immune response to the antigen by interacting with pattern recognition receptors (PRRs) on or within accessory cells.

[0220] Adjuvants include, but are not limited to: Toll Like Receptor (“TLR”) agonists (e.g., TLR9), lipopolysaccharides (LPS), and aluminum hydroxide (alum).Antigens

[0221] Compositions, methods, and devices described herein may further comprise tumor antigens, cancer cell antigens or other antigens. Antigens elicit protective immunity or generate a therapeutic immune response in a subject to whom such a device was administered. In embodiments, employed antigens may be tumor cell lysates. For example, a whole tumor or tumor biopsy sample isextracted from a human patient (or non-human animal) and digested using collagenase to degrade the extra cellular matrix. The tumor cells then undergo three cycles of a freeze-thaw process, in which the cells are frozen in liquid N2 and then thawed in a water bath. This process generates the tumor antigens, which are then loaded onto the MSRs at the same time with GM-CSF and CpG ODN. For example, a vaccine dose of tumor cell lysate antigen is the amount obtained from 1 x 1 o106tumor cells. After fabrication of the MSRs, the particles are suspended in a physiologically accepted buffer, e.g., PBS, and the recruitment compound (e.g., GM-CSF), activating compound (e.g., CpG), and antigen added to the suspension of rods. The mixture is shaken at room temperature overnight and then lyophilized for about 4 hours. Prior to administration, the rods are again suspended in buffer and the suspension loaded into a 1 ml syringe (18 gauge needle). A typical vaccine dose is 150 pl of the mixture per injection.

[0222] Exemplary cancer antigens encompassed by the compositions, methods, and devices of the present invention include, but are not limited to, tumor lysates extracted from biopsies, cancerous cell lysates, circulating cancer cell lysates, AML cell lysates, irradiated tumor cells, melanoma-associated antigens including but not limited to MAGE series of antigens (MAGE-1 is an example), MART-1 / melan-A, tyrosinase, ganglioside, gp100, GD-2, O-acetylated GD-3, GM-2, MUC-1 , Sos1 , Protein kinase C-binding protein, Reverse transcriptase protein, AKAP protein, VRK1 , KIAA1735, T7-1 , T11-3, T11 -9, Homo Sapiens telomerase ferment (hTRT), Cytokeratin-19 (CYFRA21-1), SQUAMOUS CELL CARCINOMA ANTIGEN 1 (SCCA-1), (PROTEIN T4-A), SQUAMOUS CELL CARCINOMA ANTIGEN 2 (SCCA-2), Ovarian carcinoma antigen CA125 (1A1-3B) (KIAA0049), MUCIN 1 (TUMOR-ASSOCIATED MUCIN), (CARCINOMA-ASSOCIATED MUCIN), (POLYMORPHIC EPITHELIAL MUCIN), (PEM), (PEMT), (EPISIALIN), (TUMOR-ASSOCIATED EPITHELIAL MEMBRANE ANTIGEN), (EMA), (H23AG), (PEANUT-REACTIVE URINARY MUCIN), (PUM), (BREAST CARCINOMA-ASSOCIATED ANTIGEN DF3), CTCL tumor antigen sel-1 , CTCL tumor antigen se14-3, CTCL tumor antigen se20-4, CTCL tumor antigen se20-9, CTCL tumor antigen se33-1 , CTCL tumor antigen se37-2, CTCL tumor antigen se57-1 , CTCL tumor antigen se89-1 , Prostate-specific membrane antigen, 5T4 oncofetal trophoblast glycoprotein, Orf73 Kaposi's sarcoma-associated herpesvirus, MAGE-C1 (cancer / testis antigen CT7), MAGE-B1 ANTIGEN (MAGE-XP ANTIGEN) (DAM10), MAGE-B2 ANTIGEN (DAM6), MAGE-2 ANTIGEN, MAGE-4a antigen, MAGE-4b antigen, Colon cancer antigen NY-CO-45, Lung cancer antigen NY-LU-12 variant A, Cancer associated surface antigen, Adenocarcinoma antigen ART1 , Paraneoplastic associated braintestis-cancer antigen (onconeuronal antigen MA2; paraneoplastic neuronal antigen), Neuro-oncological ventral antigen 2 (NOVA2), Hepatocellular carcinoma antigen gene 520, TUMOR-ASSOCIATED ANTIGEN CO-029, Tumor-associated antigen MAGE-X2, Synovial sarcoma, X breakpoint 2, Squamous cell carcinoma antigen recognized by T cell, Serologically defined colon cancer antigen 1 , Serologically defined breast cancer antigen NY-BR-15, Serologically defined breast cancer antigen NY-BR-16, Chromogranin A; parathyroid secretory protein 1 , DUPAN-2, CA 19-9, CA 72-4, CA 195,Carcinoembryonic antigen (CEA). Purified tumor antigens are used alone or in combination with one another.

[0223] Coupling an antigen to an MSR or other mesoporous silica material is also useful to generate an immune response to other antigens such as microbial pathogens (e.g., bacteria, viruses, fungi).Methods of Use

[0224] Accordingly, an aspect of the invention is to generate a systemic immune response through the administration of mesoporous silica rods to treat cancer through the suppression, reduction, and in certain cases elimination of cancerous tissues, cancerous cells, circulating cancer cells, cancerous lesions, tumors or other cancerous quantities.

[0225] The suppression, reduction and elimination of cancerous tissues, cancerous cells, circulating cancer cells, cancerous lesions and tumors is achieved through the action of immune cells systemically generated in response to the administration of mesoporous silica rods.

[0226] In embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods.

[0227] In yet other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods and a cytokine payload.

[0228] In other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods and a granulocyte-macrophage colony-stimulating-factor (GM-CSF).

[0229] In yet other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods and an antigen.

[0230] In embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods and a cytotoxic payload.

[0231] In other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods and a therapeutic protein.

[0232] In yet other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods and an adjuvant.

[0233] In other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a cytokine payload, and an adjuvant.

[0234] In embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, GM-CSF, and an adjuvant.

[0235] In yet other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a cytotoxic payload, and an adjuvant.

[0236] In embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a therapeutic protein, and an adjuvant.

[0237] In other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, an adjuvant and an antigen.

[0238] In yet other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a cytokine payload and an antigen.

[0239] In embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, GM-CSF and an antigen.

[0240] In other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a therapeutic protein and an antigen.

[0241] In yet other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a cytotoxic payload and an antigen.

[0242] In embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, GM-CSF, an adjuvant and an antigen.

[0243] In other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a cytokine payload, an adjuvant and an antigen.

[0244] In yet other embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a cytotoxic payload, an adjuvant and an antigen.

[0245] In embodiments, a systemic immune response is generated by the subcutaneous injection of mesoporous silica rods, a therapeutic protein, an adjuvant and an antigen.

[0246] An aspect of the invention relates to methods for treating circulating cancer cells by the subcutaneous injection of mesoporous silica rods. Circulating cancer cells can include acute myeloid leukemia (AML) cells, acute lymphoblastic leukemia cells, chronic lymphocytic leukemia cells, chronic myelogenous leukemia cells, Hodgkin lymphoma cells, non-Hodgkin lymphoma cells, Waldenstrom macroglobulinem cells, follicular lymphoma cells, B-cell lymphoma cells, cutaneous T-cell lymphoma cells, multiple myeloma cells, plasmacytoma cells, or cancer precursor cells.

[0247] It is a further aspect of the invention to alter the therapeutic window and utilization of cytokine proteins to achieve preferable outcomes in inflammation treatment through the suppression, reduction, and in certain cases elimination of an inflammatory region through the generation of cytokine proteins catalyzed by the administration of mesoporous silica rods. Conditions ameliorated by immune stimulation caused by administration of mesoporous silica rods include, for example, infectious diseases, (e.g., bacterial, fungal, viral and parasitic infectious diseases). Also, conditions associated with uncontrolled proliferation of cells (e.g., cancers) can be ameliorated by systemic immune stimulation triggered by administration of mesoporous silica rods.

[0248] It is a further aspect of the invention to alter the therapeutic window and utilization of cytokine proteins to achieve preferable outcomes in inflammation treatment through the suppression, reduction, and in certain cases elimination of an inflammatory region through the generation of cytokine proteins catalyzed by subcutaneous injection of mesoporous silica rods. Conditions ameliorated by immune stimulation caused by subcutaneous injection of mesoporous silica rods include, for example, infectious diseases, (e.g., bacterial, fungal, viral and parasitic infectious diseases). Also, conditions associated with uncontrolled proliferation of cells (e.g., cancers) can be ameliorated by systemic immune stimulation triggered by subcutaneous injection of mesoporous silica rods.

[0249] Subcutaneous injection of mesoporous silica rods, cytokine payloads and / or adjuvants and / or antigens may be accomplished through any subcutaneous delivery method or mechanism known in the art. Mesoporous silica rods, cytokine payloads and / or adjuvants may be delivered by a needled syringe of any suitable gauge and material composition. Subcutaneous injection of mesoporous silica rods, cytokine payloads and / or adjuvants may be performed through auto-injection by an autoinjector. However, any suitable administration technique or device (e.g., syringes, pen injectors, autoinjectors, robotic injectors, Al-controlled injectors) designed for subcutaneous injection known in the art is contemplated by this disclosure.

[0250] As described herein, delivery of mesoporous silica rods may be accompanied by adjuvants which serve as secondary signals for inflammasome activation. Examples of adjuvants contemplated by the present invention include but are not limited to: Toll Like Receptor (“TLR”) agonists (e.g., TLR9), lipopolysaccharides (LPS), and aluminum hydroxide (alum).

[0251] The internal geometry of the mesoporous silica rod and mesoporous silica material can vary based on application. The internal structure may be labyrinthine (maze-like) or may contain internal pores of varying diameter to allow controlled influx of inflammatory response agents generated by the organism while metering egress of any of the therapeutic payloads and / or adjuvants described above. The internal geometry may also comprise several dead ends, as an internal pore does not necessarily have to lead to an aperture opening on the surface of the mesoporous silica rod, which therefore offers a time-release mechanism for excretion of therapeutic product.

[0252] The overall geometry of the mesoporous silica material and mesoporous silica rod is also not limited to that of a cylinder. Other common and equivalent shapes are contemplated, including but not limited to spheres, tubes, icosahedrons, bars, irregular rods, pyramids, cubes, cuboids, prisms, octahedrons, dodecahedrons, triangular prisms, octagonal prisms, pentagonal prisms, ellipsoids, irregular ellipsoids, tetrahedrons, square pyramids, hexagonal pyramids, and any other geometrical equivalents and substitutes known in the art.

[0253] The mesoporous silica rods may include immune cell recruitment compounds and immune cell activating compounds attached as therapeutic payloads as disclosed herein.

[0254] Immune cell recruitment compounds include granulocyte macrophage-colony stimulating factor (GM-CSF). Other examples of recruitment compounds are disclosed in US Patent No. 1 1 ,278,604 and are incorporated herein by reference. These recruitment compounds include chemokines, e.g., a chemokine selected from the group consisting of chemokine (C-C motif) ligand 21 (CCL-21 , GenBank Accession Number: (aa) CAG29322.1 (G 1:47496599), (na) EF064765.1 (Gl:117606581), chemokine (C-C motif) ligand 19 (CCL-19, GenBank Accession Number: (aa) CAG33149.1 (Gl:48145853), (na) NM_006274.2 (Gl:22165424), incorporated herein by reference), as well as FMS-like tyrosine kinase 3 ligand (Flt3) ligand; Genbank Accession Number: (aa) AAI44040 (Gl:219519004), (na) NM_004119 (Gl: Gl:121114303).

[0255] Immune cell activating compounds include TLR agonists. Such agonists include pathogen associated molecular patterns (PAMPs), e.g., an infection-mimicking composition such as a bacterially- derived immunomodulator (a.k.a., danger signal). TLR agonists include nucleic acid or lipid compositions [e.g., monophosphoryl lipid A (MPLA)]. In one example, the TLR agonist comprises a TLR9 agonist suchas a cytosine-guanosine oligonucleotide (CpG-ODN), a poly(ethylenimine) (PEI)-condensed oligonucleotide (ODN) such as PEI-CpG-ODN, or double stranded deoxyribonucleic acid (DNA). For example, the device comprises 5 pg, 10 pg, 25 pg, 50 pg, 100 pg, 250 pg, or 500 pg of CpG-ODN. In another example, the TLR agonist comprises a TLR3 agonist such as polyinosine-polycytidylic acid (poly I :C) , PEI-poly (I :C) , polyadenylic-polyuridylic acid (poly (A:U)), PEI-poly (A:U), or double stranded ribonucleic acid (RNA). Lipopolysaccharide (LPS) is also useful for this purpose. Additional immune cell activating compounds are disclosed in US Patent No. 11 ,278,604 and are incorporated herein by reference.Artificial intelligence (Al) or Machine Learning Assisted Selection of Agents

[0256] In an embodiment, an Al algorithm enables the prediction of the most effective single-agent or multi-agent combinations for an individual patient. An Al can be one that is publicly available and can be inputted with the relevant data or one that is created to specifically identify the therapeutic treatment for a disease for a patient. By using the Al, the treatment can be optimized to increase the efficacy and minimize the adverse effects.Progress in Interventional Radiology:

[0257] In an embodiment a radiotherapeutic is used in combination with a therapeutic and a biomedical scaffold to enhance tumor delivery using interventional radiology that enables more precise and localized delivery of treatments to a tumor throughout the body. This progress improves the effectiveness of therapies and reduces systemic toxicity.Immune Response Stimulated by Mesoporous Silica Rod Treatment

[0258] Upon subcutaneous injection of the mesoporous silica rods into the patient, a systemic immune response is initiated. The subcutaneous injection of mesoporous silica rods delivers a physical insult to the patient which triggers the development of immune cells from the lymph nodes, spleen, bone marrow and blood.Innovative Approaches to Lymph Node Treatment:

[0259] In an embodiment, a treatment does not involve the resection of a local lymph node to improve immunotherapy responses. In another embodiment, a treatment by treating regional lymph nodes with localized therapies (e.g. a biomedical scaffold with a therapeutic, including a cytokine) in addition to providing a therapeutic that targets the primary tumor, which enhances overall treatment outcomes.Modular and Simple Platform:In an embodiment, the biomaterial scaffold, multiple cytokines, antigens, and immunotherapies, can be stored and then mixed or provided to a patient at a hospital pharmacy. MSRs are also stored at the site where the patient will be administered the combined therapeutic and scaffold. Upon patient diagnosis, biopsy, IHC / molecular testing, and Al analysis, a pharmacist can quickly prepare a personalized, localized tumor therapy through preparation of an aqueous admix procedure using the selected therapeutic components and MSRs.Advantages Over Current Solutions:

[0260] Time. In an embodiment, the platform of the present invention significantly reduces the time required to develop patient-specific treatments. This rapid response capability is crucial for patients with progressive diseases, where time is of the essence.

[0261] Safety. In an embodiment, the present invention does not require lymphodepleting chemotherapy prior to administration of the MSR therapeutic combination. In a further embodiment, the present invention is safer for patients. In addition, localized delivery of cytokines can minimize systemic exposure and toxicity.

[0262] Efficacy potential. In another embodiment, personalized combination therapy has a stronger efficacy signal than traditional in situ therapies. In an embodiment the stronger efficacy signal when conducted using a multi-agent exposure can signal multiple cell types.

[0263] Scalability. In an embodiment, the modular nature of the platform provides for easy expansion and adaptation to new therapies and technologies.The Process

[0264] Option 1 : This embodiment, works by using existing databases of patient data, including RNA SEQ and single cell RNA seq, to determine mechanisms of immune dysfunction in tumors In this embodiment, an Al mines the scientific literature for cytokines or immune agonists that can reverse the diseases phenotype, for example, a cancer, including a circulating cancer, by treating a patient with an MSR plus cytokine that is manufactured to be administered to a patient. In an embodiment, the MSR plus cytokine combination is administer to a tumor and / or to a tumor draining lymph node Following treatment, omics data is collected from the treated patient and used to build a database for improved future treatments to the patient and other patients with similar results.

[0265] Option 2: This embodiment works by using existing databases of patient data including RNA SEQ and single cell RNA seq to determine a mechanism of immune dysfunction in tumors, In this embodiment, an Al is used to mine patient data from a clinical database (including one managed by Attivare) for cytokines and / or immune agonists that can reverse a phenotype using an MSR plus a cytokine. In this embodiment, the MSR plus cytokine combination is injected into a tumor and / or into a tumor draining lymph node Following treatment, ‘omics data is collected from the treated patient and used to build a database for improved future treatments to the patient and other patients with similar results.

[0266] Option 3: This embodiment works with existing databases of patient data, including RNA SEQ and single cell RNA seq, to determine a mechanism of immune dysfunction in tumors This embodiment works with an Al to mine patient data from patient clinical database (including one managed by Attivare) for cytokines or immune agonists that can reverse a disease phenotype using an MSR plus cytokine Inject the MSR plus a cytokine combination. In this embodiment, the MSR plus a cytokine or immune agonist or immunotherapy combination is administered to a tumor and / or in a tumor draining lymph node. After administration, the patient to whom the combination therapeutic has been administered is monitored over time for a reduction or resolution of the disease and / or its symptoms, including a cancer. For those patients that show a reduction or resolution of the disease and / or its symptoms, biomarker data is collected. Patient ‘omics data is then stored in a database to be mined by an Al to determine improved future treatments for the patient or other patients with similar phenotypes and markers for the same or a similar disease. Omics include, but are not limited to genomics, proteomics, metabolomics, metagenomics, phenomics and transcriptom ics.

[0267] After subcutaneous injection of mesoporous silica rods, a granuloma forms near the injection site, producing inflammation and forming a macro-immune structure containing neutrophils, lymphocytes, macrophages and other immune cells in the vicinity of the injected mesoporous silica rods. Downstream of the formed granuloma, inflammatory cells cause the release of systemic cytokines. The immune response also stimulates the differentiation of myeloid cells into neutrophils, macrophages, antigen-presenting cells and / or other immune cells.

[0268] After subcutaneous injection of mesoporous silica rods into the patient, the lymph nodes produce immune cells that flood to the macro-immune structure induced near the injection site and then circulate to attack cancer cells and other cancerous tissue.

[0269] In the treatment of cancers such as acute myeloid leukemia, the immune response also stimulates the differentiation of myeloid cells, AML cells, and leukemic blasts into neutrophils, macrophages, antigen-presenting cells and / or other immune cells.

[0270] The immune response generated by subcutaneous injection of mesoporous silica rods has been shown to arise in the blood, bone marrow, spleen and lymph nodes. This results in increased production of immune cells, cytokines (e.g., IL-1 beta, IL-6, IL-8, IL-12) and cluster of differentiation cells (e.g., CD8+ T Cells, CD11 b-GR-1 cells, CD19 cells).

[0271] The macro-immune structure arises from the physical insult generated by the subcutaneous injection of mesoporous silica rods into the patient. The macro-immune structure includes neutrophils, lymphocytes, macrophages and other immune cells which form a scaffolding around the injected mesoporous silica rods. The mesoporous silica rods can carry a payload of GM-CSF which further stimulates production of the macro-immune structures.

[0272] The effectively differentiable myeloid cells can be healthy precursor cancer cells or acute myeloid leukemia cells. The production of these cells is generated by a systemic immune response stimulated by the subcutaneous injection of mesoporous silica rods and / or mesoporous silica rod conjugates.

[0273] Referring to the drawing figures, FIG. 1 depicts the formation of a macro-immune structure 10 induced by the subcutaneous injection of mesoporous silica rods 5 containing therapeutics and / or adjuvants. The subcutaneous injection of mesoporous silica rods further triggers immune cell trafficking to and from lymph nodes 15 and tumors 20. The macro-immune structure is comprised of mesoporous silica rods, tumor / cancer cells, neutrophils, lymphocytes, macrophages and other immune cells provided through the systemic immune response generated by the subcutaneous injection of mesoporous silica rods and / or mesoporous silica rod conjugates.

[0274] FIG. 2A is a graph showing survival percentages of mice challenged with 100K c1498-eGFP cells when treated with chemotherapy plus subcutaneous injection of mesoporous silica rods versus mice treated with chemotherapy plus subcutaneous injection of GM-CSF + CpG versus mice treated with chemotherapy alone versus an untreated control group. It was found that mice treated with chemotherapy and the subcutaneous injection of mesoporous silica rods presented the highest survival rates, with over 75 percent of mice surviving more than 175 days post-challenge.

[0275] FIG. 2B is a graph showing survival percentages of mice challenged with 250K WEHI-3 cells when treated with chemotherapy plus subcutaneous injection of mesoporous silica rods versus mice treated with chemotherapy plus subcutaneous injection of GM-CSF + CpG versus mice treated with chemotherapy alone versus an untreated control group. It was found that mice treated with chemotherapy and the subcutaneous injection of mesoporous silica rods presented the highest survival rates, with over 50 percent of mice surviving more than 150 days post-challenge.

[0276] FIG. 3A-3F are flow charts depicting the cascading results of a systemic immune response elicited by subcutaneous injection of mesoporous silica rods into murine subjects. The left side of each flow chart presents sites of immune cell production and displays resultant outcomes of immune cell production over time as each chart flows to the right. Each tissue or organ site depicted in FIG. 3A-3F undergoes adaptation throughout the systemic immune response generated by the subcutaneous injection of mesoporous silica rods into murine subjects during a controlled study.

[0277] It was found that the injection site beneath the skin of murine subjects typically exhibits early granuloma formation within seven days post-injection (see FIG. 3A). Within 2-3 weeks a granuloma is formed at the injection site. By 30 days post-injection, a late granuloma with lymphocyte aggregates is present. Meanwhile, it was found that over this time period, murine blood exhibited elevated neutrophil counts and eventually elevated monocyte counts in blood samples obtained over the course of the study (see FIG. 3B).

[0278] As depicted in FIG. 3C, a robust immune response was also found in murine lymphatic systems which displayed lymph node enlargement in the forms of sinus hyperplasia, follicular hyperplasia, paracortical hyperplasia, and mixed reactive hyperplasia.

[0279] As shown by FIG. 3D, the immune response affected the bone marrow of murine subjects by generating elevated CD8+ T cell counts, elevated CD11 b-Gr-1 cell counts, and generating myelocyte dense hypercellular marrow.

[0280] The spleens of murine subjects exhibited elevated CD11 b-Gr-1 cell counts (see FIG. 3E) while the tumor cells present in the bone marrow, lymph nodes and spleen were either differentiated into immune cells or were cleared by an immune-mediated mechanism (see FIG. 3F).

[0281] FIG. 4 depicts an AML blast differentiating into other types of immune cells as a result of the systemic immune response generated by the subcutaneous injection of mesoporous silica rods. Through combinatory treatment comprising chemotherapy and subcutaneous injection of mesoporous silica rods, AML blast cells can differentiate into neutrophils, macrophages, antigen-presenting cells and other immune cells. The resulting neutrophils are short-lived and cleared by the immune system shortly after generation of the systemic immune response.

[0282] FIG. 5A is a graph showing production of cluster of differentiation cells (i.e., CD11 b Ly6C, CD11 b Ly6G, CD115 Ly6C, CD45 B220, CD115 Ly6G, CD19 B220) in an unstimulated condition versus production of cluster of differentiation cells in a stimulated condition following 100ng / mL injections of GM-CSF for 5 days. In this study, in vitro differentiation of C1498 eGFP AML cells by GM-CSF was not observed.

[0283] FIG. 5B is a graph showing appearance of GFP-labeled cancer cells (i.e., GFP+ Ly6C+, GFP+ Ly6G+, GFP+ CD1 1 b+, GFP+ CD115+, GFP+ CD19+, GFP+ B220+, GFP+ CD45.2+) in an unstimulated condition versus appearance of GFP-labeled cancer cells in a stimulated condition following 100ng / mL injections of GM-CSF for 5 days. The graphical results show that treatment with GM-CSF alone did not have a significant impact on GFP+ CD cells.

[0284] FIG. 5C is a graph showing the baseline values recorded for presence of B220, CD11 b CD18, CD11 b GR-1 , and CD11 c CD86 when challenged by C1498. The graphical results show that treatment with GM-CSF alone did not have a significant impact on B220 and CD cells.Methods of Manufacturing Mesoporous Silica Rods (MSRs)

[0285] In embodiments, the product for methods of use of the present invention includes mesoporous silica rods (MSRs) loaded (via adsorption) with the cytokine GM-CSF (Leukine®) and the adjuvant CpG 7909 (CpG). The dosage form can be a lyophilized powder that is reconstituted in water (WFI) prior to administration. As described above, the drug product can be administered via subcutaneous injection.

[0286] FIG. 12 depicts an overview of mesoporous silica rod manufacture. A symmetric triblock copolymer (Pluronic P123) composed of poly (ethylene oxide) and poly (propylene oxide) is employed to form rod shaped micelles in solution. Tetraethyl Orthosilicate (TEOs) is added to the solution and the silica deposits on the micelles, creating a hexagonal pore structure. The Pluronic P123 is rinsed and calcinated (high temperature treatment) to remove the polymer, leaving a silica mesoporous structure.

[0287] The MSRs can be 70 - 100 pm long with a width of 3 - 6 pm, and the unique structure of the material provides a high pore volume and large surface area, which allows component (Leukine and CpG) loading and controlled release. Synthetic amorphous silica is known to have a good biocompatibility toxicity profile and in situ dissolution / excretion properties.

[0288] Tertraethyl orthosilicate silica (TEOS) can be used as a raw material source of silicon dioxide in the manufacturing process. TEOS serves as a precursor to silicon dioxide. Accordingly, the TEOS material can be used in the manufacturing process to fabricate the mesoporous silica. As shown below, the TEOS converts to silicon dioxide upon the addition of water.Si (OC2H5)4 + 2 H2O SiO2+ 4 C2H5OH

[0289] The rod structure of the mesoporous silica can be created using a poloxamer (e.g., pluronic P123 symmetric triblock copolymer) as a structural template on which the silicon dioxide can form a three- dimensional architecture. After the silica forms around the polymer template, the polymer can be removed using high temperature. The removal of the polymer leaves behind the longitudinal pores.

[0290] As depicted in the flow chart of FIG. 13, Pluronic P123 (P123) is heated to 40°C using an EasyMax 402® system. Water for injection (WFI) is added and the solution is mixed (450 to 600 rpm). Acid (37% Hydrochloric acid) is added along with tetraethyl orthosilicate (TEOS). The silica rod morphology will form in the solution. The solution is left for about 48 hours (i.e., aged) at 100°C.

[0291] On day three, water (WFI) is added and the solution is sieved (180 pm) and then vacuum filtered (30 pm). The product is then dried (calcinated at 550°C for 5 hours). The final product can then be sterilized. The product can be stored and shipped in lyophilized form. It remains stable at -20°C.

[0292] FIG. 15 shows an overview of a process of manufacturing mesoporous silica rod conjugates according to aspects of the invention. The mesoporous silica rods are combined with the sterile filtered components (GM-CSF, CpG) to form mesoporous silica rod conjugates and mixed well to allow adsorption. This solution is then placed in vials, frozen, lyophilized and stored at -20°C. The proposed configuration of the system will consist of a sterile vial which contains three components (MSR, GM-CSF & CpG) as a lyophilized powder, which is reconstituted with water for injection.Porosity / Volume Determination

[0293] The mesoporous silica rod material provides a large surface area to adsorb the components of the system that can facilitate controlled delivery once injected into the subcutaneous space.

[0294] The pore size distribution analysis is conducted using a static pressure (volumetric) analyzer using the gas adsorption technique. The amount of inert gas adsorbed to the surface of a sample is measured at varying relative pressures by this technique. Once the pressure is reduced incrementally (desorption), the condensed gas evaporates from the pores. From the resulting isotherm, the Barrett, Joyner, and Halenda (BJH) theory is used to determine the cylindrical equivalent pore volumes and pore areas from the amount of gas adsorbed and desorbed. The surface area can also be determined using the Brunauer-Emmett-Teller (BET) theory.Silica Purity (Thermo-Gravimetric Analysis)

[0295] The thermal mass loss profile of the samples is measured via thermo-gravimetric analysis (TGA). The instrument utilizes a microbalance encased within a furnace that utilizes a sample carrier and thermocouple combination to accurately record changes in sample mass overtime as the temperature is increased. The mass change overtime can also be observed as the sample can be held constant at a temperature of interest. As the mesoporous silica consists of a high percent of SiO2, the TGA will provide the percent of the sample that is silica. impurities (Inductively Coupled Plasma Mass Spectroscopy)

[0296] Inductively coupled plasma mass spectroscopy (ICP-MS) is an analytical technique for determining trace multi-elemental and isotopic concentrations in liquid, solid, or gaseous samples. It combines an ion-generating argon plasma source with the sensitive detection limit of mass spectrometry detection. This technique will be utilized to identify the absence of residual metals and carbon-based impurities in the mesoporous silica.Silica Dimensions (Malvern® Morphologi G3S image analyzer)

[0297] The particle size and shape analysis of the MSRs will be conducted on a Malvern® Morphologi G3S or Malvern® Morphologi 4 image analyzer. This instrument is an automated microscope that uses a series of objectives of varying magnifications, a motorized stage, and a digital camera to capture images of particles to determine particle size and shape. The analytical range for this technique is approximately 0.50 pm to 1 ,000 pm, though measurements up to 10,000 pm can be achieved for certain applications.

[0298] The instrument captures an image (see FIG. 14A and 14B) of the particle as it passes into the chosen objective's field of view. The instrument determines the size of an individual pixel for the chosen magnification and creates a projected two-dimensional image. The instrument then converts the pixels of the two-dimensional image into a circle that has the same pixel area as the two-dimensional image, thus reporting the circular equivalent (CE) diameter.

[0299] FIG. 14A and 14B are scanning electron microscope (SEM) sizing images of Standard MSR length and width as analyzed by Malvern® Morphologi G3S instrument. Standard MSRs length (left) and width (right). D[n,0.1](pm), D[n,0.5](pm) D[n,0.9](pm) = biodistribution of MSRs sized at 10% or less, 50% or less, and 90% or less respectively within the total number mean.

[0300] Important functional properties of the mesoporous silica rods are related to the physical properties, the dimensions, surface area, pore size and volume, which are included in the release criteria. In addition, the identity and purity of the material (SIO2) is achieved by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), which will identify the elemental content of the base material. Increasing thetemperature and comparing the combustion thermal transitions to pure components can reveal its chemical composition. TGA is used to determine the purity of synthesized nanomaterials by comparing them to standards.

[0301] In aspects, the following routine tests are applied to the mesoporous silica rods as shown inTable 1 below:Table 1 : Proposed Specification for Mesoporous Silica RodsA dministration

[0302] The mesoporous silica rods (MSRs) and mesoporous silica rod conjugates, which is anMSR loaded with a drug product (MSR conjugates) may be administered via subcutaneous injection intoa patient or subject. The injection of MSRs and MSR conjugates creates a physical insult that triggers a systemic immune response that acts upon cancerous cells and tissues. The MSRs and MSR conjugates can carry a cytotoxic payload, an adjuvant, or both. Further, the MSRs and MSR conjugates can be coated with any therapeutic protein or therapeutic known in the art, including a cytotoxic agent and an adjuvant. The MSRs and MSR conjugates may also be administered alone to create a physical insult to the tumor or inflammatory area, and thus spur an innate systemic immune response from the human body or organism, which in effect will lead to tumor and cancer cell regression through natural cellular processes such as phagocytosis.

[0303] Various sized molecules of therapeutics are also contemplated by the present invention and can be employed within the mesoporous silica rod or mesoporous silica material. Various sized molecules may also be applied to the outer surface of the mesoporous silica rod or mesoporous silica material as a coating. Furthermore, various molecules may be combined with any number of other therapeutic agents, adjuvants, prodrugs, buffers, agents, and the like.

[0304] The cytokine payloads of the present invention may also be directly injected into a targeted tumor area or cancerous region, injected adjacent to the targeted tumor area or cancerous region, or injected remote from the targeted tumor area or cancerous region.

[0305] The mesoporous silica rod or mesoporous silica material may contain at least 6 pores designated for excretion of therapeutic payloads. The mesoporous silica rod or mesoporous silica material may also contain at least 1 pore, at least 2 pores, at least 3 pores, at least 4 pores, at least 5 pores, at least 7 pores, at least 8 pores, at least 9 pores, at least 10 pores, at least 11 pores, at least 12 pores, at least 13 pores, at least 14 pores, at least 15 pores, at least 20 pores, at least 25 pores, at least 30 pores, at least 40 pores, at least 50 pores, at least 60 pores, at least 75 pores, at least 100 pores, at least 125 pores, at least 150 pores, at least 175 pores, at least 200 pores, at least 250 pores, at least 300 pores, at least 350 pores, at least 400 pores, at least 500 pores, at least 600 pores, at least 750 pores, at least 1 ,000 pores, at least 1 ,200 pores, at least 1 ,500 pores, at least 2,000 pores, at least 3,000 pores, at least 5,000 pores, at least 10,000 pores, at least 15,000 pores, at least 20,000 pores, at least 30,000 pores, at least 50,000 pores, at least 100,000 pores, at least 1 ,000,000 pores or more.

[0306] In embodiments, the mesoporous silica rod or mesoporous silica material may contain no more than 1 pore, no more than 2 pores, no more than 3 pores, no more than 4 pores, no more than 5 pores, no more than 6 pores, no more than 7 pores, no more than 8 pores, no more than 9 pores, no more than 10 pores, no more than 11 pores, no more than 12 pores, no more than 13 pores, no more than 14 pores, no more than 15 pores, no more than 20 pores, no more than 25 pores, no more than 30 pores, no more than 40 pores, no more than 50 pores, no more than 60 pores, no more than 75 pores, nomore than 100 pores, no more than 125 pores, no more than 150 pores, no more than 175 pores, no more than 200 pores, no more than 250 pores, no more than 300 pores, no more than 350 pores, no more than 400 pores, no more than 500 pores, no more than 600 pores, no more than 750 pores, no more than 1 ,000 pores, no more than 1 ,200 pores, no more than 1 ,500 pores, no more than 2,000 pores, no more than 3,000 pores, no more than 5,000 pores, no more than 10,000 pores, no more than 15,000 pores, no more than 20,000 pores, no more than 30,000 pores, no more than 50,000 pores, no more than 100,000 pores, or no more than 1 ,000,000 pores.

[0307] The pores of the mesoporous silica rod or mesoporous silica material may possess uniform diameters, or in other aspects possess varying diameters. In yet other aspects, the pores of the mesoporous silica rod may possess an array of diameters arranged in a pattern of alternating diameters and dimensions. In yet other aspects, the varying pore diameter of the mesoporous silica rod may be created and determined randomly either by human input, an algorithm, a recursive algorithm, a computer program, by nano-machining, by nano-engineering, by 3-D printing, by 3-D printing programs, through chemical processes, and all other processes and methods of design suitable for the purposes of the present invention and well known and recognized by the art.

[0308] The mesoporous silica rods of the present invention may be 100 microns (micrometers) in length, but myriad other lengths are also contemplated. The mesoporous silica rods may be about 1 micron in length, about 2 microns in length, about 3 microns in length, about 4 microns in length, about 5 microns in length, about 6 microns in length, about 7 microns in length, about 8 microns in length, about 9 microns in length, about 10 microns in length, about 11 microns in length, about 12 microns in length, about 13 microns in length, about 14 microns in length, about 15 microns in length, about 16 microns in length, about 17 microns in length, about 18 microns in length, about 19 microns in length, about 20 microns in length, about 22 microns in length, about 25 microns in length, about 27 microns in length, about 30 microns in length, about 32 microns in length, about 35 microns in length, about 38 microns in length, about 40 microns in length, about 43 microns in length, about 45 microns in length, about 48 microns in length, about 50 microns in length, about 55 microns in length, about 60 microns in length, about 65 microns in length, about 70 microns in length, about 75 microns in length, about 80 microns in length, about 85 microns in length, about 90 microns in length, about 95 microns in length, about 100 microns in length, about 110 microns in length, about 120 microns in length, about 130 microns in length, about 140 microns in length, about 150 microns in length, about 160 microns in length, about 170 microns in length, about 180 microns in length, about 190 microns in length, about 200 microns in length, about 215 microns in length, about 225 microns in length, about 240 microns in length, about 250 microns in length, about 275 microns in length, about 300 microns in length, about 325 microns in length, about 350 microns in length, about 375 microns in length, about 400 microns in length, about 425 microns in length, about 450 microns in length about 475 microns in length, about 500 microns in length or more.

[0309] The mesoporous silica rods of the present invention may also be no more than 1 micron in length, no more than 2 microns in length, no more than 3 microns in length, no more than 4 microns in length, no more than 5 microns in length, no more than 6 microns in length, no more than 7 microns in length, no more than 8 microns in length, no more than 9 microns in length, no more than 10 microns in length, no more than 11 microns in length, no more than 12 microns in length, no more than 13 microns in length, no more than 14 microns in length, no more than 15 microns in length, no more than 16 microns in length, no more than 17 microns in length, no more than 18 microns in length, no more than 19 microns in length, no more than 20 microns in length, no more than 22 microns in length, no more than 25 microns in length, no more than 27 microns in length, no more than 30 microns in length, no more than 32 microns in length, no more than 35 microns in length, no more than 38 microns in length, no more than 40 microns in length, no more than 43 microns in length, no more than 45 microns in length, no more than 48 microns in length, no more than 50 microns in length, no more than 55 microns in length, no more than 60 microns in length, no more than 65 microns in length, no more than 70 microns in length, no more than 75 microns in length, no more than 80 microns in length, no more than 85 microns in length, no more than 90 microns in length, no more than 95 microns in length, no more than 100 microns in length, no more than 110 microns in length, no more than 120 microns in length, no more than 130 microns in length, no more than 140 microns in length, no more than 150 microns in length, no more than 160 microns in length, no more than 170 microns in length, no more than 180 microns in length, no more than 190 microns in length, no more than 200 microns in length, no more than 215 microns in length, no more than 225 microns in length, no more than 240 microns in length, no more than 250 microns in length, no more than 275 microns in length, no more than 300 microns in length, no more than 325 microns in length, no more than 350 microns in length, no more than 375 microns in length, no more than 400 microns in length, no more than 425 microns in length, no more than 450 microns in length no more than 475 microns in length, or no more than 500 microns in length.

[0310] The mesoporous silica rods of the present invention may also be at least 1 micron in length, at least 2 microns in length, at least 3 microns in length, at least 4 microns in length, at least 5 microns in length, at least 6 microns in length, at least 7 microns in length, at least 8 microns in length, at least 9 microns in length, at least 10 microns in length, at least 11 microns in length, at least 12 microns in length, at least 13 microns in length, at least 14 microns in length, at least 15 microns in length, at least 16 microns in length, at least 17 microns in length, at least 18 microns in length, at least 19 microns in length, at least 20 microns in length, at least 22 microns in length, at least 25 microns in length, at least 27 microns in length, at least 30 microns in length, at least 32 microns in length, at least 35 microns in length, at least 38 microns in length, at least 40 microns in length, at least 43 microns in length, at least 45 microns in length, at least 48 microns in length, at least 50 microns in length, at least 55 microns in length, at least 60 microns in length, at least 65 microns in length, at least 70 microns in length, at least75 microns in length, at least 80 microns in length, at least 85 microns in length, at least 90 microns in length, at least 95 microns in length, at least 100 microns in length, at least 110 microns in length, at least 120 microns in length, at least 130 microns in length, at least 140 microns in length, at least 150 microns in length, at least 160 microns in length, at least 170 microns in length, at least 180 microns in length, at least 190 microns in length, at least 200 microns in length, at least 215 microns in length, at least 225 microns in length, at least 240 microns in length, at least 250 microns in length, at least 275 microns in length, at least 300 microns in length, at least 325 microns in length, at least 350 microns in length, at least 375 microns in length, at least 400 microns in length, at least 425 microns in length, at least 450 microns in length at least 475 microns in length, or at least 500 microns in length.

[0311] The mesoporous silica rods of the present invention may have a cross-sectional area of 10 nm2, however, myriad cross-sectional areas and configurations are contemplated. The cross-sectional area of the mesoporous silica rods may be about 1 nm2, about 2 nm2, about 3 nm2, about 4 nm2, about 5 nm2, about 6 nm2, about 7 nm2, about 8 nm2, about 9 nm2, about 11 nm2, about 12 nm2, about 13 nm2, about 14 nm2, about 15 nm2, about 16 nm2, about 17 nm2, about 18 nm2, about 19 nm2, about 20 nm2, about 22 nm2, about 25 nm2, about 30 nm2, about 35 nm2, about 40 nm2, about 45 nm2, about 50 nm2, about 60 nm2, about 70 nm2, about 80 nm2, about 90 nm2, about 100 nm2, about 110 nm2, about 125 nm2, about 150 nm2, about 175 nm2, about 200 nm2, about 225 nm2, about 250 nm2, about 275 nm2, about 300 nm2, about 325 nm2, about 350 nm2, about 375 nm2, about 400 nm2, about 425 nm2, about 450 nm2, about 475 nm2, about 500 nm2, about 550 nm2, about 600 nm2, about 650 nm2, about 700 nm2, about 750 nm2, about 800 nm2, about 850 nm2, about 900 nm2, about 950 nm2, about 1000 nm2, about 1 pm2, about2 pm2, about 3 pm2, about 4 pm2, about 5 pm2, about 6 pm2, about 7 pm2, about 8 pm2, about 9 pm2, about 10 pm2, about 11 pm2, about 12 pm2, about 13 pm2, about 14 pm2, about 15 pm2, about 16 pm2, about 17 pm2, about 18 pm2, about 19 pm2, about 20 pm2, about 25 pm2, about 30 pm2, about 35 pm2, about 40 pm2, about 45 pm2, about 50 pm2, about 55 pm2, about 60 pm2, about 65 pm2, about 70 pm2, about 75 pm2, about 80 pm2, about 85 pm2, about 90 pm2, about 95 pm2, about 100 pm2, or more.

[0312] In further aspects, the cross-sectional area of the mesoporous silica rods may be at least 1 nm2, at least 2 nm2, at least 3 nm2, at least 4 nm2, at least 5 nm2, at least 6 nm2, at least 7 nm2, at least 8 nm2, at least 9 nm2, at least 11 nm2, at least 12 nm2, at least 13 nm2, at least 14 nm2, at least 15 nm2, at least 16 nm2, at least 17 nm2, at least 18 nm2, at least 19 nm2, at least 20 nm2, at least 22 nm2, at least 25 nm2, at least 30 nm2, at least 35 nm2, at least 40 nm2, at least 45 nm2, at least 50 nm2, at least 60 nm2, at least 70 nm2, at least 80 nm2, at least 90 nm2, at least 100 nm2, at least 110 nm2, at least 125 nm2, at least 150 nm2, at least 175 nm2, at least 200 nm2, at least 225 nm2, at least 250 nm2, at least 275 nm2, at least 300 nm2, at least 325 nm2, at least 350 nm2, at least 375 nm2, at least 400 nm2, at least 425 nm2, at least 450 nm2, at least 475 nm2, at least 500 nm2, at least 550 nm2, at least 600 nm2, at least 650 nm2, at least 700 nm2, at least 750 nm2, at least 800 nm2, at least 850 nm2, at least 900 nm2, at least 950 nm2, atleast 1000 nm2, at least 1 pm2, at least 2 pm2, at least 3 pm2, at least 4 pm2, at least 5 pm2, at least 6 pm2, at least 7 pm2, at least 8 pm2, at least 9 pm2, at least 10 pm2, at least 11 pm2, at least 12 pm2, at least 13 pm2, at least 14 pm2, at least 15 pm2, at least 16 pm2, at least 17 pm2, at least 18 pm2, at least 19 pm2, at least 20 pm2, at least 25 pm2, at least 30 pm2, at least 35 pm2, at least 40 pm2, at least 45 pm2, at least 50 pm2, at least 55 pm2, at least 60 pm2, at least 65 pm2, at least 70 pm2, at least 75 pm2, at least 80 pm2, at least 85 pm2, at least 90 pm2, at least 95 pm2, at least 100 pm2, or more.

[0313] In yet further aspects, the cross-sectional area of the mesoporous silica rods may be no more than 1 nm2, no more than 2 nm2, no more than 3 nm2, no more than 4 nm2, no more than 5 nm2, no more than 6 nm2, no more than 7 nm2, no more than 8 nm2, no more than 9 nm2, no more than 11 nm2, no more than 12 nm2, no more than 13 nm2, no more than 14 nm2, no more than 15 nm2, no more than 16 nm2, no more than 17 nm2, no more than 18 nm2, no more than 19 nm2, no more than 20 nm2, no more than 22 nm2, no more than 25 nm2, no more than 30 nm2, no more than 35 nm2, no more than 40 nm2, no more than 45 nm2, no more than 50 nm2, no more than 60 nm2, no more than 70 nm2, no more than 80 nm2, no more than 90 nm2, no more than 100 nm2, no more than 110 nm2, no more than 125 nm2, no more than 150 nm2, no more than 175 nm2, no more than 200 nm2, no more than 225 nm2, no more than 250 nm2, no more than 275 nm2, no more than 300 nm2, no more than 325 nm2, no more than 350 nm2, no more than 375 nm2, no more than 400 nm2, no more than 425 nm2, no more than 450 nm2, no more than 475 nm2, no more than 500 nm2, no more than 550 nm2, no more than 600 nm2, no more than 650 nm2, no more than 700 nm2, no more than 750 nm2, no more than 800 nm2, no more than 850 nm2, no more than 900 nm2, no more than 950 nm2, no more than 1000 nm2, no more than 1 pm2, no more than 2 pm2, no more than 3 pm2, no more than 4 pm2, no more than 5 pm2, no more than 6 pm2, no more than 7 pm2, no more than 8 pm2, no more than 9 pm2, no more than 10 pm2, no more than 11 pm2, no more than 12 pm2, no more than 13 pm2, no more than 14 pm2, no more than 15 pm2, no more than 16 pm2, no more than 17 pm2, no more than 18 pm2, no more than 19 pm2, no more than 20 pm2, no more than 25 pm2, no more than 30 pm2, no more than 35 pm2, no more than 40 pm2, no more than 45 pm2, no more than 50 pm2, no more than 55 pm2, no more than 60 pm2, no more than 65 pm2, no more than 70 pm2, no more than 75 pm2, no more than 80 pm2, no more than 85 pm2, no more than 90 pm2, no more than 95 pm2, or no more than 100 pm2.

[0314] Therapeutic payloads include all of the payloads described above, as well as any other art- recognized equivalent or any therapeutic known in the art.

[0315] The mesoporous silica rod or mesoporous silica material of the present invention may also dissolve within the target area and / or remain localized to the site of treatment.

[0316] The mesoporous silica rod or mesoporous silica material may carry or be coated with therapeutic payloads, and these therapeutic payloads may be a combination of all of the substances, drugs, adjuvants, and proteins described herein as well as all art-known equivalents and therapeutic equivalents recognized in the art.

[0317] In certain aspects of the present invention, the pore size of the mesoporous silica material and mesoporous silica rod disclosed herein can range from 1 nm to 50 nm, from 1 nm to 40 nm, from 1 nm to 30 nm, from 1 nm to 20 nm, from 1 nm to 15 nm, from 1 nm to 10 nm, from 1 nm to 7 nm, from 1 nm to 5 nm, from 1 nm to 4 nm, from 1 nm to 3 nm, from 1 nm to 2 nm, from 2 nm to 3 nm, from 2 nm to 4 nm, from 2 nm to 5 nm, from 2 nm to 10 nm, from 2 nm to 15 nm, from 2 nm to 20 nm, from 2 nm to 25 nm, from 2 nm to 30 nm, from 2 nm to 35 nm, from 2 nm to 40 nm, from 2 nm to 45 nm, from 2 nm to 50 nm, from 5 nm to 10 nm, from 5 nm to 15 nm, from 5 nm to 20 nm, from 5 nm to 25 nm, from 5 nm to 30 nm, from 5 nm to 35 nm, from 5 nm to 40 nm, from 5 nm to 45 nm, from 5 nm to 50 nm, from 10 nm to 15 nm, from 10 nm to 20 nm, from 10 nm to 25 nm, from 10 nm to 30 nm, from 10 nm to 35 nm, from 10 nm to 40 nm, from 10 nm to 45 nm, from 10 nm to 50 nm, from 15 nm to 20 nm, from 15 nm to 25 nm, from 15 nm to 30 nm, from 15 nm to 35 nm, from 15 nm to 40 nm, from 15 nm to 45 nm, from 15 nm to 50 nm, from 20 nm to 25 nm, from 20 nm to 30 nm, from 20 nm to 35 nm, from 20 nm to 40 nm, from 20 nm to 45 nm, from 20 nm to 50 nm, from 25 nm to 30 nm, from 25 nm to 35 nm, from 25 nm to 40 nm, from 25 nm to 45 nm, from 25 nm to 50 nm, from 30 nm to 35 nm, from 30 nm to 40 nm, from 30 nm to 45 nm, from 30 nm to 50 nm, from 35 nm to 40 nm, from 35 nm to 45 nm, from 35 nm to 50 nm, from 40 nm to 45 nm, from 40 nm to 50 nm, or from 45 nm to 50 nm.

[0318] In certain aspects of the present invention, the pore size of the mesoporous silica material and mesoporous silica rod disclosed herein can be about 1 nm, about 2 nm, about 3 nm, about 4 nm, about 5 nm, about 6 nm, about 7 nm, about 8 nm, about 9 nm, about 10 nm, about 11 nm, about 12 nm, about 13 nm, about 14 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, about 45 nm, about 50 nm, about 55 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 110 nm, about 120 nm, about 130 nm, about 140 nm, about 150 nm, about 160 nm, about 175 nm, about 190 nm, about 200 nm, about 215 nm, about 225 nm, about 250 nm, about 275 nm, about 300 nm or more.

[0319] In certain aspects of the present invention, the pore size of the mesoporous silica material and mesoporous silica rod disclosed herein can be no more than 1 nm, no more than 2 nm, no more than 3 nm, no more than 4 nm, no more than 5 nm, no more than 6 nm, no more than 7 nm, no more than 8 nm, no more than 9 nm, no more than 10 nm, no more than 11 nm, no more than 12 nm, no more than 13 nm, no more than 14 nm, no more than 15 nm, no more than 20 nm, no more than 25 nm, no more than 30 nm, no more than 35 nm, no more than 40 nm, no more than 45 nm, no more than 50 nm, no morethan 55 nm, no more than 60 nm, no more than 70 nm, no more than 80 nm, no more than 90 nm, no more than 100 nm, no more than 1 10 nm, no more than 120 nm, no more than 130 nm, no more than 140 nm, no more than 150 nm, no more than 160 nm, no more than 175 nm, no more than 190 nm, no more than 200 nm, no more than 215 nm, no more than 225 nm, no more than 250 nm, no more than 275 nm, no more than 300 nm, no more than 325 nm, no more than 350 nm, no more than 500 nm.

[0320] In certain aspects of the present invention, the pore size of the mesoporous silica material and mesoporous silica rod disclosed herein can be at least 1 nm, at least 2 nm, at least 3 nm, at least 4 nm, at least 5 nm, at least 6 nm, at least 7 nm, at least 8 nm, at least 9 nm, at least 10 nm, at least 1 1 nm, at least 12 nm, at least 13 nm, at least 14 nm, at least 15 nm, at least 20 nm, at least 25 nm, at least 30 nm, at least 35 nm, at least 40 nm, at least 45 nm, at least 50 nm, at least 55 nm, at least 60 nm, at least 70 nm, at least 80 nm, at least 90 nm, at least 100 nm, at least 110 nm, at least 120 nm, at least 130 nm, at least 140 nm, at least 150 nm, at least 160 nm, at least 175 nm, at least 190 nm, at least 200 nm, at least 215 nm, at least 225 nm, at least 250 nm, at least 275 nm, at least 300 nm, at least 325 nm, at least 350 nm, at least 500 nm.

[0321] In certain aspects of the present invention, the pore size of the mesoporous silica material and mesoporous silica rod disclosed herein can range from 1 nm to 55 nm, from 1 nm to 60 nm, from 1 nm to 65 nm, from 1 nm to 70 nm, from 1 nm to 75 nm, from 1 nm to 80 nm, from 1 nm to 85 nm, from 1 nm to 90 nm, from 1 nm to 95 nm, from 1 nm to 100 nm, from 1 nm to 105 nm, from 1 nm to 110 nm, from 1 nm to 1 15 nm, from 1 nm to 120 nm, from 2 nm to 55 nm, from 2 nm to 60 nm, from 2 nm to 65 nm, from 2 nm to 70 nm, from 2 nm to 75 nm, from 2 nm to 80 nm, from 2 nm to 85 nm, from 2 nm to 90 nm, from 2 nm to 95 nm, from 2 nm to 100 nm, from 2 nm to 105 nm, from 2 nm to 110 nm, from 2 nm to 115 nm, from 2 nm to 120 nm, from 5 nm to 55 nm, from 5 nm to 60 nm, from 5 nm to 65 nm, from 5 nm to 70 nm, from 5 nm to 75 nm, from 5 nm to 80 nm, from 5 nm to 85 nm, from 5 nm to 90 nm, from 5 nm to 95 nm, from 5 nm to 100 nm, from 5 nm to 105 nm, from 5 nm to 110 nm, from 5 nm to 115 nm, from 5 nm to 120 nm, from 10 nm to 55 nm, from 10 nm to 60 nm, from 10 nm to 65 nm, from 10 nm to 70 nm, from 10 nm to 75 nm, from 10 nm to 80 nm, from 10 nm to 85 nm, from 10 nm to 90 nm, from 10 nm to 95 nm, from 10 nm to 100 nm, from 10 nm to 105 nm, from 10 nm to 1 10 nm, from 10 nm to 115 nm, from 10 nm to 120 nm, from 20 nm to 55 nm, from 20 nm to 60 nm, from 20 nm to 65 nm, from 20 nm to 70 nm, from 20 nm to 75 nm, from 20 nm to 80 nm, from 20 nm to 85 nm, from 20 nm to 90 nm, from 20 nm to 95 nm, from 20 nm to 100 nm, from 20 nm to 105 nm, from 20 nm to 1 10 nm, from 20 nm to 115 nm, from 20 nm to 120 nm, from 30 nm to 55 nm, from 30 nm to 60 nm, from 30 nm to 65 nm, from 30 nm to 70 nm, from 30 nm to 75 nm, from 30 nm to 80 nm, from 30 nm to 85 nm, from 30 nm to 90 nm, from 30 nm to 95 nm, from 30 nm to 100 nm, from 30 nm to 105 nm, from 30 nm to 1 10 nm, from 30 nm to 115 nm, from 30 nm to 120 nm, from 40 nm to 55 nm, from 40 nm to 60 nm, from 40 nm to 65 nm, from 40 nm to 70 nm, from 40 nm to 75 nm, from 40 nm to 80 nm, from 40 nm to 85 nm, from 40 nm to 90 nm, from 40 nm to 95 nm,from 40 nm to 100 nm, from 40 nm to 105 nm, from 40 nm to 1 10 nm, from 40 nm to 115 nm, from 40 nm to 120 nm, from 50 nm to 55 nm, from 50 nm to 60 nm, from 50 nm to 65 nm, from 50 nm to 70 nm, from 50 nm to 75 nm, from 50 nm to 80 nm, from 50 nm to 85 nm, from 50 nm to 90 nm, from 50 nm to 95 nm, from 50 nm to 100 nm, from 50 nm to 105 nm, from 50 nm to 1 10 nm, from 50 nm to 115 nm, from 50 nm to 120 nm, from 60 nm to 65 nm, from 60 nm to 70 nm, from 60 nm to 75 nm, from 60 nm to 80 nm, from 60 nm to 85 nm, from 60 nm to 90 nm, from 60 nm to 95 nm, from 60 nm to 100 nm, from 60 nm to 105 nm, from 60 nm to 110 nm, from 60 nm to 115 nm, from 60 nm to 120 nm, from 70 nm to 75 nm, from 70 nm to 80 nm, from 70 nm to 85 nm, from 70 nm to 90 nm, from 70 nm to 95 nm, from 70 nm to 100 nm, from 70 nm to 105 nm, from 70 nm to 110 nm, from 70 nm to 1 15 nm, from 70 nm to 120 nm, from 80 nm to 85 nm, from 80 nm to 90 nm, from 80 nm to 95 nm, from 80 nm to 100 nm, from 80 nm to 105 nm, from 80 nm to 110 nm, from 80 nm to 115 nm, from 80 nm to 120 nm, from 90 nm to 95 nm, from 90 nm to 100 nm, from 90 nm to 105 nm, from 90 nm to 110 nm, from 90 nm to 115 nm, from 90 nm to 120 nm, from 100 nm to 105 nm, from 100 nm to 110 nm, from 100 nm to 115 nm, from 100 nm to 120 nm, from 110 nm to 115 nm, from 110 nm to 120 nm.

[0322] In certain aspects of the present invention, the pore size of the mesoporous silica material and mesoporous silica rod disclosed herein can range from about 1 nm to about 50 nm, from about 1 nm to about 40 nm, from about 1 nm to about 30 nm, from about 1 nm to about 25 nm, from about 1 nm to about 20 nm, from about 1 nm to about 15 nm, from about 1 nm to 10 nm, from about 1 nm to 5 nm, from about 2 nm to about 50 nm, from about 2 nm to about 5 nm, from about 2 nm to about 10 nm, from about 2 nm to about 15 nm, from about 2 nm to about 20 nm, from about 2 nm to about 30 nm, from about 2 nm to about 40 nm, from about 5 nm to about 10 nm, from about 5 nm to about 15 nm, from about 5 nm to about 20 nm, from about 5 nm to about 30 nm, from about 5 nm to about 40 nm, from about 5 nm to about 50 nm, from about 10 nm to about 15 nm, from about 10 nm to about 20 nm, from about 10 nm to about 25 nm, from about 10 nm to about 30 nm, from about 10 nm to about 40 nm, from about 10 nm to about 50 nm, from about 15 nm to about 20 nm, from about 15 nm to about 25 nm, from about 15 nm to about 30 nm, from about 15 nm to about 40 nm, from about 15 nm to about 50 nm, from about 20 nm to about 25 nm, from about 20 nm to about 30 nm, from about 20 nm to about 35 nm, from about 20 nm to about 40 nm, from about 20 nm to about 50 nm, from about 25 nm to about 30 nm, from about 25 nm to about 35 nm, from about 25 nm to about 40 nm, from about 25 nm to about 50 nm, from about 30 nm to about 35 nm, from about 30 nm to about 40 nm, from about 30 nm to about 45 nm, from about 30 nm to about 50 nm, from about 35 nm to about 40 nm, from about 35 nm to about 45 nm, from about 35 nm to about 50 nm, from about 40 nm to about 45 nm, from about 40 nm to about 50 nm, from about 45 nm to about 50 nm, from about 46 nm to about 50 nm, from about 47 nm to about 50 nm, from about 48 nm to about 50 nm, from about 49 nm to about 50 nm.

[0323] In certain aspects of the present invention, the pore size of the mesoporous silica material and mesoporous silica rod disclosed herein can range from about 1 nm to about 50 nm, from about 1 nm to about 55 nm, from about 1 nm to about 60 nm, from about 1 nm to about 65 nm, from about 1 nm to about 70 nm, from about 1 nm to about 80 nm, from about 1 nm to about 100 nm, from about 1 nm to about 120 nm, from about 1 nm to about 150 nm, from about 1 nm to about 200 nm, from about 2 nm to about 50 nm, from about 2 nm to about 55 nm, from about 2 nm to about 60 nm, from about 2 nm to about 65 nm, from about 2 nm to about 65 nm, from about 2 nm to about 80 nm, from about 2 nm to about 100 nm, from about 2 nm to about 120 nm, from about 2 nm to about 150 nm, from about 2 nm to about 200 nm, from about 5 nm to about 55 nm, from about 5 nm to about 60 nm, from about 5 nm to about 65 nm, from about 5 nm to about 80 nm, from about 5 nm to about 100 nm, from about 5 nm to about 120 nm, from about 5 nm to about 150 nm, from about 5 nm to about 200 nm, from about 10 nm to about 55 nm, from about 10 nm to about 60 nm, from about 10 nm to about 65 nm, from about 10 nm to about 80 nm, from about 10 nm to about 100 nm, from about 10 nm to about 120 nm, from about 10 nm to about 150 nm, from about 10 nm to about 200 nm, from about 15 nm to about 55 nm, from about 15 nm to about 60 nm, from about 15 nm to about 80 nm, from about 15 nm to about 100 nm, from about 15 nm to about 120 nm, from about 15 nm to about 150 nm, from about 15 nm to about 200 nm, from about 20 nm to about 55 nm, from about 20 nm to about 60 nm, from about 20 nm to about 65 nm, from about 20 nm to about 80 nm, from about 20 nm to about 100 nm, from about 20 nm to about 100 nm, from about 20 nm to about 120 nm, from about 20 nm to about 150 nm, from about 20 nm to about 200 nm, from about 30 nm to about 55 nm, from about 30 nm to about 80 nm, from about 30 nm to about 100 nm, from about 30 nm to about 120 nm, from about 30 nm to about 150 nm, from about 30 nm to about 200 nm, from about 40 nm to about 55 nm, from about 40 nm to about 80 nm, from about 40 nm to about 100 nm, from about 40 nm to about 120 nm, from about 40 nm to about 150 nm, from about 40 nm to about 200 nm, from about 50 nm to about 60 nm, from about 50 nm to about 65 nm, from about 50 nm to about 80 nm, from about 50 nm to about 100 nm, from about 50 nm to about 120 nm, from about 50 nm to about 150 nm, from about 50 nm to about 200 nm, from about 60 nm to about 65 nm, from about 60 nm to about 70 nm, from about 60 nm to about 80 nm, from about 60 nm to about 100 nm, from about 60 nm to about 120 nm, from about 60 nm to about 150 nm, from about 60 nm to about 200 nm, from about 70 nm to about 75 nm, from about 70 nm to about 80 nm, from about 70 nm to about 100 nm, from about 70 nm to about 110 nm, from about 70 nm to about 120 nm, from about 70 nm to about 130 nm, from about 70 nm to about 140 nm, from about 70 nm to about 150 nm, from about 70 nm to about 200 nm, from about 80 nm to about 85 nm, from about 80 nm to about 90 nm, from about 80 nm to about 100 nm, from about 80 nm to about 120 nm, from about 80 nm to about 150 nm, from about 80 nm to about 200 nm, from about 90 nm to about 100 nm, from about 90 nm to about 110 nm, from about 90 nm to about 120 nm, from about 90 nm to about 150 nm, from about 90 nm to about 200 nm, from about 100 nm to about 110 nm, from about 100 nm to about 120 nm, from about 100 nm to about 130 nm, from about 100 nm to about 140 nm, from about 100 nm to about 150 nm, from about 100 nm to about 200 nm, from about 1 10nm to about 120 nm, from about 110 nm to about 140 nm, from about 110 nm to about 150 nm, from about 110 nm to about 200 nm, from about 120 nm to about 135 nm, from about 120 nm to about 150 nm, from about 120 nm to about 200 nm, from about 130 nm to about 140 nm, from about 130 nm to about 150 nm, from about 130 nm to about 200 nm, from about 140 nm to about 145 nm, from about 140 nm to about 150 nm, from about 140 nm to about 200 nm, or from about 150 nm to about 200 nm.

[0324] The dosage of payload, silica material, cytokine protein, inflammasome, interleukin, adjuvant, pharmaceutical composition, therapeutic, prodrug, conjugate and other equivalents of the present invention can range from 1 ng (nanogram) to about 100 ng, from 1 ng to about 500 ng, from 1 ng to about 1 pg (microgram), from 1 ng to about 2 pg, from about 1 ng to about 3 pg, from about 1 ng to about 4 pg, from about 1 ng to about 5 pg, from about 1 ng to about 6 pg, from about 1 ng to about 7 pg, from about 1 ng to about 8 pg, from about 1 ng to about 9 pg, from about 1 ng to about 10 pg, from about 1 ng to about 12 pg, from about 1 ng to about 15 pg, from about 1 ng to about 17 pg, from about 1 ng to about 20 pg, from about 1 ng to about 25 pg, from about 1 ng to about 30 pg, from about 1 ng to about 35 pg, from about 1 ng to about 40 pg, from about 1 ng to about 50 pg, from about 1 ng to about 60 pg, from about 1 ng to about 75 pg, from about 1 ng to about 100 pg, from about 1 ng to about 1 mg, from about 1 ng to about 1 g.

[0325] In other aspects, the dosage of payload, silica material, cytokine protein, inflammasome, interleukin, adjuvant, pharmaceutical composition, therapeutic, prodrug, conjugate and other equivalents of the present invention can range from about 1 pg to about 2 pg, from about 1 pg to about 3 pg, from about 1 pg to about 4 pg, from about 1 pg to about 5 pg, from about 1 pg to about 6 pg, from about 1 pg to about 8 pg, from about 1 pg to about 10 pg, from about 1 pg to about 15 pg, from about 1 pg to about 20 pg, from about 1 pg to about 30 pg, from about 1 pg to about 40 pg, from about 1 pg to about 50 pg, from about 1 pg to about 100 pg, from about 1 pg to about 500 pg, from about 1 pg to about 1 mg, from about 1 pg to about 5 mg, from about 1 pg to about 10 mg, from about 1 pg to about 20 mg, from about 1 pg to about 50 mg, from about 1 pg to about 100 mg, from about 1 pg to about 200 mg, from about 1 pg to about 500 mg, from about 1 pg to about 750 mg, from about 1 pg to about 1 g.

[0326] The dosage of payload, silica material, cytokine protein, inflammasome, interleukin, adjuvant, pharmaceutical composition, therapeutic, prodrug, conjugate and other equivalents of the present invention can be about 1 ng, about 2 ng, about 3 ng, about 4 ng, about 5 ng, about 6 ng, about 7 ng, about 8 ng, about 9 ng, about 10 ng, about 11 ng, about 12 ng, about 13 ng, about 14 ng, about 15 ng, about 16 ng, about 17 ng, about 19 ng, about 20 ng, about 25 ng, about 30 ng, about 35 ng, about 40 ng, about 45 ng, about 50 ng, about 55 ng, about 60 ng, about 65 ng, about 70 ng, about 75 ng, about 80 ng, about 85 ng, about 90 ng, about 95 ng, about 100 ng, about 1 10 ng, about 120 ng, about 130 ng, about 140 ng, about 150 ng, about 160 ng, about 175 ng, about 190 ng, about 200 ng, about 225 ng,about 250 ng, about 275 ng, about 300 ng, about 325 ng, about 350 ng, about 375 ng, about 400 ng, about 425 ng, about 450 ng, about 475 ng, about 500 ng, about 550 ng, about 600 ng, about 650 ng, about 700 ng, about 750 ng, about 800 ng, about 850 ng, about 900 ng, about 950 ng, about 1 ,000 ng, about 1 ,200 ng, about 1 ,500 ng, about 2,000 ng, about 3,500 ng, about 5,000 ng.

[0327] The dosage of payload, silica material, cytokine protein, inflammasome, interleukin, adjuvant, pharmaceutical composition, therapeutic, prodrug, conjugate and other equivalents of the present invention can be about 1 pg, about 2 pg, about 3 pg, about 4 pg, about 5 pg, about 6 pg, about 7 pg, about 8 pg, about 9 pg, about 10 pg, about 1 1 pg, about 12 pg, about 13 pg, about 14 pg, about 15 pg, about 16 pg, about 17 pg, about 18 pg, about 19 pg, about 20 pg, about 21 pg, about 22 pg, about 23 pg, about 24 pg, about 25 pg, about 30 pg, about 32 pg, about 35 pg, about 37 pg, about 40 pg, about 42 pg, about 45 pg, about 48 pg, about 50 pg, about 55 pg, about 60 pg, about 65 pg, about 70 pg, about 75 pg, about 80 pg, about 85 pg, about 90 pg, about 95 pg, about 100 pg, about 1 10 pg, about 120 pg, about 130 pg, about 140 pg, about 150 pg, about 160 pg, about 170 pg, about 180 pg, about 190 pg, about 200 pg, about 215 pg, about 230 pg, about 250 pg, about 265 pg, about 275 pg, about 300 pg, about 325 pg, about 350 pg, about 375 pg, about 400 pg, about 425 pg, about 450 pg, about 475 pg, about 500 pg, about 550 pg, about 600 pg, about 650 pg, about 700 pg, about 750 pg, about 800 pg, about 850 pg, about 900 pg, about 950 pg, about 1 mg, about 1 .5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 4 mg, about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg, about 7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 22 mg, about 24 mg, about 25 mg, about 27 mg, about 29 mg, about 30 mg, about 33 mg, about 35 mg, about 37 mg, about 40 mg, about 43 mg, about 45 mg, about 48 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 220 mg, about 240 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, about 1 g, about 1 .5 g, about 2 g, about 2.5 g, about 3 g, about 3.5 g, about 4 g, about 4.5 g, about 5 g, about 5.5 g, about 6 g, about 6.5 g, about 7 g, about 7.5 g, about 8 g, about 8.5 g, about 9 g, about 9.5 g, about 10 g, about 11 g, about 12 g, about 13 g, about 14 g, about 15 g, about 16 g, about 17 g, about 18 g, about 19 g, about 20 g, about 22 g, about 24 g, about 25 g, about 27 g, about 28 g, about 30 g, about 33 g, about 35 g, about 38 g, about 40 g, about 43 g, about 45 g, about 48 g, about 50 g, about 55 g, about 60 g, about 65 g, about 70 g, about 75 g, about 80 g, about 85 g, about 90 g, about 95 g, about 100 g.

[0328] The dosage of payload, silica material, cytokine protein, inflammasome, interleukin, adjuvant, pharmaceutical composition, therapeutic, prodrug, conjugate and other equivalents of the present invention can be at least 1 ng, at least 2 ng, at least 3 ng, at least 4 ng, at least 5 ng, at least 6 ng, at least 7 ng, at least 8 ng, at least 9 ng, at least 10 ng, at least 11 ng, at least 12 ng, at least 13 ng, at least 14 ng, at least 15 ng, at least 16 ng, at least 17 ng, at least 19 ng, at least 20 ng, at least 25 ng, at least 30 ng, at least 35 ng, at least 40 ng, at least 45 ng, at least 50 ng, at least 55 ng, at least 60 ng, at least 65 ng, at least 70 ng, at least 75 ng, at least 80 ng, at least 85 ng, at least 90 ng, at least 95 ng, at least 100 ng, at least 110 ng, at least 120 ng, at least 130 ng, at least 140 ng, at least 150 ng, at least 160 ng, at least 175 ng, at least 190 ng, at least 200 ng, at least 225 ng, at least 250 ng, at least 275 ng, at least 300 ng, at least 325 ng, at least 350 ng, at least 375 ng, at least 400 ng, at least 425 ng, at least 450 ng, at least 475 ng, at least 500 ng, at least 550 ng, at least 600 ng, at least 650 ng, at least 700 ng, at least 750 ng, at least 800 ng, at least 850 ng, at least 900 ng, at least 950 ng, at least 1 ,000 ng, at least 1 ,200 ng, at least 1 ,500 ng, at least 2,000 ng, at least 3,500 ng, at least 5,000 ng.

[0329] The dosage of payload, silica material, cytokine protein, inflammasome, interleukin, adjuvant, pharmaceutical composition, therapeutic, prodrug, conjugate and other equivalents of the present invention can be at least 1 pg, at least 2 pg, at least 3 pg, at least 4 pg, at least 5 pg, at least 6 pg, at least 7 pg, at least 8 pg, at least 9 pg, at least 10 pg, at least 1 1 pg, at least 12 pg, at least 13 pg, at least 14 pg , at least 15 pg , at least 16 pg , at least 17 pg , at least 18 pg , at least 19 pg , at least 20 pg , at least 21 pg, at least 22 pg, at least 23 pg, at least 24 pg, at least 25 pg, at least 30 pg, at least 32 pg, at least 35 pg, at least 37 pg, at least 40 pg, at least 42 pg, at least 45 pg, at least 48 pg, at least 50 pg, at least 55 pg, at least 60 pg, at least 65 pg, at least 70 pg, at least 75 pg, at least 80 pg, at least 85 pg, at least 90 pg, at least 95 pg, at least 100 pg, at least 1 10 pg, at least 120 pg, at least 130 pg, at least 140 pg, at least 150 pg, at least 160 pg, at least 170 pg, at least 180 pg, at least 190 pg, at least 200 pg, at least 215 pg, at least 230 pg, at least 250 pg, at least 265 pg, at least 275 pg, at least 300 pg, at least 325 pg, at least 350 pg, at least 375 pg, at least 400 pg, at least 425 pg, at least 450 pg, at least 475 pg, at least 500 pg, at least 550 pg, at least 600 pg, at least 650 pg, at least 700 pg, at least 750 pg, at least 800 pg, at least 850 pg, at least 900 pg, at least 950 pg, at least 1 mg, at least 1 .5 mg, at least 2 mg, at least 2.5 mg, at least 3 mg, at least 3.5 mg, at least 4 mg, at least 4.5 mg, at least 5 mg, at least 5.5 mg, at least 6 mg, at least 6.5 mg, at least 7 mg, at least 7.5 mg, at least 8 mg, at least 8.5 mg, at least 9 mg, at least 9.5 mg, at least 10 mg, at least 11 mg, at least 12 mg, at least 13 mg, at least 14 mg, at least 15 mg, at least 16 mg, at least 17 mg, at least 18 mg, at least 19 mg, at least 20 mg, at least 22 mg, at least 24 mg, at least 25 mg, at least 27 mg, at least 29 mg, at least 30 mg, at least 33 mg, at least 35 mg, at least 37 mg, at least 40 mg, at least 43 mg, at least 45 mg, at least 48 mg, at least 50 mg, at least 55 mg, at least 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, at least 100 mg, at least 110 mg, at least 120 mg, at least 130 mg, at least 140 mg, at least 150 mg, at least 160 mg, at least 170 mg, at least 180 mg, at least 190 mg, at least 200 mg, at least220 mg, at least 240 mg, at least 250 mg, at least 275 mg, at least 300 mg, at least 325 mg, at least 350 mg, at least 375 mg, at least 400 mg, at least 425 mg, at least 450 mg, at least 475 mg, at least 500 mg, at least 550 mg, at least 600 mg, at least 650 mg, at least 700 mg, at least 750 mg, at least 800 mg, at least 850 mg, at least 900 mg, at least 950 mg, at least 1 g, at least 1 .5 g, at least 2 g, at least 2.5 g, at least 3 g, at least 3.5 g, at least 4 g, at least 4.5 g, at least 5 g, at least 5.5 g, at least 6 g, at least 6.5 g, at least 7 g, at least 7.5 g, at least 8 g, at least 8.5 g, at least 9 g, at least 9.5 g, at least 10 g, at least 11 g, at least 12 g, at least 13 g, at least 14 g, at least 15 g, at least 16 g, at least 17 g, at least 18 g, at least 19 g, at least 20 g, at least 22 g, at least 24 g, at least 25 g, at least 27 g, at least 28 g, at least 30 g, at least 33 g, at least 35 g, at least 38 g, at least 40 g, at least 43 g, at least 45 g, at least 48 g, at least 50 g, at least 55 g, at least 60 g, at least 65 g, at least 70 g, at least 75 g, at least 80 g, at least 85 g, at least 90 g, at least 95 g, at least 100 g.

[0330] The dosage of payload, silica material, cytokine protein, inflammasome, interleukin, adjuvant, pharmaceutical composition, therapeutic, prodrug, conjugate and other equivalents of the present invention can be no more than 1 ng, no more than 2 ng, no more than 3 ng, no more than 4 ng, no more than 5 ng, no more than 6 ng, no more than 7 ng, no more than 8 ng, no more than 9 ng, no more than 10 ng, no more than 1 1 ng, no more than 12 ng, no more than 13 ng, no more than 14 ng, no more than 15 ng, no more than 16 ng, no more than 17 ng, no more than 19 ng, no more than 20 ng, no more than 25 ng, no more than 30 ng, no more than 35 ng, no more than 40 ng, no more than 45 ng, no more than 50 ng, no more than 55 ng, no more than 60 ng, no more than 65 ng, no more than 70 ng, no more than 75 ng, no more than 80 ng, no more than 85 ng, no more than 90 ng, no more than 95 ng, no more than 100 ng, no more than 110 ng, no more than 120 ng, no more than 130 ng, no more than 140 ng, no more than 150 ng, no more than 160 ng, no more than 175 ng, no more than 190 ng, no more than 200 ng, no more than 225 ng, no more than 250 ng, no more than 275 ng, no more than 300 ng, no more than 325 ng, no more than 350 ng, no more than 375 ng, no more than 400 ng, no more than 425 ng, no more than 450 ng, no more than 475 ng, no more than 500 ng, no more than 550 ng, no more than 600 ng, no more than 650 ng, no more than 700 ng, no more than 750 ng, no more than 800 ng, no more than 850 ng, no more than 900 ng, no more than 950 ng, no more than 1 ,000 ng, no more than 1 ,200 ng, no more than 1 ,500 ng, no more than 2,000 ng, no more than 3,500 ng, no more than 5,000 ng.

[0331] The dosage of payload, silica material, cytokine protein, inflammasome, interleukin, adjuvant, pharmaceutical composition, therapeutic, prodrug, conjugate and other equivalents of the present invention can be no more than 1 pg, no more than 2 pg, no more than 3 pg, no more than 4 pg, no more than 5 pg, no more than 6 pg, no more than 7 pg, no more than 8 pg, no more than 9 pg, no more than 10 pg, no more than 1 1 pg, no more than 12 pg, no more than 13 pg, no more than 14 pg, no more than 15 pg, no more than 16 pg, no more than 17 pg, no more than 18 pg, no more than 19 pg, no more than 20 pg, no more than 21 pg, no more than 22 pg, no more than 23 pg, no more than 24 pg, nomore than 25 pg, no more than 30 pg, no more than 32 pg, no more than 35 pg, no more than 37 pg, no more than 40 pg, no more than 42 pg, no more than 45 pg, no more than 48 pg, no more than 50 pg, no more than 55 pg, no more than 60 pg, no more than 65 pg, no more than 70 pg, no more than 75 pg, no more than 80 pg, no more than 85 pg, no more than 90 pg, no more than 95 pg, no more than 100 pg, no more than 110 pg, no more than 120 pg, no more than 130 pg, no more than 140 pg, no more than 150 pg, no more than 160 pg, no more than 170 pg, no more than 180 pg, no more than 190 pg, no more than 200 pg, no more than 215 pg, no more than 230 pg, no more than 250 pg, no more than 265 pg, no more than 275 pg, no more than 300 pg, no more than 325 pg, no more than 350 pg, no more than 375 pg, no more than 400 pg, no more than 425 pg, no more than 450 pg, no more than 475 pg, no more than 500 pg, no more than 550 pg, no more than 600 pg, no more than 650 pg, no more than 700 pg, no more than 750 pg, no more than 800 pg, no more than 850 pg, no more than 900 pg, no more than 950 pg, no more than 1 mg, no more than 1 .5 mg, no more than 2 mg, no more than 2.5 mg, no more than 3 mg, no more than 3.5 mg, no more than 4 mg, no more than 4.5 mg, no more than 5 mg, no more than 5.5 mg, no more than 6 mg, no more than 6.5 mg, no more than 7 mg, no more than 7.5 mg, no more than 8 mg, no more than 8.5 mg, no more than 9 mg, no more than 9.5 mg, no more than 10 mg, no more than 11 mg, no more than 12 mg, no more than 13 mg, no more than 14 mg, no more than 15 mg, no more than 16 mg, no more than 17 mg, no more than 18 mg, no more than 19 mg, no more than 20 mg, no more than 22 mg, no more than 24 mg, no more than 25 mg, no more than 27 mg, no more than 29 mg, no more than 30 mg, no more than 33 mg, no more than 35 mg, no more than 37 mg, no more than 40 mg, no more than 43 mg, no more than 45 mg, no more than 48 mg, no more than 50 mg, no more than 55 mg, no more than 60 mg, no more than 65 mg, no more than 70 mg, no more than 75 mg, no more than 80 mg, no more than 85 mg, no more than 90 mg, no more than 95 mg, no more than 100 mg, no more than 110 mg, no more than 120 mg, no more than 130 mg, no more than 140 mg, no more than 150 mg, no more than 160 mg, no more than 170 mg, no more than 180 mg, no more than 190 mg, no more than 200 mg, no more than 220 mg, no more than 240 mg, no more than 250 mg, no more than 275 mg, no more than 300 mg, no more than 325 mg, no more than 350 mg, no more than 375 mg, no more than 400 mg, no more than 425 mg, no more than 450 mg, no more than 475 mg, no more than 500 mg, no more than 550 mg, no more than 600 mg, no more than 650 mg, no more than 700 mg, no more than 750 mg, no more than 800 mg, no more than 850 mg, no more than 900 mg, no more than 950 mg, no more than 1 g, no more than 1 .5 g, no more than 2 g, no more than 2.5 g, no more than 3 g, no more than 3.5 g, no more than 4 g, no more than 4.5 g, no more than 5 g, no more than 5.5 g, no more than 6 g, no more than 6.5 g, no more than 7 g, no more than 7.5 g, no more than 8 g, no more than 8.5 g, no more than 9 g, no more than 9.5 g, no more than 10 g, no more than 11 g, no more than 12 g, no more than 13 g, no more than 14 g, no more than 15 g, no more than 16 g, no more than 17 g, no more than 18 g, no more than 19 g, no more than 20 g, no more than 22 g, no more than 24 g, no more than 25 g, no more than 27 g, no more than 28 g, no more than 30 g, no more than 33 g, no more than 35 g, no more than 38 g, no more than 40 g, no more than 43 g, no more than 45 g, no more than 48 g, no more than 50 g, no more than 55g, no more than 60 g, no more than 65 g, no more than 70 g, no more than 75 g, no more than 80 g, no more than 85 g, no more than 90 g, no more than 95 g, no more than 100 g.

[0332] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of a circulating cancer by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0333] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of a circulating cancer by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0334] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of a circulating cancer by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0335] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of circulating cancer cells by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0336] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of circulating cancer cells by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0337] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of circulating cancer cells by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0338] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of acute myeloid leukemia by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0339] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of acute myeloid leukemia by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0340] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of acute myeloid leukemia by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0341] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of a lymphatic cancer by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0342] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of a lymphatic cancer by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0343] In an embodiment, treatment with an MSR conjugate disclosed herein reduces the signs and symptoms of a lymphatic cancer by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0344] In an embodiment, treatment with an MSR disclosed herein reduces inflammation (e.g., signs and symptoms) by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0345] In an embodiment, treatment with an MSR conjugate disclosed herein reduces inflammation (e.g., signs and symptoms) by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0346] In an embodiment, treatment with an MSR conjugate disclosed herein reduces inflammation (e.g., signs and symptoms) by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0347] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor proliferation (e.g., size and / or number of tumors) by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0348] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor proliferation (e.g., size and / or number of tumors) by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0349] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor proliferation (e.g., size and / or number of tumors) by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0350] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor circumference by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0351] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor circumference by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0352] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor circumference by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0353] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor diameter by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0354] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor diameter by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0355] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor diameter by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0356] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor volume by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0357] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor volume by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0358] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor volume by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0359] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor mass by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0360] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor mass by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0361] In an embodiment, treatment with an MSR conjugate disclosed herein decreases tumor mass by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0362] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of circulating cancer cells by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0363] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of circulating cancer cells by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0364] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of circulating cancer cells by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0365] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of a lymphatic cancer by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0366] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of a lymphatic cancer by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0367] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of a lymphatic cancer by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0368] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of acute myeloid leukemia by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0369] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of acute myeloid leukemia by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0370] In an embodiment, treatment with an MSR conjugate disclosed herein decreases the proliferation of acute myeloid leukemia by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0371] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion proliferation (e.g., size and / or number) by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0372] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion proliferation (e.g., size and / or number) by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0373] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion proliferation (e.g., size and / or number) by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0374] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion diameter by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0375] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion diameter by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0376] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion diameter by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0377] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion size by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0378] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion size by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0379] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion size by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0380] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion mass by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0381] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion mass by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0382] In an embodiment, treatment with an MSR conjugate disclosed herein decreases lesion mass by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0383] In an embodiment, treatment with an MSR conjugate disclosed herein decreases cancer cell count by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0384] In an embodiment, treatment with an MSR conjugate disclosed herein decreases cancer cell count by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0385] In an embodiment, treatment with an MSR conjugate disclosed herein decreases cancer cell count by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0386] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst circumference by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0387] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst circumference by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0388] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst circumference by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0389] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst diameter by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0390] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst diameter by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0391] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst diameter by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0392] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst volume by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0393] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst volume by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0394] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst volume by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0395] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst mass by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.

[0396] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst mass by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0397] In an embodiment, treatment with an MSR conjugate disclosed herein decreases sebaceous cyst mass by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%.

[0398] In an embodiment, treatment with an MSR conjugate disclosed herein increases the production of lymph node immune cells by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 110%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0399] In an embodiment, treatment with an MSR conjugate disclosed herein increases the production of lymph node immune cells by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0400] In an embodiment, treatment with an MSR conjugate disclosed herein increases the production of bone marrow immune cells by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 110%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0401] In an embodiment, treatment with an MSR conjugate disclosed herein increases the production of bone marrow immune cells by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0402] In an embodiment, treatment with an MSR conjugate disclosed herein increases the production of spleen immune cells by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 110%, about 125%,about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0403] In an embodiment, treatment with an MSR conjugate disclosed herein increases the production of spleen immune cells by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0404] In an embodiment, treatment with an MSR conjugate disclosed herein increases the production of cluster of differentiation (CD) cells by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 110%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0405] In an embodiment, treatment with an MSR conjugate disclosed herein increases the production of cluster of differentiation (CD) cells by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0406] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of neutrophils in the blood by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 1 10%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0407] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of neutrophils in the blood by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0408] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of monocytes in the blood by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 1 10%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0409] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of monocytes in the blood by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0410] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of CD8+ T cells in the bone marrow by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 110%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0411] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of CD8+ T cells in the bone marrow by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0412] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of CD11 b-Gr-1 cells in the bone marrow by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 110%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0413] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of CD11 b-Gr-1 cells in the bone marrow by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0414] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of CD11 b-Gr-1 cells in the spleen by, e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 110%, about 125%, about 150%, about 175%, about 200%, about 250%, about 300%, about 350%, about 400%, about 500% or more.

[0415] In an embodiment, treatment with an MSR conjugate disclosed herein increases the count of CD11 b-Gr-1 cells in the spleen by, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 125%, at least 150%, at least 175%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 500% or more.

[0416] The present specification also provides a pharmaceutical composition for the administration to a subject. The pharmaceutical composition disclosed herein may further include a pharmaceutically acceptable carrier, excipient, or diluent. As used herein, the term "pharmaceutically acceptable" means that the composition is sufficient to achieve the therapeutic effects without deleterious side effects, and may be readily determined depending on the type of the diseases, the patient's age, body weight, health conditions, gender, and drug sensitivity, administration route, administration mode, administration frequency, duration of treatment, drugs used in combination or coincident with the composition disclosed herein, and other factors known in medicine.

[0417] The composition may be used by blending with a variety of pharmaceutically acceptable carriers such as physiological saline or organic solvents. In order to increase the stability or absorptivity, carbohydrates such as glucose, sucrose or dextrans, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers may be used.

[0418] The administration dose and frequency of the pharmaceutical composition disclosed herein are determined by the type of active ingredient, together with various factors such as the disease to be treated, administration route, patient's age, gender, and body weight, and disease severity.

[0419] The total effective dose of the compositions disclosed herein may be administered to a patient in a single dose or may be administered for a long period of time in multiple doses according to a fractionated treatment protocol. In the pharmaceutical composition disclosed herein, the content of active ingredient may vary depending on the disease severity. However, the effective dose of the compositions disclosed are determined considering various factors including patient's age, body weight, health conditions, gender, disease severity, diet, and secretion rate, in addition to administration route and treatment frequency of the pharmaceutical composition. In view of this, those skilled in the art may easily determine an effective dose suitable for the particular use of the pharmaceutical composition disclosed herein. The pharmaceutical composition disclosed herein is not particularly limited to the formulation, and administration route and mode, as long as it shows suitable effects.

[0420] In various embodiments, a formulation can include one or more preservatives and / or additives known in the art. Similarly, a formulation can further be formulated, without limitation, into any of various known delivery formulations. For example, in an embodiment, a formulation can include, surfactants, adjuvant, biodegradable polymers, hydrogels, etc., such optional components, their chemical and functional characteristics are known in the art. Similarly known in the art are formulations that facilitate rapid, sustained or delayed release of the bioactive agents after administration. A formulation as described can be produced to include these or other formulation components known in the art.

[0421] The composition can therefore be administered as a single dose, or as two or more doses (which may or may not contain the same amount of the desired molecule) over time, or as a continuous infusion via an implantation device or catheter. Further refinement of the appropriate dosage is routinely made by those of ordinary skill in the art and is within the ambit of tasks routinely performed by them. Appropriate dosages may be ascertained through use of appropriate dose-response data. In various embodiments, the bioactive agents in formulations described herein can, without limitation, be administered to patients throughout an extended time period, such as chronic administration for a chronic condition.

[0422] Packaging and instruments for administration may be determined by a variety of considerations, such as, without limitation, the volume of material to be administered, the conditions for storage, whether skilled healthcare practitioners will administer or patient self-compliance, the dosage regime, the geopolitical environment (e.g., exposure to extreme conditions of temperature for developing nations), and other practical considerations.

[0423] Injection devices include pen injectors, auto injectors, safety syringes, injection pumps, infusion pumps, glass prefilled syringes, plastic prefilled syringes and needle free injectors that may be prefilled with liquid, or may be dual chambered, for example, for use with lyophilized material. An example of a syringe for such use is the Lyo-Ject™, a dual-chamber pre-filled lyosyringe available from Vetter GmbH, Ravensburg, Germany. Another example is the LyoTip which is a prefilled syringe designed to conveniently deliver lyophilized formulations available from LyoTip, Inc., Camarillo, California, U.S.A. Administration by injection may be, without limitation intravenous, intramuscular, intraperitoneal, or subcutaneous, as appropriate. Administrations by non-injection route may be, without limitation, nasal, oral, ocular, cochlear, dermal, or pulmonary, as appropriate. The above injection device may be used in conjunction with catheters, cannulas, ports, shunts and the like.

[0424] In certain embodiments, kits can include one or more single or multi-chambered syringes (e.g., liquid syringes and lyosyringes) for administering one or more formulations described herein. In various embodiments, the kit can comprise formulation components for parenteral, subcutaneous, intramuscular or IV administration, sealed in a vial under partial vacuum in a form ready for loading into a syringe and administration to a subject. In this regard, the composition can be disposed therein under partial vacuum. In all of these embodiments and others, the kits can contain one or more vials in accordance with any of the foregoing, wherein each vial contains a single unit dose for administration to a subject.

[0425] The kits can comprise lyophilates, disposed as herein, that upon reconstitution provide compositions in accordance therewith. In various embodiments the kits can contain a lyophilate and a sterile diluent for reconstituting the lyophilate.

[0426] Also described herein are methods for treating a subject in need of therapy, comprising administering to the subject an effective amount of a formulation as described herein. The therapeutically effective amount or dose of a formulation will depend on the disease or condition of the subject and actual clinical setting.

[0427] In an embodiment, a formulation as described herein can be administered by any suitable route, specifically by parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. It will also be appreciated that the preferred route will vary with the condition and age of the recipient, and the disease being treated. Methods of determining the most effective means and dosage of administration are known to those of skill in the art and will vary, without limitation, with the composition used for therapy, the purpose of the therapy, and the subject being treated. Single or multiple administrations can be carried out, without limitation, of the dose level and pattern being selected by thetreating physician. Suitable dosage formulations and methods of administering the agents are known in the art.

[0428] The formulations as described herein can be used in the manufacture of medicaments and for the treatment of humans and other animals by administration in accordance with conventional procedures.

[0429] Compositions in accordance with embodiments described herein have desirable properties, such as desirable solubility, viscosity, syringeability and stability. Lyophilates in accordance with embodiments described herein have desirable properties, as well, such as desirable recovery, stability and reconstitution.

[0430] In an embodiment, the pH of the pharmaceutical formulation is at least about 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11 , 11.25, 1 1.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, or 14.

[0431] In an embodiment, the pH of the pharmaceutical formulation is from about 3 to about 9, about 4 to about 9, about 5 to about 9, about 6 to about 8, about 6 to about 7, about 6 to about 9, about 5 to about 6, about 5 to about 7, about 5 to about 8, about 4 to about 9, about 4 to about 8, about 4 to about 7, about 4 to about 6, about 4 to about 5, about 3 to about 8, about 3 to about 7, about 3 to about 6, about 3 to about 5, about 3 to about 4, about 7 to about 8, about 7 to about 9, about 7 to about 10, about 7 to about 11 , about 8 to about 9, about 8 to about 10, about 8 to about 11 , about 8 to about 12, about 8 to about 13, about 8 to about 14, about 3 to about 10, about 3 to about 11 , about 3 to about 12, about 3 to about 13, about 3 to about 14, about 5 to about 10, about 5 to about 12, about 5 to about 13, about 5 to about 14, about 6 to about 10, about 6 to about 1 1 , about 6 to about 12, about 6 to about 13, about 6 to about 14, about 7 to about 12, about 7 to about 13, about 7 to about 14.EXAMPLES

[0432] The compositions and methods described herein will be further understood by reference to the following examples, which are intended to be purely exemplary. The compositions and methods described herein are not limited in scope by the exemplified embodiments, which are intended as illustrations of single aspects only. Any methods that are functionally equivalent are within the scope of the invention. Various modifications of the compositions and methods described herein in addition to those expressly described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications fall within the scope of the invention.EXAMPLE 1Immunological Response of AML Tumor Cells to Chemo and MSR Treatment

[0433] A study was conducted to determine the immunological response of AML tumor cells to chemotherapy treatment and subsequent treatment with mesoporous silica rods (ATT-01). Murine C1498- eGFP and WEHI-3 cell lines were cultured and exposed to cytarabine and / or doxorubicin for 3 days. After chemotherapy treatment, cells were exposed to mesoporous silica rods for 1 , 3 and 7 days. Supernatant and cells were collected by ELISA for multiplex analysis and flow cytometry assay. The ELISA cytokine assays focused on inflammatory immune responses (TNFa, IFNg, IL-6, GM-CSF, IL-1 b, IL-2).

[0434] Inflammatory immune responses were compared across the following eight conditions for both C1948-eGFP and WEHI-3 cell lines: 100K cells untreated, 100K cells treated with chemotherapy, 100K cells treated with mesoporous silica rods (ATT-01), 100K cells treated with both chemotherapy and ATT-01 , 500K cells untreated, 500K cells treated with chemotherapy, 500K cells treated with ATT-01 , and 500K cells treated with both chemotherapy and ATT-01 .

[0435] As shown in FIG. 6A, two graphs depict the resultant tumor cell death analysis of C1498 cells after the first day of MSR exposure. The results are illustrated for the eight aforementioned treatment scenarios. It was found that cell lines treated with ATT-01 exhibited dramatically higher tumor cell death than cell lines that were untreated or treated with chemotherapy alone. The percentage of dead tumor cells found after the first day of ATT-01 exposure displayed similar ratios in both the treatment of 100K cells and the treatment of 500K cells. In both conditions, cell lines treated with ATT-01 exhibited greater than 30% tumor cell death and cell lines treated with ATT-01 combined with chemotherapy showed greater than 20% tumor cell death.

[0436] As shown in FIG. 6B, two graphs depict the resultant percentage of CD19+ B220+ present in C1498 cells after the first day of MSR exposure. The results are illustrated for the eight aforementioned treatment scenarios. Cell lines treated with ATT-01 exhibited a higher percentage of CD19+ B220+ present in C1498 cells than in cell lines that were untreated or treated with chemotherapy alone. 100K cell lines and 500K cell lines treated with ATT-01 showed greater than 5% CD19+ B220+ cell content. While cell lines treated with ATT-01 and chemotherapy showed 3% CD19+ B220+ cell content in the 100K cell treatment and 2% CD19+ B220+ cell content in the 500K cell treatment. These results demonstrate that leukemic cells are differentiating into B cells after treatment with chemotherapy and ATT-01 .

[0437] As shown in FIG. 6C, four graphs depict the percentage of CD11 b+ CD86+ cells present (top row) and percentage of CD11 b+ GR-1 + cells present (bottom row) in the eight aforementionedtreatment scenarios. Cell lines treated with ATT-01 showed increased proportion of CD11 b+ GR-1 cells, while chemotherapy contributed to the increased presence of CD86+ cells. These results demonstrate that the leukemic cell line is differentiating into other cell types after treatment with chemotherapy and ATT-01 .

[0438] The results of the study demonstrated the increased immunomodulatory effect of MSR treatment and MSR treatment coupled with chemotherapy in murine subjects. Treatments with MSR were shown to achieve greater tumor cell death and greater immune cell response than the untreated control groups. Combined treatments of MSR and chemotherapy were shown to achieve greater tumor cell death than the untreated control groups.EXAMPLE 2Immune Response to MSR Vaccine in Porcine Model

[0439] A Yorkshire pig female was vaccinated on both flanks with 700 uL of mesoporous silica rod conjugate by subcutaneous injection with an 18G needle. Implant site measurements of temperature, complete blood cell count (CBC) and chemistry serum collection were performed on days 0 (baseline), 3, 7, 14, and 21 of the study. After the study organs and implants were collected for histology.

[0440] FIG. 7 displays the obtained cytokine levels measured in the collected pig serum. IL-6 was originally present in high concentration prior to immunization while various spikes in cytokine concentration occurred post-immunization. IL-8 was found in high concentration at day 7 postimmunization, as well as markedly increased levels of IL-12p40 and IL-10. A dramatic increase in IL-1 b was found in collected pig serum 21 -days post-immunization.

[0441] The porcine model demonstrated that vaccination with MSR by subcutaneous injection increases immune activity shortly after administration. Indicating elevated levels of cytokines resulting from subcutaneous injection of MSR.EXAMPLE 3In Vivo Immune Response to MSR Vaccine in Murine Subjects

[0442] A study (Study AT024) was conducted to determine and characterize the in vivo immune response to MSR vaccine dosing under antigen specific vaccines (WT-1) and ATT-01 .

[0443] MSR + / - GM-CSF and CPG + / - WT-1 were inoculated into the sub-q flank of four groups of non-tumor bearing murine subjects. Group 1 (murine subjects 1-15) served as the control group,receiving no inoculation. Group 2 (murine subjects 16-30) were injected with GM-CSF and CPG. Group 3 (murine subjects 31-45) were injected with MSR, GM-CSF and CPG. Group 4 (murine subjects 46-60) were injected with MSR, GM-CSF, CPG and WT-1 . Group 5 (murine subjects 61-75) were injected with only MSR. The change in volume of the treatment site was measured, and samples of blood, spleen, lymph node and skin at treatment site were collected for analysis of immune infiltration response.

[0444] A total of 75 murine subjects were studied and a histopathology assessment was performed for all subjects. Resultant histology samples were collected and placed onto slides. FIG. 8A-8I display selected histology samples collected from murine injection sites and lymph nodes used for evaluating immune infiltration response. More particularly, FIG. 8A displays a histology slide of a fragment of hairbearing skin from murine subject number 40. Shown on the slide is a granuloma comprised of aggregated epithelioid histiocytes and multinucleated giant cells encircling an aggregate of MSR. Surrounding the granuloma is a rim of neutrophils, lymphocytes and plasma cells. No malignancy or subcutaneous lymph nodes are identified.

[0445] FIG. 8B displays a histology slide of a fragment of hair-bearing skin from murine subject 37 (MSR+GM-CSF+CpG). Shown on the slide is a granuloma comprised of aggregated epithelioid histiocytes and multinucleated giant cells encircling an aggregate of MSR. Surrounding the granuloma is a rim of neutrophils, lymphocytes and plasma cells. No malignancy or subcutaneous lymph nodes are identified.

[0446] FIG. 8C displays a histology slide of a fragment of hair-bearing skin from murine subject 34 (MSR+GM-CSF+CpG). Shown on the slide is a lobular panniculitis with abundant neutrophils, lymphocytes and foamy histiocytes present. No MSR is present and no vasculitis is identified.

[0447] FIG. 8D displays a histology slide of a fragment of hair-bearing skin from murine subject 32 (MSR+GM-CSF+CpG). Shown on the slide is a mature granuloma comprised of aggregated epithelioid histiocytes and multinucleated giant cells interspersed with cholesterol clefts. No MSR is present. The tissue surrounding the granuloma shows no significant inflammation. No malignancy nor subcutaneous lymph nodes are identified.

[0448] FIG. 8E displays a histology slide of a lymph node from murine subject 25 (GM-CSF+CpG).

[0449] FIG. 8F (GM-CSF+CpG) displays a histology slide from murine subject 23. An activated lymph node is shown.

[0450] FIG. 8G displays a histology slide from murine subject 60 (MSR+GM-CSF+CpG+antigen).Shown on the slide is a lymph node with follicular hyperplasia.

[0451] FIG. 8H displays a histology slide of a lymph node from murine subject 8 (PBS) with sinus hyperplasia.

[0452] FIG. 8I displays a histology slide of a lymph node from murine subject 62 (MSR). Mixed reactive hyperplasia was observed.

[0453] Overall, Control Group 1 (PBS) and Group 2 (GM-CSF and CpG) showed minimal histologic changes. At least 50% of subjects in Groups 3 (MSR+GM-CSF+CpG), 4 (MSR+GM-CSF+antigen), and 5 (MSR alone) showed granulomatous inflammation or florid panniculitis at the injection site. No granulomatous inflammation was found in subjects not treated with MSR (Groups 1 and 2).

[0454] Granulomatous inflammation present in Day 45 subjects (M32 and M46) showed late / resolving changes consistent with expected time course; no MSR identified in these subjects suggested MSR has been successfully digested by the histiocytes.

[0455] Overall, six lymph nodes (LNs) showed sheets of lymphoid cells. All were from subjects at early time points: Day 4 (4x) or Day 6 (2x). These may represent early follicular hyperplasia or minimal / non-reactive lymph nodes. However, since four of the six lymph nodes were noted to be very small, the histologic sections may not be representative of the entire lymph nodes sampled.

[0456] Eight lymph nodes showed a mixed pattern with signs of follicular and paracortical hyperplasia. All were from subjects at later time points: Day 20 (1x) and Day 45 (6x). This pattern likely represents the expected time course of lymph node exposed to antigenic stimuli. Among lymph nodes showing a mixed pattern, four of the eight were from Group 5.

[0457] Groups 1 and 2 had the highest number of lymph nodes showing minimal or no reaction: Group 1 displayed minimal or no reaction in 7 of 11 murine subjects. Group 2 displayed minimal or no reaction in 7 of 15 murine subjects.

[0458] Follicular hyperplasia and mixed FH / PH patterns were consistent with antigen-stimulation and antibody production, which marks the switch from innate to adaptive immunity. Groups receiving MSR showed a higher number of cases with follicular hyperplasia and mixed FH / PH pattern. Group 3 displayed FH in 7 of 14 subjects. Group 4 displayed FH in 8 of 14 subjects with mixed results in 1 of 14subjects. Group 5 displayed FH in 7 of 14 subjects with mixed results in 4 of 14 subjects. A summary of findings is reproduced in Table 2 below.Table 2: Summary of AT024 Study Findings

[0459] The study found that granulomas formed quickly in murine subjects treated with MSR and in murine subjects treated with MSR alone and MSR coupled with GM-CSF and / or CPG (Groups 3-5). The study further showed that the immune response was systemic in the majority of subjects in those groups, showing that treatment with MSR and MSR coupled with GM-CSF and / or CPG resulted in hyperplasia signifying increased immune cell production in the lymphatic system.EXAMPLE 4MSR Vaccine Effect on AML Cells and Immunodepleting Antibodies

[0460] Female mice (C57 / BL6 - 8 weeks old) were administered immunodepleting antibodies (NK 1.1 , CD4, CD8) and were given intravenous injections of C1498-eGFP murine AML cells via a 29-gauge needle (AML challenge). Following the AML challenge Group A was untreated, Group B was treated with chemotherapy 7 days after the AML challenge, and Group C was treated with chemotherapy 7 days after the AML challenge and subcutaneously injected MSR vaccine on both flanks 14 days after the AML challenge.

[0461] Mice in Groups B and C were administered cytarabine at 100 mg / kg for five days and Doxorubicin at 3 mg / kg for three days. Complete blood cell (CBC) counts for all groups were sampled days 7, 14, 21 , 28, 35, 42, and 63 post-immunization.

[0462] FIG. 9 is four graphs showing complete blood cell (CBC) count results across three conditions: untreated (Group A), treatment with chemotherapy, MSR, GM-CSF and CpG (Group B), and treatment with chemotherapy plus GM-CSF and CpG(Group C). Over the course of 63 days postimmunization, white blood cell counts generally increased in subjects treated with chemotherapy plus mesoporous silica rods and in subjects treated with chemotherapy plus GM-CSF and CpG. During this same time period, lymphocyte counts generally increased in subjects treated with chemotherapy plus mesoporous silica rods and in subjects treated with chemotherapy plus GM-CSF and CpG. Monocyte counts showed greatest increase in subjects treated with chemotherapy plus mesoporous silica rods, GM-CSF, and CpG. At 21 and 28 days post-immunization, monocyte counts in Group C increased threefold compared to samples drawn at other milestones. The bottom right of FIG. 9 displays a graph showingneutrophil counts taken during this post-immunization time period. Subjects in Group C exhibited elevated neutrophil counts 21 and 28 days post-immunization, while subjects in Group B exhibited elevated neutrophil counts 21 and 63 days post-immunization.

[0463] The combination of chemotherapy and an MSR vaccine was shown to elevate immune cell production over a prolonged period following the AML challenge and administration of immunodepleting antibodies. Untreated subjects (Group A) showed marked depletion of immune cell counts during this same period subject to the same conditions.EXAMPLE 5MSR Vaccine Effect After AML Challenge

[0464] Female mice (C57 / BL6 - 8 weeks old) were administered an AML challenge through intravenous injection of C1498-eGFP murine AML cells with a 29-gauge needle. Following the AML challenge Group A was untreated, Group B was treated with chemotherapy 7 days after the AML challenge, Group C was treated with chemotherapy 7 days after the AML challenge and subcutaneously injected MSR vaccine on both flanks 14 days after the AML challenge. Group D was treated with chemotherapy 7 days after the AML challenge, subcutaneously injected MSR vaccine on both flanks 14 days after the AML challenge and administered GM-CSF and CpG.

[0465] Mice in Groups B, C and D were administered cytarabine at 100 mg / kg for five days and Doxorubicin at 3 mg / kg for three days.

[0466] Bone marrow and implant site samples were collected 7 and 20-days post vaccination. Flow cytometry analysis was conducted to measure percentage T cells, macrophages, monocytes, neutrophils and AML cells present in the bone marrow and implant sites. Blood was collected for complete blood cell count measurements. Spleen samples were measured for Treg (FOXP3+ CD4+) / CD8+ ratio by flow cytometry and IFNg+ T cells were assayed by ELISPOT.

[0467] FIG. 10A shows graphs depicting the percentage of GFP+ positive cells in the bone marrow, at the injection site, in the lymph nodes and in splenocytes 7 days post-treatment with MSR-based therapy. Mice in Group D generally exhibited the highest percentage of GFP+ positive cells in bone marrow. Mice in Group C showed presence of GFP+ positive cells at the injection site. Mice in Groups C and D exhibited a slightly greater percentage of GFP+ positive cells in lymph nodes compared to Groups A and B. Day 7 splenocyte samples showed no relative difference in GFP+ positive cell percentage across Groups A-D.

[0468] FIG. 10B shows graphs depicting percentage of GFP+ positive cells in the bone marrow, at the implant site, in the lymph nodes and in the spleen 22 days post-immunization. Mice in Group B generally exhibited the highest percentage of GFP+ positive cells in bone marrow. Mice in Group C showed presence of GFP+ positive cells at the injection site. Mice in Groups C and D exhibited a markedly lower percentage of GFP+ positive cells in lymph nodes compared to Groups A and B. Day 22 splenocyte samples showed no relative difference in GFP+ positive cell percentage across Groups A-D.

[0469] FIG. 10C shows graphs depicting percentage of GFP+ positive cells in the blood, ovaries, and in the liver 22 days post-immunization. Mice in Group B generally exhibited the highest percentage of GFP+ positive cells in blood. Mice in Group B also showed the highest percentage of GFP+ positive cells in the ovaries. Mice in Group D exhibited a greater percentage of GFP+ positive cells in the liver compared to Groups A, B and C.

[0470] FIG. 11 A shows graphs depicting immune cell populations at the subcutaneous injection site of mesoporous silica rods on day 7 post immunization for Group C. Percentage CD11 b+ GR-1 cells were shown to be the most elevated at the injection site, ranging from 40-80%.

[0471] FIG. 11 B shows graphs depicting immune cell populations in the bone marrow on day 7 post immunization. Mice in Group D generally exhibited the highest percentage of CD4 helper T cells. Mice in Group C generally exhibited the highest percentage of CD8 T-cells and the highest percentage of CD11 b+ GR-1 cells.

[0472] FIG. 11 C shows graphs depicting immune cell populations in the bone marrow on day 22 post immunization. Mice in Group B generally exhibited the highest percentage of CD4 helper T cells and CD11 b+ GR-1 cells. Mice in Group D generally exhibited the highest percentage of CD8 T-cells.

[0473] FIG. 11 D shows graphs depicting immune cell populations in spleens on day 7 post immunization. Mice in Groups C and D generally exhibited the highest percentage of CD11 c+ GR-1 cells. Mice in Group C also generally exhibited the highest percentage of CD11 b+ GR-1 cells.

[0474] FIG. 11 E shows graphs depicting immune cell populations in spleens on day 22 post immunization. Mice in Groups C and D generally exhibited the highest percentage of CD4 helper T-cells. Mice in Group B generally exhibited the highest percentage of CD3+ cells plus NK 1.1 antibodies and the highest percentage of CD11 b+ CD86+ cells.

[0475] FIG. 11 F shows graphs depicting immune cell infiltration in draining lymph nodes on day 7 post immunization. Mice in Groups B and C generally exhibited the highest percentage of CD4 helper T- cells and CD8 T-cells, but it was not significant.

[0476] FIG. 11 G shows graphs depicting immune cell infiltration in draining lymph nodes on day 22 post immunization. Mice in Group D generally exhibited the highest percentage of CD4 helper T-cells and CD3+ NK1 .1 , but it was not significant. Mice in Groups B and C generally exhibited the highest percentage of CD11 b+ CD86+ cells.

[0477] FIG. 11 H shows graphs depicting CD86+ cells infiltration in organs on day 7 post immunization. Mice in Groups B and C generally exhibited the highest percentage CD86+ cells in lymph nodes. Group C also generally exhibited the highest percentage of CD86+ cells in the bone marrow and at the injection site, but it was not significant.

[0478] The MSR vaccine effect after AML challenge on murine subjects was significant in terms of survival and resulted in elevated localization of key immune cell populations at the injection site, within the bone marrow, lymph nodes and spleens of murine subjects.EXAMPLE 6Synthesis of Mesoporous Silica Rods

[0479] Methods of producing Mesoporous Silica Rods (MSRs) are known in the art. The size, morphology, pore size, and pore structure of MSNs can be rationally designed and the synthesis process can be freely controlled. Particle size and shape can also influence their blood circulation, cellular uptake and tumor penetration, being determinant parameters to achieve therapeutic effects. Regarding particle size, a diameter range from 50 to 300 nm can favor an optimal cellular uptake, long circulation time, high drug loading and high accumulation in tumors.

[0480] In aspects, five ingredients are used in a simple manufacturing process with low cost and minimal complex chemistry.1) Poly (ethylene glycol)-block-poly (propylene glycol)-block- poly (ethylene glycol), Pluronic®(P123)2) Water for injection (WFI)3) Hydrochloric acid 37% acid-fuming4) Tetraethyl orthosilicate (TEOS)5) Pure 70% Ethanol

[0481] The mesoporous materials can vary in their structural arrangement and pore size. For example, MCM-48 has a cubic arrangement whereas MCM-50 has a lamella-like arrangement. Non-ionic triblock copolymers like alkyl polyethylene oxide) (PEO) oligomeric surfactants and poly(alkylene oxide) block copolymers have also been used for synthesis of SBA. The ratio of ethylene oxide to propylene oxide can be varied to achieve a desired symmetry of mesoporous materials: (SBA-11 (cubic), SBA-12 (3-d hexagonal), SBA-15 (hexagonal) and SBA-16 (cubic cage-structured). This is different from MCM in that they possess larger pores of 4.6 - 30 nm and thicker silica walls. FSM-16, that is, folded sheets of mesoporous materials are another type of mesoporous materials, which can be synthesized using quaternary ammonium surfactant as a template and layered polysilicate kanemite.

[0482] The longitudinal pores in mesoporous silica rods, are essentially long, tubular channels that run along the length of the rods. These pores are hexagonally ordered and have diameters ranging from 2-15 nm. In an embodiment, a pore is 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 11 nm, 12 nm, 13 nm, 14 nm, 15 nm or larger in diamerter. The structure provides a high surface area and uniform pore size, which allows the mesoporous silica to absorb and then release substances useful for drug delivery.

[0483] The number of tubular channels in mesoporous silica depends on the specific synthesis conditions and dimensions of the material. Typically, these materials have a hexagonal arrangement of the channels, and the number of channels can vary. Each mesoporous silica rod particle has hundreds, thousands, tens of thousands or hundeds or thousands of these channels running parallel to one another. In an embodiment, a mesoporous silica rod particle has at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least 1500, at least1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 3000, at least 4000, at least 5000, at least 6000, at least 7000, at least 8000, at least 9000, at least 10,000, at least 20,000, at least 30,000, at least 40,000, at least 50,000, at least 60,000, at least 70,000, at least 80,000, at least 90,000, at least 100,000, at least 200,000, at least 300,000 or more channels running parallel to one another.Vaccine Manufacture

[0484] Methods of producing mesoporous silica rods vaccines for humoral response are also known in the art. In this example, lyophilized MSRs are suspended in PBS that contains IL-12 (1 - 60 pg per mg of MSR) and gently mixed to make ATT-02. The IL-12 is allowed to adsorb to the surface of the MSRs for between 1 - 3 hours at 37°C. Next, the contents were pulled into a syringe and injected directly into the tumor. The content and the release profile of the IL-12 is confirmed using in vitro methods.EXAMPLE 7MSR Therapy for Treating Acute Myeloid Leukemia (AML)

[0485] Patients with AML generally present with a high burden of disease at the time of diagnosis and require effective and tolerable systemic therapy. Chemotherapy can rapidly induce remission; however, relapse occurs in the majority of patients, highlighting the difficulty in eradicating all AML cells. Therapeutic vaccines have the potential to achieve a lasting AML-specific immune response that is capable of eradicating the residual disease, as demonstrated by graft vs. host disease. The development of an effective immune response typically requires T-cell activation resulting from effective presentation of one or more tumor antigens in the context of co-stimulation. Recent studies have demonstrated that an injectable biomaterial system can create a local, controlled immunological microenvironment that serves as a site for modulating the immune response against AML. The biomaterial system recruits immune cells and circulating tumor cells that work in concert with immunoregulatory factors to evoke a potent and durable response against AML. In the first demonstration, a murine model of established AML, treated with a cryogel version of the system when used following induced chemotherapy (iCt), resulted in lower disease burden of leukemia cells, improved survival, and conferred transferable immunity without the direct incorporation of tumor antigens. Using this concept, Applicants utilize mesoporous silica rods (MSRs) biomaterial as the scaffold (which are more easily manufactured) and coated with GM-CSF and CpG, to generate similar results in an alternative syngeneic murine AML model.

[0486] ATT-01 is a product that includes mesoporous silicon dioxide (silica) with GM-CSF (Leukine®) and unmethylated cytosine-guanine dinucleotide (CpG 7909). ATT-01 demonstrated a robust immunogenic response in animal models following treatment with standard chemotherapy regimens by creating a localized microenvironment, which can recruit circulating immune cells and colocalize them with other inflammatory cells, priming the immune cells for long-term anti-tumor and anticancer efficacy. This leads to a sustained anti-tumor and anti-cancer immune response that may significantly extend patient survival with minimal added toxicity.

[0487] ATT-01 for the treatment of AML is made by using an ad / mix process in which solutions of Leukine® and CpG 7909 are first added to a suspension of Mesoporous Silica Rods (MSRs) and then mixed for a brief period to allow adsorption, then placed in vials, frozen, and lyophilized. The final product is a fine white powder that is resuspended in WFI and injected subcutaneously with a 23-gauge needle. Once subcutaneously injected, a systemic immune response created by the biomaterial and components recruit numerous immune cells into the three-dimensional structure created as the MSRs settle. In addition to the immune cells, the biomaterial recruits circulating AML cells; in the case of ATT-01 , postinduction apoptotic AML cells are recruited, which then serve as the antigenic target for the immune system.

[0488] In this example, a patient visits her oncologist to discuss AML treatment options. The oncologist suggests using ATT-01 .

[0489] The agent is provided as a reconstituted lyophilized powder. It is dissolved in sterile water for subcutaneous injection (23g Needle). A single dose can include 2, 4 or 8 injections. After treatment, the patient is monitored periodically for signs / symptoms of AML. Blood, bone marrow, and injection site samples are periodically obtained to measure immune cell infiltration. During a follow-up appointment eight weeks later, the oncologist discovered that the patient exhibited no outward symptoms of AML. A bone marrow biopsy was conducted and showed reduced incidence of blast cell clusters and collections.EXAMPLE 8

[0490] The following describes the compositions used in the Examples set forth below. ATT-02 comprises an MSR and IL-12. OVA is Ovalbumin.

[0491] The study groups described below in the Examples are:MSR (1 mg) + IL-12 (20ug). Route: [i.t.]. Volume injected:70uL; MSR (1 mg) + IL-12 (20ug) + OVA (100ug). Route: [i.t.]. Volume injected: 70uL;IL-12 (20ug) Route: [i.t.]. Volume injected: 70uL;IL-12 (20ug) + OVA (100ug). Route: [i.t.] Volume injected:70uL; MSR (1 mg) + IL-12 (20ug) + OVA (100ug). Route: [s.c.]. Volume injected: 70uL;IL-12 (20ug) + OVA (100ug). Route: [s.c.] Volume injected: 70uL.

[0492] Tumor cell lines (B16F10 [melanoma] and EMT-6) were cultured with an MSR, MSR+IL-12, or an MSR+IL-12+OVA, The results show that after 24 or 48 hours, tumor cells express myeloid markers, including cd11 b, cd11 c, MHC II, and / or CD86. (See Fig. 25). Following treatment, some tumor cell lines become more macrophage like, others become more neutrophil, and / or dendritic cell like.EXAMPLE 9

[0493] Therapeutic effects of a single treatment of ATT-02 and ATT-02 OVA in the primary tumor. Randomization on primary tumor: 70mm3.C57 / BI6 mice (untreated n = 9 mice / group, MSR intratumorally n = 9 mice / group, ATT-02 intratumorally n = 9 mice / group, ATT-02 OVA intratumorally. n = 9 mice / group, IL-12 intratumorally n = 9 mice / group, IL-12 OVA intratumorally n = 9 mice / group, ATT-02 OVA subcutaneous n = 9 mice / group, and IL-12 OVA subcutaneous n = 9 mice / group) wereinoculated s.c. with 10A6 and 0.25 x 1 OA6 B16F10-OVA melanoma tumor cells on the left and right flanks, respectively. At day 11 , the left flank tumor was treated intratumorally or subcutaneously with PBS (a), MSR (1 mg) (b), ATT-02 (1 mg MSR 20ug IL-12) (f), ATT-02 OVA (1 mg MSR 20ug IL-12 and 100ug OVA) (g, h), IL-12 (20ug IL-12) (c), IL-12 OVA (20ug IL-12 and 100ug OVA) (d, e) either intratumorally or subcutaneously. Shown in Figure 18 is the individual tumor growth curves for the treated. Black arrows indicate timing of treatment.EXAMPLE 10

[0494] Abscopal effects of single treatment of ATT-02 and ATT-02 OVA in the secondary tumor. Shown in Figure 19 is the individual tumor growth curves for the untreated (a - h).EXAMPLE 11

[0495] IL-12 and IFNgamma detection in the serum of mice (n=9) receiving single dose of PBS,MSR, ATT-02, ATT-02 OVA, 11-12, and IL-12 OVA either i.t. or s.c. Serum samples were collected and analyzed 1 -, 3-, 7-, and 14-days post treatment. IL-12 (a) and IFN-gamma (b) levels were measured by ELISA kit (Life Technologies). Both cytokines were detected at baseline levels in the MSR IL-12 group as shown in Figure 20.EXAMPLE 12

[0496] Mice were inoculated with 1 million cells on their left-flank (primary) and 0.25 million cells on their right-flank (secondary) with B16F10 tumor s.c. Shown are the quantification of total and percentage of CD8+ T cells and the antigen specific T- cells by SIINFEKL tetramer staining after PBS, MSR, ATT-02, ATT-02 OVA, IL-12 and IL-12 OVA treatments either i.t. or s.c. in treated and untreated tumors. Data shown in Figure 21 represent the mean ± SD of each group.EXAMPLE 13

[0497] Immune cells analysis by quantification of total and percentage of myeloid cells after PBS, MSR, ATT-02, ATT-02 OVA, IL-12 and IL-12 OVA treatments either i.t. or s.c. in in treated and untreated tumors. Data shown in Figure 22 represent the mean ± SD of each group.EXAMPLE 14

[0498] Immune cells were analyzed by quantification of total and percentage of macrophages after PBS, MSR, ATT-02, ATT-02 OVA, IL-12 and IL-12 OVA treatments either i.t. or s.c. in in treated (a -d) and untreated tumors. Data shown in Figure 23 represent the mean ± SD of each group.EXAMPLE 15

[0499] Mice were inoculated with 1 million cells on left-flank (primary) and 0.25 million cells on right-flank (secondary) with B16F10 tumor s.c. Shown are the quantification of total and percentage of CD8+ T cells and the antigen specific T- cells by SIINFEKL tetramer staining after PBS, MSR, ATT-02, ATT-02 OVA, IL-12 and IL-12 OVA treatments either i.t. or s.c. in lymph nodes of treated and untreated animals. Data shown in Figure 24 represent the mean ± SD of each group.EXAMPLE 16

[0500] Phenotypic changes in tumor cells exposed to MSR are shown in Fig. 25A. Immune cell marker quantification after PBS, MSR, ATT-02, ATT-02 OVA, IL-12 and IL-12 OVA treatments of tumor cells in vitro show phenotypic changes. THP-1 (human cells, leukemic cell line) were treated as described with MSR therapies as described in figure 26, 27, and 28 and show changes in activation and differentiation state.

[0501] Certain embodiments of the present invention are described herein, including the best mode known to the inventors for carrying out the invention. Of course, variations on these described embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventor expects skilled artisans to employ such variations as appropriate, and the inventors intend for the present invention to be practiced otherwise than specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the abovedescribed embodiments in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[0502] Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability.When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0503] Unless otherwise indicated, all numbers expressing a characteristic, item, quantity, parameter, property, term, and so forth used in the present specification and claims are to be understood as being modified in all instances by the term “about.” As used herein, the term “about” means that the characteristic, item, quantity, parameter, property, or term so qualified encompasses a range of plus or minus ten percent above and below the value of the stated characteristic, item, quantity, parameter, property, or term. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numericalindication should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and values setting forth the broad scope of the invention are approximations, the numerical ranges and values set forth in the specific examples are reported as precisely as possible. Any numerical range or value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Recitation of numerical ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate numerical value falling within the range. Unless otherwise indicated herein, each individual value of a numerical range is incorporated into the present specification as if it were individually recited herein.

[0504] The terms “a,” “an,” “the” and similar referents used in the context of describing the present invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein is intended merely to better illuminate the present invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the present specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0505] Specific embodiments disclosed herein may be further limited in the claims using consisting of or consisting essentially of language. When used in the claims, whether as filed or added per amendment, the transition term “consisting of’ excludes any element, step, or ingredient not specified in the claims. The transition term “consisting essentially of’ limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel characteristic(s).Embodiments of the present invention so claimed are inherently or expressly described and enabled herein.

[0506] Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0507] All patents, patent publications, and other publications referenced and identified in the present specification are individually and expressly incorporated herein by reference in their entirety for the purpose of describing and disclosing, for example, the compositions and methodologies described in such publications that might be used in connection with the present invention. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regardshould be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

[0508] In closing, it is to be understood that although aspects of the present specification are highlighted by referring to specific embodiments, one skilled in the art will readily appreciate that these disclosed embodiments are only illustrative of the principles of the subject matter disclosed herein. Therefore, it should be understood that the disclosed subject matter is in no way limited to a particular methodology, protocol, and / or reagent, etc., described herein. As such, various modifications or changes to or alternative configurations of the disclosed subject matter can be made in accordance with the teachings herein without departing from the spirit of the present specification. Lastly, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which is defined solely by the claims. Accordingly, the present invention is not limited to that precisely as shown and described.

Claims

CLAIMSWhat is claimed is:1 . A method of treating a disease in a subject in need thereof, the method comprising:(a) preparing mesoporous silica rods, wherein the mesoporous silica rods comprise a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(b) administering treatment to a subject by subcutaneously injecting the mesoporous silica rods into the subject at a treatment site;(c) wherein the treatment results in a localized inflammatory reaction near the treatment site, thereby treating the disease.

2. The method of claim 1 , wherein the cytokine payload is a payload comprising one or more of interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating-factor (GM-CSF).

3. The method of claim 1 , wherein the adjuvant is one or more of aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

4. The method of claim 1 , wherein the mesoporous silica rods are substantially cylindrical.

5. The method of claim 1 , wherein the mesoporous silica rods each have an internal labyrinthine structure.

6. A method of treating a cancer in a subject in need thereof, the method comprising:(a) preparing mesoporous silica rods, wherein the mesoporous silica rods comprise a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(b) administering treatment to a subject by subcutaneously injecting the mesoporous silica rods into the subject at a treatment site;(c) wherein the treatment results in a localized inflammatory reaction near the treatment site, thereby treating the cancer.

7. The method of claim 6, wherein the cytokine payload is a payload comprising one or more of interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating-factor (GM-CSF).

8. The method of claim 6, wherein the adjuvant is one or more of aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

9. The method of claim 6, wherein the mesoporous silica rods are substantially cylindrical.

10. The method of claim 6, wherein the mesoporous silica rods each have an internal labyrinthine structure.

11. A method of treating a tumor in a subject in need thereof, the method comprising:(a) preparing mesoporous silica rods, wherein the mesoporous silica rods comprise a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(b) administering treatment to a subject by subcutaneously injecting the mesoporous silica rods into the subject at a treatment site adjacent to a tumor;(c) wherein the treatment results in a localized inflammatory reaction near the treatment site, thereby treating the tumor.

12. The method of claim 11 , wherein the cytokine payload is a payload comprising one or more of interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating-factor (GM-CSF).

13. The method of claim 11 , wherein the adjuvant is one or more of aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

14. The method of claim 11 , wherein the mesoporous silica rods are substantially cylindrical.

15. The method of claim 11 , wherein the mesoporous silica rods each have an internal labyrinthine structure.

16. A method of treating a disease in a subject in need thereof, the method comprising:(a) preparing mesoporous silica rods, wherein the mesoporous silica rods comprise a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(b) administering treatment to a subject by subcutaneously injecting the mesoporous silica rods into the subject;(c) wherein the treatment results in a systemic innate inflammatory response, thereby treating the disease.

17. The method of claim 16, wherein the cytokine payload is a payload comprising one or more of interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating-factor (GM-CSF).

18. The method of claim 16, wherein the adjuvant is one or more of aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

19. The method of claim 16, wherein the mesoporous silica rods are substantially cylindrical.

20. The method of claim 16, wherein the mesoporous silica rods each have an internal labyrinthine structure.

21. The method of claim 16, wherein the systemic innate inflammatory response comprises granulopoiesis.

22. The method of claim 21 , wherein the granulopoiesis is steady-state granulopoiesis.

23. The method of claim 21 , wherein the granulopoiesis is emergency granulopoiesis.

24. A method of treating cancer cells in a subject in need thereof, the method comprising:(a) preparing mesoporous silica rods, wherein the mesoporous silica rods comprise a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(b) administering treatment to a subject by subcutaneously injecting the mesoporous silica rods into the subject;(c) wherein the treatment results in a systemic innate inflammatory response, thereby reducing the number of cancer cells within the subject.

25. The method of claim 24, wherein the cytokine payload is a payload comprising one or more of interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating-factor (GM-CSF).

26. The method of claim 24, wherein the adjuvant is one or more of aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

27. The method of claim 24, wherein the mesoporous silica rods are substantially cylindrical.

28. The method of claim 24, wherein the mesoporous silica rods each have an internal labyrinthine structure.

29. The method of claim 24, wherein the systemic innate inflammatory response comprises granulopoiesis.

30. The method of claim 24, wherein the granulopoiesis is steady-state granulopoiesis.31 . The method of claim 24, wherein the granulopoiesis is emergency granulopoiesis.

32. The method of claim 24, wherein the cancer cells are one or more of: acute myeloid leukemia (AML) cells, acute lymphoblastic leukemia cells, chronic lymphocytic leukemia cells, chronic myelogenous leukemia cells, Hodgkin lymphoma cells, non-Hodgkin lymphoma cells, Waldenstrom macroglobulinem cells, follicular lymphoma cells, B-cell lymphoma cells, cutaneous T-cell lymphoma cells, multiple myeloma cells, plasmacytoma cells, cancer stem cells or cancer precursor cells.

33. The method of claim 24, wherein the cancer cells are acute myeloid leukemia (AML) cells.

34. The method of claim 24, wherein the number of cancer cells in the subject is reduced by at least 15%.

35. The method of claim 24, wherein the number of cancer cells in the subject is reduced by at least 25%.

36. The method of claim 24, wherein the number of cancer cells in the subject is reduced by at least 50%.

37. The method of claim 24, wherein the number of cancer cells in the subject is reduced by at least 80%.

38. A method of treating acute myeloid leukemia (AML) in a subject in need thereof, the method comprising:(a) preparing mesoporous silica rods, wherein the mesoporous silica rods comprise a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(b) administering treatment to a subject by subcutaneously injecting the mesoporous silica rods into the subject;(c) wherein the treatment results in a systemic innate inflammatory response, thereby reducing the number of AML cells within the subject.

39. The method of claim 38, wherein the cytokine payload is a payload comprising one or more of interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating-factor (GM-CSF).

40. The method of claim 38, wherein the adjuvant is one or more of aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

41. The method of claim 38, wherein the mesoporous silica rods are substantially cylindrical.

42. The method of claim 38, wherein the mesoporous silica rods each have an internal labyrinthine structure.

43. The method of claim 38, wherein the systemic innate inflammatory response comprises granulopoiesis.

44. The method of claim 38, wherein the granulopoiesis is steady-state granulopoiesis.

45. The method of claim 38, wherein the granulopoiesis is emergency granulopoiesis.

46. The method of claim 38, wherein the number of AML cells in the subject is reduced by at least 15%.

47. The method of claim 38, wherein the number of AML cells in the subject is reduced by at least 25%.

48. The method of claim 38, wherein the number of AML cells in the subject is reduced by at least 50%.

49. The method of claim 53, wherein the number of AML cells in the subject is reduced by at least 80%.

50. A method of treating a cancer or heme malignancy in a subject in need thereof, the method comprising:(a) preparing mesoporous silica rods, wherein the mesoporous silica rods comprise a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(b) administering treatment to a subject by subcutaneously injecting the mesoporous silica rods into the subject;(c) wherein the treatment results in differentiation of tumor cells into other cell types, thereby treating the cancer within the subject.

51. The method of claim 50, wherein the differentiated cell types are one of: neutrophils, monocytes, macrophages, antigen presenting cells and dendritic cells.

52. The method of claim 50 wherein the differentiated cell types are antigen presenting cells and generate a vaccine-like response against the tumor.

53. The method of claim 50, wherein the differentiated cell types are one or more of: neutrophils, monocytes, macrophages, antigen presenting cells and dendritic cells.

54. The method of claim 50, wherein the differentiated cell types are cleared from the immune system of the subject within 48 hours.

55. The method of claim 50, wherein the differentiated cell types are cleared from the immune system of the subject within 24 hours.

56. The method of claim 50, wherein the differentiated cell types are cleared from the immune system of the subject within weeks.

57. The method of claim 50, wherein the differentiated cell types are cleared from the immune system of the subject within months.

58. The method of claim 50, wherein the cytokine payload is a payload comprising one or more of interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating-factor (GM-CSF).

59. The method of claim 50, wherein the adjuvant is one or more of aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

60. The method of claim 50, wherein the mesoporous silica rods are substantially cylindrical.

61. The method of claim 50, wherein the mesoporous silica rods each have an internal labyrinthine structure.

62. A method of treating a leukemia in a subject in need thereof, the method comprising:(a) preparing mesoporous silica rods, wherein the mesoporous silica rods comprise a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(b) administering treatment to a subject by subcutaneously injecting the mesoporous silica rods into the subject;(c) wherein the treatment results in an adaptive immune response, thereby treating the leukemia within the subject.

63. The method of claim 62, wherein the adaptive immune response comprises differentiation of leukemic blasts.

64. The method of claim 62, wherein the adaptive immune response comprises production of leukemic antigens on neutrophil extracellular traps (NETs).

65. The method of claim 62, wherein the cytokine payload is a payload comprising one or more of interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating-factor (GM-CSF).

66. The method of claim 62, wherein the adjuvant is one or more of aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

67. The method of claim 62, wherein the mesoporous silica rods are substantially cylindrical.

68. The method of claim 62, wherein the mesoporous silica rods each have an internal labyrinthine structure.

69. A mesoporous silica rod (MSR) for subcutaneous injection into a subject, wherein the mesoporous silica rod comprises a cytokine payload and an adjuvant for time- released excretion of the cytokine payload and the adjuvant.

70. The mesoporous silica rod of claim 69, wherein the cytokine payload is selected from interleukin- 12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating factor (GM-CSF).71 . The mesoporous silica rod of claim 69, wherein the adjuvant is selected from aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

72. The mesoporous silica rod of claim 69, wherein the mesoporous silica rod is substantially cylindrical.

73. The mesoporous silica rod of claim 69, wherein the mesoporous silica rod has an internal labyrinthine structure.

74. The mesoporous silica rod of claim 69, wherein the cytokine payload is in the amount of 3 pg.

75. The mesoporous silica rod of claim 69, wherein the cytokine payload is in the amount of 6 pg.

76. The mesoporous silica rod of claim 69, wherein the cytokine payload is in the amount of 20 pg.

77. A method of administering a mesoporous silica rod (MSR) to a subject with cancer, the method comprising:(a) pretreating the subject with a predetermined course of chemotherapy;(b) preparing a mesoporous silica rod, wherein the mesoporous silica rod comprises a therapeutically effective amount of one or more of the following: a. a cytokine payload; b. an adjuvant;(c) subcutaneously injecting the mesoporous silica rod into the subject;(d) wherein following the injection of the mesoporous silica rod the number of cancer cells in the subject is reduced.

78. The method of claim 77, wherein the cytokine payload is selected from interleukin-12 (IL-12), interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating factor (GM-CSF).

79. The method of claim 77, wherein the adjuvant is selected from aluminum hydroxide (alum), a lipopolysaccharide (LPS), cytosine guanosine dinucleotide (CpG) and a toll like receptor agonist (TLR agonist).

80. The method of claim 77, wherein the mesoporous silica rod is substantially cylindrical.

81. The method of claim 77, wherein the mesoporous silica rod has an internal labyrinthine structure.

82. The method of claim 77, wherein the cytokine payload is in the amount of 3 pg.

83. The method of claim 77, wherein the cytokine payload is in the amount of 6 pg.

84. The method of claim 77, wherein the cytokine payload is in the amount of 20 pg.

85. The method of claim 77, wherein the cytokine payload is in the amount of 40 pg.

86. The method of claim 77, wherein the number of cancer cells in the subject is reduced by at least 15%.

87. The method of claim 77, wherein the number of cancer cells in the subject is reduced by at least 25%.

88. The method of claim 77, wherein the number of cancer cells in the subject is reduced by at least 50%.

89. The method of claim 77, wherein the number of cancer cells in the subject is reduced by at least 80%.

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