Patent Application
20230272101
Radiation therapy is commonly used as a treatment for cancer. Radiation therapy uses ionizing radiation to damage the genetic material of the targeted cells, resulting in death and damage of the affected cells. However, in many circumstances, radiation therapy is not sufficient to eradicate all remnants of a cancer and/or tumor, or prevent distal metastases of a cancer.
Dendritic cells are antigen-presenting cells which process antigenic material and present it on the cell surface to the T-cells of the immune system. When T-cells are presented with tumor specific antigens by dendritic cells, the T-cells are then able to play a critical role in the immune system’s ability to target and kill tumor cells. This disclosure describes uses of dendritic cell activating molecules that improve the effectiveness of radiation treatment, and establish systemic anti-cancer/tumor immunity. In these methods, the dendritic cell activating molecule is administered after radiation treatment. This results in improved treatment of the cancer or tumor compared to treatment with radiation or dendritic cell activating molecule alone, as well as when compared to simultaneous treatment with radiation and a dendritic cell activating molecule.
Described herein in one aspect is a method of treating a tumor or a cancer in an individual, the method comprising administering to the individual a dose of a radiation therapy and a dendritic cell activating molecule, wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy is administered. Also described is a method of treating a tumor or a cancer in an individual, the method comprising administering to the individual a dendritic cell activating molecule, wherein the individual has received a dose of a radiation therapy, and wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy has been administered. In certain embodiments, the dendritic cell activating molecule is administered at least two days after the radiation therapy is administered. In certain embodiments, the dendritic cell activating molecule is administered at least three days after the radiation therapy is administered. In certain embodiments, the dose of the radiation therapy comprises a plurality of doses of radiation therapy. In certain embodiments, the radiation therapy is external beam radiation therapy. In certain embodiments, the external beam radiation therapy is selected from the list consisting of: three-dimensional conformal radiation therapy, intensity modulated radiation therapy, image guided radiation therapy, stereotactic radiation therapy, intraoperative radiation therapy, proton beam therapy, neutron beam therapy, and combinations thereof. In certain embodiments, the dose of radiation therapy comprises at least about 2 Gy. In certain embodiments, the dose of radiation therapy comprises at least about 2 Gy and no more than about 20 Gy. In certain embodiments, the dendritic cell activating molecule is administered at least three days after the dose of the radiation therapy. In certain embodiments, the dendritic cell activating molecule is administered at least five days after the dose of the radiation therapy. In certain embodiments, the dendritic cell activating molecule is administered at least seven days after the dose of the radiation therapy. In certain embodiments, the dendritic cell activating molecule induces maturation of an immature dendritic cell. In certain embodiments, the dendritic cell activating molecule activates dendritic cell activation through a toll-like receptor, a NOD-like receptor, a RIG-1 or MDA-5 receptor, a C-type lectin receptor, a costimulatory molecule, a cytokine receptor, or a STING pathway. In certain embodiments, the dendritic cell activating molecule is a toll-like receptor agonist selected from the list consisting of CpG oligonucleotide, SD-101, LFX453, imiquimod, Bacillus Calmette-Guérin (BCG), monophosphoryl lipid A, Poly ICLC, GSK1795091, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a NOD-like receptor agonist selected from the list consisting of bacterial peptidoglycan, an acylated derivative of iE-DAP (C12-iE-DAP), D-gamma-Glu-mDAP (iE-DAP), L-Ala-gamma-D-Glu-mDAP (Tri-DAP), muramyl dipeptide (MDP), muramyl tripeptide, L18-MDP, M-TriDAP, murabutide, PGN-ECndi, PGN-ECndss, PGN-SAndi, N-glycolylated muramyl dipeptide, murabutide, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a RIG-1 or MDA-5 receptor agonist selected from the list consisting of poly(I:C), Poly(dA:dT), Poly(dG:dC), 3p-hpRNA, 5′ppp-dsRNA, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a C-type lectin receptor agonist selected from the list consisting of Beta-1,3-glucan, zymosan, Heat-killed C. albicans, cord factor, and Trehalose-6,6-dibehenate, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a costimulatory molecule agonist selected from the list consisting of a CD40 agonist, aCD80 agonist, a CD86 agonist, an OX40 agonist, and combinations thereof. In certain embodiments, the CD40 agonist is an anti-CD40 agonistic antibody. In certain embodiments, the anti-CD40 agonistic antibody comprises dacetuzumab, CP-870,893, ADC-1013, 2141-v11, APX005M, Chi Lob 7/4, BG9588 (NIAMS), CFZ533, PG10, BMS-986004, lucatumumab, HCD122, JNJ-64457107, selicrelumab, ASKP1240, or SEA-CD40. In certain embodiments, the dendritic cell activating molecule is a cytokine selected from the list consisting of granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-15 (IL-15), tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a STING agonist selected from the list consisting of 2′,3′-cGAMP (CAS Number, 1441190-66-4), 4-[(2-Chloro-6-fluorophenyl)methyl]-N-(furan-2-ylmethyl)-3-oxo-1,4-benzothiazine-6-carboxamide, MK-1454, ADU-S100/MIW815, SRCB-0074, SYNB1891, E-7766, or SB11285, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is administered to a tumor being treated with the dose of the radiation therapy. In certain embodiments, the tumor or cancer is a solid tissue tumor or cancer. In certain embodiments, the solid tumor or cancer is of breast, prostate, or a melanoma.
Described herein in one aspect is a method of treating a tumor or a cancer in an individual, the method comprising administering to the individual a dose of an energy-based therapy and a dendritic cell activating molecule, wherein the dose of the energy-based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy. Also described is a method of treating a tumor or a cancer in an individual, the method comprising administering to the individual a dendritic cell activating molecule, wherein the individual has been administered a dose of an energy-based therapy, wherein the dose of the energy-based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy. In certain embodiments, the dose of the energy base therapy comprises a plurality of doses of energy-based therapy. In certain embodiments, the energy-based therapy is Irreversible Electroporation (IRE). In certain embodiments, the energy-based therapy is microwave therapy In certain embodiments, the energy-based therapy is Low-Intensity Focused Ultrasound (LOFU). In certain embodiments, the LOFU is administered at an intensity of between 10 and 1000 W/cm2 in the area of treatment. In certain embodiments, the energy-based therapy is High-Intensity Focused Ultrasound (HIFU). In certain embodiments, the energy-based therapy is cryotherapy. In certain embodiments, the dendritic cell activating molecule is administered at least three days after the dose of the energy-based therapy. In certain embodiments, the dendritic cell activating molecule is administered at least five days after the dose of the energy-based therapy. In certain embodiments, the dendritic cell activating molecule is administered at least seven days after the dose of the energy-based therapy. In certain embodiments, the dendritic cell activating molecule activates maturation of an immature dendritic cell. In certain embodiments, the dendritic cell activating molecule activates dendritic cell activation through a toll-like receptor, a NOD-like receptor, a RIG-1 or MDA-5 receptor, a C-type lectin receptor, a costimulatory molecule, a cytokine receptor, or a STING pathway. In certain embodiments, the dendritic cell activating molecule is a toll-like receptor agonist selected from the list consisting of CpG oligonucleotide, SD-101, LFX453, imiquimod, Bacillus Calmette-Guérin (BCG), monophosphoryl lipid A, Poly ICLC, GSK1795091, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a NOD-like receptor agonist selected from the list consisting of bacterial peptidoglycan, an acylated derivative of iE-DAP (C12-iE-DAP), D-gamma-Glu-mDAP (iE-DAP), L-Ala-gamma-D-Glu-mDAP (Tri-DAP), muramyl dipeptide (MDP), muramyl tripeptide, L18-MDP, M-TriDAP, murabutide, PGN-ECndi, PGN-ECndss, PGN-SAndi, N-glycolylated muramyl dipeptide, murabutide, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a RIG-1 or MDA-5 receptor agonist selected from the list consisting of poly(I:C), Poly(dA:dT), Poly(dG:dC), 3p-hpRNA, 5′ppp-dsRNA, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a C-type lectin receptor agonist selected from the list consisting of Beta-1,3-glucan, zymosan, Heat-killed C. albicans, cord factor, and Trehalose-6,6-dibehenate, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a costimulatory molecule agonist selected from the list consisting of a CD40 agonist, aCD80 agonist, a CD86 agonist, an OX40 agonist, and combinations thereof. In certain embodiments, the CD40 agonist is an anti-CD40 agonistic antibody. In certain embodiments, the anti-CD40 agonistic antibody comprises dacetuzumab, CP-870,893, ADC-1013, 2141-v11, APX005M, Chi Lob 7/4, BG9588 (NIAMS), CFZ533, PG10, BMS-986004, lucatumumab, HCD122, JNJ-64457107, selicrelumab, ASKP1240, or SEA-CD40. In certain embodiments, the dendritic cell activating molecule is a cytokine selected from the list consisting of granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-15 (IL-15), tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a STING agonist selected from the list consisting of 2′,3′-cGAMP (CAS Number, 1441190-66-4), 4-[(2-Chloro-6-fluorophenyl)methyl]-N-(furan-2-ylmethyl)-3-oxo-1,4-benzothiazine-6-carboxamide, MK-1454, ADU-S100/MIW815, SRCB-0074, SYNB1891, E-7766, or SB11285, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is administered to a tumor being treated with the dose of the energy-based therapy. In certain embodiments, the tumor or cancer is a solid tissue tumor or cancer. In certain embodiments, the solid tumor or cancer is of breast, prostate, or a melanoma.
In one aspect described herein is a method of increasing T cell infiltration into a tumor distal to a tumor being treated in an individual, the method comprising administering to the individual a dose of a radiation therapy and a dendritic cell activating molecule, wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy is administered. In certain embodiments, the dendritic cell activating molecule is administered at least two days after the radiation therapy is administered. In certain embodiments, the dendritic cell activating molecule is administered at least three days after the radiation therapy is administered. In certain embodiments, the dose of the radiation therapy comprises a plurality of doses of radiation therapy. In certain embodiments, the radiation therapy is external beam radiation therapy. In certain embodiments, the external beam radiation therapy is selected from the list consisting of: three-dimensional conformal radiation therapy, intensity modulated radiation therapy, image guided radiation therapy, stereotactic radiation therapy, intraoperative radiation therapy, proton beam therapy, neutron beam therapy, and combinations thereof. In certain embodiments, the dose of radiation therapy comprises at least about 2 Gy. In certain embodiments, the dose of radiation therapy comprises at least about 2 Gy and no more than about 20 Gy. In certain embodiments, the dendritic cell activating molecule is administered at least three days after the dose of the radiation therapy. In certain embodiments, wherein the dendritic cell activating molecule is administered at least five days after the dose of the radiation therapy. In certain embodiments, the dendritic cell activating molecule is administered at least seven days after the dose of the radiation therapy. In certain embodiments, the dendritic cell activating molecule activates maturation of an immature dendritic cell. In certain embodiments, wherein the dendritic cell activating molecule activates dendritic cell activation through a toll-like receptor, a NOD-like receptor, a RIG-1 or MDA-5 receptor, a C-type lectin receptor, a costimulatory molecule, a cytokine receptor, or a STING pathway. In certain embodiments, the dendritic cell activating molecule is a toll-like receptor agonist selected from the list consisting of CpG oligonucleotide, SD-101, LFX453, imiquimod, Bacillus Calmette-Guérin (BCG), monophosphoryl lipid A, Poly ICLC, GSK1795091, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a NOD-like receptor agonist selected from the list consisting of bacterial peptidoglycan, an acylated derivative of iE-DAP (C12-iE-DAP), D-gamma-Glu-mDAP (iE-DAP), L-Ala-gamma-D-Glu-mDAP (Tri-DAP), muramyl dipeptide (MDP), muramyl tripeptide, L18-MDP, M-TriDAP, murabutide, PGN-ECndi, PGN-ECndss, PGN-SAndi, N-glycolylated muramyl dipeptide, murabutide, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a RIG-1 or MDA-5 receptor agonist selected from the list consisting of poly(I:C), Poly(dA:dT), Poly(dG:dC), 3p-hpRNA, 5′ppp-dsRNA, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a C-type lectin receptor agonist selected from the list consisting of Beta-1,3-glucan, zymosan, Heat-killed C. albicans, cord factor, and Trehalose-6,6-dibehenate, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a costimulatory molecule agonist selected from the list consisting of a CD40 agonist, aCD80 agonist, a CD86 agonist, and combinations thereof. In certain embodiments, the CD40 agonist is an anti-CD40 agonistic antibody. In certain embodiments, the anti-CD40 agonistic antibody comprises dacetuzumab, CP-870,893, ADC-1013, 2141-v11, APX005M, Chi Lob 7/4, BG9588 (NIAMS), CFZ533, PG10, BMS-986004, lucatumumab, HCD122, JNJ-64457107, selicrelumab, ASKP1240, or SEA-CD40. In certain embodiments, the dendritic cell activating molecule is a cytokine selected from the list consisting of granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-15 (IL-15), tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), and combinations thereof. In certain embodiments, the dendritic cell activating molecule is a STING agonist selected from the list consisting of 2′,3′-cGAMP (CAS Number, 1441190-66-4), 4-[(2-Chloro-6-fluorophenyl)methyl]-N-(furan-2-ylmethyl)-3-oxo-1,4-benzothiazine-6-carboxamide, MK-1454, ADU-S100/MIW815, SRCB-0074, SYNB1891, E-7766, or SB11285, and combinations thereof. In certain embodiments, the dendritic cell activating molecule is administered to a tumor being treated with the dose of the radiation therapy. In certain embodiments, the tumor is a solid tumor. In certain embodiments, the solid tumor is a breast tumor, a prostate tumor, or a melanoma.
Disclosed herein is a method of treating a tumor or a cancer in an individual by administering a dendritic cell activating molecule to an individual at least one day after treatment with either radiation therapy or an energy therapy. Both radiation therapy and energy therapies treat tumors and cancers in individuals by killing or damaging the cancer cells. The addition of administering a dendritic cell activating molecule activates the dendritic cells of the individual’s immune system and aids in treating the tumor or cancer.
In one aspect described herein is a method comprising administering to the individual a dose of a radiation therapy and a dendritic cell activating molecule, wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy is administered. In another aspect described herein is a method comprising administering to the individual a dendritic cell activating molecule, wherein the individual has received a dose of a radiation therapy, and wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy has been administered.
In one aspect described herein is a method comprises administering to the individual a dose of an energy-based therapy and a dendritic cell activating molecule, wherein the dose of the energy-based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy. In another aspect described herein is a method comprising administering to the individual a dendritic cell activating molecule, wherein the individual has been administered a dose of an energy-based therapy, wherein the dose of the energy based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy.
This application also discloses a method of increasing T cell infiltration into a tumor distal to a tumor being treated in an individual. In one aspect described herein is a method comprising administering to the individual a dose of a radiation therapy and a dendritic cell activating molecule, wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy is administered. In another aspect described herein is a method comprising administering to the individual a dose of an energy-based therapy and a dendritic cell activating molecule, wherein the dose of the energy-based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy.
In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments. However, one skilled in the art will understand that the embodiments provided may be practiced without these details. Unless the context requires otherwise, throughout the specification and claims which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to.” As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the content clearly dictates otherwise. It should also be noted that the term “or” is generally employed in its sense including “and/or” unless the content clearly dictates otherwise. Further, headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed embodiments.
“Consisting essentially of” when used to define compositions and methods, shall mean excluding other elements of any essential significance to the combination for the stated purpose. Thus, a composition consisting essentially of the elements as defined herein would not exclude other materials or steps that do not materially affect the basic and novel characteristic(s) of the claimed invention. Compositions for treating or preventing a given disease can consist essentially of the recited active ingredient, exclude additional active ingredients, but include other non-material components such as excipients, carriers, or diluents. “Consisting of” shall mean excluding more than trace elements of other ingredients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this disclosure.
As used herein the term “about” refers to an amount that is near the stated amount by 10%.
As used herein the terms “individual,” “patient,” or “subject” are used interchangeably and refer to individuals diagnosed with, suspected of being afflicted with, or at-risk of developing at least one disease for which the described compositions and method are useful for treating. In certain embodiments the individual is a mammal. In certain embodiments, the mammal is a mouse, rat, rabbit, dog, cat, horse, cow, sheep, pig, goat, llama, alpaca, or yak. In certain embodiments, the individual is a human.
As used herein the term “treat” or “treating” refers to interventions to a physiological or disease state of an individual designed or intended to ameliorate at least one sign or symptom associated with said physiological or disease state. The skilled artisan will recognize that given a heterogeneous population of individuals afflicted with a disease, not all individuals will respond equally, or at all, to a given treatment.
The terms “polypeptide” and “protein” are used interchangeably to refer to a polymer of amino acid residues, and are not limited to a minimum length. Polypeptides, including the provided antibodies and antibody chains and other peptides, e.g., linkers and binding peptides, may include amino acid residues including natural and/or non-natural amino acid residues. The terms also include post-expression modifications of the polypeptide, for example, glycosylation, sialylation, acetylation, phosphorylation, and the like. In some aspects, the polypeptides may contain modifications with respect to a native or natural sequence, as long as the protein maintains the desired activity. These modifications may be deliberate, as through site-directed mutagenesis, or may be accidental, such as through mutations of hosts which produce the proteins or errors due to PCR amplification.
The term “radiotherapy” or “radiation therapy” means the treatment of an individual with ionizing radiation. Exemplary types of radiation therapy include without limitation three-dimensional conformal radiation therapy, intensity modulated radiation therapy, image guided radiation therapy, stereotactic radiation therapy, intraoperative radiation therapy, proton beam therapy, and neutron beam therapy.
The term “energy-based therapy” means the treatment of an individual with a form of energy, including without limitations electrical currents, electromagnetic waves, and temperature. Exemplary types of energy-based therapy include without limitation Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy.
The term “immune cell” refers to a cell that plays a role in the immune response and originates from a hematopoietic precursor. Without limitation, immune cells include lymphocytes, such as B cells and T cells; natural killer cells; and myeloid cells, such as monocytes, macrophages, eosinophils, mast cells, basophils, dendritic cells, and granulocytes.
The term “dendritic cell” refers to an antigen-presenting cell of the immune system of hematopoietic origin. Dendritic cells can be characterized by the expression of class II MHC, CD11c and CD86. Dendritic cells include without limitation activated dendritic cells, non-activated dendritic cells, mature dendritic cells, and immature dendritic cells.
The term “dendritic cell activating molecule” refers to a molecule that increases the immunological activity of dendritic cells as compared to the dendritic cell activity prior to exposure to the activating agent. Changes in the immunological activity of dendritic cells may include without limitation changes to antigen presentation, migration to lymph nodes, interaction with T cells and B cells, T-cell priming, cytokine release, and chemokine release. Examples of dendritic cell activating molecules include, without limitation, CD40L, an anti-CD40 agonist antibody, a TLR activator, a NOD-like receptor agonist, a RIG-1 receptor agonist, an MDA-5 receptor agonist, a C-type lectin receptor agonist, a STING activator, a costimulatory molecule or a cytokine receptor. Other suitable activating molecules useful in the practice of the methods described herein include a RANKL peptide, TNF peptide, IL-1 peptide, CpG-rich DNA sequences, lipopolysaccharide (LPS), RIG1 helicase ligand, RNA, dsDNA or variations thereof (e.g., polypeptides or DNA sequences comprising one or more insertions, substitutions, or deletions).
The term “antibody” as used herein refers to polypeptides comprising at least one antibody derived antigen binding site (e.g., VH/VL region or Fv, or CDR), and includes whole antibodies and any antigen binding fragments (i.e., “antigen-binding portions” or antigen binding fragments thereof) or single chains thereof. Antibodies include known forms of antibodies. For example, the antibody can be a human antibody, a humanized antibody, a bispecific antibody, or a chimeric antibody. A “whole antibody” refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, in which each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region; and each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. An “antigen-binding fragment” includes without limitations Fab, Fab′, F(ab′)2, scFv, Fv, recombinant IgG, and heavy chain antibodies.
The term “tumor,” or “cancer” as used herein, and unless otherwise specified, refers to a neoplastic cell growth, and includes pre-cancerous and cancerous cells and tissues. Tumors usually present as a lesion or lump. As used herein, “treating” a tumor means that one or more symptoms of the disease, such as the tumor itself, vascularization of the tumor, or other parameters by which the disease is characterized, are reduced, ameliorated, inhibited, placed in a state of remission, or maintained in a state of remission. “Treating” a tumor also means that one or more hallmarks of the tumor may be eliminated, reduced or prevented by the treatment. Non-limiting examples of such hallmarks include uncontrolled degradation of the basement membrane and proximal extracellular matrix, migration, division, and organization of the endothelial cells into new functioning capillaries, and the persistence of such functioning capillaries.
The methods described herein comprise or consist essentially of administering a radiation therapy and a dendritic cell activator to an individual in need thereof. Any of the radiation therapies described herein can be administered either alone or in combination. Radiation therapies described herein can be administered either singly or as plurality of doses.
In general, radiation therapy, radio-immunotherapy or pre-targeted radioimmunotherapy are used for the treatment of diseases of oncological nature. “Radiotherapy” or radiation therapy means the treatment of cancer and other diseases with ionizing radiation. Ionizing radiation deposits energy that injures or destroys cells in the area being treated (the target tissue) by damaging their genetic material, making it impossible for these cells to continue to grow. Radiotherapy may be used to treat localized solid tumors, such as cancers of the skin, tongue, larynx, brain, breast, lung, liver, kidney, pancreas, or uterine cervix. It can also be used to treat leukemia and lymphoma, i.e. cancers of the blood-forming cells and lymphatic system, respectively. In certain aspects of the methods disclosed herein, radiation therapy is used to treat a tumor.
Ionizing radiation is widely used for the treatment of solid tumors. Several types of ionizing radiation can be used, including X-rays and gamma rays. Radiotherapy can be applied using a machine to focus the radiation on the tumor, or by placing radioactive implants directly into the tumor or in a nearby body cavity. Moreover, radiolabeled antibodies can be used to target tumor cells. Other radiotherapy techniques may also be used in the methods described herein, including intraoperative irradiation, particle beam radiation, as well as the use of radiosensitizers to make tumor cells more sensitive to radiation, or radioprotectants to protect normal cells.
One type of radiation therapy commonly used involves photons, e.g. X-rays. Depending on the amount of energy they possess, the rays can be used to destroy cancer cells on the surface of or deeper in the body. The higher the energy of the x-ray beam, the deeper the x-rays can go into the target tissue. Linear accelerators and betatrons are machines that produce x-rays of increasingly greater energy. The use of machines to focus radiation (such as x-rays) on a cancer site is called external beam radiotherapy. In one embodiment of the methods, external beam radiotherapy is used.
Three-dimensional conformal radiation therapy, intensity-modulated radiation therapy, and image-guided radiation therapy are methods of external beam radiotherapy that allow for more precise targeting of the tumor while avoiding more of the surrounding healthy issue. The increased precision allows for higher levels of radiation, which is more effective in shrinking and killing tumors. In three -dimensional conformal radiation therapy, targeting information is used to shape the radiation beam to the shape of the tumor. In image-guided radiation therapy, computer-controlled linear accelerators are used to target specific areas within a tumor. This method allows the radiation dose to more closely match the shape of the tumor by controlling the intensity of the beam in multiple small volumes. Image-guided radiation therapy uses imaging during the radiation therapy to improve the precision and accuracy of treatment. Imaging methods include but are not limited to fiducial markers, ultrasound, MRI, x-ray images, CT-scan, 3-D body surface mapping, electromagnetic transponders, or colored tattoos. Image-guided radiation therapy is especially useful in tumors located in areas of the body that move, such as the lungs. In one embodiment of the methods, three-dimensional conformal radiation therapy is used. In another embodiment of the methods, intensity-modulated radiation therapy is used. In another embodiment of the methods, image-guided radiation therapy is used.
High dose radiotherapy, such as stereotactic ablative radiotherapy (SABR) or stereotactic body radiation therapy (SBRT), is another method of external beam radiation radiotherapy. Higher doses, in the range of 15 to 20 Gy are used than in convention radiotherapy. One type of SABR is stereotactic radiosurgery (SRS), which has been used for small intracranial tumors that was made possible by technology allowing for submillimeter delivery precision and steep dose gradients beyond the tumor target. SABR (or SBRT) has been developed for use on tumors outside of the brain and includes tumors of practically every major body site (e.g., lung tumors). In one embodiment, the external beam radiation therapy is stereotactic ablative radiotherapy.
Another method of external beam radiotherapy is intraoperative irradiation, in which a large dose of external radiation is directed at the tumor and surrounding tissue during surgery. In one embodiment, the external beam radiation is intraoperative irradiation.
Gamma rays are another form of photons used in radiotherapy. Gamma rays are produced spontaneously as certain elements (such as radium, uranium, and cobalt 60) release radiation as they decompose or decay. In one embodiment, the external beam radiation is gamma ray radiation.
Another approach is particle beam radiation therapy. This type of therapy differs from photon radiotherapy in that it involves the use of fast-moving subatomic particles to treat localized cancers. This includes, but is not limited to, proton beam therapy, neutron beam therapy, pion beam therapy, and heavy ion beam therapy. Some particles (neutrons, pions, and heavy ions) deposit more energy along the path they take through tissue than do x-rays or gamma rays, thus causing more damage to the cells they hit. This type of radiation is often referred to as high linear energy transfer (high LET) radiation. Radio-sensitizers make the tumor cells more likely to be damaged, and radio-protectors protect normal tissues from the effects of radiation. In one embodiment, the external beam radiation is selected from the list consisting of proton beam therapy, neutron beam therapy, pion beam therapy, and heavy ion beam therapy. In one embodiment the external beam radiation used is proton beam therapy. In another embodiment, the external beam therapy used is neutron beam therapy. In another embodiment, the external beam therapy used is pion beam therapy. In another embodiment, the external beam therapy used is heavy ion beam therapy. In one embodiment, the external beam radiation therapy is selected from the list consisting of: three-dimensional conformal radiation therapy, intensity modulated radiation therapy, image guided radiation therapy, stereotactic radiation therapy, intraoperative radiation therapy, proton beam therapy, neutron beam therapy, and combinations thereof.
Another technique for delivering radiation to cancer cells is to place radioactive implants directly in a tumor or body cavity. This is called internal radiotherapy. Brachytherapy, interstitial irradiation, and intracavitary irradiation are types of internal radiotherapy. In this treatment, the radiation dose is concentrated in a small area, and the patient stays in the hospital for a few days. Internal radiotherapy is frequently used for cancers of the tongue, uterus, and cervix. In one embodiment, internal radiotherapy is used. In another embodiment, the internal radiotherapy is selected from the list comprising brachytherapy, interstitial irradiation, and intracavitary irradiation, or combinations thereof.
In certain cases, the total irradiation dose can be spread over several sessions (i.e., dose fractionation) and can be spaced by at least 6 hours, days, or even weeks. Conventional definitive radiation treatment involves multiple treatments, generally 20-40, with low doses (<2- 3 Gy) stretching over weeks. In certain cases, such as high doses radiotherapy discussed above, the dose is greater than 15-20 Gy and is given is up to 5 treatments.
Certain aspects of the method disclosed herein comprise treating a patient with radiotherapy. In one embodiment, the method includes a plurality of doses of radiation therapy. In one embodiment, the method includes at least 2 doses of radiation therapy. In another embodiment, the method includes at least 3 doses of radiation therapy. In another embodiment, the method includes at least 4 doses of radiation therapy. In another embodiment, the method includes at least 5 doses of radiation therapy. In another embodiment the method includes at least 6 doses of radiation therapy. In another embodiment, the method includes at least 7 doses of radiation therapy. In another embodiment, the method includes at least 8 doses of radiation therapy. In another embodiment, the method includes at least 9 doses of radiation therapy. In another embodiment, the method includes at least 10 doses of radiation therapy. In another embodiment, the method includes at least 11 doses of radiation therapy. In another embodiment, the method includes at least 12 doses of radiation therapy. In another embodiment, the method includes at least 13 doses of radiation therapy. In another embodiment, the method includes at least 14 doses of radiation therapy. In another embodiment, the method includes at least 15 doses of radiation therapy. In another embodiment, the method includes at least 20 doses of radiation therapy. In another embodiment, the method includes at least 25 doses of radiation therapy. In another embodiment, the method includes at least 30 doses of radiation therapy. In another embodiment, the method includes at least 35 doses of radiation therapy. In another embodiment, the method includes at least 40 doses of radiation therapy. In another embodiment, the method includes at least 45 doses of radiation therapy. In another embodiment, the method includes at least 50 doses of radiation therapy.
In one aspect of the methods described herein, the radiation therapy uses ionizing radiation for treating cancer in a subject. In one embodiment, the dose of radiation therapy is at least about 2 Gy. In another embodiment, the dose of radiation therapy is at least about 3 Gy. In another embodiment, the dose of radiation therapy is at least about 4 Gy. In another embodiment, the dose of radiation therapy is at least about 5 Gy. In another embodiment, the dose of radiation therapy is at least about 6 Gy. In another embodiment, the dose of radiation therapy is at least about 7 Gy. In another embodiment, the dose of radiation therapy is at least about 8 Gy. In another embodiment, the dose of radiation therapy is at least about 9 Gy. In another embodiment, the dose of radiation therapy is at least about 10 Gy. In another embodiment, the dose of radiation therapy is at least about 15 Gy. In another embodiment, the dose of radiation therapy is at least about 20 Gy. In another embodiment, the dose of radiation therapy is at least about 25 Gy. In another embodiment, the dose of radiation therapy is at least about 30 Gy. In another embodiment, the dose of radiation therapy is at least about 40 Gy. In another embodiment, the dose of radiation therapy is at least about 50 Gy. In another embodiment, the dose of radiation therapy is at least about 60 Gy. In another embodiment, the dose of radiation therapy is at least about 70 Gy. In another embodiment, the dose of radiation therapy is at least about 80 Gy. In another embodiment, the dose of radiation therapy is at least about 90 Gy. In another embodiment, the dose of radiation therapy is at least about 100 Gy.
In one embodiment, the total radiation dose for a cycle of treatment is between 5 and 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 10 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 20 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 30 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 40 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 50 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 60 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 70 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 80 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 90 and about 100 Gy. In another embodiment, the total radiation dose for a cycle of treatment is about 100 Gy.
In one embodiment, the total radiation dose for a cycle of treatment is between about 20 to about 50 Gy. In one embodiment, the total radiation dose for a cycle of treatment is between about 20 to about 50 Gy on one occasion. In another embodiment, the total radiation dose for a cycle of treatment is between about 20 to about 50 Gy on each of two occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 10 to about 30 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 10 to about 30 Gy on one occasion. In another embodiment, the total radiation dose for a cycle of treatment is between about 10 to about 30 Gy on each of two occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 10 to about 30 Gy on each of three occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 10 to about 30 Gy on each of four occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 10 to about 30 Gy on each of five occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 10 to about 30 Gy on each of two to four occasions.
In another embodiment, the total radiation dose for a cycle of treatment is between about 5 and about 20 Gy. In another embodiment, the total radiation dose for a cycle of treatment is between about 5 and about 20 Gy on one occasion. In another embodiment, the total radiation dose for a cycle of treatment is between about 5 and about 20 Gy on each of two occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 5 and about 20 Gy on each of three occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 5 and about 20 Gy on each of four occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 5 and about 20 Gy on each of five occasions. In a certain embodiments, the total radiation dose for a cycle of treatment is between about 20 and about 50 Gy on one occasion, between about 10 and about 30 Gy on each of two to four occasions, or between about 5 and about 20 Gy on each of 5 occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 30 to about 40 Gy on one occasion. In another embodiment, the total radiation dose for a cycle of treatment is between about 30 to about 40 Gy on each of two occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 15 to about 20 Gy on one occasion. In another embodiment, the total radiation dose for a cycle of treatment is between about 15 to about 20 Gy on each of two occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 15 to about 20 Gy on each of three occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 15 to about 20 Gy on each of four occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 8 to about 12 Gy on one occasion. In another embodiment, the total radiation dose for a cycle of treatment is between about 8 to about 12 Gy on each of two occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 8 to about 12 Gy on each of three occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 8 to about 12 Gy on each of four occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 8 to about 12 Gy on each of five occasions. In another embodiment, the total radiation dose for a cycle of treatment is between about 8 to about 12 Gy on each of six occasions. In a certain embodiments, the total radiation dose for a cycle of treatment is between about 30 to about 40 Gy on one occasion, about 15 to about 20 Gy on each of three occasions, or about 8 to about 12 Gy on each of 5 occasions.
The methods described herein may also be combined with post-ablation modulation (PAM) after high dose radiation. PAM can be administered from about 0.1 Gy to about 2 Gy, from about 0.1 Gy to about 1 Gy, from about 0.2 to about to about 2 Gy, from about 0.1 to about to about 0.8 Gy, from about 0.1 to about to about 0.6 Gy, from about 0.2 to about to about 0.6 Gy, from about 0.4 to about to about 0.6 Gy. PAM can be administered at about 0.1, 0.2, 0.4, 0.5, 0.6, 0.8, or 1.0 Gy. PAM can be administered for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more doses.
The methods described herein comprise or consist essentially of administering an energy-based therapy and a dendritic cell activator to an individual in need thereof. Any of the energy based therapies described herein can be administered either alone or in combination. Energy-based therapies described herein can be administered either singly or a plurality of times.
A variety of energy-based therapies can be administered to treat cancer. These methods use electromagnetic waves, electromagnetic currents or temperature to kill or damage cancer or tumor cells. These include, but are not limited to, Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy. In one aspect of the methods disclosed herein, the dose of the energy-based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy.
Certain aspects of the methods disclosed herein involve treating a patient with an energy-based therapy. In one embodiment, the dose of the energy-based therapy comprises a plurality of doses of energy-based therapy. In one embodiment, the dose of the energy-based therapy may comprise at least 2 doses. In another embodiment, the dose of the energy-based therapy may comprise at least 3 doses. In another embodiment, the dose of the energy-based therapy may comprise at least 4 doses. In another embodiment, the dose of the energy-based therapy may comprise at least 5 doses. In another embodiment, the dose of the energy-based therapy may comprise at least 6 doses. In another embodiment, the dose of the energy-based therapy may comprise at least 7 doses. In another embodiment, the dose of the energy-based therapy may comprise at least 8 doses. In another embodiment, the dose of the energy-based therapy may comprise at least 9 doses. In another embodiment, the dose of the energy-based therapy may comprise at least 10 doses. In another embodiment, the dose of the energy-based therapy may comprise more than 10 doses.
Irreversible Electroporation (IRE) is a method of treating a tumor that uses electrical currents to damage and destroy cancer cells. Electrodes are placed around the tumor and a current is delivered through the electrodes. The application of the current results in permeabilization of the cell membrane, resulting in apoptosis of the cancer cells. In one embodiment, the energy-based therapy is Irreversible Electroporation (IRE).
Treatment of localized tumors by focused ultrasound (FUS) is an image guided minimally invasive therapy that uses a range of input energy for in situ tumor ablation. The application of FUS to biological tissues is associated with the generation of thermal and cavitation effects, causing changes in target cell physiology, depending on the energy delivered. High intensity focused ultrasound (HIFU) has been used clinically to thermally ablate localized tumors. The substantial thermal energy generated by that modality of FUS treatment causes rapid coagulative necrosis of the tissue at the targeted focal spots. Though several studies have reported some immunomodulatory effects, including increased lymphocyte infiltration, generation of IFNγ producing tumor-specific T cells in lymphoid organs and dendritic cell maturation and migration into tumors, the thermally induced coagulative necrosis resulting from HIFU treatment can also attenuate the release of immunostimulatory molecules within the tumor microenvironment. Thus, although able to halt the progression of established primary tumors, HIFU might fail to protect against local and distant metastases arising from the surviving tumor cells. In one embodiment, the energy-based therapy is High-Intensity Focused Ultrasound (HIFU).
In some embodiments, HIFU is administered with an intensity of about 100 to about 10000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 1000 to about 2000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 2000 to about 3000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 3000 to about 4000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 4000 to about 5000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 5000 to about 6000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 6000 to about 7000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 7000 to about 8000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 8000 to about 9000 W/cm2 in the area of treatment. In some embodiments, HIFU is administered with an intensity of about 9000 to about 10000 W/cm2 in the area of treatment.
Low energy non-ablative focused ultrasound, or LOFU is an ultrasound treatment, generated using a concave transducer to focus the ultrasound in a treatment zone. Methods and systems for treatment of cancer with LOFU are described in US 202003/98084 and U.S. 10,974,077, which are herein incorporated by reference. LOFU produces mild mechanical and thermal stress in tumor cells, while avoiding cavitation and coagulative necrosis both of which result in tissue damage. A non-ablative “sonic” stress response is induced in the tumor that increases the expression of heat shock proteins without actually killing them directly. In one embodiment, the energy-based therapy is Low-Intensity Focused Ultrasound (LOFU).
In some embodiments, LOFU involves the application of ultrasound at an acoustic power between 10 and 1000 W/cm2 spatial peak temporal average intensity (Ispta) in a treatment zone, with the ultrasound applied continuously for a time in the range of 0.5 to 5 seconds, wherein the frequency is in the range of 0.01 to 10 MHz and the mechanical index is less than 4. Mechanical Index (MI) is the rarefaction pressure in units of MPa over the square root of the central frequency in units of MHz. The energy and intensity of ultrasound applied is intended to fall between energies and intensities of ultrasound that either induce primarily ablative effects or primarily diagnostic effects.
In some embodiments, the LOFU includes a transducer that generates acoustic power between 10 and 1000 W/cm2 spatial peak temporal average intensity (Ispta) in a treatment zone. The ultrasound is applied continuously for a time in the range of 0.5 to 5 seconds or pulsed with pulse durations of 1 to 100 ms, wherein the frequency is in the range of 0.01 to 10 MHz. In some embodiments the frequency is in the range of 0.05 to 5 MHz. In some embodiments the frequency range is from 0.1 to 2 MHz. In some embodiments the minimum diameter of any ultrasound beam in the treatment zone is about 1 cm. In an embodiment, the LOFU is administered at 10 to 1000 W/cm2 in the area of treatment. In an embodiment, the LOFU is administered at 10 to 100 W/cm2 Ispta in the area of treatment. In an embodiment, the LOFU is administered at 100 to 200 W/cm2 Ispta in the area of treatment. In an embodiment, the LOFU is administered at 300 to 400 W/cm2 Ispta in the area of treatment. In an embodiment, the LOFU is administered at 400 to 500 W/cm2 Ispta in the area of treatment. In an embodiment, the LOFU is administered at 500 to 600 W/cm2 Ispta in the area of treatment. In an embodiment, the LOFU is administered at 600 to 700 W/cm2 Ispta in the area of treatment. In an embodiment, the LOFU is administered at 700 to 800 W/cm2 Ispta in the area of treatment. In an embodiment, the LOFU is administered at 800 to 900 W/cm2 Ispta in the area of treatment. In an embodiment, the LOFU is administered at 900 to 1000 W/cm2 Ispta in the area of treatment. In an embodiment, the ultrasound is applied for a time in the range of 0.5 to 1 second. In an embodiment, the ultrasound is applied for a time in the range of 1 to 2 seconds. In an embodiment, the ultrasound is applied for a time in the range of 2 to 3 seconds. In an embodiment, the ultrasound is applied for a time in the range of 4 to 5 seconds. In embodiment, the ultrasound is applied at a frequency of 0.01 to 1 MHz. In embodiment, the ultrasound is applied at a frequency of 1 to 2 MHz. In embodiment, the ultrasound is applied at a frequency of 2 to 3 MHz. In embodiment, the ultrasound is applied at a frequency of 3 to 4 MHz. In embodiment, the ultrasound is applied at a frequency of 4 to 5 MHz. In embodiment, the ultrasound is applied at a frequency of 5 to 6 MHz. In embodiment, the ultrasound is applied at a frequency of 6 to 7 MHz. In embodiment, the ultrasound is applied at a frequency of 7 to 8 MHz. In embodiment, the ultrasound is applied at a frequency of 8 to 9 MHz. In embodiment, the ultrasound is applied at a frequency of 9 to 10 MHz.
Both microwave therapy and radiofrequency therapy are methods that create localized heat regions to destroy tumors. In radiofrequency therapy, high frequency electrical currents are passed through an electrode placed in a tumor. This creates a small region of heat. In microwave therapy, a needle placed in the tumor creates microwaves which then create a small region of heat. In both treatment methods, the cancer cells within the localized heat region are damaged or destroyed. In one embodiment, the energy-based therapy is microwave therapy. In another embodiment, the energy-based therapy is radiofrequency therapy.
In contrast, cryotherapy is an energy-based therapy uses extreme cold to destroy cancer tissue. Intense cold is created, usually by applying either liquid nitrogen or pressurized argon gas to a localized site. Cells and tissues that encounter the cold are killed. This method can be used on both internal and external tumors. In one embodiment, the energy-based therapy is cryotherapy.
The methods described herein comprise or consist essentially of administering: (a) a radiation therapy, an energy based therapy, or a combination thereof; and (b) a dendritic cell activator to an individual in need thereof. Any of the radiation therapies or energy-based therapies described herein can be administered either alone or in combination. Radiation or energy-based therapies described herein can be administered either singly or a plurality of times.
Administration of the dendritic cell activating therapy may be administered at such time as the T cells associated with a with a radiation or energy treated tumor have recovered from the effects of the treatment. Without being bound by theory administration of radiation or energy based therapies disproportionately harms rapidly dividing cells, such as immune cells, and an interval between the administration of a radiation or energy based therapy and a dendritic cell activator may be beneficial to subsequent immune response.
With regard to the timing of a subsequent administration the radiation or energy-based therapy is considered administered on day 0, with the next day after the treatment comprising 1 day after the therapy. Additionally, the amount of days after administration is calculated from the temporally most recent doe of the therapy. Therefore, for example, if an individual is administered a plurality of doses of radiation or energy-based therapy the interval for administration of a dendritic cell activating therapy is calculated based upon the last dose of the plurality before the dendritic cell activating therapy is administered.
In one aspect of the methods disclosed herein, the methods comprise administering a dendritic cell activating molecule after radiation therapy. In one embodiment, the dendritic cell activating molecule is administered at least 1 day after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 2 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 3 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 4 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 5 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 6 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 7 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 8 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 9 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 10 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 11 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 12 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 13 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered at least 14 days after radiation therapy. In another embodiment, the dendritic cell activating molecule is administered more than 14 days after radiation therapy.
In one embodiment, the dendritic cell activating molecule is administered between 1 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 2 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 3 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 4 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 5 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 6 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 7 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 8 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 9 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 10 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 11 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 12 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 13 and 14 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 1 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 2 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 3 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 4 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 5 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 6 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 7 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 8 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 9 and 10 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 1 and 7 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 2 and 7 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 3 and 7 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 4 and 7 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 5 and 7 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 6 and 7 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 1 and 5 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 2 and 5 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 3 and 5 days after radiation treatment. In another embodiment, the dendritic cell activating molecule is administered between 4 and 5 days after radiation treatment.
In one aspect of the methods disclosed herein, the methods comprise administering the dendritic cell activating molecules after a dose of an energy-based therapy. In one embodiment, the dendritic cell activating molecule is administered at least 1 day after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 2 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 3 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 4 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 5 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 6 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 7 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 8 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 9 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 11 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 12 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 13 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered at least 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered more than 14 days after a dose of the energy-based therapy.
In one embodiment, the dendritic cell activating molecule is administered between 1 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 2 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 3 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 4 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 5 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 6 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 7 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 8 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 9 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 10 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 11 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 12 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 13 and 14 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 1 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 2 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 3 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 4 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 5 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 6 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 7 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 8 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 9 and 10 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 1 and 7 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 2 and 7 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 3 and 7 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 4 and 7 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 5 and 7 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 6 and 7 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 1 and 5 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 2 and 5 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 3 and 5 days after a dose of the energy-based therapy. In another embodiment, the dendritic cell activating molecule is administered between 4 and 5 days after a dose of the energy-based therapy. In
Dendritic cells play a critical role in the immune system’s ability to target and kill tumor cells, but are relatively rare in most tissues. Dendritic cell activating molecules increase the total number of dendritic cells, activate the antigen presenting function of dendritic cells, increase costimulatory molecule expression, cytokine secretion, or otherwise increase their ability to prime adaptive T-cell immunity. Dendritic cell activating molecules are useful in the methods described herein. Increasing the total number of dendritic cells or activating their immunostimulatory function by administering a dendritic cell activating molecule after radiation or energy treatment can improve the ability of an individual’s immune system to target and kill cancer cells, as described in the examples.
Cancer cells can keep dendritic cells in immature states to prevent them from acting against the cancer. Immature dendritic cells can facilitate tolerance towards cancer cells while mature dendritic cells can strongly promote anticancer immunity. Promotion of maturation of dendritic cells can result in increased apoptosis in cancer cells. In one embodiment, the dendritic cell activating molecule activates maturation of an immature dendritic cell. In one embodiment, the dendritic cell activating molecule increase expression of one or more dendritic cell costimulatory molecules selected from CD70, CD80, CD86, CD40, OX40, 4-1BBL and combinations thereof. In one embodiment, the dendritic cell activating molecule increase expression or secretion of one or more dendritic cell cytokines selected from IL-12, IL-4, IL-15, or IL-17, TNFα, and combinations thereof.
A dendritic cell activator according to the methods of this disclosure can be a pathogen-associated molecular pattern (PAMP) or a synthetic version. PAMPs are small molecules conserved within a class of microbes and include without limitation glycans, glycol-conjugations, bacterial flagellin, lipoteichoic acid, peptidoglycan, and double stranded RNA. PAMPs activate of variety of innate immune receptors, known as pattern recognition receptors, expressed in antigen presenting cells and initiate adaptive immune response attributable to B and T cells. Dendritic cells express a variety of pattern recognition receptors and are activated in response to their binding to PAMPs. Pattern recognition receptors include, without limitation, toll-like receptors, NOD-like receptors, RIG-1 receptors, MDA-5 receptors, and the STING pathway. In one embodiment, the dendritic cell activating molecule activates dendritic cell activation through a toll-like receptor, a NOD-like receptor, a RIG-1 or MDA-5 receptor, a C-type lectin receptor, or a STING pathway.
Toll-like receptors are a class of receptors that are involved in the innate immune system. They are present on dendritic cells and activation of toll-like receptors with a toll-like receptor agonist or a synthetic version results in activation of the dendritic cell. In one embodiment, the dendritic cell activating molecule activates dendritic cell activation through a toll-like receptor. In another embodiment, the dendritic cell activating molecule is a toll-like receptor agonist from the list consisting of a CpG oligonucleotide, SD-101, LFX453, imiquimod, Bacillus Calmette-Guérin (BCG), monophosphoryl lipid A, Poly ICLC, GSK1795091, and combinations thereof.
NOD-like receptors are a class of pattern recognition receptors found intracellularly in dendritic cells that bind PAMPs and play a role in the innate immune system. In one embodiment, the dendritic cell activating molecule activates dendritic cell activation through a NOD-like receptor. In another embodiment, the dendritic cell activating molecule is a NOD-like receptor agonist selected from the list consisting of bacterial peptidoglycan, an acylated derivative of iE-DAP (C12-iE-DAP), D-gamma-Glu-mDAP (iE-DAP), L-Ala-gamma-D-Glu-mDAP (Tri-DAP), muramyl dipeptide (MDP), muramyl tripeptide, L18-MDP, M-TriDAP, murabutide, PGN-ECndi, PGN-ECndss, PGN-SAndi, N-glycolylated muramyl dipeptide, murabutide, and combinations thereof.
RIG-1 and MDA-5 receptors also recognize PAMPs. Specifically, both RIG-1 receptors and MDA-5 receptors are involved in the recognition of viruses by the innate immune system. RIG-1 receptors generally bind to single or double stranded RNA strands less than 2000 base pairs, while MDA-5 receptors generally bind to virally-derived single or double RNA strands greater than 2000 base pairs. When activated, these receptors promote interferon signaling and other responses of the innate immune system. In one embodiment, the dendritic cell activating molecule activates dendritic cell activation through a RIG-1 or MDA5 receptor. In another embodiment, the dendritic cell activating molecule is a RIG-1 or MDA-5 receptor agonist selected from the list consisting of poly(I:C), Poly(dA:dT), Poly(dG:dC), 3p-hpRNA, 5′ppp-dsRNA, and combinations thereof.
C-type lectin receptors are involved in recognition of PAMPs, particularly those derived from fungi and mycobacteria. When a PAMP binds to a C-type lectin receptor, the innate immune system is activated. In one embodiment, the dendritic cell activating molecules activates dendritic cell activation through a C-type lectin receptor. In another embodiment, the dendritic cell activating molecule is a C-type lectin receptor agonist selected from the list consisting of Beta-1,3-glucan, zymosan, heat-killed C. albicans, cord factor, and Trehalose-6,6-dibehenate, and combinations thereof.
The STING pathway is involved in innate immunity and the detection of PAMPs. Activation of the STING pathway results in expression of type I interferon. In one embodiment, the dendritic cell activating molecule activates dendritic cell activity through a STING pathway. In another embodiment, the dendritic cell activating molecule is a STING agonist selected from the list consisting of 2′,3′-cGAMP (CAS Number, 1441190-66-4), 4-[(2-Chloro-6-fluorophenyl)methyl]-N-(furan-2-ylmethyl)-3-oxo-1,4-benzothiazine-6-carboxamide, MK-1454, ADU-S100/MIW815, SRCB-0074, SYNB 1891, E-7766, or SB11285, and combinations thereof.
Co-stimulatory molecules are cell surface molecules present on antigen presenting cells including dendritic cells that can amplify or otherwise affect the activating signals that T cells receive when they interact with an antigen/MHC complex. They can affect T-cell fate and differentiation. In one embodiment, the dendritic cell activating molecule activates dendritic cell activation through a costimulatory molecule. In one embodiment, the dendritic cell activating molecule is a costimulatory molecule agonist selected from the list consisting of a CD40 agonist, aCD80 agonist, a CD86 agonist, an OX40 agonist, and combinations thereof.
CD40 is a TNF-family receptor expressed on dendritic cells. CD40 signaling results in expression of costimulatory ligands, cytokines, enhanced antigen presentation, and trafficking to the draining lymph node. In one embodiment, the CD40 agonistic is a CD40 agonistic antibody. Examples of CD40 agonist antibodies include, but are not limited to, dacetuzumab (also known as SGN-40, Seattle Genetics), CP-870,893 (University of Pennsylvania/Hoffmann-LaRoche), ADC-1013 (Alligator Bioscience AB), 2141-v11 (Rockefeller University), APX005M (Apexigen, Inc), Chi Lob 7/4 (Cancer research UKK), BG9588 (NIAMS), CFZ533 (Novartis), PG10 (PanGenetics UK Limited), BMS-986004 (Bristol-Myer Squibbs), lucatumumab (also known as HCD122, Novartis), HCD122 (Novartis), JNJ-64457107 (Janssen Research & Development), selicrelumab (also known as RO7009789), Hoffman-La Roche), ASKP1240 (Astellas Pharma Global Development), CDX-1140, and SEA-CD40 (Seattle Genetics).
Antibodies including CD40 agonistic antibodies can be administered directly to or near the tumor being treated. In some embodiments anti-CD40 agonist antibodies can be administered at or near a tumor being treated by an energy-based or radiation-based therapyat a dose about 0.1 milligrams to about 5 milligrams. In some embodiments anti-CD40 agonist antibodies can be administered at or near a tumor being treated by an energy-based or radiation-based therapyat a dose about 0.1 milligrams to about 0.2 milligrams, about 0.1 milligrams to about 0.5 milligrams, about 0.1 milligrams to about 1 milligram, about 0.1 milligrams to about 2 milligrams, about 0.1 milligrams to about 3 milligrams, about 0.1 milligrams to about 4 milligrams, about 0.1 milligrams to about 5 milligrams, about 0.2 milligrams to about 0.5 milligrams, about 0.2 milligrams to about 1 milligram, about 0.2 milligrams to about 2 milligrams, about 0.2 milligrams to about 3 milligrams, about 0.2 milligrams to about 4 milligrams, about 0.2 milligrams to about 5 milligrams, about 0.5 milligrams to about 1 milligram, about 0.5 milligrams to about 2 milligrams, about 0.5 milligrams to about 3 milligrams, about 0.5 milligrams to about 4 milligrams, about 0.5 milligrams to about 5 milligrams, about 1 milligram to about 2 milligrams, about 1 milligram to about 3 milligrams, about 1 milligram to about 4 milligrams, about 1 milligram to about 5 milligrams, about 2 milligrams to about 3 milligrams, about 2 milligrams to about 4 milligrams, about 2 milligrams to about 5 milligrams, about 3 milligrams to about 4 milligrams, about 3 milligrams to about 5 milligrams, or about 4 milligrams to about 5 milligrams. In some embodiments anti-CD40 agonist antibodies can be administered at or near a tumor being treated by an energy-based or radiation-based therapy at a dose about 0.1 milligrams, about 0.2 milligrams, about 0.5 milligrams, about 1 milligram, about 2 milligrams, about 3 milligrams, about 4 milligrams, or about 5 milligrams. In some embodiments anti-CD40 agonist antibodies can be administered at or near a tumor being treated by an energy-based or radiation-based therapy at a dose at least about 0.1 milligrams, about 0.2 milligrams, about 0.5 milligrams, about 1 milligram, about 2 milligrams, about 3 milligrams, or about 4 milligrams. In some embodiments anti-CD40 agonist antibodies can be administered at or near a tumor being treated by an energy-based or radiation-based therapy at a dose at most about 0.2 milligrams, about 0.5 milligrams, about 1 milligram, about 2 milligrams, about 3 milligrams, about 4 milligrams, or about 5 milligrams. Individuals may be administered at anti CD40 agonistic antibodies at a dose of between 0.01 to 5 mg/kg, 0.1 to 5 mg/kg, 0.01 to 2 mg/kg, 0.01 to 5 mg/kg, 0.01 to 1 mg/kg, 0.01 to 1 mg/kg, by intravenous administration.
Dendritic cells both produce cytokines and can be activated by cytokines. Cytokines can control the maturation of immature dendritic cells and activate dendritic cells. In one embodiment, the dendritic cell activating molecule activates dendritic cell activity through a cytokine receptor. In another embodiment, the dendritic cell activating molecule is a cytokine selected from the list consisting of granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-15 (IL-15), tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), and combinations thereof.
The dendritic cell activating molecule may be applied directly to the site of the tumor that received either the radiation treatment or the energy treatment. In one embodiment, the dendritic cell activating molecule is administered to a tumor being treated with the dose of the radiation therapy. In another embodiment, the dendritic cell activating molecule is administered to a tumor being treated with the dose of the energy therapy. Dendritic cell activators may also be administered systemically by intravenous or subcutaneous administration.
The methods described herein are useful for treating cancers and/or tumors. In certain embodiments the tumor is a solid tumor. In certain embodiments, the cancer is a blood cancer. In an embodiment, the tumor is a prostrate tumor. In an embodiment, the tumor is a melanoma. In an embodiment, the tumor is an immunotherapy resistant tumor. In an embodiment, the tumor is an immunotherapy-resistant melanoma. In an embodiment the tumor is a metastatic cancer. In an embodiment, the tumor is a metastatic breast cancer. In an embodiment of the methods, the tumor is a tumor of the prostate, breast, nasopharynx, pharynx, lung, bone, brain, sialaden, stomach, esophagus, testes, ovary, uterus, endometrium, liver, small intestine, appendix, colon, rectum, bladder, gall bladder, pancreas, kidney, urinary bladder, cervix, vagina, vulva, prostate, thyroid or skin, head or neck, glioma or soft tissue sarcoma. In an embodiment of the methods, the tumor is a prostate cancer. In an embodiment, the tumor is a malignant neoplasm.
In one embodiment, the cancer is leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, myeloblasts promyelocyte myelomonocytic monocytic erythroleukemia, chronic leukemia, chronic myelocytic (granulocytic) leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, primary central nervous system lymphoma, Burkitt’s lymphoma and marginal zone B cell lymphoma, Polycythemia vera Lymphoma, Hodgkin’s disease, non-Hodgkin’s disease, multiple myeloma, Waldenstrom’s macroglobulinemia, heavy chain disease, solid tumors, sarcomas, and carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chrondrosarcoma, osteogenic sarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing’s tumor, leiomyosarcoma, rhabdomyosarcoma, colon sarcoma, colorectal 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 tumor, lung carcinoma, small cell lung carcinoma, non-small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma, retinoblastoma, nasopharyngeal carcinoma, esophageal carcinoma, basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancers, connective tissue cancer, cancer of the digestive system, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, gastric cancer, intraepithelial neoplasm, kidney cancer, larynx cancer, liver cancer, lung cancer (small cell, large cell), melanoma, neuroblastoma; oral cavity cancer (for example lip, tongue, mouth and pharynx), ovarian cancer, pancreatic cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer; cancer of the respiratory system, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, and cancer of the urinary system.
Also described herein are methods using combinations of radiation and/or energy based therapies and dendritic cell activating molecules are methods of treating cancers or tumors that are resistant to checkpoint inhibitor therapies. Current checkpoint inhibitor therapies target PD-1, PD-L1, PD-L2, or CTLA4, using antibodies such as pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, durvalumab, ipilimumab.
Also described herein are uses of dendritic cell activating molecules in a method of treating a cancer or tumor in an induvial, wherein the individual has received a dose of radiation or energy-based therapy.
Also described herein are dendritic cell activating molecules for the manufacture of a medicament for treating a cancer or tumor in an induvial, wherein the individual has received a dose of radiation or energy-based therapy.
In one aspect described here in is a method of increasing T cell infiltration into a tumor distal to a tumor being treated in an individual, the method comprising administering to the individual a dose of a radiation therapy and a dendritic cell activating molecule, wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy is administered. In certain embodiments, the dendritic cell activating molecule is an antiCD40 agonistic antibody.
In one aspect described herein is a method of increasing T cell infiltration into a tumor distal to a tumor being treated in an individual, the method comprising administering to the individual a dose of an energy-based therapy and a dendritic cell activating molecule, wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy is administered. In certain embodiments, the dendritic cell activating molecule is an antiCD40 agonistic antibody. In certain embodiments, the dose of the energy-based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy.
In one aspect described herein is a method of reversing T cell exhaustion in a tumor distal to a tumor being treated in an individual, the method comprising administering to the individual a dose of a radiation therapy and a dendritic cell activating molecule, wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy is administered. In certain embodiments, the dendritic cell activating molecule is an antiCD40 agonistic antibody.
In one aspect described herein is a method of reversing T cell exhaustion in a tumor distal to a tumor being treated in an individual, the method comprising administering to the individual a dose of an energy-based therapy and a dendritic cell activating molecule, wherein the dendritic cell activating molecule is administered at least one day after the radiation therapy is administered. In certain embodiments, the dendritic cell activating molecule is an antiCD40 agonistic antibody. In certain embodiments, the dose of the energy-based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy.
In an embodiment, treating a tumor by the methods described herein reduces the size or volume of the tumor by about 10%, 20%, 25%, 30%, 40%, 50% or more. In an embodiment, treating a tumor by the methods described herein reduces the size or volume of a tumor that is not the tumor treated with radiation or energy-based therapy by about 10%, 20%, 25%, 30%, 40%, 50% or more. In an embodiment, treating a tumor by the methods described herein prevents metastasis of a tumor or cancer described herein.
In certain embodiments the dendritic cell activating molecule of the current disclosure is included in a pharmaceutical composition comprising one or more pharmaceutically acceptable excipients, carriers, and diluents. In certain embodiments, the dendritic cell activating molecule of the current disclosure is administered suspended in a sterile solution. In certain embodiments, the solution comprises about 0.9% NaCl or about 5% dextrose. In certain embodiments, the solution further comprises one or more of: buffers, for example, acetate, citrate, histidine, succinate, phosphate, bicarbonate and hydroxymethylaminomethane (Tris); surfactants, for example, polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), and poloxamer 188; polyol/disaccharide/polysaccharides, for example, glucose, dextrose, mannose, mannitol, sorbitol, sucrose, trehalose, and dextran 40; amino acids, for example, glycine or arginine; antioxidants, for example, ascorbic acid, methionine; or chelating agents, for example, EDTA or EGTA.
In certain embodiments, the dendritic cell activating molecule of the current disclosure is shipped/stored lyophilized and reconstituted before administration. In certain embodiments, lyophilized antibody formulations comprise a bulking agent such as, mannitol, sorbitol, sucrose, trehalose, dextran 40, or combinations thereof. The lyophilized formulation can be contained in a vial comprised of glass or other suitable non-reactive material. The dendritic cell activating molecule when formulated, whether reconstituted or not, can be buffered at a certain pH, generally less than 7.0. In certain embodiments, the pH can be between 4.5 and 6.5, 4.5 and 6.0, 4.5 and 5.5, 4.5 and 5.0, or 5.0 and 6.0.
Numbered embodiment 1 comprises a method of increasing T cell infiltration into a tumor distal to a tumor being treated in an individual, the method comprising administering to the individual a dose of an energy-based therapy and a dendritic cell activating molecule, wherein the dose of the energy-based therapy is selected from the list consisting of Irreversible Electroporation (IRE), Microwave, Low-Intensity Focused Ultrasound (LOFU), High-Intensity Focused Ultrasound (HIFU), Radiofrequency energy, and cryotherapy. Numbered embodiment 2 comprises the method of embodiment 1, wherein the dose of the energy base therapy comprises a plurality of doses of energy-based therapy. Numbered embodiment 3 comprises the method of embodiments 1 or 2, wherein the energy-based therapy is Irreversible Electroporation (IRE). Numbered embodiment 4 comprises the method of embodiments 1 or 2, wherein the energy-based therapy is microwave therapy. Numbered embodiment 5 comprises the method of embodiments 1 or 2, wherein the energy-based therapy is Low-Intensity Focused Ultrasound (LOFU Numbered embodiment 6 comprises the method of embodiments 1 or 2, wherein the energy-based therapy is High-Intensity Focused Ultrasound (HIFU). Numbered embodiment 7 comprises the method of embodiments 1 or 2, wherein the energy-based therapy is cryotherapy. Numbered embodiment 8 comprises the method of any one of embodiments 1 to 7, wherein the dendritic cell activating molecule is administered at least three days after the dose of the energy-based therapy. Numbered embodiment 9 comprises the method of any one of embodiments 1 to 7, wherein the dendritic cell activating molecule is administered at least five days after the dose of the energy-based therapy. Numbered embodiment 10 comprises the method of any one of embodiments 1 to 7, wherein the dendritic cell activating molecule is administered at least seven days after the dose of the energy-based therapy. Numbered embodiment 11 comprises the method of any one of embodiments 1 to 10, wherein the dendritic cell activating molecule activates maturation of an immature dendritic cell. Numbered embodiment 12 comprises the method of any one of embodiments 1 to 10, wherein the dendritic cell activating molecule activates dendritic cell activation through a toll-like receptor, a NOD-like receptor, a RIG-1 or MDA-5 receptor, a C-type lectin receptor, a costimulatory molecule, a cytokine receptor, or a STING pathway. Numbered embodiment 13 comprises the method of any one of embodiments 1 to 10, wherein the dendritic cell activating molecule is a toll-like receptor agonist selected from the list consisting of CpG oligonucleotide, SD-101, LFX453, imiquimod, Bacillus Calmette-Guérin (BCG), monophosphoryl lipid A, Poly ICLC, GSK1795091, and combinations thereof. Numbered embodiment 14 comprises the method of any one of embodiments 1 to 10, wherein the dendritic cell activating molecule is a NOD-like receptor agonist selected from the list consisting of bacterial peptidoglycan, an acylated derivative of iE-DAP (C12-iE-DAP), D-gamma-Glu-mDAP (iE-DAP), L-Ala-gamma-D-Glu-mDAP (Tri-DAP), muramyl dipeptide (MDP), muramyl tripeptide, L18-MDP, M-TriDAP, murabutide, PGN-ECndi, PGN-ECndss, PGN-SAndi, N-glycolylated muramyl dipeptide, murabutide, and combinations thereof. Numbered embodiment 15 comprises the method of any one of embodiments 1 to 10, wherein the dendritic cell activating molecule is a RIG-1 or MDA-5 receptor agonist selected from the list consisting of poly(I:C), Poly(dA:dT), Poly(dG:dC), 3p-hpRNA, 5′ ppp-dsRNA, and combinations thereof. Numbered embodiment 16 comprises the method of any one of embodiments 1 to 10, wherein the dendritic cell activating molecule is a C-type lectin receptor agonist selected from the list consisting of Beta-1,3-glucan, zymosan, Heat-killed C. albicans, cord factor, and Trehalose-6,6-dibehenate, and combinations thereof. Numbered embodiment 17 comprises the method of any one of embodiments 1 to 10, wherein the dendritic cell activating molecule is a costimulatory molecule agonist selected from the list consisting of a CD40 agonist, aCD80 agonist, a CD86 agonist, an OX40 agonist, and combinations thereof. Numbered embodiment 18 comprises the method of embodiment 17, wherein the CD40 agonist is an anti-CD40 agonistic antibody. Numbered embodiment 19 comprises the method of embodiment 17, wherein the anti-CD40 agonistic antibody comprises dacetuzumab, CP-870,893, ADC-1013, 2141-v11, APX005M, Chi Lob 7/4, BG9588 (NIAMS), CFZ533, PG10, BMS-986004, lucatumumab, HCD122, JNJ-64457107, selicrelumab, ASKP1240, or SEA-CD40. Numbered embodiment 20 comprises the method of any one of embodiments 1 to 19, wherein the dendritic cell activating molecule is a cytokine selected from the list consisting of granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-15 (IL-15), tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), and combinations thereof. Numbered embodiment 21 comprises the method of any one of embodiments 1 to 19, wherein the dendritic cell activating molecule is a STING agonist selected from the list consisting of 2′,3′-cGAMP (CAS Number, 1441190-66-4), 4-[(2-Chloro-6-fluorophenyl)methyl]-N-(furan-2-ylmethyl)-3-oxo-1,4-benzothiazine-6-carboxamide, MK-1454, ADU-S100/MIW815, SRCB-0074, SYNB 1891, E-7766, or SB11285, and combinations thereof. Numbered embodiment 22 comprises the method of any one of embodiments 1 to 21, wherein the dendritic cell activating molecule is administered to a tumor being treated with the dose of the energy-based therapy.
The following illustrative examples are representative of embodiments of compositions and methods described herein and are not meant to be limiting in any way.
A non-metastatic human PSA expressing TPSA murine implanted tumor model was used to assess the effect of administering αCD40 concurrently with radiation treatment (RT). Some test groups were also treated with low intensity focused ultrasound (LOFU).
Mice were injected with 0.9×106 tumor cells on the right flank. On day 14-17, mice with palpable tumors were randomly segregated into different treatment groups. The treatment groups were control (un-irradiated), RT (10Gy×2), RT (10Gy×2) + αCD40, RT (10Gy×2)+LOFU and LOFU+RT+αCD40. Mice were treated with RT and LOFU (5W 99.5%) on day 14 and day 16 with concurrent αCD40 therapy (day 14, day 16, and day 18; 3×100µg/ per mice), as depicted in
In this example, the effect of administering αCD40 concurrently with radiation treatment (RT) in PSA transgenic mice was assessed. Some test groups were also treated with low intensity focused ultrasound (LOFU).
The experimental treatment is depicted in
The growth of tumor volume per treatment is depicted in
This example assessed the effect of treating immunotherapy resistant melanoma cells with αCD40 administered after radiation treatment consisting of ionizing radiation (IR).
The treatment schedule of the mice is depicted in
When administered sequentially, αCD40 effectively enhanced the long-term survival and cure in the RES499 tumor bearing mice. As seen in
At day 120, the tumor-free mice were re-challenged with RES499 cells, as depicted in
This example showed that treatment with αCD40 enhanced the radiotherapy associated survival and cure in mice with immunotherapy resistant tumors. The re-challenge experiment showed an increased adaptive memory response against immunotherapy resistant tumors.
This example assessed the ability of αCD40 administration following radiation (IR) to retard the abscopal tumor growth of tumors resistant to radiotherapy and αCTLA-4 therapy.
The RES499 tumor line was developed from tumors which were non-responsive to the systemic effects of combined radiotherapy and αCTLA-4 therapy, as depicted in
C57BL/6 mice were injected subcutaneously with 0.2×106 RES499 melanoma cells in the right flank (index tumor; irradiated) on day 0 and 0.1×106 RES499 cells in the left flank (abscopal tumor; non-irradiated) on day 4. On days 7-9, when primary tumors were palpable, animals were randomly assigned to the different treatment groups. For treatment, mice were irradiated with 3 fractions (1 fraction every day) of 20 Gy each from day 7-9. αCD40 (3×100ug) was administered on day 12, day 14, and day 18, as depicted in
This experiment showed that a combination of IR and αCD40 treatment significantly reduced the growth of both primary and abscopal tumors in an immunotherapy-resistant tumor line.
This example assessed the effect of the systemic αCD40 therapy in combination with radiation (IR) on tumor-infiltrating host cells.
Three days after the second dose of αCD40, tumors were excised and digested postmortem using a cocktail of collagenase type IV and DNase. After digestion at 37° C. for 30 minutes, cells were passed through a 70-µm filter. Cells were stained for cell surface and cytosolic proteins. Cells were then analyzed by flow cytometry and zombie IR (Thermo Fisher) was used as a viability dye.
There was a significant increase (p<0.5) in the co-stimulatory markers (4-1BBL, CD40 and CD86) and type 1 inflammation (TNF-α) in the tumor infiltrating CD103+ dendritic cells (DC) derived from mice that had received a combination of αCD40 and IR when compared to the IR treated group, as depicted in
The agonist CD40 antibody also affected the immature suppressor cells of myeloid origin (Ly6C high CD1lb+). The myeloid derived suppressor cells (MDSC) showed an increase in co-stimulatory markers CD80 and 4-1BBL when derived from mice treated with both radiation and αCD40, as depicted in
Inducible nitric oxide synthetase (INOS) is a cell-killing effector of the myeloid and DCs. Treatment with αCD40 significantly increased INOS levels in the CD103+DCs, MDSCs, and total pool of the myeloid cells, compared to treatment with radiation alone (
This example assessed the effect of the systemic αCD40 therapy in combination with radiation (IR) on infiltrating host cells in the draining lymph node (DLN).
Three days after the second dose of αCD40, DLNs were harvested and cells were passed through a 40-µm filter. Cells were stained for cell surface and cytosolic proteins. Cells were then analyzed by flow cytometry and zombie IR (Thermo Fisher) was used as a viability dye.
In the gross CD11b+ leucocytes and its subpopulations, activation associated costimulatory molecules CD86 and CD40 were increased (p<0.01-0.001), as depicted in
The granulocytic MSDCs (PM-MDSCs) showed a decrease in the percent of CD11b+ cells in the DLN after treatment with both radiation and αCD40, depicted in
This example assessed the effect of sequential αCD40 treatment on CD8 effector function.
Characterization of T cells in the αCD40 +IR treated lungs showed that there was an increase in the frequency and the functional competence of the effector cytotoxic CD8 T cells when compared with the IR alone treated group.
The CD8 proportion in the tumor was assessed by measuring the frequency of CD8 cells and CD4/CD8 ratio which is a marker of an effective anti-tumor immune response. αCD40 treatment affected the CD8 proportion in the tumor. There was a significant decrease in CD8 numbers in the tumor derived from mice that had received a combination treatment when compared to mice that had received radiation alone. This was both an increased seen as both an increase in CD8 frequency as well as in a reduced CD4/CD8 ratio (
Functions of CD8 cells were assessed using both the frequency of the functional IFNγ+ cells and the increased proliferating cells. αCD40 treatment increased both the percent of IFNγ+ CD8 cells and the mean fluorescent intensity (MFI) of IFNγ+ CD8 cells, as depicted in
This example assessed the effect of αCD40 treatment combined with radiation on CD8 effector function in the abscopal tumor draining lymph node (DLN).
The effect of αCD40 and radiation treatment on CD8 effector function in mice was measured in the abscopal tumor DLN. The CD4/CD8 ratio was decreased in mice that had received both αCD40 and radiation, compared to mice that had received radiation alone, as illustrated in
This was due to the highly activated state of the cells, which was determined by both the increase in Ki67+ cells and CD44+ cells depicted in
This example assessed the effects of αCD40 administration combined with radiation treatment of metastatic cancer using the murine orthotropic breast tumor cell line 4T1. Radiation treatments include ionizing radiation (IR), and post ablation modulation (PAM, 4 doses of 0.5 Gy IR dose every day).
0.2×106 4T1 cells were injected in the mammary fat pad of BALB/c mice (syngeneic to 4T1). At day 7, mice with palpable tumors were randomly segregated into 5 groups: control (un-irradiated), IR (20Gy×3) + PAM (0.5Gy×4), and IR (20Gy×3) + PAM+ αCD40. Mice were irradiated on day 7-9 and αCD40 was given post IR (Day 10, 14 and 18). 0.5Gy×4 doses (PAM) were given on days 10-13. Tumor volumes and survival was recorded at multiple times. The treatment protocol is depicted in
When combined with radiotherapy, αCD40 significantly inhibited the metastatic events and improved overall mouse survival, as seen in
This example showed that sequential treatment with radiation and αCD40 can effectively treat metastatic disease and inhibit death in a metastatic model.
This example assessed the effects of αCD40 administration combined with radiation treatment on abscopal (non -irradiated) tumor growth using the murine melanoma lines B16F10 and RES499 (checkpoint resistant line). Radiation treatments include ionizing radiation (IR).
C57BL/6 mice were injected subcutaneously with 0.2×106 RES499 and B16 melanoma cells in the right flank (index tumor; irradiated) on day 0 and 0.1×106 RES499 cells in the left flank (abscopal tumor; non-irradiated) on day 4. On days 7-9, when primary tumors were palpable, animals were randomly assigned to the different treatment groups. For treatment, mice were irradiated with 3 fractions (1 fraction every day) of 20 Gy each from day 7-9. αCD40 (3×100ug) was administered on day 12, day 14, and day 18, as depicted in
Mice treated with αCD40 showed lower tumor volumes and higher rates of survival than mice treated with radiation alone (
This example showed that sequential treatment with radiation and αCD40 therapy can effectively inhibit tumor growth in a melanoma model and in checkpoint resistant tumors.
Patients with cancer may follow the disease and treatment progression shown in
This example assessed the effect of anti-CD40 therapy on the exhaustion of tumor-infiltrating cells. The experimental protocol is depicted in
IR has been shown to induce exhaustion of T cells during the radiotherapy. IR alone group showed minimal population of the functional subtype (GrB+KI67high) in the early exhausted cells (PDlintEomeshi). In the IR+anti-CD40 group, functional subtype of the exhausted population was significantly increased (p<0.05). Early exhaustion is marked by the PD1 intermediate and EOMES low CD8 cells (PDlintEomeshi). Anti-CD40 +IR combination group increased the Ki67 (proliferating) high GRZ+ (granzyme secreting) population in the pool suggesting a reversal of the exhausted phenotype (
Depletion experiments were performed to investigate the role of different subsets of the immune cells in mediating the therapeutic effect of the anti-CD40 and IR combination. The experimental protocol is depicted in
For immune-phenotyping studies, on 17th day post tumor inoculation tumors were excised postmortem and dissociated using a cocktail of collagenase type IV and DNase. After digestion at 37° C. for 30 minutes, cells were passed through a 70-µm filter. Cells were stained for cell surface and cytosolic proteins and analyzed by flow cytometry as previously and zombie IR (Thermo Fisher) was used as a viability dye.
To investigate the role of the CD8 T cells in the therapeutic efficacy of IR+ anti-CD40 combination group, anti-CD8 antibodies were used to deplete CD8 cells in the C57BL6 mice. Tumor growth delay in the IR+anti-CD40 combination was partially reversed in the anti-CD8 depleted mice, as depicted in
Homozygous athymic nude mice lack T cells and suffer from a lack of cell-mediated immunity. Homozygous nude mice also show partial defect in B cell development. Similar results were also observed in the nude mice experiments where the effect of combination was not significant compared to the IR alone group, as depicted in
To further look at which antigen presentation and processing population pool contributed to the therapeutic benefits of the combination group (IR+antiCD40), the LY6C and CD11b population was depleted in the C57BL6 mice. Ly6C high myeloid cells are known to be critical cross presenting APCs along with the dendritic cells. While tumor growth delay observed in the IgG control groups was partially reversed in the CD11b depleted mice, Ly6c depletion completely reversed (p<0.05) the tumor growth delay (
While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention.
All publications, patent applications, issued patents, and other documents referred to in this specification are herein incorporated by reference as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions that are contained in text incorporated by reference are excluded to the extent that they contradict definitions in this disclosure.
This application claims the benefit of U.S. Provisional Application No. 63/062,185, filed Aug. 6, 2020, which application is incorporated herein by reference herein in its entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US2021/044856 | 8/5/2021 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 63062185 | Aug 2020 | US |
