Composition for preventing or treating cancer having distal effect, and method for preventing or treating cancer using same

Abstract

The present invention relates to a method and a composition for inducing at least one immunostimulation for the treatment of primary cancer and cellular carcinoma dispersed in the human body and its metastases, in particular for inducing and promoting the immune system of the patient's own to identify and kill cancer cells and establish memory to prevent the recurrence of cancer diseases.

Description

Technical Field

[0001] The present invention relates to a composition for preventing or treating cancer having a distal effect, and a method for treating cancer using the composition, and more particularly to a method for enhancing the distal effect for treating cancer and tumors, and a composition for inducing the distal effect.

[0002] The present invention claims priority to Korean Patent Application No. 10-2022-0117466 filed on September 16, 2022 and Korean Patent Application No. 10-2023-0119122 filed on September 7, 2023, the entire contents of the specifications and drawings of which are incorporated herein by reference. Background Art

[0003] Generally, it is known that radiotherapy impairs immune function. However, as more is learned about the role of T cells in radiotherapy, new aspects of the immune response related to radiotherapy, such as immunomodulation, are also gradually becoming known. As representative effects exhibited at the cellular level after radiation exposure, the activity of natural killer (NK) cells increases, the infiltration of T cells (especially CD8 T cells) increases, the antigen presentation of dendritic cells increases, the production of immunostimulatory cytokines increases, and so on.

[0004] The distal effect is particularly well-known for its immunomodulatory effect, which refers to the anti-cancer treatment effect of inhibiting cancer growth in areas outside the tumor tissue within the region irradiated locally with radiation, including metastatic tumors in areas not irradiated with radiation. That is, this is a situation where radiotherapy sensitizes tumor cells to respond more quickly to cancer treatment.

[0005] According to the distal effect, radiotherapy affects the human immune system so that it attacks not only cancer cells in a specific part of the body but also cancer cells throughout the body. The immune effect can eliminate not only cancer cells in the area exposed to radiation but also cancer cells in other distant parts of the body. Radiotherapy causes dendritic cells to release specific markers extracellularly, send danger signals using HMGB1 or ATP, and enhance CD8 + T cells to induce an anti-cancer response. Therefore, during radiotherapy, not only cancer in the irradiated area but also cancer in other non-irradiated areas is cleared. It has recently been found that high-dose ablative radiotherapy (20 Gy in a single dose) can increase CD8 +The activity of cytotoxic T cells, thereby controlling cancer not only at the treatment site but also at untreated distant metastatic sites. Recently, it has been reported that when a vaccine (Flt3L) that inhibits TLR9 is injected into low-grade malignant lymphoma and the cancer at a specific site is irradiated with radiation, the cancer at untreated distant sites also shows a reduced response. In addition, when patients with metastatic lung cancer are injected with granulocyte-macrophage colony-stimulating factor (GM-CSF) while receiving radiotherapy, a reduced response is observed in some cancers in the areas not receiving radiotherapy.

[0006] Meanwhile, the MHC for antigen presentation on cancer cells leads to CD8 + Cytotoxic T lymphocytes secrete interferon γ, which promotes the production of the PD-1 ligand on tumor cells. This PD-1 ligand binds to PD-1 on T cells, thereby weakening the activity of T cells. Therefore, antibodies that can inhibit this ligand (for example, anti-PD-1 antibodies or bispecific T cell-engaging antibodies; CD133xCD3 antibodies) can be used for cancer treatment.

[0007] Multiple research reports show that immune checkpoint inhibitors can effectively treat cancer. Clinically, ipilimumab, an immune checkpoint inhibitor of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), and nivolumab, which acts on programmed death-1 (PD-1), have been reported to improve the survival rate of patients with advanced melanoma. Another anti-PD-1 antibody, pembrolizumab, is also effective against gastric cancer, nasopharyngeal cancer, lung cancer, etc. However, as a single therapy, the effectiveness of immunotherapy is limited to 10%-30%.

[0008] In summary, multiple studies are underway to explore methods to enhance the distal effect. However, no method has been proposed that can effectively maximize the distal effect. SUMMARY OF THE INVENTION

[0009] TECHNICAL PROBLEM

[0010] Therefore, the present invention is made in view of the above problems. It has been confirmed that tumor tissues or cancer cells isolated from cancer patients, after being treated by radiation irradiation or cancer immunotherapy drugs to inhibit the activity of cancer cells or induce their death and then administered to cancer patients, can activate the anti-cancer immune function of the body, further improve the anti-cancer treatment effect, thereby inhibiting the growth, recurrence, metastasis, etc. of cancer, thus realizing the present invention. In particular, it has been confirmed that cancer cells induced to inhibit activity or death as described above can exert a distal effect when administered to cancer patients.

[0011] In addition, it has been confirmed that when the above-mentioned cancer cells whose activity or death is induced are used in combination with radiotherapy and / or cancer immunotherapy drugs, the anti-cancer effect can be further maximized, anti-cancer immunity can be enhanced, and a powerful distal effect can be achieved.

[0012] Therefore, an object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, the pharmaceutical composition comprising tumor tissue or cancer cells as an active ingredient, which are isolated from a cancer patient and subjected to one or more treatments selected from the group consisting of the following (a) and (b):

[0013] (a) Radiation exposure; and

[0014] (b) Cancer immunotherapy drugs.

[0015] In addition, the present invention provides a kit for preventing or treating cancer comprising the composition.

[0016] In addition, the present invention provides a method for preparing the composition.

[0017] More specifically, an object of the present invention is to provide a composition for preventing or treating cancer and a method for preventing or treating cancer using the composition, which are used for invasive or non-invasive treatment of primary cancer and its metastatic cancer in patients with cancer or tumors, and can enhance the anti-cancer effect through radiotherapy, treatment with anti-cancer drugs or immunopotentiators, or exert a distal effect to prevent cancer recurrence after treatment.

[0018] Another object of the present invention is to provide a composition for preventing or treating cancer and a method for preventing or treating cancer using the composition, the composition can be used as a tumor vaccine to prevent immune escape of cancer cells by making the immune system more effectively recognize cancer cells. In particular, the composition according to the present invention can exert a distal effect to prevent cancer recurrence in patients at risk of recurrence after cancer treatment, and thus has high practicality as a tumor vaccine.

[0019] Another object of the present invention is to provide a composition and a method for preventing or treating cancer using the composition, the composition can provide memory and systemic immune responses to cancer cells (especially disseminated cells of micrometastatic cancer) through a distal effect, can be used as a vaccine therapy, and can be used to prevent recurrence after general cancer and tumor treatment.

[0020] Another object of the present invention is to provide a composition and a method for preventing or treating cancer using the composition, which can increase the distal effects for treating cancer and tumors in a subject, but can inhibit immune system damage, local or systemic damage and burden, undesirable cancer and tumor mutations or increased drug resistance, etc., such as side effects and burden of radiation exposure and anticancer agents on the subject, and side effects and burden of metabolites caused by immunomodulators.

[0021] Another object of the present invention is to reduce or delay the likelihood of developing cancer by administering the composition according to the present invention to a subject without cancer, thereby enabling the immune system to learn and remember cancer and enhancing immunity against cancer. Therefore, the composition according to the present invention can achieve the effect of preventing cancer in subjects at high risk of developing cancer due to genetic and environmental factors.

[0022] Another object of the present invention is to achieve a more effective anti-cancer effect by combining the composition according to the present invention with other anti-cancer treatments. For example, when the composition according to the present invention is used in combination with radiotherapy and / or cancer immunotherapy drugs, the growth of primary tumors (or primary tumors) can be more effectively inhibited, and the growth of secondary tumors can also be inhibited through distal effects, thereby more effectively inhibiting cancer metastasis. In addition, the combination therapy can also enhance the anti-cancer immune function, thereby achieving a synergistic anti-cancer effect.

[0023] For this purpose, the present invention provides a method of extracting an appropriate amount of cancer and tumor tissues from a subject in vitro, inducing the extracted tissues or cancer cells to weaken or die by one or more physical, chemical, and medical methods (such as radiation exposure or immunomodulators), and then re-administering them to the subject to induce distal effects in the subject's body.

[0024] Method for solving technical problems

[0025] The present invention provides a pharmaceutical composition for preventing or treating cancer, which comprises tumor tissues or cancer cells as active ingredients, which are isolated from cancer patients and subjected to one or more treatments selected from the group consisting of the following (a) and (b):

[0026] (a) Radiation exposure; and

[0027] (b) Cancer immunotherapy drugs.

[0028] The cancer here includes primary cancer, secondary tumors, metastatic cancer, and recurrent cancer.

[0029] In addition, the present invention provides a kit for preventing or treating cancer comprising the composition.

[0030] In addition, the present invention provides the use of the composition for preventing or treating cancer.

[0031] In addition, the present invention provides a method for preventing or treating cancer, comprising administering the composition to a subject in need thereof. Preferably, the subject is the subject from whom the tumor tissue or cancer cells are isolated, or an allogeneic subject of the subject. In one embodiment of the present invention, the method for preventing or treating cancer may further comprise the step of subjecting the subject to anti-cancer treatment. The anti-cancer treatment may be two or more types of anti-cancer treatment (i.e., different types of anti-cancer treatment). In addition, the anti-cancer treatment may be carried out simultaneously with or sequentially to the administration of the composition. There is no limitation on the order of implementation.

[0032] In addition, the present invention provides the use of the composition for the preparation of a medicament for treating cancer.

[0033] In addition, the present invention provides the use of the composition in combination with anti-cancer treatment.

[0034] In addition, the present invention provides the use of the composition for the preparation of a preparation that can be used in combination with anti-cancer treatment.

[0035] In addition, the present invention provides a method for preventing or treating cancer, comprising: step (i) administering the composition to a subject in need thereof; and step (ii) administering a second anti-cancer treatment to the subject. The second anti-cancer treatment may be two or more types of anti-cancer treatment (i.e., different types of anti-cancer treatment). Step (i) and step (ii) may be carried out simultaneously or sequentially, and the order of execution is not limited. In addition, when two or more types of anti-cancer treatment are carried out, the order of execution of each anti-cancer treatment is not limited, and it may be carried out simultaneously with the administration of the composition or sequentially.

[0036] In one embodiment of the present invention, the tumor tissue or cancer cells can meet one or more characteristics selected from the group consisting of the following i) to iii) through treatment, but are not limited thereto:

[0037] i) The activity of cancer cells is weakened;

[0038] ii) The activity of cancer cells is stopped; and

[0039] iii) Cancer cells are killed.

[0040] In another embodiment of the present invention, the composition comprises autologous tumor tissue or cancer cells of a cancer patient, and can be administered to the cancer patient, or can be administered to a cancer patient allogeneic to the cancer patient, but is not limited thereto.

[0041] In another embodiment of the present invention, the composition can be administered to a subject without cancer to prevent cancer, but is not limited thereto. The subject may be a subject at high risk of developing cancer genetically or environmentally.

[0042] In another embodiment of the present invention, when the treatment is (a) radiation exposure, the radiation may satisfy one or more characteristics selected from the group consisting of the following i) and ii), but is not limited thereto:

[0043] i) The radiation is one or more selected from the group consisting of gamma-rays, X-rays, ultraviolet rays, laser rays, and infrared rays; and

[0044] ii) The irradiation dose of the radiation is 1 to 500 Gray.

[0045] In another embodiment of the present invention, the cancer immunotherapy drug may be one or more selected from the group consisting of immune checkpoint inhibitors, co-stimulatory molecule preparations, cytokine therapeutics, CAR-T cell therapeutics, and autologous CD8 + T immune cell therapeutics, but is not limited thereto.

[0046] In another embodiment of the present invention, the immune checkpoint inhibitor may be one or more selected from the group consisting of the following inhibitors: PD-L1 inhibitor, PD-1 inhibitor, CTLA-4 inhibitor, LAG3 inhibitor, TIM3 inhibitor, 4-1BB inhibitor, LAG-3 inhibitor, B7-H4 inhibitor, HVEM inhibitor, TIM4 inhibitor, GAL9 inhibitor, VISTA inhibitor, KIR inhibitor, TIGIT inhibitor, and BTLA inhibitor, but is not limited thereto.

[0047] In another embodiment of the present invention, when the composition is administered to a cancer patient, it may satisfy one or more characteristics selected from the group consisting of the following i) to iv), but is not limited thereto:

[0048] i) Exert a distal effect;

[0049] ii) Inhibit cancer metastasis;

[0050] iii) Reduce tumor burden; and

[0051] iv) Inhibit the proliferation of cancer cells.

[0052] In another embodiment of the present invention, the composition is for single or multiple administrations, but is not limited thereto.

[0053] In another embodiment of the present invention, the composition may include two or more kinds of tumor tissues or cancer cells, and the two or more kinds of tumor tissues or cancer cells may be each subjected to the same treatment or different treatments, but is not limited thereto.

[0054] In another embodiment of the present invention, the composition may be a mixture of two or more tumor tissues or cancer cells, but is not limited thereto.

[0055] In another embodiment of the present invention, the composition can be prepared by separately formulating two or more tumor tissues or cancer cells and can be administered simultaneously, separately, or sequentially, but is not limited thereto.

[0056] In another embodiment of the present invention, the composition can be used in combination with anti-cancer treatment, but is not limited thereto.

[0057] In another embodiment of the present invention, the anti-cancer treatment can be one or more selected from the group consisting of radiotherapy, chemotherapy, targeted anti-cancer therapy, and cancer immunotherapy, but is not limited thereto.

[0058] In another embodiment of the present invention, the composition can be used in combination with radiotherapy and cancer immunotherapy, but is not limited thereto.

[0059] In another embodiment of the present invention, the composition can be administered simultaneously, separately, or sequentially with anti-cancer treatment, but is not limited thereto.

[0060] In another embodiment of the present invention, the targeted anti-cancer agent can be one or more selected from the group consisting of tyrosine kinase inhibitors, PARP inhibitors, angiogenesis inhibitors, and CDK4 / 6 inhibitors, but is not limited thereto.

[0061] In another embodiment of the present invention, the cancer immunotherapy drug can be at least one selected from the group consisting of immune checkpoint inhibitors, costimulatory molecule agents, cytokine therapy agents, CAR-T cell therapy agents, and autologous CD8 + T immune cell therapy agents, but is not limited thereto.

[0062] In another embodiment of the present invention, the cancer checkpoint inhibitor can be one or more selected from the group consisting of the following inhibitors: PD-L1 inhibitors, PD-1 inhibitors, CTLA-4 inhibitors, LAG3 inhibitors, TIM3 inhibitors, 4-1BB inhibitors, LAG-3 inhibitors, B7-H4 inhibitors, HVEM inhibitors, TIM4 inhibitors, GAL9 inhibitors, VISTA inhibitors, KIR inhibitors, TIGIT inhibitors, and BTLA inhibitors, but is not limited thereto.

[0063] In another embodiment of the present invention, the tumor tissue can be ground and diluted or dissolved in a solution, but is not limited thereto.

[0064] In another embodiment of the present invention, the composition may include two or more tumor tissues, where the two or more tumor tissues may have the same or different sizes of ground particles, but are not limited thereto.

[0065] In another embodiment of the present invention, the cancer may be one or more selected from the group consisting of blood cancer and solid cancer, but is not limited thereto.

[0066] In another embodiment of the present invention, the cancer may be one or more selected from the group consisting of: breast cancer, colorectal cancer, lung cancer, head and neck cancer, small cell lung cancer, gastric cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, cervical cancer, cutaneous melanoma, uveal melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, colon cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney cancer, ureteral cancer, renal cell carcinoma, renal pelvic cancer, central nervous system tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma, but is not limited thereto.

[0067] In addition, the present invention provides a method for preparing the composition, the method comprising: (S1) isolating tumor tissues or cancer cells from a cancer patient; and

[0068] (S2) applying one or more treatments selected from the group consisting of the following (a) and (b) to the isolated tumor tissues or cancer cells,

[0069] (a) radiation exposure; and

[0070] (b) cancer immunotherapy drugs.

[0071] In one embodiment of the present invention, the method may further include grinding the tumor tissue after step (S1) and diluting or dissolving it in a solution, but is not limited thereto.

[0072] In another embodiment of the present invention, the composition may include autologous tumor tissues or cancer cells of a cancer patient and may be administered to the cancer patient or a cancer patient allogeneic to the cancer patient, but is not limited thereto.

[0073] In another embodiment of the present invention, the composition may be administered to a subject without cancer to prevent cancer, but is not limited thereto. The subject may be a subject having a high risk of developing cancer due to genetic or environmental factors.

[0074] In addition, the present invention provides an anti-cancer vaccine composition or vaccine, comprising tumor tissue or cancer cells as an active ingredient, which are isolated from cancer patients and treated with one or more selected from the group consisting of the following (a) and (b):

[0075] (a) Radiation irradiation; and

[0076] (b) Cancer immunotherapy drugs.

[0077] In one embodiment of the present invention, the vaccine composition or vaccine can be administered to subjects who have not had cancer or have been cured of cancer to prevent cancer, but is not limited thereto.

[0078] In addition, the present invention provides a method for preventing cancer, comprising administering to a subject in need tumor tissue or cancer cells treated with one or more of the above-mentioned treatments.

[0079] In addition, the present invention provides the use of tumor tissue or cancer cells treated with one or more of the above-mentioned treatments for preventing cancer.

[0080] In addition, the present invention provides the use of tumor tissue or cancer cells treated with one or more of the above-mentioned treatments for manufacturing cancer prevention preparations (such as cancer prevention vaccines).

[0081] Advantages of the Invention

[0082] When irradiating a patient with radiation to induce a distal effect, there are problems that normal tissues are damaged and necrotic due to radiation irradiation. There is a risk that radiation irradiation of certain cancer tissues may induce cancer mutations, leading to the development of other forms of radiation-resistant or transformed cancer tissues. The present invention is designed to solve these problems. It relates to a composition that can inhibit the growth, recurrence, and metastasis of cancer, etc., by exerting an excellent distal effect, thereby enhancing the body's immune function against tumors and further improving the anti-cancer effect of anti-cancer treatment. The present invention is characterized in that cancer tissue or cancer cells are extracted from cancer patients, and the cancer cells are induced to weaken or apoptosis by radiation irradiation or cancer immunotherapy drug treatment, and then administered to cancer patients. The re-administered cancer cells or tumor tissue can enhance the sensitivity of the immune system to tumors, thereby maximizing the anti-cancer effect of anti-cancer treatment through the distal effect and minimizing the risk of damage, necrosis, or mutation of normal tissues. In addition, the present invention also has the effects of reducing the minimal residual disease of cancer and tumors, improving the remission of cancer or tumors, prolonging the remission period, reducing the recurrence rate of cancer or tumors, preventing metastasis, reducing the metastasis rate, or achieving a combination of two or more of the above. In addition, when the composition according to the present invention is used in combination with other anti-cancer treatments, a stronger tumor growth inhibitory effect and distal effect can be obtained. In particular, the levels of immune cells and interferons with anti-cancer activity can be increased to enhance the anti-cancer immune function, thereby obtaining a synergistic anti-cancer effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0083] - Figure 1 is a block diagram showing the application sequence and expected positive effects of the present invention;

[0084] - Figures 2A and 2B show the results of observing the radiation-induced sensitization effect by MTT assay after irradiating cancer cells with high-dose radiation (Figure 2A, microscopic observation results; Figure 2B, cell viability comparison results) (C, control group; RT, radiotherapy group).

[0085] - Figure 3 shows the comparison results of primary tumor growth after using IVAM and radiation alone or in combination in a tumor animal model to confirm the anti-cancer effect of combining IVAM treatment and radiotherapy according to the present invention.

[0086] - Figure 4 shows the comparison results of secondary tumor (unirradiated tumor in the left abdomen) growth after using IVAM and radiation alone or in combination in a tumor animal model to confirm the distal effect of combining IVAM treatment and radiotherapy according to the present invention.

[0087] - Figures 5A and 5B show the results of observing the primary tumor (right hind limb) and secondary tumor (left abdomen) by IVIS imaging after using IVAM and radiation alone or in combination in a tumor animal model to confirm the anti-cancer effect of combining IVAM treatment and radiotherapy according to the present invention (Figure 5A), and to quantify the degree of tumor growth (Figure 5B).

[0088] - Figure 6 shows the measurement results of body weight change over time in a tumor animal model treated with IVAM and / or radiation to determine the toxicity of IVAM treatment and radiotherapy according to the present invention.

[0089] - Figure 7 shows a schematic diagram of an animal experiment to confirm the combined effect of IVAM treatment, cancer immunotherapy drug, and radiotherapy according to the present invention.

[0090] - Figure 8 shows the observation results of the growth of the primary tumor (tumor in the irradiated right hind limb) over time after administering the IVAM therapy, cancer immunotherapy drug (a-PD-L1), and radiotherapy (RT) according to the present invention to a tumor animal model alone or in a dual combination or triple combination.

[0091] - Figure 9Shows the observation results of the growth of secondary tumors (unirradiated tumors in the left abdomen) over time after administering the IVAM therapy, cancer immunotherapy drugs, and radiotherapy according to the present invention to a tumor animal model alone or in dual or triple combinations.

[0092] - Figures 10A to 10C show the results of observing the degree of tumor growth by IVIS imaging after administering the IVAM therapy, cancer immunotherapy drugs, and radiotherapy according to the present invention to a tumor animal model alone or in dual or triple combinations.

[0093] - Figure 11 Shows the observation results of the change in the body weight of mice over time when the IVAM therapy, cancer immunotherapy drugs, and radiotherapy according to the present invention are administered to a tumor animal model alone or in dual or triple combinations.

[0094] - Figures 12A and 12B show the results of comparing the degree of lung metastasis by measuring the number of metastatic nodules in lung tissue after administering the IVAM therapy, cancer immunotherapy drugs, and radiotherapy according to the present invention to a tumor animal model alone or in dual or triple combinations.

[0095] - Figures 13A to 13C show the observation results of the distribution changes of immune cells (CD8 + T cells, Tregs, and CD8 + effector memory T cells) in the tumor microenvironment of primary tumors obtained from a tumor animal model. Figures 13A and 13B represent one graph as a whole.

[0096] - Figures 14A and 14B show the changes in various immune cell populations observed in the spleen obtained from a tumor animal model. Figures 14A and 14B represent one graph as a whole.

[0097] - Figure 15 Shows the observation results of the changes in the levels of interferons (IFN-β and IFN-γ) in the serum obtained from a tumor animal model.

[0098] - Figure 16 Shows a schematic diagram of an animal experiment for confirming the long-term survival rate and toxicity of combination therapy by IVAM treatment, cancer immunotherapy drugs, and radiotherapy according to the present invention.

[0099] - Figures 17A and 17B show the observation results of the long-term survival rate of mice (Figure 17A) and the change in the body weight of mice over time (Figure 17B) by combination therapy of IVAM treatment, cancer immunotherapy drugs, and radiotherapy according to the present invention.

[0100] - Figure 18It shows a schematic diagram of an animal experiment to confirm the anti-cancer effect and toxicity of multiple administrations of IVAM according to the present invention.

[0101] - Figure 19 It shows the observation results of the degree of tumor growth by multiple administrations of IVAM according to the present invention.

[0102] - Figure 20 It shows the results of observing the degree of tumor growth by IVIS imaging after multiple administrations of IVAM according to the present invention.

[0103] - Figure 21 It shows the observation results of the change in the body weight of mice over time after multiple administrations of IVAM according to the present invention.

[0104] Best Mode

[0105] The present invention relates to a composition that can enhance the anti-tumor immune function of the body through a distal effect and improve the anti-cancer effect of chemotherapy. It has been confirmed that when tumor tissues or cancer cells isolated from cancer patients are specially treated to induce the inhibition of cancer cell activity or the induction of cancer cell death and then administered to cancer patients, the anti-cancer immune function of the body is activated, and the anti-cancer effect of anti-cancer treatment is significantly enhanced, thereby inhibiting the growth, recurrence, and metastasis of cancer, thus completing the present invention. In particular, it has been confirmed that cancer cells induced to lose activity or be killed as described above can exert a distal effect when administered to cancer patients. In addition, it has also been confirmed that when the composition according to the present invention is used in combination with other chemotherapy, a stronger tumor growth inhibitory effect and distal effect can be obtained, especially the levels of immune cells and interferons with anti-cancer activity can be increased, thereby enhancing the anti-cancer immune function. Therefore, by combining the composition of the present invention with other anti-cancer treatments, a synergistic anti-cancer effect can be obtained.

[0106] Therefore, the main object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, including tumor tissues or cancer cells as active ingredients, which are isolated from cancer patients and are subjected to one or more treatments selected from the group consisting of the following (a) and (b):

[0107] (a) Radiation irradiation; and

[0108] (b) Cancer immunotherapy drugs.

[0109] In this specification, tumor tissues or cancer cells subjected to one or more of the above treatments; or a pharmaceutical composition containing the tumor tissues or cancer cells can be referred to as in vitro distal method (IVAM).

[0110] In addition, the composition can not only achieve the effect of self-preventing or treating cancer by enhancing the anti-cancer immune function of the body, but also further enhance the anti-cancer effect of known anti-cancer treatments. Therefore, the composition of the present invention can be used for the purpose of preventing or treating cancer, and inhibiting the occurrence or progression of metastatic cancer or recurrent cancer. For example, the composition according to the present invention can be used as a vaccine composition for preventing cancer, or can be used to manufacture a vaccine for preventing cancer.

[0111] Generally, when irradiating a patient with radiation for a distal effect, the problem of damage and necrosis of normal tissues caused by radiation irradiation inevitably occurs. In particular, the radiation irradiation of some cancer tissues may cause cancer mutations, which can lead to the development of other forms of cancer tissues that are radiation-resistant or transformed. However, the composition of the present invention has the following advantages: by extracting cancer tissues from cancer patients and subjecting the extracted cancer tissues to physical, chemical, and / or medical treatments (such as radiation irradiation) in vitro to weaken or kill cancer cells or tumor tissues, problems such as normal tissue damage, necrosis, and mutations can be prevented. As described above, when the tumor tissues or cancer cells weakened or killed by radiation irradiation are re-administered to cancer patients, the body's immune system recognizes, attacks, and / or preys on the killed or inactivated cancer cells through heat treatment. Through the experience of attacking the killed cancer cells, the body becomes more sensitive to other cancer cells, enabling it to more sensitively recognize, attack, and kill them. That is, according to the present invention, a distal effect can be expected through the tumor tissues or cancer cells subjected to radiation irradiation. In particular, the inventors of the present invention have confirmed that when the above-mentioned cancer cells irradiated in vitro are re-administered to a tumor animal model and then used in combination with radiotherapy, not only is the anti-cancer effect of radiotherapy further improved, but also metastatic cancer is effectively inhibited.

[0112] In addition, the present invention is characterized in that cancer tissues extracted from cancer patients are treated with cancer immunotherapy drugs to induce the death or weakening of cancer cells or tumor tissues in vitro, and then re-administered to cancer patients. The immune system of cancer patients can easily recognize, attack, and / or prey on the re-administered tumor tissues or cancer cells. Through the experience of attacking the killed cancer cells, the immune system becomes more sensitive to other cancer cells in the body, and thus can more sensitively recognize, attack, and kill them. That is, according to the present invention, a distal effect can be expected through the tumor tissues or cancer cells treated with cancer immunotherapy drugs.

[0113] The tumor tissues or cancer cells that have undergone the above-mentioned special treatment, when re-administered to the cancer patients from whom they originated, will stimulate the immune system and induce the same effects as immunostimulants. That is, if the tumor tissues or cancer cells are re-introduced into the body in a weakened or killed state through special treatment, even if they were not previously recognized by the immune system, the immune system can attack them and acquire a learning effect, enabling it to recognize cancer cells and metastatic cells that were not previously recognized. Therefore, the immune system of a subject that has acquired the ability to recognize cancer cells through learning can attack malignant cells throughout the body.

[0114] That is, since the administration of the inoculated tissue cells of the present invention that induce distal effects triggers an immune response and activates the immune system, the administration of the inoculated tissue cells of the present invention can be used to treat primary cancers and their metastases, as well as prevent the recurrence of cancer diseases in patients with a weak or suppressed immune system.

[0115] The order of administering the composition according to the present invention to a subject, its distal effects, and its anti-cancer effects are schematically shown in Figure 1 as follows.

[0116] The term "cancer" as used herein refers to a disease characterized by uncontrolled cell growth. Due to this abnormal cell growth, cell masses called tumors are formed, which infiltrate the surrounding tissues and, in severe cases, also metastasize to other organs of the body. Academically, it is also called a neoplasm. Cancer is an incurable chronic disease that, in many cases, cannot be fundamentally cured even through surgery, radiation, and chemotherapy, causing pain to patients and ultimately leading to death. The causes of cancer are diverse, but are mainly divided into internal and external factors. The exact mechanism by which normal cells transform into cancer cells is not yet clear, but it is known that a large number of cancers are caused by external factors such as environmental factors. Internal factors include genetic factors, immunological factors, etc., while external factors include chemicals, radiation, viruses, etc. Genes involved in cancer development include oncogenes and tumor suppressor genes, and cancer appears when the balance between genes is disrupted by the above internal or external factors. In the present invention, cancer includes all primary cancers, cancers treated with radiation, cancers not treated with radiation, metastatic cancers, and recurrent cancers. In addition, the composition according to the present invention may be particularly suitable for the treatment of non-immunogenic tumors or cancers.

[0117] In addition, in the present invention, cancer may include all types of solid tumors and all types of blood cancers. As a non-limiting example, the cancers of the present invention may be selected from the group consisting of: adenocarcinoma, choroidal melanoma, acute leukemia, acoustic neuroma, ampullary cancer, anal cancer, astrocytoma, basal cell carcinoma, pancreatic cancer, desmoid tumor, bladder cancer, bronchial cancer, non-small cell lung cancer (NSCLC), breast cancer, Burkitt lymphoma, gastric body cancer, cancer of unknown primary (CUP) syndrome, colorectal cancer, small intestine cancer, small intestine tumors, ovarian cancer, endometrial cancer, ependymoma, epithelial cancer types, Ewing's tumor, gastrointestinal tumors, gastric cancer, gallbladder cancer, gallbladder carcinoma, uterine cancer, cervical cancer, cervix, glioblastoma, gynecological tumors, ENT tumors, hematological tumors, hairy cell leukemia, urethral cancer, skin cancer, skin testicular cancer, brain tumors (gliomas), brain metastases, testicular cancer, pituitary tumors, carcinoid, Kaposi sarcoma, laryngeal cancer, germ cell tumor, bone cancer, colorectal cancer, head and neck tumors (tumors of the ear, nose and throat region), colon cancer, craniopharyngioma, oral cancer (cancer of the oral cavity and lips), central nervous system cancer, liver cancer, liver metastases, leukemia, eyelid tumors, lung cancer, lymph node cancer (Hodgkin / non-Hodgkin), lymphoma, gastric cancer, malignant melanoma, malignant tumors, gastrointestinal malignancies, breast cancer, rectal cancer, medulloblastoma, melanoma, meningioma, Hodgkin's disease, mycosis fungoides, nasal cancer, schwannoma, neuroblastoma, kidney cancer, renal cell carcinoma, non-Hodgkin lymphoma, oligodendroglioma, esophageal cancer, osteolytic and osteoblastic carcinoma, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, plasmacytoma, head and neck squamous cell carcinoma (SCCHN), prostate cancer, throat cancer, rectal cancer, retinoblastoma, vaginal cancer, thyroid cancer, Schnberg disease, esophageal cancer, spinalioms, T-cell lymphoma (Mycobacterium tuberculosis), thymoma, ductal carcinoma, eye tumors, urethral cancer, urinary system tumors, urothelial carcinoma, vulvar cancer, verrucous appearance, soft tissue tumors, soft tissue sarcoma, Wilms tumor, cervical cancer, and tongue cancer. For example, it may be selected from the group consisting of astrocytoma, glioblastoma, and renal cell carcinoma.

[0118] The present invention can also be regarded as inducing vaccination by irradiating and / or immunologically treating isolated tumor tissues or cancer cells of cancer patients. Thus, in one embodiment of the present invention, a vaccine composition for preventing cancer can be manufactured, which comprises tumor tissues or cancer cells that have been treated therewith. This vaccine composition can enhance the function of the immune system of cancer patients, enabling them to recognize cancer cells that are not recognized as clearance targets because they are judged to be their own body tissues (i.e., increasing the sensitivity to cancer). In addition, the vaccine composition can also enable the immune system to recognize metastatic cancer when it occurs and generate memory for cancer, so as to recognize new cancer cells that will still recur even after the successful cancer treatment of the patient. The composition of the present invention can fully play the above roles, thereby achieving personalized tumor and cancer vaccination in the subject. In other words, the composition of the present invention can be used as an immune stimulant to enhance the ability of the immune system of the subject to recognize and eliminate cancer cells, eradicate metastatic cancer, and recognize and eliminate new cancer cells that recur after successful cancer treatment.

[0119] In one embodiment of the present invention, the tumor tissue or cancer cells can be tumor tissues or cancer cells isolated from an individual in need of preventing or treating cancer. That is, the tumor tissue or cancer cells can be autologous tumor tissues or cancer cells of a subject in need of cancer prevention or treatment. In another embodiment of the present invention, the tumor tissue or cancer cells can be tumor tissues or cancer cells isolated from a subject different from the subject in need of cancer prevention or treatment. That is, the tumor tissue or cancer cells can be allogeneic tumor tissues or cancer cells of a subject in need of cancer prevention or treatment. Therefore, the pharmaceutical composition according to the present invention can be administered to a cancer patient from whom the tumor tissue or cancer cells serving as the active ingredient are isolated (i.e., re-administering killed or attenuated autologous tumor tissues or cancer cells), or can be administered to a subject different from the cancer patient from whom the tumor tissue or cancer cells are isolated (i.e., administering killed or attenuated allogeneic tumor tissues or cancer cells).

[0120] In particular, the composition according to the present invention can be administered to a subject without cancer (i.e., a normal subject) to prevent cancer. For example, the composition according to the present invention can be characterized by containing tumor tissues or cancer cells that are isolated from a cancer patient, weakened or killed by the treatment according to the present invention, and administered to a subject allogeneic to the cancer patient, where the subject can be a subject without cancer. Thus, even if a subject is likely to develop cancer due to genetic or environmental reasons, the anti-cancer ability of the immune system can be enhanced by receiving allogeneic tumor tissues or cancer tissues that are weakened or killed by the treatment according to the present invention, enabling the immune system to experience and understand cancer, thereby preventing or delaying the occurrence of cancer.

[0121] The vaccinated tumor tissue or cancer cells of the present invention can act on the whole body through distal effects, etc. That is, the present invention can achieve systemic treatment only by extracting local cancer tissue. Since the composition according to the present invention can act on the whole body, tumor antigens that cannot be detected by existing anticancer drugs or radiotherapy can be discovered, and cancer can be treated without artificial non-invasive or invasive measures that impose a burden on the body. In addition, this immune induction function can even be induced in frail patients without any additional physiological burden.

[0122] As used herein, the term "distal effect" refers to a systemic anti-cancer effect that can inhibit metastatic tumors in areas not directly irradiated by radiation, in addition to the local area directly irradiated by radiation. Therefore, the distal effect causes the shrinkage of tumors distributed in other parts of the body beyond the scope of local tumor treatment. However, it cannot be said that only radiation exposure induces the distal effect; the distal effect can also be induced by treatments such as immunotherapy.

[0123] In the present invention, the substance that induces the distal effect, i.e., the substance that induces antibodies against cancer cells such as tumor antigens, is broadly regarded as part of the vaccine. Therefore, the distal effect-inducing substance can be expressed as "distal effect-inducing vaccine".

[0124] In particular, the present invention plays an equally important role as an immune stimulant in treating primary cancers and their metastatic cancers in a dispersed cell state that cannot be detected by imaging methods, and in preventing the recurrence of cancer diseases. This immune stimulation method can cause a systemic vaccine effect that activates the immune system, and is particularly suitable for treating metastatic cancers and primary cancers that cannot be detected by existing technical methods, and is particularly suitable for preventing the recurrence of cancers that have been successfully treated.

[0125] Through the present invention, the treatment of the following primary cancer types can be achieved, the treatment of metastatic cancers of the following cancer types can be achieved, and the recurrence of the following cancer types can be prevented:

[0126] Adenocarcinoma, choroidal melanoma, acute leukemia, acoustic neuroma, ampullary cancer, anal cancer, astrocytoma, basal cell carcinoma, pancreatic cancer, desmoid tumor, bladder cancer, bronchial cancer, non-small cell lung cancer (NSCLC), breast cancer, Burkitt lymphoma, gastric body cancer, carcinoma of unknown primary (CUP) syndrome, colorectal cancer, small intestine cancer, small intestine tumor, ovarian cancer, endometrial cancer, ependymoma, epithelial cancer type, Ewing's tumor, gastrointestinal tumor, gastric cancer, gallbladder cancer, gallbladder cancer, uterine cancer, cervical cancer, cervix, glioblastoma, gynecological tumor, ENT tumor, hematological tumor, hairy cell leukemia, urethral cancer, skin cancer, skin testicular cancer, brain tumor (glioma), brain metastasis, testicular cancer, pituitary tumor, carcinoid, Kaposi sarcoma, laryngeal cancer, germ cell tumor, bone cancer, colorectal cancer, head and neck tumor (tumor in the ear, nose and throat area), colon cancer, craniopharyngioma, oral cancer (cancer in the oral cavity and lips), central nervous system cancer, liver cancer, liver metastasis, leukemia, eyelid tumor, lung cancer, lymph node cancer (Hodgkin / non-Hodgkin), lymphoma, gastric cancer, malignant melanoma, malignant tumor, gastrointestinal malignant tumor, breast cancer, rectal cancer, medulloblastoma, melanoma, meningioma, Hodgkin's disease, mycosis fungoides, nasal cancer, schwannoma, neuroblastoma, kidney cancer, renal cell carcinoma, non-Hodgkin lymphoma, oligodendroglioma, esophageal cancer, osteolytic and osteoblastic cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, plasmacytoma, head and neck squamous cell carcinoma (SCCHN), prostate cancer, throat cancer, rectal cancer, retinoblastoma, vaginal cancer, thyroid cancer, Schnberg disease, esophageal cancer, spinal tumor (spinalioms), T-cell lymphoma (Mycobacterium tuberculosis), thymoma, ductal carcinoma, eye tumor, urethral cancer, urinary system tumor, urothelial carcinoma, vulvar cancer, verrucous appearance, soft tissue tumor, soft tissue sarcoma, Wilms tumor, cervical cancer and tongue cancer. For example, it can be particularly suitable for treating astrocytoma, glioblastoma, pancreatic cancer, bronchial cancer, breast cancer, colorectal cancer, ovarian cancer, gastric cancer, laryngeal cancer, malignant melanoma, esophageal cancer, cervical cancer, liver cancer, bladder cancer and renal cell carcinoma.

[0127] In other words, the composition according to the present invention enables the patient's immune system to recognize cancer cells of the above cancer types that are different from the normal cells of the subject and kill them.

[0128] In addition, the composition according to the present invention only performs radiation irradiation, anti-cancer treatment, etc. on tumor tissues or cancer cells isolated from cancer patients in vitro, and the treatment does not directly act on cancer patients, so there is no need to worry about serious side effects caused by the treatment. For example, the tissues or cancer cells extracted from the patient will be completely killed, or injected into the patient in a weakened state, which makes it unable to grow and proliferate, so it cannot exist as other mutant cancer cells. Therefore, the application of the present invention is suitable for combination with standard treatments involving one or more of the standard treatments (surgery, radiotherapy, anti-cancer treatment) developed so far and general cancer treatment methods (such as immunotherapy).

[0129] In the present invention, the treatment can be radiation irradiation. In the present invention, the radiation can be selected from but not limited to alpha rays, beta rays, gamma rays, X-rays, ultraviolet rays, infrared rays, near-infrared rays, hadron rays, deuteron rays, etc.

[0130] When radiation acts on tumor tissues or cancer cells extracted from cancer patients, the following mechanism can be expected: normal cancer cells will hide factors that can be recognized by the immune system as much as possible, but when irradiated, various MHC antigens will be highly expressed. Cancer cells killed by radiation will produce a large number of factors that activate the innate immune system, such as HMGB-1 or ATP. For example, irradiated cancer cells have a high level of ICAM-1 expression on their surface, making it easier for immune cells to recognize and attack them. In this way, cancer cells killed or weakened by radiation not only present obvious markers (Fas receptor), enabling immune cells to better recognize them (calreticulin) and kill them, but also emit more battle signals (i.e., activation signals (NKG2D)) to arm immune cells. In addition, irradiated cancer cells will produce more chemokines that recruit immune cells. Therefore, when a composition containing irradiated tumor tissues or cancer cells according to the present invention is administered to a cancer patient, immune cells will gather around the administered tumor tissues or cancer cells, thus increasing the possibility of immune cells recognizing cancer cells. Therefore, the vaccine effect induced by the present invention helps to activate the immune system in the subject. In addition, since the radiation irradiation of the present invention is carried out in vitro, different from the above-mentioned existing methods, it can prevent the possibility of cancer tissues in the subject's body (in vivo) mutating due to radiation.

[0131] In the present invention, the dose of the irradiation radiation can be 1 to 500 Gray, 1 to 400 Gray, 1 to 300 Gray, 1 to 200 Gray, 1 to 100 Gray, 1 to 80 Gray, 1 to 50 Gray, 1 to 30 Gray, 1 to 10 Gray, 5 to 10 Gray, 10 to 200 Gray, 10 to 100 Gray, 10 to 80 Gray, 20 to 60 Gray, 30 to 60 Gray or 40 to 60 Gray, but not limited thereto.

[0132] The radiation can be administered in a single dose or multiple doses. For example, multiple dose equivalents can be irradiated instead of a single dose of radiation.

[0133] In the present invention, the treatment can be a cancer immunotherapy drug.

[0134] The term "anticancer agent" used in the specification generally refers to substances used for treating malignant tumors. Most anticancer drugs are drugs that interfere with various metabolic pathways of cancer cells, mainly inhibiting nucleic acid synthesis or exhibiting anticancer activity. Currently, anticancer drugs used for treating cancer can be classified into six categories according to their biochemical mechanisms of action: alkylating agents, antimetabolites, antibiotics, mitotic inhibitors (vinca alkaloids), hormones, and others. However, the anticancer drug according to the present invention may not be included in these categories.

[0135] "Cancer immunotherapy" using a cancer immunotherapy drug is a cancer treatment that activates the body's immune system to fight cancer cells. Cancer immunotherapy drugs exert their anticancer effects by enhancing the specificity, memory, and adaptability of the immune system. That is, by utilizing the human immune system, it precisely attacks only cancer cells with fewer side effects, and by taking advantage of the memory and adaptability of the immune system, patients responsive to cancer immunotherapy drugs can see continuous anticancer effects. Preferably, the cancer immunotherapy drug can be selected from the group consisting of immune checkpoint inhibitors, costimulatory molecule preparations, cytokine therapeutics, CAR-T cell therapeutics, and autologous CD8 +One or more of a group consisting of T cell therapy agents, but not limited thereto. Immune checkpoint inhibitors refer to agents that can inhibit immune checkpoints involved in the immune escape mechanism of cancer cells. Some cancer cells evade the immune system by using immune checkpoints of immune cells. Immune checkpoint inhibitors bind to the binding sites of cancer cells and T cells, block immune escape signals, and prevent the formation of immune synapses. Therefore, T cells not blocked by immune escape have a mechanism to eliminate cancer cells. Immune checkpoint inhibitors can be at least one selected from the group consisting of PD-L1 inhibitors, PD-1 inhibitors, CTLA-4 inhibitors, LAG3 inhibitors, TIM3 inhibitors, 4-1BB inhibitors, LAG-3 inhibitors, B7-H4 inhibitors, HVEM inhibitors, TIM4 inhibitors, GAL9 inhibitors, VISTA inhibitors, KIR inhibitors, TIGIT inhibitors, and BTLA inhibitors, but not limited thereto. The inhibitors are not limited to specific types and include, but are not limited to, any substance that can inhibit the function or expression of the target (immune checkpoint protein). As specific examples, there are its antibodies or fragments (Fab, Fab', F(ab')2, scFv, (scFv)2, Fv, dsFv, diabodies, nanobodies, Fd, and Fd', etc.), compounds, other peptides, etc. Examples of commercially available immune checkpoint inhibitors include, but are not limited to: pembrolizumab (Keytruda), ipilimumab (Yervoy), nivolumab (Opdivo), atezolizumab (Tecentriq), cemiplimab (Libtayo), atezolizumab (Tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), tremelimumab (Imjuno), relatlimab, nivolumab, etc.

[0136] In addition to cancer immunotherapy drugs, immunotherapy can also include immune cell therapy. Genetic modification of immune cells is well-known as cancer immune cell therapy. These immune cell therapies are based on manipulating autologous or allogeneic immune cells and administering them to subjects in need. Immune cell-based therapies include natural killer cell therapy, dendritic cell therapy, and T cell immunotherapy (including naive T cells, effector T cells (also known as T helper cells), cytotoxic T cells, regulatory T cells (Tregs), etc.).

[0137] Genetically modified immune cells can be T cells. In another embodiment, the T cells are naive T cells. In another embodiment, the T cells are naive CD4 + T cells. In another embodiment, the T cells are naive T cells. In another embodiment, the T cells are naive CD8 +T cells. In another embodiment, the genetically modified immune cell is a natural killer (NK) cell. In another embodiment, the genetically modified immune cell is a dendritic cell. In another embodiment, the genetically modified T cell is a cytotoxic T lymphocyte (CTL cell). In another embodiment, the genetically modified T cell is a regulatory T cell (Treg). In another embodiment, the genetically modified T cell is a chimeric antigen receptor (CAR) T cell. In another embodiment, the genetically modified T cell is a genetically modified T cell receptor (TCR) cell.

[0138] Immunotherapy involves cytokines. Cytokines include granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukins (such as IL-2) and / or interferons (such as IFN-α). Other methods of enhancing tumor-targeted immune responses include additional immune checkpoint inhibition. Immune checkpoint inhibitors include anti-CTLA4, anti-PD-1, anti-PD-L1, anti-PD-L2, anti-TIM-3, anti-LAG-3, anti-A2aR or anti-KIR antibodies. Immunotherapy includes co-stimulatory receptor agonists (e.g., anti-OX40 antibody, anti-GITR antibody, anti-CD137 antibody, anti-CD40 antibody and anti-CD27 antibody). Immunotherapy involves inhibiting T regulatory cells (Tregs), myeloid-derived suppressor cells (MDSCs) and cancer-associated fibroblasts (CAFs). Immunotherapy includes stimulating innate immune cells (e.g., natural killer (NK) cells, macrophages and dendritic cells). Other immune-stimulating therapies include IDO inhibitors, TGF-β inhibitors, IL-10 inhibitors, stimulator of interferon genes (STING) agonists, toll-like receptor (TLR) agonists (such as TLR7, TLR8 or TLR9), tumor vaccines (such as whole tumor cell vaccines, peptide and recombinant tumor-associated antigen vaccines), adoptive cell therapy (ACT) (such as T cells, natural killer cells, TILs and LAK cells) and ACT with genetically engineered receptors (such as chimeric antigen receptors (CARs) and T cell receptors (TCRs)). Combinations of these agents (e.g., combinations of immune checkpoint inhibitors), checkpoint inhibition + T cell co-stimulatory receptor agonist effects and checkpoint inhibition + TIL ACT can be used. Other anti-cancer treatments include combinations of immune checkpoint inhibitors (such as avelumab), 4-1BB (CD-137) agonists (such as utomilumab) and OX40 (TNFRS4) agonists.

[0139] The composition according to the invention may contain only one type of tumor cell or cancer cell that has been specifically treated as an active ingredient, or may contain two or more types of tumor tissues or cancer cells. Here, the two or more types of tumor tissues or cancer cells may have been treated with the same type of treatment, but may also have been treated with different types of treatment.

[0140] When the composition according to the present invention comprises two or more types of tumor tissues or cancer cells, the composition may be in the form of a mixture in which two or more types of tumor tissues or cancer cells are mixed. That is, two or more types of tumor tissues or cancer cells may be administered simultaneously.

[0141] The composition may be in a form in which two or more types of tumor tissues or cancer cells are formulated separately and administered simultaneously or sequentially. In this case, the composition may be a pharmaceutical composition for combined administration for simultaneously or sequentially administering tumor tissues or cancer cells. In the case of sequential administration, there is no limitation on the administration order, and the administration regimen may be appropriately adjusted according to the patient's condition and the like.

[0142] In addition, the composition according to the present invention can be used in combination with other chemotherapy. The composition according to the present invention can enhance the sensitivity of a subject to chemotherapy through a distal effect, and regulate the activities and levels of immune cells and immunomodulatory factors involved in the immune response, thereby maximizing the anti-cancer effect of anti-cancer drugs. Therefore, the composition of the present invention can enhance the anti-cancer effect of combined anti-cancer therapy and reduce side effects. In addition, studies have confirmed that when the composition according to the present invention is used in combination with radiotherapy and cancer immunotherapy drugs, tumor growth can be more effectively inhibited, the distal effect of radiotherapy can be further enhanced, and cancer metastasis can be reduced. In addition, it has also been confirmed that the excellent anti-cancer effect of the combination therapy is the result of an increase in anti-cancer immune cells in tumors and the spleen and an increase in the level of anti-cancer interferon in the serum. In addition, it has also been confirmed that the composition according to the present invention can maintain the survival rate without causing side effects such as toxicity even during a long period of 50 days when used in combination with anti-cancer therapy.

[0143] According to one embodiment of the present invention, it has been confirmed that the composition according to the present invention can maintain the survival rate without showing toxicity when administered once a day or 1 to 3 times, 1 to 2 times, or 2 to 3 times at intervals of 3 to 4 days. Therefore, the composition according to the present invention may be administered 1 to 3 times, but is not limited thereto.

[0144] The term "combination" as used herein can be achieved by simultaneously, sequentially, or separately administering (treating) the individual components of a treatment regimen. By simultaneously, sequentially, or alternately performing two or more anti-cancer treatment processes at fixed or non-fixed intervals, a combined treatment effect can be obtained. Combination therapy can be defined as a treatment that provides a synergistic effect and is therapeutically superior to the efficacy that can be obtained by administering one or another component of the combination therapy at a conventional dose (e.g., but not limited to, the efficacy measured by the degree of response, response rate, time to disease progression, or survival time).

[0145] The composition according to the present invention can be used in combination with one or more selected from the group consisting of radiotherapy, chemotherapy, targeted anti-cancer therapy, and cancer immunotherapy, but is not limited thereto.

[0146] In the present invention, the "targeted anti-cancer agent" refers to an agent that exerts an anti-cancer effect by targeting proteins or genes that have specific changes in cancer cells or cancer tissues and interfering with molecular activities involved in cancer growth and occurrence. The targeted anti-cancer agent can be one or more selected from the group consisting of tyrosine kinase inhibitors (TKIs), poly-ADP ribose polymerase inhibitors (PARP inhibitors), angiogenesis inhibitors, cyclin-dependent kinase 4 / 6 inhibitors (CDK4 / 6 inhibitors), hormone therapy drugs, and antibody-drug conjugates, but is not limited thereto.

[0147] The term "chemotherapeutic agent" used in the specification refers to the first-generation anti-cancer agents, also known as "cytotoxic anti-cancer agents" or "chemical anti-cancer agents".

[0148] The composition according to the present invention is characterized by being used in combination with radiotherapy and cancer immunotherapy. The detailed description of cancer immunotherapy drugs and cancer immunotherapy has been described above, so it is omitted.

[0149] The composition according to the present invention can be administered simultaneously, separately, or sequentially with chemotherapy. In addition, each chemotherapy can be repeated.

[0150] For example, the composition according to the present invention can be administered 1 to 3 days, 1 to 5 days, 1 to 7 days, 1 to 10 days, 1 to 20 days, 1 to 30 days, 5 to 15 days, 5 to 20 days, 10 minutes to 2 hours, 10 minutes to 1 hour, 10 minutes to 50 minutes, or 10 minutes to 30 minutes before anti-cancer treatment, can be administered simultaneously with anti-cancer treatment, and can be administered 5 minutes to 10 minutes, 30 minutes to 1 hour, 6 hours to 12 hours, 12 hours to 24 hours, 1 day to 3 days, 1 day to 5 days, 1 day to 7 days, 1 day to 10 days, 1 day to 20 days, 1 day to 30 days, 5 days to 15 days, or 5 days to 20 days after anti-cancer treatment. In addition, the composition according to the present invention can be administered before or after anti-cancer treatment.

[0151] The anti-cancer treatment is not limited and can include any anti-cancer treatment known in the art. As non-limiting examples, there are surgical therapy, chemotherapy (e.g., administration of protein kinase inhibitors or EGFR-targeted therapy), embolization therapy, chemoembolization therapy, radiotherapy, cryotherapy, hyperthermia, phototherapy, radiofrequency ablation, hormone therapy, immunotherapy, small molecule therapy, receptor kinase inhibitor therapy, anti-angiogenesis therapy, cytokine therapy or biotherapy, monoclonal antibodies, siRNA, miRNA, antisense oligonucleotides, ribozymes, or gene therapy, etc.

[0152] In the treatment using anti-cancer drugs, actinomycin D, aminoglutethimide, amsacrine, anastrozole, antagonists of purine and pyrimidine bases, anthracyclines, aromatase inhibitors, asparaginase, anti-estrogens, bexarotene, bleomycin, buserelin, busulfan, camptothecin derivatives, capecitabine, carboplatin, carmustine, chlorambucil, cladribine, cyclophosphamide, cytarabine, cytosine arabinoside, alkylating cytostatic agents, dacarbazine, actinomycin D, daunorubicin, docetaxel, doxorubicin (adriamycin), doxorubicin liposome (Lipo-Dox), epirubicin, estramustine, etoposide, exemestane, fludarabine, fluorouracil, folic acid antagonists, formestane, gemcitabine, glucocorticoids, goserelin, hormone antagonists, hexamethylmelamine, hydroxyurea, idarubicin, ifosfamide, imatinib, irinotecan, letrozole, leuprorelin, lomustine, melphalan, mercaptopurine, methotrexate, miltefosine, mitomycin, mitotic inhibitors, mitoxantrone, nimustine, oxaliplatin, paclitaxel, pentostatin, procarbazine, tamoxifen, temozolomide, teniposide, testolactone, thiotepa, thioguanine, topoisomerase inhibitors, topotecan, treosulfan, retinoic acid, triptorelin, trofosfamide, vinblastine, vincristine, vindesine, vinorelbine, antibiotics with cytotoxic activity, etc. can be used.

[0153] Treatment using immunotherapeutics generally relies on the use of immune effector cells and molecules to target and destroy cancer cells. For example, the immune effector can be an antibody specific to certain markers on the surface of tumor cells. The antibody itself can serve as the effector of the treatment or can recruit other cells that actually affect cell death. The antibody can also be conjugated with a drug or toxin (chemotherapeutic drugs, radionuclides, ricin A chain, cholera toxin, pertussis toxin, etc.) and act only as a targeting agent. Alternatively, the effector can also be a lymphocyte that delivers surface molecules that directly or indirectly interact with cell targets. Various effector cells include cytotoxic T cells and NK cells, as well as genetically engineered variants of these cell types that have been modified to express chimeric antigen receptors.

[0154] Other complementary immunotherapies, such as GM-CSF that increases the number of innate immune system cells derived from the bone marrow, low-dose cyclophosphamide or PI3K inhibitors that eliminate T regulatory cells that suppress innate and adaptive immunity, and 5FU, PI3K inhibitors or histone deacetylase inhibitors that eliminate myeloid-derived suppressor cells, can also be added to the above-mentioned therapies to further improve the efficacy.

[0155] Immunotherapy can also include the administration of interleukins (such as IL-2) or interferons (such as INFα).

[0156] Examples of immunotherapies that can be combined with p53, ADP, and / or MDA-7 gene therapies, as well as CD122 / CD132 agonists, include immune adjuvants (e.g., Mycobacterium bovis, Plasmodium falciparum, dinitrochlorobenzene, and aromatic compounds), cytokine therapy, gene therapy, and monoclonal antibodies. Thus, the use of one or more anti-cancer therapies in combination with the p53, ADP, and / or MDA-7 gene therapies described in this application can be considered.

[0157] Other immunotherapies that can be combined with p53, ADP, and / or MDA-7 gene therapies, as well as CD122 / CD132 agonists, include immune checkpoint inhibitors, co-stimulatory receptor agonists, innate immune cell stimulants, or innate immune activators. In certain aspects, the immune checkpoint inhibitor is an inhibitor of CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, BTLA, B7H3, B7H4, TIM3, KIR, or A2aR. In certain aspects, at least one immune checkpoint inhibitor is an anti-killer cell immunoglobulin-like receptor (KIR) antibody.

[0158] In some aspects, at least one immune checkpoint inhibitor is a human programmed cell death 1 (PD-1) axis-binding antagonist. In certain aspects, the PD-1 axis-binding antagonist is selected from the group consisting of PD-1-binding antagonists, PDL1-binding antagonists, and PDL2-binding antagonists. In some aspects, the PD-1 axis-binding antagonist is a PD-1-binding antagonist. In certain aspects, the PD-1-binding antagonist inhibits the binding of PD-1 to PDL1 and / or PDL2. In particular, the PD-1-binding antagonist is a monoclonal antibody or an antigen-binding fragment thereof.

[0159] Co-stimulatory receptor agonists can be anti-OX40 antibodies, anti-GITR antibodies, anti-CD137 antibodies, anti-CD40 antibodies, or anti-CD27 antibodies. Stimulants of innate immune cells include, but are not limited to, KIR monoclonal antibodies, cytotoxic inhibitory receptor inhibitors, and toll-like receptors. Activators of innate immune cells (such as natural killer (NK) cells, macrophages, and dendritic cells) include IDO inhibitors, TGF inhibitors, and IL-10 inhibitors. Indoximod is a typical innate immune activator.

[0160] Immunotherapy can include inhibiting T regulatory cells, myeloid-derived suppressor cells (MDSC), and cancer-associated fibroblasts (CAF). In some embodiments, the immunotherapy is a tumor vaccine (e.g., whole tumor cell vaccine, dendritic cell vaccine, DNA and / or RNA expression vaccine, peptide and recombinant tumor-associated antigen vaccine), or adoptive cell therapy (ACT) (e.g., T cells, natural killer cells, TIL, and LAK cells). T cells and / or natural killer cells can be engineered with a chimeric antigen receptor (CAR) or a T cell receptor (TCR) to a specific tumor antigen. The chimeric antigen receptor (or CAR) used in the present invention can refer to any specifically engineered receptor against an antigen of interest, which, when expressed in a T cell or a natural killer cell, can confer the specificity of the CAR to the T cell or the natural killer cell. Once T cells or natural killer cells expressing the chimeric antigen receptor are generated using standard molecular techniques, they can be introduced into a patient through techniques such as adoptive cell transfer.

[0161] The immunotherapy can be a cancer vaccine comprising one or more cancer antigens, particularly a protein or an immunogenic fragment thereof, DNA or RNA encoding a cancer antigen, particularly a protein or an immunogenic fragment thereof, a cancer cell lysate, and / or a protein preparation of tumor cells. The cancer antigen used in the specification is an antigenic substance present in cancer cells. In principle, any protein that is produced in cancer cells, is upregulated in cancer cells compared to normal cells, or has an abnormal structure due to mutation can serve as a cancer antigen. In principle, the cancer antigen can be the product of a mutated or overexpressed oncogene and tumor suppressor gene, the product of other mutated genes, a cell protein that is overexpressed or abnormally expressed, a cancer antigen produced by an oncogenic virus, a carcinoembryonic antigen, an altered cell surface glycolipid and glycoprotein, or a cell type-specific differentiation antigen. Examples of cancer antigens include the abnormally or overexpressed products of the ras and p53 genes. Other examples include tissue differentiation antigens, mutant protein antigens, oncogenic virus antigens, cancer-testis antigens, and vascular or stroma-specific antigens. Tissue differentiation antigens refer to antigens that are specific to a particular type of tissue. Mutant protein antigens may be more specific to cancer cells because normal cells should not contain these proteins. Normal cells display normal protein antigens on their MHC molecules, while cancer cells display mutant protein antigens. Some viral proteins are associated with the formation of cancer, so some viral antigens are also cancer antigens.

[0162] The immunotherapy can be an antibody, for example, part of a polyclonal antibody preparation, or a monoclonal antibody. The antibody can be a humanized antibody, a chimeric antibody, an antibody fragment, a bispecific antibody or a single-chain antibody. The antibodies disclosed herein include antibody fragments such as, but not limited to, Fab, Fab', and F(ab')2, Fd, single-chain Fv (scFv), single-chain antibody, disulfide-linked Fv (sdfv), and fragments comprising VL or VH domains. In some aspects, the antibody or its fragment specifically binds to epidermal growth factor receptor (EGFR1, Erb-Bl), HER2 / neu (Erb-B2), CD20, vascular endothelial growth factor (VEGF), insulin-like growth factor receptor (IGF-1R), TRAIL-receptor, epithelial cell adhesion molecule, carcinoembryonic antigen, prostate-specific membrane antigen, mucin-1, CD30, CD33 or CD40.

[0163] Examples of monoclonal antibodies that can be used in combination with the compositions provided herein include, but are not limited to, trastuzumab (anti-HER2 / neu antibody); pertuzumab (anti-HER2 monoclonal antibody); cetuximab (chimeric monoclonal antibody to epidermal growth factor receptor EGFR); panitumumab (anti-EGFR antibody); nimotuzumab (anti-EGFR antibody); zalutumumab (anti-EGFR monoclonal antibody); necitumumab (anti-EGFR monoclonal antibody); MDX-210 (humanized anti-HER-2 bispecific antibody); MDX-210 (humanized anti-HER-2 bispecific antibody); MDX-447 (humanized anti-EGF receptor bispecific antibody); rituximab (chimeric mouse / human anti-CD20 monoclonal antibody); obinutuzumab (anti-CD20 monoclonal antibody); ofatumumab (anti-CD20 monoclonal antibody); tositumomab-Il3l (anti-CD20 monoclonal antibody); ibritumomab tiuxetan (anti-CD20 monoclonal antibody); bevacizumab (anti-VEGF monoclonal antibody); ramucirumab (anti-VEGFR2 monoclonal antibody); ranibizumab (anti-VEGF monoclonal antibody); aflibercept (extracellular domains of VEGFR1 and VEGFR2 fused to IgGl Fc); AMG386 (angiopoietin-1 and -2 binding peptide fused to IgGl Fc); daratumumab (anti-IGF-lR monoclonal antibody); gemtuzumab ozogamicin (anti-CD33 monoclonal antibody); alemtuzumab (anti-Campath-l / CD52 monoclonal antibody); brentuximab vedotin (anti-CD30 monoclonal antibody); catumaxomab (bispecific mAh targeting epithelial cell adhesion molecule and CD3); naprotumab (anti-5T4 monoclonal antibody); gimatecan (anti-carbonic anhydrase ix); or farletuzumab (anti-folate receptor). Other examples also include antibodies such as PanorexTM (17-1A) (murine monoclonal antibody); Panorex (@(17-1A)) (chimeric murine monoclonal antibody); BEC2 (anti-idiotypic monoclonal antibody, mimicking GD epitope) (with BCG); Oncolym (Lym-1 monoclonal antibody); SMART M195 antibody, humanized 13'1LYM-1 (Oncolym), Ovarex (B43.13, anti-idiotypic murine monoclonal antibody); 3622W94 mAb that can bind to EGP40 (17-1A) pancreatic cancer antigen on adenocarcinoma; Zenapax (SMART anti-Tac (IL-2 receptor); SMART M195 antibody, humanized antibody, humanized); NovoMAb-G2 (pancreatic cancer-specific antibody); TNT (histone antigen chimeric mAb); TNT (histone antigen chimeric mAb); Gliomab-H (monoclonal - humanized antibody); GNI-250 monoclonal antibody; EMD-72000 (chimeric - EGF antagonist); ipilimumab (humanized IL.L.2 antibody); and MDX-260 bispecific, targeting GD-2, ANA antibody, SMART IDIO antibody, SMART ABL 364 antibody or ImmuRAIT-CEA.

[0164] Other examples of antibodies also include: zalutumumab (anti-CD4 monoclonal antibody); keliximab (anti-CD4 monoclonal antibody); ipilimumab (MDX-101; anti-CTLA-4 monoclonal antibody); tremelimumab (anti-CTLA-4 monoclonal antibody); daclizumab (anti-CD25 / IL-2R monoclonal antibody); basiliximab (anti-CD25 / IL-2R monoclonal antibody); MDX-1106 (anti-PDl monoclonal antibody); GITR antibody; GC1008 (anti-TGF-b antibody); metelimumab / CAT-192 (anti-TGF-b antibody); lededumab / CAT-152 (anti-TGF-b antibody); ID11 (anti-TGF-b antibody); denosumab (anti-RANKL monoclonal antibody); BMS-663513 (humanized anti-4-1BB monoclonal antibody); SGN-40 (humanized anti-CD40 monoclonal antibody); CP870,893 (human anti-CD40 monoclonal antibody); infliximab (chimeric anti-TNF monoclonal antibody); adalimumab (human anti-TNF monoclonal antibody); certolizumab (humanized Fab anti-TNF); golimumab (anti-TNF); etanercept (extracellular domain of TNFR fused to IgG1 Fc); belatacept (extracellular domain of CTLA-4 fused to Fc); abatacept (extracellular domain of CTLA-4 fused to Fc); belimumab (anti-B lymphocyte stimulator); muromonab-CD3 (anti-CD3 monoclonal antibody); ortelizumab (anti-CD3 monoclonal antibody); teplizumab (anti-CD3 monoclonal antibody); tocilizumab (anti-IL6R monoclonal antibody); REGN88 (anti-IL6R monoclonal antibody); ustekinumab (anti-IL-12 / 23 monoclonal antibody); brodalumab (anti-IL-12 / 23 monoclonal antibody); natalizumab (anti-a4 integrin); vedolizumab (anti-a4 b7 integrin monoclonal antibody); T1h (anti-CD6 monoclonal antibody); epratuzumab (anti-CD22 monoclonal antibody), efalizumab (anti-CDlla monoclonal antibody); and atacicept (extracellular domain of transmembrane activator and calcium modulator ligand interactor fused to Fc).

[0165] The grinding methods include physical impact, crushing, dry and wet grinding, decomposition, cryogenic and cryogenic grinding, cutting, etc. The grinding methods include physical impact, crushing, dry and wet grinding, disintegration, cryogenic and cryogenic grinding, cutting, etc. The solution can be water or a liquid, buffer solution, isotonic solution, salt solution with water as the main component, etc., but is not limited to a specific type.

[0166] In addition, when the composition includes two or more types of tumor tissues, the two or more types of tumor tissues may have the same or different sizes of ground particles.

[0167] Furthermore, the present invention provides a kit for preventing or treating cancer, which contains the composition. In addition to the above composition, the kit according to the present invention may include other ingredients, compositions, solutions, devices, etc. that are commonly required for preventing or treating cancer, without limitation. In particular, the kit according to the present invention may include instructions for the correct use and storage of the composition according to the present invention.

[0168] The content of tumor tissues or cancer cells in the composition of the present invention can be appropriately adjusted according to disease symptoms, the degree of symptom progression, the patient's condition, etc. For example, based on the total weight of the composition, it can be 0.0001 - 99.9 wt% or 0.001 - 50 wt%, but is not limited thereto. The content ratio is a value based on the dry weight after removing the solvent.

[0169] The pharmaceutical composition according to the present invention may further include suitable carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions. For example, the excipient can be one or more selected from the group consisting of diluents, binders, disintegrants, glidants, adsorbents, humectants, film-forming materials, and controlled-release additives.

[0170] The pharmaceutical composition according to the present invention can be formulated and used in the form of external preparations according to traditional methods. External preparations such as powders, granules, sustained-release granules, enteric-coated granules, liquids, eye drops, ovules, emulsions, suspensions, alcohols, tablets, aromatic waters, lemonades, pills, sustained-release pills, enteric-coated pills, sublingual pills, hard capsules, soft capsules, sustained-release capsules, enteric-coated capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, irrigation solutions, cautery agents, lotions, pastes, sprays, inhalants, patches, sterile injection solutions, or aerosols. The external preparation can be formulated and used in the form of external preparations (such as emulsions, gels, patches, sprays, ointments, cautery agents, lotions, liniments, pastes, or cataplasms).

[0171] Examples of carriers, excipients, and diluents that can be included in the pharmaceutical composition according to the present invention may include lactose, glucose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl paraben, propyl paraben, talc, magnesium stearate, mineral oil, etc.

[0172] During formulation, commonly used diluents or excipients are used, such as fillers, bulking agents, binders, wetting agents, disintegrants, or surfactants.

[0173] According to the present invention, as additives for tablets, powders, granules, capsules, pills and tablets, excipients, binders, disintegrants and lubricants can be used. Excipients such as corn starch, potato starch, wheat starch, lactose, white sugar, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium hydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, anhydrous lanolin, microcrystalline cellulose, dextrin, sodium alginate, methyl cellulose, sodium carboxymethyl cellulose, kaolin, urea, colloidal silica, hydroxypropyl starch, hydroxypropyl methyl cellulose (HPMC), HPMC1928, HPMC2208, HPMC2906, HPMC2910, propylene glycol, casein, calcium lactate and Primojel; Binders such as gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose, purified water, sodium caseinate, glycerol, stearic acid, sodium carboxymethyl cellulose, sodium methyl cellulose, methyl cellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethyl cellulose, purified shellac, starch, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl alcohol and polyvinylpyrrolidone; Disintegrants such as hydroxypropyl methyl cellulose, corn starch, agar powder, methyl cellulose, bentonite, hydroxypropyl starch, sodium carboxymethyl cellulose, sodium alginate, calcium carboxymethyl cellulose, calcium citrate, sodium lauryl sulfate, silicon anhydride, 1-hydroxypropyl cellulose, dextran, ion exchange resin, polyvinyl acetate, casein and gelatin treated with formaldehyde, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, starch gel, gum arabic, amylopectin, pectin, sodium polyphosphate, ethyl cellulose, white sugar, magnesium aluminum silicate, di-sorbitol solution and light anhydrous silicic acid; Lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc powder, lycopodium powder, kaolin, petrolatum, sodium stearate, cocoa butter, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (lubricating wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, polyethylene glycol, synthetic aluminum silicate, silicon anhydride, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine and light anhydrous silicic acid.

[0174] As additives for the liquid according to the present invention, water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitan fatty acid ester (Tween ester), polyoxyethylene monoalkyl ether, lanolin ether, lanolin ester, acetic acid, hydrochloric acid, ammonia water, ammonium carbonate, potassium hydroxide, sodium hydroxide, gliadin, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.

[0175] In the syrup according to the present invention, a white sugar solution, other sugars or sweeteners, etc. can be used, and when necessary, flavoring agents, coloring agents, preservatives, stabilizers, suspending agents, emulsifying agents, thickening agents, etc. can also be used.

[0176] In the emulsion according to the present invention, purified water can be used, and when necessary, emulsifying agents, preservatives, stabilizers, flavoring agents, etc. can be used.

[0177] In the suspension according to the present invention, suspending agents can be used, such as gum arabic, tragacanth gum, methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, microcrystalline cellulose, sodium alginate, hydroxypropyl methyl cellulose (HPMC), HPMC1828, HPMC2906, HPMC2910, etc., and when necessary, surfactants, preservatives, stabilizers, coloring agents and flavoring agents can be used.

[0178] The injection according to the present invention may include: solvents such as distilled water for injection, 0.9% sodium chloride solution, Ringer's solution, glucose solution, glucose + sodium chloride solution, PEG, lactated Ringer's solution, ethanol, propylene glycol, non-volatile oils - sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate and phenyl benzoate; cosolvents such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, phenylbutazone, propylene glycol, Tween series, nicotinamide, hexamine and dimethylacetamide; buffers such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptone and gums; isotonic agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen (N2) and ethylenediaminetetraacetic acid; sulfiding agents such as 0.1% sodium hydrosulfide, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate and sodium acetone bisulfite, etc.; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose and calcium gluconate; and suspending agents such as sodium CMC, sodium alginate, Tween 80 and aluminum monostearate.

[0179] In the suppository according to the present invention, a base can be used, and the base is, for example, cocoa butter, lanolin, Witepsol, polyethylene glycol, glycerogel, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cocoa butter + cholesterol, lecithin, Lanette wax, glyceryl monostearate, Tween or Span, imhausen, monolan (propylene glycol monostearate), glycerol, solid animal fat, buytyrum Tego - G, cebesPharma 16, hexalide base 95, hydrogenated cottonseed oil, Hydrokote (SP, S - 70 - XXA, S - 70 - XX75(S - 70 - XX95), Hydrokote 25, Hydrokote 711, idropostal, massa estrarium (A, AS, B, C, D, E, I, T), masa - MF, masupol, masupol - 15, neosuppostal - N, paramount - B, supposiro (OSI, OSIX, A, B, C, D, H, L), suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), suppostal (N, Es), Wecoby (W, R, S, M, Fs) and tegester triglyceride substances (TG - 95, MA, 57).

[0180] Oral solid preparations include tablets, pills, powders, granules, capsules, etc., and such solid preparations are formulated by mixing an extract with at least one excipient (such as starch, calcium carbonate, sucrose, lactose, and gelatin, etc.). In addition to simple excipients, lubricants such as magnesium stearate and talcum powder can also be used.

[0181] Examples of liquid preparations for oral use include suspensions, oral liquids, emulsions, syrups, etc. These liquid preparations can include various types of excipients in addition to simple common diluents (such as water and liquid paraffin), such as wetting agents, sweeteners, flavoring agents, preservatives, etc. Preparations for parenteral administration include sterile aqueous solutions, non - aqueous solvents, suspensions, emulsions, freeze - dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils (such as olive oil), and injectable esters (such as ethyl oleate) can be used as non - aqueous solvents and suspensions.

[0182] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The effective dose level can be determined according to various factors, including the type of disease of the patient, the severity of the disease, the drug activity, the sensitivity to the drug, the administration time, the administration route, the excretion rate, the treatment period, the drugs used simultaneously, and other factors well known in the medical field.

[0183] The pharmaceutical composition according to the present disclosure can be administered as a single therapeutic agent, can be administered in combination with other therapeutic agents, can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered once or multiple times. Considering all the above factors, it is important that the administered amount can achieve the maximum effect with the minimum amount and without producing side effects, which can be easily determined by those skilled in the art to which the present disclosure pertains.

[0184] The pharmaceutical composition of the present disclosure can be administered to a subject by various routes. All administration methods can be considered, such as oral administration, subcutaneous injection, intraperitoneal injection, intravenous injection, intramuscular injection, paravertebral space (intrathecal) injection, sublingual administration, buccal administration, rectal insertion, vaginal insertion, ocular administration, otic administration, nasal administration, inhalation, nasal and oral spray, cutaneous administration, transdermal administration, etc.

[0185] The pharmaceutical composition of the present disclosure is determined according to the type of drug as the active ingredient and multiple relevant factors (such as the disease to be treated, the administration route, the age, sex, weight of the patient, and the severity of the disease). Specifically, the effective dose of the composition according to the present invention can vary according to the age, sex, and weight of the patient, and is generally taken at 0.001 to 150 mg, preferably 0.01 to 100 mg per kg of body weight per day, every other day, every 3 to 4 days, or divided into 1 to 3 times a day. However, the dose can be increased or decreased according to the administration route, disease severity, sex, weight, age, etc., so the above dose does not limit the scope of the present invention in any way.

[0186] In the present invention, the term "subject" refers to a subject in need of treating a disease. More specifically, it can be a mammal such as a human or a non-human primate, mouse, rat, dog, cat, horse, cow, etc., but is not limited thereto.

[0187] In the present invention, the term "administer" refers to providing the predetermined composition of the present invention to a subject by any suitable method.

[0188] In the present invention, the term "prevention" refers to all actions that inhibit or delay the onset of a target disease, the term "treatment" refers to all actions that improve or beneficially alter a target disease and its metabolic abnormalities by administering a pharmaceutical composition according to the present invention, and the term "improvement" refers to any action that reduces a parameter (such as the degree of symptoms) associated with a target disease by administering a composition according to the present invention.

[0189] In addition, the present invention provides a method for preparing a composition according to the present invention, the method comprising: step (S1): isolating tumor tissue or cancer cells from a cancer patient; and

[0190] step (S2): applying one or more treatments selected from the group consisting of the following (a) and (b) to the isolated tumor tissue or cancer cells:

[0191] (a) radiation irradiation; and

[0192] (b) cancer immunotherapy drugs.

[0193] The tumor tissue or cancer cells can be obtained by removing them, or the tumor tissue or cancer cells removed during a treatment or diagnosis process can be utilized. For example, by using a tissue specimen for a biopsy to determine whether there is a malignant tumor in the body, rather than discarding the specimen after confirmation by microscopic examination or other tissue examinations, the pain brought to the patient by removing cancerous tissue can be reduced.

[0194] In addition, if the amount of tissue removed or reused after a biopsy is small, the tissue can be cultured to increase the required amount. In cases where culturing is not easy, the small amount of excised tissue can be ground, diluted with water and increased in volume for in vitro treatment.

[0195] Therefore, the method may further comprise the step of grinding the tumor tissue and diluting or dissolving it in a solution after step (S1).

[0196] The extracted tissue can be separated into the amount required for in vitro distal induction treatment, labeled and then treated.

[0197] Preferred embodiments are described below to help understand the present invention. However, the following embodiments provided are only for helping to more easily understand the present invention, and the content of the present invention is not limited by the following embodiments. Examples

[0198] Example 1. Preparation of a composition with a distal effect by radiation irradiation

[0199] As the first method for achieving the distal effect of cancer or tumor treatment, tumor tissue is isolated from a subject, and then the extracted tissue is irradiated and then administered again. Specifically, when irradiating the extracted tissue specimen, a single dose of radiation can be irradiated. The irradiation dose at this time can be in the range of 1 Gray to 500 Gray, and can be administered in one or multiple doses. Alternatively, irradiation and mixing of multiple dose equivalents can be performed, or irradiation and mixing of two or more total doses can be performed instead of single-dose irradiation. In this case, the excised tissue specimens are classified and irradiated, and then the irradiated tissues are mixed to produce a composition for administration.

[0200] For example, two (A, B) radiation irradiation dose mixtures can be prepared under the following conditions:

[0201] - Tissue A: 1 to 50 Gray / total irradiation dose

[0202] - Tissue B: 50 to 500 Gray / total irradiation dose

[0203] Three (A, B, C) radiation irradiation dose mixtures can be produced under the following conditions:

[0204] - Tissue A: 1 to 30 Gray / total irradiation dose

[0205] - Tissue B: 30 to 100 Gray / total irradiation dose

[0206] - Tissue C: 100 to 1000 Gray / total irradiation dose

[0207] The above examples give 2 to 3 total dose mixtures, and if necessary, more total dose mixture compositions can also be made.

[0208] Example 2. Preparation of a composition with a distal effect by immunological treatment

[0209] In this example, by isolating tumor tissue from a subject and then immunologically treating the extracted tissue, a composition capable of exerting a distal effect is produced. When immunologically treating the tissue specimen extracted from the subject, the tissue specimen can be treated as a single drug, or it can be separated into two or more tissue specimens and then treated with two or more immunological agents.

[0210] When immunologically treating the isolated tissue specimen, the isolated tissue specimen is made such that the tissue cells are killed or weakened so that they cannot grow or proliferate, and preferably the immune cells can easily recognize the tissue cells of the immunologically treated composition.

[0211] Immunological treatment includes using immune effector cells and molecules to target and destroy the extracted cancer and tumor tissue cells.

[0212] For example, the immune effector can be a specific antibody against certain markers on the surface of tumor cells. The antibody can be used alone as an effector for treatment, or it can recruit other cells to achieve actual cell death. The antibody can also be conjugated with a drug or toxin (radionuclide, ricin A chain, cholera toxin, pertussis toxin, etc.) and mainly used as a targeting agent. Another example is that the effector can be a surface molecule carried by lymphocytes that can directly or indirectly interact with target tumor cells. Various effector cells include cytotoxic T cells, NK cells, etc. It also includes the use of apoptotic cells in immunotherapy.

[0213] The immunopreparation can be administered by mixing one or more previously developed preparations.

[0214] For example, when two (A, B) extracted specimens are treated with an immunotherapeutic agent, apoptosis or attenuation of the cancer tissue in the extracted tissue specimen A is induced by the treatment with the immunotherapeutic agent, thereby producing a composition. The treatment of the excised tissue specimen B is carried out by an immunotherapy other than the immunotherapy of specimen A, thereby inducing apoptosis or attenuation of the cancer cells in the specimen B tissue, resulting in the production of a composition. The compositions of the treated specimens A and B are administered to the subject separately or as a mixture.

[0215] When there are three or more immunological methods to be treated, the treatment is carried out in the same segmentation method as the two immunotherapy methods, and the prepared compositions are administered to the subject separately or as a mixture.

[0216] In another embodiment of the present invention, each composition in the extracted cancer tissue treated by at least one method selected from the group consisting of radiation irradiation treatment, immunotherapy, etc., or a composition prepared by integrating the compositions, can be administered into the subject. If necessary, each composition prepared by treating the tissue by one or more methods, or a composition prepared by mixing two or more compositions, can be administered to the patient.

[0217] The administration time is not specified separately. When administered together with an immunological reagent, it can be administered 30 minutes to 2 hours before the administration of the immunological reagent, or simultaneously with the immunological reagent, or 30 minutes to 1 week after the administration of the immunological reagent.

[0218] In the present invention, "cancer" includes all types of solid tumors and all types of blood cancers.

[0219] To apply the present invention to blood cancers, the extracted blood is separated by centrifugation, mesh, filtration, etc., and only the cancer tissue is separated and treated in the same manner as solid cancer tissue, thereby producing a distal vaccine.

[0220] Example 3. Confirmation of the anti-cancer effect of a composition (IVAM) with a distal effect produced by radiation treatment

[0221] The in vitro distal method (IVAM) of the present invention is expected to activate the anti-tumor immunity of a subject and induce a distal effect by administering irradiated tumor cells. Therefore, the inventors of the present invention confirmed the effectiveness of the present invention using a breast cancer animal model with low immunogenicity and an immunosuppressive tumor microenvironment.

[0222] 3-1. Experimental method

[0223] Specifically, the in vitro experiment was conducted as follows: 4T1 murine triple-negative breast cancer cells were seeded in 60-mm culture dishes at a density of 2×10 5 cells and in 24-well plates at a density of 1×10 4 cells, and then irradiated (50 Gy) the next day. Cell morphology was observed under a microscope on the 2nd, 4th, and 7th days after irradiation, and cell viability was measured by running an MTT assay.

[0224] The in vivo experiment was conducted in accordance with the animal ethics regulations of the IACUC animal experiment plan (BA-2112-333-009-01) of Seoul National University Bundang Hospital. First, to establish a tumor animal model, 4T1 murine triple-negative breast cancer cells (6×10 5 cells in the right hind limb or 1×10 5 cells in the left abdomen) were injected into the hind limbs and abdomen of 6-week-old immunocompetent BALB / c mice. In the present invention, 4T1 cells can be injected with 4T1-Luc cells expressing luciferase (the same as in the following example), and when luciferin is injected into the mouse, the expressed luciferase (enzyme) emits light using it. This light can be detected by in vivo bioluminescence imaging to monitor the size and location of tumors in the mouse. Seven days after injection, when stable tumor growth was confirmed, the experimental groups were assigned. The experimental groups were divided into 4 groups, with 5 mice assigned to each group: a control group; an IVAM administration group; a radiation treatment group; an IVAM + radiation combination treatment group. Specifically, 4T1 cells irradiated with 50 Gy for one week were cultured, and then cell supernatant (containing dead cells) was obtained. 200 μl of the cell supernatant was intravenously injected once (on the 10th day) to perform IVAM administration. In addition, radiation treatment was performed using a 6-MV electron beam three times a week (on the 10th, 12th, and 14th days), 8 Gy each time, for a total of 24 Gy.

[0225] The tumor size and body weight of each experimental group were measured and recorded every Monday, Wednesday, and Friday, and the tumor growth inhibition effect and the distal effect of radiation were compared and analyzed using IVIS imaging on the 4th and 28th days after tumor implantation.

[0226] 3-2. Sensitization effect of high-dose radiation in the 4T1 murine triple-negative breast cancer cell line

[0227] To measure the cell viability of 4T1 cells exposed to high-dose radiation (50 Gy), a cytotoxicity test (MTT assay) was performed, and the changes in cell shape were observed under a microscope.

[0228] As a result, as shown in Figures 2A and 2B, compared with the control group, damage to the cell membrane of cancer cells receiving radiation exposure was observed, and the proportion of surviving cells was also significantly lower than that of the control group, confirming the sensitization effect of radiation (p < 0.0001).

[0229] 3-3. Combining IVAM and radiation confirmed the synergistic anti-cancer effect and distant effect

[0230] By observing tumor growth for 28 days after tumor inoculation, it was confirmed that compared with the control group, the tumor growth in the group treated with IVAM alone was significantly delayed (p < 0.05). In addition, when treated with a combination of radiation and IVAM, a synergistic effect was confirmed, in which compared with the control group ( Figure 3 ), the tumor growth delay was more obvious (p < 0.01)( Figure 3 ).

[0231] In addition, compared with the control group, in the group treated with radiation alone or the group treated with a combination of radiation + IVAM, the growth of tumors (secondary tumors) in the non-irradiated left abdomen was significantly (p > 0.05, p < 0.01) inhibited. The group treated with IVAM alone showed a slight but statistically insignificant reduction in the left abdominal tumors( Figure 4 ).

[0232] In addition, IVIS imaging using the luciferase system was used to compare and analyze the results on the 4th and 28th days after tumor implantation. By directly measuring the size of the tumors, it was confirmed that when IVAM and radiation were used in combination, a synergistic anti-cancer effect was exerted, similar to the previously confirmed results. In addition, it was also confirmed that according to the combined use of IVAM and radiation, a synergistic tumor inhibitory effect was also exerted in the secondary tumors that were not irradiated, confirming the occurrence of the distant effect (Figures 5A and 5B).

[0233] The results showed that when the IVAM of the present invention was used in combination with radiation, the anti-cancer effect was significantly increased compared with each single therapy, and an even better anti-cancer effect could be achieved through the distant effect.

[0234] 3-4. Confirming the toxicity of the combination of IVAM and radiation

[0235] When measuring the body weight of mice for 28 days, no significant weight loss caused by IVAM and radiation was observed compared with the control group( Figure 6)。The results showed that the combined use of IVAM and radiation in the present invention could only exert excellent anti-cancer effects and distal effects without causing side effects such as toxicity to the subjects.

[0236] Example 4. Confirming the combined effects of IVAM, radiotherapy, and cancer immunotherapy drugs

[0237] In this example, the combined effects of other anti-cancer treatments according to the present invention and IVAM were confirmed. Specifically, the anti-cancer effects were compared by combining IVAM with radiotherapy and / or cancer immunotherapy drugs.

[0238] 4-1. Experimental method

[0239] The animal experiment plan according to this example was as Figure 7 shown. All animal experiments were conducted in accordance with the animal ethics regulations of the IACUC animal experiment plan (BA-2112-333-009-01) of Seoul National University Bundang Hospital.

[0240] Specifically, the experiment was conducted as follows: 4T1 murine triple-negative breast cancer cells (6×10 5 cells in the right hind limb or 1×10 5 cells in the left abdomen) were injected into the hind limbs and abdomen of 6-week-old immunocompetent BALB / c mice. Seven days after injection, when the stable growth of the tumor was confirmed, the experimental groups were divided into eight groups, with five mice assigned to each group: ① control group, ② radiotherapy (RT) group, ③ IVAM administration group, ④ IVAM administration group + radiotherapy combined treatment group, ⑤ a-PD-L1 administration group, ⑥ a-PD-L1 + radiotherapy combined treatment group, ⑦ IVAM administration group + a-PD-L1 administration group, and ⑧ IVAM administration group + a-PD-L1 administration group + radiotherapy triple combined treatment group. At this time, the implementation process of each anti-cancer treatment was as follows:

[0241] 1) IVAM: ① The 4T1 cells were cultured for one week after irradiation with 5, 25, 50, and 100 Gray, and then the cell supernatants were collected in a ratio of 1:1:1:1 (1x10 5 dead cells each time), and 200 μl of the cell supernatant was intravenously injected once (on the 10th day).

[0242] 2) Radiotherapy (RT): Using the X-RAD320 device of Precision X-ray Co., it was administered three times a week (on the 11th, 12th, and 13th days), 8 Gray each time, and a total of 24 Gray was finally administered. Radiation was only applied to the right hind limb of the mice.

[0243] 3) a-PD-L1 (anti-programmed death ligand 1 antibody): Concentration 5 mg / kg, 100 μl each time, intraperitoneally injected twice (on the 11th day and the 18th day).

[0244] Measure and record the tumor size and body weight of each group of mice on Mondays, Wednesdays, and Fridays. On the 7th and 28th days after tumor implantation, use IVIS imaging to compare and analyze the tumor growth inhibitory effect of each treatment and the distal effect of radiation. On the 19th day after tumor implantation, collect orbital blood from the mice, and the centrifuged serum is used to measure the levels of interferon-γ (IFN-γ) and interferon-β (IFN-β) by multiplex immunoassay. All mice were euthanized on the 31st day after tumor implantation. At this time, primary (tumor in the irradiated right hind limb) and secondary (tumor in the non-irradiated left abdomen) tumor tissues, as well as spleen, draining lymph nodes (dLN), and lung tissues, were extracted from the hind limbs and abdomen of the mice, respectively. A part of each tumor tissue and lung tissue was immediately fixed with 4% paraformaldehyde after extraction, and the other tumor and spleen tissues were used for flow cytometry analysis (FACS) after single-cell isolation. The number of metastatic lung nodules observed in the fixed lung tissues was counted using a dissecting microscope. Statistical analysis was performed using one-way ANOVA (Tukey's multiple comparison test) and unpaired two-tailed Student's t-test of PRISM statistical analysis and graphing software (GraphPad 8).

[0245] 4-2. Observe the inhibitory effect of IVAM, a-PD-L1, and radiotherapy combination on tumor growth

[0246] The results of observing the tumor growth in the irradiated right hind limb (primary tumor) are as Figure 8 shown. After the tumor was inoculated into the mice, the tumor growth was observed for 28 days. First, in the irradiated right hind limb (primary tumor), it was confirmed that the tumor growth in the triple combination (IVAM + a-PD-L1 + radiation) group was significantly delayed compared with the control group (p < 0.0001). In addition, it was also confirmed that in each of the RT single-treatment group (p < 0.001), the IVAM + RT group, the IVAM + a-PD-L1 group, and the a-PD-L1 + RT group (all three groups p < 0.0001), the tumor size was slightly larger than that in the triple combination group, but the tumor growth was significantly inhibited compared with the control group. Compared with the control group, a slight delay in tumor growth was observed in the IVAM single-treatment group, but there was no significant difference.

[0247] Compared with the IVAM single-treatment group, it was confirmed that the tumor growth in the RT single-treatment group (p ≤ 0.01), the IVAM + RT group, the IVAM + a-PD-L1 group, the a-PD-L1 + RT group (all three groups p < 0.001), and the triple combination group (p < 0.0001) was significantly delayed.

[0248] When the a-PD-L1 single treatment group was compared with the IVAM + a-PD-L1 group, there was no significant difference (p = 0.6380), but the tumor size tended to be smaller when combined treatment was used. However, there was no difference in tumor size between the RT single treatment group and the IVAM + RT group.

[0249] Therefore, comparing the results of single treatment, dual combination, and triple combination of IVAM, a-PD-L1, and RT confirmed that the triple combination group combining all three treatments had the strongest anti-cancer treatment effect.

[0250] The results of observing the growth of secondary tumors are as Figure 9 shown. As shown in the figure, when comparing the growth of tumors (secondary tumors) in the left abdomen that did not receive radiation with the control group, the tumor growth in the IVAM + a-PD-L1 + RT triple combination group was significantly delayed, confirming the distal effect (p < 0.05). There was no significant difference in tumor size between different treatment groups.

[0251] In addition, on the 4th and 28th days after tumor implantation, the tumors were compared and analyzed using IVIS imaging with the luciferase system. The results confirmed that the pattern of tumor growth inhibition effect (Figures 10A to 10A) was similar to the previously confirmed results of tumor growth inhibition effect comparison ( Figure 8 and Figure 9 ). That is, compared with the control group, the tumor growth in all groups was inhibited, and in particular, the anti-cancer effect of the IVAM + a-PD-L1 + RT triple combination was confirmed to be the strongest.

[0252] In summary, regarding the tumor growth of primary tumors, compared with the control group, a decrease in tumor size was observed in the IVAM single treatment group, and a further tendency for the tumor size to decrease was observed in the IVAM + a-PD-L1 combination treatment group compared with the a-PD-L1 single treatment group. In particular, it was confirmed that the tumor growth in the IVAM + a-PD-L1 + radiation triple combination group was most strongly inhibited. In addition, regarding secondary tumors that did not directly receive radiation, a distal effect was observed in the triple combination group. These results demonstrate that the combination therapy of IVAM, radiotherapy, and cancer immunotherapy drugs according to the present invention can not only achieve a synergistic anti-cancer effect, but also exert a more powerful anti-cancer effect through the distal effect. In addition, by measuring the changes in the immune cell population of the animal model, it was confirmed that the inhibitory effect on the growth of primary tumors in the triple combination was due to the increase in CD8 + T cells and CD8 + effector memory T cells in the tumor microenvironment, and the enhancement of anti-tumor immunity caused by the decrease in Tregs. In particular, through the triple combination, Ki67 + CD8 + T cells and Ki67+ CD4 + The increase in CD4 T cells was also confirmed. In addition, an increase in IFN-γ, which contributes to anti-tumor immune effects, was observed in the mice treated with the triple combination therapy, which is an important clue to the distal effects of the triple combination therapy.

[0253] 4-3. Toxicity of the combination therapy of IVAM, a-PD-L1, and radiation

[0254] To determine whether there are any side effects of the combination of IVAM therapy, a-PD-L1, and radiation according to the present invention, the change in the body weight of mice over time was examined during the treatment process. By measuring the body weight of the mice for 31 days, it was confirmed that there was no significant weight loss in any group treated with IVAM, a-PD-L1, and / or radiation compared with the control group ( Figure 11 ). This result indicates that the combined use of IVAM, the anti-cancer immunotherapy drug, and radiation of the present invention can exert excellent anti-cancer and distal effects without causing side effects such as toxicity to the subject.

[0255] 4-4. Confirmation of the lung metastasis inhibitory effect of the combination therapy of IVAM, a-PD-L1, and radiation

[0256] To confirm the metastasis inhibitory effect of the triple combination of IVAM, the cancer immunotherapy drug, and radiation according to the present invention, mice were inoculated with tumors, and lung tissues were extracted from the mice euthanized on the 31st day to measure the metastatic lung nodules observed on the tissue surface. As a result, a very large number of nodules were observed in the control group, confirming the occurrence of lung metastasis. On the other hand, it was confirmed that the metastasis was significantly reduced in the a-PD-L1 single treatment group, the a-PD-L1+RT double combination group, the IVAM+a-PD-L1 double combination group, and the triple combination group (all P<0.0001). In the IVAM+RT double combination group (p<0.001) and the RT single treatment group (p<0.01), the metastasis was significantly reduced, but there was no significant difference compared with the IVAM single treatment group (Figures 12A and 12B).

[0257] The largest difference was observed in the triple combination group compared with the IVAM single treatment group (P<0.0001), and differences were also found in the a-PD-L1+RT double combination group and the IVAM+a-PD-L1 double combination group (both P<0.01) and the a-PD-L1 single treatment group (P<0.05). Thus, it was confirmed that even when only two or more of the three therapies of IVAM, RT, and a-PD-L1 of the present invention are combined, the lung metastasis inhibitory effect will be significantly increased, and especially when all the therapies are combined, a synergistic metastasis inhibitory effect will be shown.

[0258] 4 - 5. Confirm the changes in the distribution of immune cells in the primary tumor according to the combined treatment of IVAM, a - PD - L1, and radiation

[0259] In the tumor microenvironment of the primary tumor (hind limb tumor), the results of flow cytometry analysis of the population changes of three immune cells (CD8 + T cells, Tregs, and CD8 + effector memory T cells) are shown in Figures 13A to 13C.

[0260] First, it was confirmed that compared with the control group, in the triple combination group of IVAM + a - PD - L1 + radiation, the number of CD8 + T cells attacking tumor cells increased significantly (p < 0.0001), and in the single RT treatment group (p < 0.05), the IVAM + RT double combination group (p < 0.01), the a - PD - L1 + RT double combination group (p < 0.01), and the IVAM + a - PD - L1 double combination group (p < 0.001), the CD8 + T cells increased significantly.

[0261] Previous research results showed that immunosuppressive regulatory T cells (T regs ) that inhibit the cytotoxic activity of CD8+ T cells increase due to radiation. Consistent with this, compared with the control group, the number of T regs increased in the single RT treatment group, and compared with the single IVAM treatment group, the number of T regs increased in the IVAM + RT group. On the other hand, compared with the single treatment group without RT (IVAM + a - PD - L1 group), a decrease in T regs was observed in the triple combination group.

[0262] In addition, in the case of CD8 + effector memory T cells (which enable memory T cells with memory for specific antigens to attack tumor cells with the same antigen), a significant increase was observed in each of the single RT treatment group, the IVAM + a - PD - L1 double combination group (p < 0.01), the IVAM + RT double combination group, the a - PD - L1 + RT double combination group, and the triple combination group (p < 0.001) compared with the control group.

[0263] The results prove that the triple combination of IVAM + cancer immunotherapy drug + radiation can increase the number of CD8 + T cells with anti - cancer activity and reduce the number of T + that inhibit the activity of CD8 regs T cells, thereby enhancing the anti - cancer immune effect.

[0264] In summary, it can be seen that in the triple combination group, CD8+ T cells and CD8 + The number of effector memory T cells increases, and the number of T regs decreases, thereby enhancing the anti-tumor immune effect.

[0265] 4 - 6. Confirm the changes in the distribution of immune cells in the spleen according to the combined treatment of IVAM, a-PD-L1, and radiation

[0266] The spleen contains various immune cells, including lymphocytes such as T cells and monocytes that later differentiate into macrophages and dendritic cells, and thus plays an important role in regulating the systemic immune system. Therefore, the effect of the combined use of IVAM, cancer immunotherapeutic drugs, and / or radiation according to the present invention on the distribution of immune cells in the spleen was confirmed.

[0267] The results are shown in FIGS. 14A and 14B. First, when observing the population changes of CD8 + and CD4 + T cells, no significant differences were observed in each group. However, compared with the control group, it was found that Ki67 + CD8 + T cells expressing Ki670 (a cell proliferation marker) and Ki67 + CD4 + T cell clusters were significantly increased in the triple combination group. This result indicates that the distal effect of the triple combination of IVAM, cancer immunotherapeutic drugs, and radiation according to the present invention is related to the changes in the distribution of immune cells in the spleen.

[0268] Regarding T regs , compared with the control group, the number of T regs increased in the RT single treatment group; compared with the IVAM + RT combination group, the number of T regs increased in the IVAM single treatment group; compared with the a-PD-L1 single treatment group, the number of T regs increased in the a-PD-L1 + RT group, as shown in the patterns of FIGS. 13A to 13C. However, in the triple combination group, a slight decrease in T regs was observed compared with the IVAM + a-PD-L1 double combination group.

[0269] 4 - 7. Confirm the changes in cytokines in the serum according to the combined treatment of IVAM, a-PD-L1, and radiation

[0270] The effect of the combined use of IVAM, cancer immunotherapeutic drugs, and / or radiation according to the present invention on the cytokine levels in the serum was confirmed. For this purpose, serum was collected by collecting orbital blood from mice on the 19th day after tumor inoculation, and the levels of two interferons, IFN-β and IFN-γ, in the serum were measured.

[0271] As a result, a significant increase in IFN-γ was observed in the triple combination group (IVAM + a-PD-L1 + radiation) compared to the control group (p < 0.05), and an upward trend in IFN-β was observed in all groups receiving RT compared to the control group, as Figure 15 shown.

[0272] The results indicate that radiotherapy can increase the level of interferon, which is consistent with the known immune-enhancing and interferon-increasing effects of RT in previous studies. In particular, with respect to IFN-γ, a greater increase was found in the triple combination group applying the combination of IVAM and a-PD-L1 with RT treatment. In addition, the overall increase in IFN-γ in vivo by triple therapy is considered to be the basis for explaining the growth retardation effect and distal effect of triple therapy on primary tumors.

[0273] Example 5. Confirmation of long-term survival test results by treatment with a combination of IVAM, radiotherapy, and cancer immunotherapy drugs

[0274] In this example, the effect of using a combination of IVAM and other anticancer treatments according to the present invention was confirmed over a long period of time. Specifically, IVAM was combined with radiotherapy and / or cancer immunotherapy drugs to compare the anticancer effects.

[0275] 5-1. Experimental method

[0276] The animal trial plan for the long-term survival rate and toxicity confirmation experiment according to this example is as Figure 16 shown. All animal experiments were conducted in accordance with the animal ethics regulations of the IACUC animal experiment plan (BA-2112-333-009-01) of Seoul National University Bundang Hospital.

[0277] Specifically, the experiment was conducted as follows: 4T1 murine triple-negative breast cancer cells (6x10 5 cells in the right hind limb or 1x10 5 cells in the left abdomen) were injected into the hind limbs and abdomen of 6-week-old immunocompetent BALB / c mice. Seven days after injection, when stable tumor growth was confirmed, the experimental groups were divided into 7 groups with 5 mice in each group: ① control group, ② radiotherapy treatment (RT) group, ③ IVAM administration group, ④ IVAM administration group + radiation combination treatment group, ⑤ a-PD-L1 administration group, ⑥ IVAM administration group + a-PD-L1 administration group, ⑦ IVAM administration group + a-PD-L1 administration group + radiation triple combination group. Here, each chemotherapy was performed as follows:

[0278] 1) IVAM: ① 4T1 cells were irradiated with 5, 20, 50, and 100 gray and cultured for one week, and then the cell supernatants were collected at a ratio of 1:1:1:1 (1x10 for each injection5 For dead cells, 200 μl of cell supernatant was intravenously injected once (on the 10th day).

[0279] 2) Radiotherapy (RT): Using the X-RAD320 device of Precision X-ray Co., 8 Gy was administered twice (on the 11th and 12th days), for a total of 16 Gy in the end. Radiation was only applied to the right hind limb of the mice.

[0280] 3) a-PD-L1: 100 μl was intraperitoneally administered four times (on the 11th, 14th, 17th, and 20th days) at a concentration of 5 mg / kg.

[0281] The body weights of each group of mice were measured and recorded every Monday, Wednesday, and Friday. All mice were euthanized on the 50th day after tumor implantation, and the survival rate of each group of mice at 50 days was measured. Statistical analysis was performed using one-way ANOVA (Tukey's multiple comparison test) and unpaired two-tailed Student's t-test with the PRISM statistical analysis and graphing software (GraphPad 8).

[0282] 5-2. Confirm long-term survival rate and toxicity according to the combined treatment of IVAM, a-PD-L1, and radiation

[0283] To determine the long-term survival rate and side effects of the combined treatment of IVAM therapy, a-PD-L1, and radiation according to the present invention, the survival rate and body weight changes of mice over time during the treatment were measured.

[0284] As the measurement result of the survival rate of mice at 50 days, the survival rate of the control group decreased to 0% after 40 days, while the survival rate of the combined treatment of IVAM, a-PD-L1, and radiation remained at 100% even after 50 days (p < 0.001). In the single radiation treatment group, the survival rate after 50 days also remained above 80% (p < 0.05) (Figure 17A).

[0285] In addition, by measuring the body weight changes of mice at 50 days, it was confirmed that there was no significant weight loss in any group treated with IVAM, a-PD-L1, and / or radiation compared with the control group (Figure 17B).

[0286] The results show that the combined use of IVAM, a cancer immunotherapy drug, and radiation according to the present invention can exert excellent anti-cancer and distal effects and will not cause side effects such as toxicity to the subject even for a long time.

[0287] Example 6. Confirm anti-cancer effect and toxicity by multiple IVAM administrations

[0288] In this example, the anti-cancer effect of multiple IVAM administrations was confirmed by tumor size and IVIS imaging.

[0289] 6-1. Experimental Methods

[0290] The animal experiment plan for confirming the anti-cancer effect and toxicity through multiple IVAM administrations according to this embodiment is as Figure 18 shown. All animal experiments were conducted in accordance with the animal ethics regulations of the IACUC animal experiment plan (BA-2112-333-009-01) of Seoul National University Bundang Hospital. 4T1 murine triple-negative breast cancer cells (6x10 5 cells in the right hind limb or 1x10 5 cells in the left abdomen) were injected into the hind limbs and abdomen of 6-week-old immunocompetent BALB / c mice. Seven days after injection, the stable growth of tumors was confirmed, and the experimental groups were assigned. The experimental groups were divided into 4 groups, with 5 mice assigned to each group: control group; IVAM 1-dose group (IVAM-1); IVAM 2-dose group (IVAM-2); IVAM 3-dose group (IVAM-3).

[0291] Specifically, the IVAM administration was carried out as follows: 4T1 cells were seeded in 60pi dishes at a density of 2.5x10 5 cells / dish. The next day, they were irradiated with 5, 20, 50, and 100 gray respectively, and then cultured for one week. The cell supernatants were collected at a ratio of 1:1:1:1 (1.5x10 5 dead cells each time) and intravenously injected three times (on the 10th, 13th, and 17th days), 200 μl each time.

[0292] The body weights of the mice in each group were measured and recorded every Monday, Wednesday, and Friday. On the 7th and 21st days after tumor implantation, the tumor growth inhibition effect and the distal effect of radiation were compared and analyzed using IVIS imaging. All mice were euthanized on the 31st day after tumor implantation, and statistical analysis was performed using one-way ANOVA (Tukey multiple comparison test) and unpaired two-tailed Student's t-test with PRISM statistical analysis and graphing software (GraphPad 8).

[0293] 6-2. Confirming the Tumor Growth Inhibition Effect and Toxicity According to Multiple IVAM Administrations

[0294] After tumor inoculation, tumor growth was observed for 24 days. The results showed that compared with the control group, in the IVAM administration groups, the growth of the irradiated right hind limb tumors (primary tumors) and the non-irradiated left abdominal tumors (secondary tumors) was delayed. Compared with the IVAM 1-dose and 2-dose groups, the growth in the 3-dose group was further delayed, but there was no statistical significance( Figure 19 ).

[0295] In addition, IVIS imaging using the luciferase system was used to compare and analyze the results on the 7th and 21st days after tumor implantation. By directly measuring the tumor size, compared with the control group, in the IVAM administration group, a tumor suppression effect was observed in the tumor (primary tumor) of the right hind limb irradiated with radiation. Compared with the IVAM 1-dose and 2-dose groups, a higher suppression effect was observed in the 3-dose group, similar to the results confirmed above ( Figure 20 ).

[0296] In addition, by measuring the weight change of the mice for 24 days, it was confirmed that compared with the control group, there was no significant weight loss in all groups treated with IVAM once, twice, and three times ( Figure 21 ).

[0297] This result indicates that even if the IVAM of the present invention is administered three times, it can show an anti-cancer effect and a distal effect without causing side effects such as toxicity to the subject.

[0298] Through the above examples, the present inventors confirmed that when tumor tissues or cancer cells isolated from cancer patients are treated with radiation and / or immunotherapy to inhibit the activity of cancer cells or kill cancer cells, and then administered to cancer patients, the body's immune system can activate the anti-cancer immune function, effectively inhibit the progression or growth of primary cancer, and even can exert a tumor-killing function on other tumors in the body. In particular, it has been confirmed that when the tumor cells inhibited or killed by radiation are administered again, in combination with radiotherapy, not only can the anti-cancer effect on the primary tumor be further enhanced, but also a distal effect can be exerted to inhibit the growth of secondary tumors. In addition, the triple combination therapy of radiotherapy and immune checkpoint inhibitor treatment together with IVAM is expected to produce a significant anti-cancer effect and is also expected to achieve the activation function of immune cells. Therefore, the compositions and treatment methods according to the present invention are expected to be used for the treatment of various cancers because they can enhance the body's immune function against tumors through relatively simple treatments and further enhance the anti-cancer effects of other chemotherapies.

[0299] Other objects, features, and advantages of the present invention will become apparent from the following detailed description. However, it should be understood that the provided detailed description is for illustrative purposes only, and various changes and modifications from the following detailed description are obvious to those skilled in the art within the spirit and scope of the present invention.

[0300] Industrial Applicability

[0301] The composition according to the present invention has a tumor growth inhibitory effect, an anti-cancer immune function enhancing effect, and a distal effect, and thus is expected to be used for preventing or treating cancer, and thus has industrial applicability.

Claims

1. A pharmaceutical composition for preventing or treating cancer, which comprises tumor tissue or cancer cells isolated from a cancer patient as an active ingredient, and the tumor tissue or cancer cells are subjected to one or more treatments selected from the group consisting of the following (a) and (b): (a) Radiation exposure; and (b) Cancer immunotherapy drugs.

2. The pharmaceutical composition according to claim 1, wherein the tumor tissue or cancer cells satisfy one or more characteristics selected from the group consisting of the following i) to iii) through the treatment: i) The activity of cancer cells is weakened; ii) The activity of cancer cells is stopped; And iii) Cancer cells are killed.

3. The pharmaceutical composition according to claim 1, wherein the composition comprises autologous tumor tissue or cancer cells of a cancer patient and is administered to the cancer patient or a cancer patient allogeneic to the cancer patient.

4. The pharmaceutical composition according to claim 1, wherein the composition is administered to a subject without cancer to prevent cancer.

5. The pharmaceutical composition according to claim 1, wherein, When the treatment is (a) radiation exposure, the radiation satisfies one or more characteristics selected from the group consisting of the following i) and ii): i) The radiation is one or more selected from the group consisting of gamma rays, X-rays, ultraviolet rays, laser rays, and infrared rays; and ii) The irradiation dose of the radiation is 1 to 500 Gray.

6. The pharmaceutical composition according to claim 1, wherein the cancer immunotherapy drug is one or more selected from the group consisting of immune checkpoint inhibitors, costimulatory molecule reagents, cytokine therapeutics, CAR-T cell therapeutics, and autologous CD8 + T immunocyte therapeutics.

7. The pharmaceutical composition according to claim 6, wherein the immune checkpoint inhibitor is one or more selected from the group consisting of PD-L1 inhibitors, PD-1 inhibitors, CTLA-4 inhibitors, LAG3 inhibitors, TIM3 inhibitors, 4-1BB inhibitors, LAG-3 inhibitors, B7-H4 inhibitors, HVEM inhibitors, TIM4 inhibitors, GAL9 inhibitors, VISTA inhibitors, KIR inhibitors, TIGIT inhibitors, and BTLA inhibitors.

8. The pharmaceutical composition according to claim 1, wherein the composition satisfies one or more characteristics selected from the group consisting of the following i) to iv) when administered to a cancer patient: i) Exerts a distal effect; ii) Inhibits cancer metastasis iii) Reduces tumor burden; and iv) Inhibits the proliferation of cancer cells.

9. The pharmaceutical composition according to claim 1, wherein the composition is for single or multiple administrations.

10. The pharmaceutical composition according to claim 1, wherein the composition comprises two or more kinds of tumor tissue or cancer cells, and the two or more kinds of tumor tissue or cancer cells are respectively subjected to the same treatment or different treatments.

11. The pharmaceutical composition according to claim 10, wherein the composition is a mixture of two or more kinds of tumor tissue or cancer cells.

12. The pharmaceutical composition according to claim 10, wherein the composition is prepared by separately formulating two or more kinds of tumor tissue or cancer cells and is administered simultaneously, separately, or sequentially.

13. The pharmaceutical composition according to claim 1, wherein the composition is used in combination with anti-cancer treatment.

14. The pharmaceutical composition according to claim 13, wherein the anti-cancer treatment is one or more selected from the group consisting of radiotherapy, chemotherapy, targeted anti-cancer therapy, and cancer immunotherapy.

15. The pharmaceutical composition according to claim 13, wherein the composition is used in combination with radiotherapy and cancer immunotherapy.

16. The pharmaceutical composition according to claim 13, wherein the composition is administered simultaneously, separately, or sequentially with the anti-cancer treatment.

17. The pharmaceutical composition according to claim 14, wherein the targeted anti-cancer agent is one or more selected from the group consisting of tyrosine kinase inhibitors, PARP inhibitors, angiogenesis inhibitors, and CDK4 / 6 inhibitors.

18. The pharmaceutical composition according to claim 14, wherein the cancer immunotherapy drug is one or more selected from the group consisting of immune checkpoint inhibitors, costimulatory molecule reagents, cytokine therapeutics, CAR-T cell therapeutics, and autologous CD8 + T immune cell therapeutics.

19. The pharmaceutical composition according to claim 18, wherein the immune checkpoint inhibitor is one or more selected from the group consisting of PD-L1 inhibitors, PD-1 inhibitors, CTLA-4 inhibitors, LAG3 inhibitors, TIM3 inhibitors, 4-1BB inhibitors, LAG-3 inhibitors, B7-H4 inhibitors, HVEM inhibitors, TIM4 inhibitors, GAL9 inhibitors, VISTA inhibitors, KIR inhibitors, TIGIT inhibitors, and BTLA inhibitors.

20. The pharmaceutical composition according to claim 1, wherein the tumor tissue is ground and diluted or dissolved in a solution.

21. The pharmaceutical composition according to claim 20, wherein the composition comprises two or more tumor tissues, and the two or more tumor tissues have the same or different ground particle sizes.

22. The pharmaceutical composition according to claim 1, wherein the cancer is one or more selected from the group consisting of blood cancer and solid cancer.

23. The pharmaceutical composition according to claim 1, wherein the cancer is one or more selected from the group consisting of: breast cancer, colorectal cancer, lung cancer, head and neck cancer, small cell lung cancer, gastric cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, neck cancer, cutaneous melanoma, uveal melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, colon cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney cancer, ureteral cancer, renal cell carcinoma, renal pelvic cancer, central nervous system tumors, primary central nervous system lymphoma, spinal cord tumors, brainstem glioma, and pituitary adenoma.

24. A kit for preventing or treating cancer, comprising the pharmaceutical composition according to claim 1.

25. A method for preparing the composition according to claim 1, the method comprising: (S1) isolating tumor tissue or cancer cells from a cancer patient; and (S2) applying one or more treatments selected from the group consisting of the following (a) and (b) to the isolated tumor tissue or cancer cells, (a) radiation irradiation; and (b) cancer immunotherapy drugs.

26. The method according to claim 25, wherein the method further comprises, after step (S1), grinding the tumor tissue and diluting or dissolving it in a solution.

27. The method according to claim 25, wherein the composition comprises autologous tumor tissue or cancer cells of a cancer patient and is administered to the cancer patient or a cancer patient allogeneic to the cancer patient.

28. The method according to claim 25, wherein the composition is administered to a subject without cancer for cancer prevention.

29. An anti-cancer vaccine composition comprising tumor tissue or cancer cells as an active ingredient, which are isolated from a cancer patient and subjected to one or more treatments selected from the group consisting of the following (a) and (b): (a) Radiation exposure; and (b) Cancer immunotherapy drugs.

30. The anti-cancer vaccine composition according to claim 29, wherein the vaccine composition is administered to a subject without cancer or a subject cured of cancer for cancer prevention.

31. A method for preventing or treating cancer, the method comprising administering to a subject in need a composition comprising tumor tissue or cancer cells as an active ingredient, which are isolated from a cancer patient and subjected to one or more treatments selected from the group consisting of the following (a) and (b): (a) Radiation exposure; and (b) Cancer immunotherapy drugs.

32. Use of a composition for preventing or treating cancer, the composition comprising tumor tissue or cancer cells as an active ingredient, which are isolated from a cancer patient and subjected to one or more treatments selected from the group consisting of the following (a) and (b): (a) Radiation exposure; and (b) Cancer immunotherapy drugs.

33. Use of tumor tissue or cancer cells isolated from a cancer patient and subjected to one or more treatments selected from the group consisting of the following (a) and (b) for the preparation of a preparation for preventing or treating cancer: (a) Radiation exposure; and (b) Cancer immunotherapy drugs.

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