WO2023095929A1 - Agent for treatment of malignant tumors - Google Patents

Abstract

Provided are: an agent for treatment of malignant tumors, the agent being characterized by containing an immune checkpoint inhibitor and by being used in combination with a boron neutron capture therapy; (i) an immune checkpoint inhibitor selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, an anti-TIM-3 antibody, an anti-TIGIT antibody, an anti-KIR antibody, a PD-L1/TGFβ trap inhibition fusion protein, and an anti-PD-1/CTLA-4 bispecific antibody; and (ii) a combination drug for treatment of malignant tumors, which is for patients who do not respond to the treatments with the immune checkpoint inhibitors containing a boron compound for use in a boron neutron capture therapy.

Description

Malignant tumor drug

The present invention relates to a therapeutic drug for malignant tumors that is effective even for patients who do not respond to immunotherapy (immunotherapy alone does not work).

In the field of cancer treatment, surgery, radiotherapy, and chemotherapy were the three major treatments, but the immune checkpoint inhibitor Opdivo (nivolumab), which was launched in 2014, opened the door to a fourth treatment, immunotherapy. Joined and changed a lot. Anti-PD-1 antibodies, which are immune checkpoint inhibitors, release the brakes on cancer cells by inhibiting the binding of PD-L1 on the surface of cancer cells and PD-1 on the surface of immune cells. as a drug that allows immune cells to attack cancer cells. Immune checkpoint inhibitors currently approved in Japan include anti-PD-1/PD-L1 antibodies and anti-CTLA-4 antibodies.

On the other hand, there was no standard treatment for advanced stage malignant melanoma (melanoma), and the prognosis in advanced stage was extremely poor. Various target diseases of immune checkpoint inhibitors have been reported, but the first target disease approved was melanoma, which is cited as a disease for which immunotherapy is highly effective. The reason for this is still largely unknown, but one of the reasons is that the number of cancer gene mutations is thought to be involved in responsiveness to immunotherapy. When ranked, melanoma is reported to have the most mutations.

The large number of mutations in melanoma is attributed to the induction of mutations in genes by UV radiation, as it is believed that ultraviolet rays (UV) have a large effect on the occurrence of melanoma. It is believed that there are Moreover, higher genetic mutations correlated with higher cancer antigen abundance, which correlated with immunotherapeutic response rates; They match (Non-Patent Document 1). In other words, melanoma was the earliest clinical application of cancer immunotherapy using immune checkpoint inhibitors in patients, and it is one of the malignant tumors with the highest response rate to immunotherapy in clinical cancer. (Non-Patent Document 2).

Furthermore, in countries around the world, combination therapy has been studied to increase the response rate of immunotherapy. Methods of treatment with -4 antibodies have been disclosed.

Boron neutron capture therapy (BNCT) administers a boron drug, irradiates the same site with neutrons at the timing when the boron drug accumulates in the cancer site, uses the nuclear reaction (α decay) between boron and neutrons, and cancer cells It is a treatment that kills Since the extent of cell killing is at the cellular level, damage to neighboring normal cells is very low. Clinical research has been conducted in nuclear reactors so far, and in March 2020, the accelerator neutron source installed in the hospital received medical device approval, and the boron drug received drug approval, and from June 2020, recurrent head and neck cancer Medical insurance coverage has started.

BNCT is positioned in the field of cancer particle radiotherapy and is a powerful local treatment method. The key to the success of BNCT relies heavily on tumor-specific boron drug uptake. The current insurance-approved BPA (borofarane ( ¹⁰ B): 4-[( ¹⁰ B)borono]-L-phenylalanine) is taken up malignant-specifically via the amino acid transporter LAT1.

Japanese translation of PCT publication No. 2018-514550

As described in Non-Patent Document 2, the therapeutic response rate of immune checkpoint inhibitors for melanoma is about 30%, and there is currently no treatment for the remaining 70% of melanoma patients who are unresponsive to immunotherapy. is. Also, in Patent Document 1, the subject is a patient who is negative for a specific checkpoint inhibitor, and there is no description of a patient who does not respond to other checkpoint inhibitors. In addition, the response rate of Opdivo for various solid cancers such as advanced hepatocellular carcinoma and ovarian cancer is about 10 to 25% (Non-Patent Document 3, Non-Patent Document 4), and further immunotherapy and chemotherapy. Various combination therapies such as viral therapy have been investigated, but no reports have been made based on clear evidence.

Therefore, an object of the present invention is to provide a therapeutic drug for malignant tumors that is effective even for patients who do not respond to immunotherapy (immunotherapy is not successful).

As a result of examining the above problems, the present inventor found that by combining BNCT with an immune checkpoint inhibitor, it was possible to treat malignant tumors even in patients who did not respond to immunotherapy, and completed the present invention.

That is, the present invention
[1] A therapeutic drug for malignant tumors, which contains an immune checkpoint inhibitor and is used in combination with boron neutron capture therapy;
[2] immune checkpoint inhibitors are anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-KIR antibody, PD- The drug for treating malignant tumors according to [1] above, which is selected from the group consisting of an L1/TGFβ trap inhibitory fusion protein and an anti-PD-1/CTLA-4 bispecific antibody;
[3] The therapeutic drug for malignant tumors according to [2] above, wherein the immune checkpoint inhibitor is a humanized monoclonal antibody;
[4] The drug for treating malignant tumors according to [2] or [3] above, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, or semiplimab;
[5] The drug for treating malignant tumors according to [2] or [3] above, wherein the anti-PD-L1 antibody is avelumab, atezolizumab, or durvalumab;
[6] The therapeutic agent for malignant tumor according to [2] or [3] above, wherein the anti-CTLA-4 antibody is ibilimumab or tremelimumab;
[7] The therapeutic drug for malignant tumors according to any one of [1] to [6] above, wherein the boron compound used in boron neutron capture therapy is borofaran ( ¹⁰ B);
[8] Any one of the above [1] to [7], wherein the subject is a patient who does not respond to an immune checkpoint inhibitor, and the immune checkpoint inhibitor is an immune checkpoint inhibitor to which the patient did not respond. of malignant tumors,
[9] The malignant tumor according to any one of [1] to [8] above, wherein the malignant tumor is selected from the group consisting of carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma, leukemia, glioma, malignant melanoma, and lymphoma. malignant tumor drug,
[10] The therapeutic agent for malignant tumor according to [9] above, wherein the malignant tumor is malignant melanoma;
[11] The therapeutic agent for malignant tumor according to [10] above, wherein the malignant melanoma is advanced stage malignant melanoma;
[12] (i) anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-KIR antibody, PD-L1/TGFβ trap an immune checkpoint inhibitor selected from the group consisting of an inhibitory fusion protein and an anti-PD-1/CTLA-4 bispecific antibody; and (ii) said immune checkpoint comprising a boron compound for use in boron neutron capture therapy. combination drugs for the treatment of malignancies in patients who do not respond to treatment with inhibitors;
[13] The combination drug for treating malignant tumor according to [12] above, wherein the malignant tumor is selected from the group consisting of carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma, leukemia, glioma, malignant melanoma, and lymphoma,
[14] The combination drug for treatment of malignant tumor according to [13] above, wherein the malignant tumor is malignant melanoma, and [15] The treatment for malignant tumor according to [14] above, wherein the malignant melanoma is advanced stage melanoma. It relates to combination medicaments for use.

According to the present invention, by combining treatment with an immune checkpoint inhibitor with BNCT, it is possible to treat malignant tumors even in patients who do not respond to immunotherapy (immunotherapy does not work).

It is a schematic diagram explaining the experimental method of neutron irradiation. 4 is a graph showing changes in tumor volume of a tumor site in the right femoral muscle, which is an irradiation site. It is a graph which shows the change of the tumor volume of the left flank subcutaneous tumor part which is a shielding part. 2 is a graph of flow cytometry results showing the ratio of tumor tissue infiltrating lymphocytes (TIL) in neutron-irradiated tumors. It is the data of the fluorescence microscope photograph which shows the result of the immunohistochemical staining in the neutron irradiation part tumor. FIG. 10 is a graph of flow cytometry results showing the percentage of TILs in neutron shielding tumors. FIG. It is the data of the fluorescence micrograph which shows the result of the immunohistochemical staining in the neutron shielding part tumor. 1 is a graph showing concentrations of HMGB1 in serum. FIG. 2 is a graph of flow cytometry results showing the percentage of effector memory T cells (Tem) in neutron-irradiated tumors. FIG. FIG. 2 is a graph of flow cytometry results showing the percentage of effector memory T cells (Tem) within neutron shielding tumors. 4 is a graph showing the ratio of TILs in the spleen in neutron irradiation experiments. FIG. 10 is a graph showing the ratio of TILs in the spleen in experiments without neutron irradiation. FIG.

The present invention relates to a therapeutic agent for malignant tumors, which contains an immune checkpoint inhibitor and is characterized by being used in combination with boron neutron capture therapy (BNCT), and combines immunotherapy and BNCT (BNCT immune combination therapy: By Boron-Neutron Immuno Therapy (B-NIT), BNCT can enhance responsiveness to immunotherapy and provide effective treatment to patients who are unresponsive to immunotherapy alone. In addition, since BNCT is a high-dose particle beam therapy for local treatment, it is usually not possible to perform simultaneous irradiation to multiple sites or whole-body irradiation, so it is not indicated for advanced cancer with lesions outside the neutron irradiation site. However, the BNCT immune combination therapy of the present invention utilizes BNCT as a local treatment for patients who are unresponsive to immunotherapy, and by enhancing immunotherapy, not only local malignant tumors but also advanced malignant tumors. It is an epoch-making product that can be used for systemic treatment of tumors.

Thus, the present invention demonstrates that combined therapy with BNCT treatment, which exhibits direct effects, and immunotherapy using an immune checkpoint inhibitor was effective against remote sites (shielded sites) that are not irradiated with neutrons. is shown. BNCT, in combination therapy with immunotherapy, thus has therapeutic efficacy against advanced malignancies at distant sites as well.

In addition, BNCT can generally be completed in one day, and can be combined with immunotherapy performed every 2-3 weeks without affecting it.

As used herein, in the treatment of malignant tumors, the expressions "non-responsive to immunotherapy", "non-responsive to immunotherapy", and "unsuccessful to immunotherapy" are all used in the treatment of malignant tumors. When immunotherapy using a point inhibitor is given, it means a state in which the treatment does not effectively reduce the malignant tumor and does not show remission. This includes cases such as the growth of the immune checkpoint inhibitors, and re-expansion of major tissues and growth of metastatic foci during continuous use of immune checkpoint inhibitors can be confirmed by reexamination of images such as CT, MRI, and RET, and blood and tumor tissue. can be evaluated by measuring and examining tumor markers, biomarkers, etc. These expressions are also referred to as "resistant to immunotherapy" and can be used interchangeably. The same applies to the case of "not responding to treatment with immune checkpoint inhibitors."

The malignant tumor to which the present invention is applied is selected from the group consisting of solid cancer, carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma, leukemia, malignant brain tumor (such as glioma), neuroma, malignant melanoma, and lymphoma. The selected malignant tumor is preferably, but not particularly limited to, a malignant tumor already known to partially respond to an immune checkpoint inhibitor. Specific examples of squamous cell carcinoma include those occurring in the cervix, eyelids, conjunctiva, vagina, lungs, oral cavity, skin, bladder, tongue, trachea, bronchi, larynx, pharynx, esophagus, and the like. Specific examples of adenocarcinoma include those occurring in the prostate, small intestine, endometrium, cervix, large intestine, lung, pancreas, esophagus, rectum, uterus, stomach, breast, ovary, and the like. Specific examples of sarcoma include myogenic sarcoma. Specific examples of malignant brain tumors include glioma, glioblastoma, malignant meningioma, primary central nervous system malignant lymphoma, and the like. In yet another embodiment, the malignant tumor is from hepatocellular carcinoma, colon cancer, urothelial cancer, nasopharyngeal cancer, non-small cell lung cancer, ovarian cancer, esophageal cancer, anal cancer, etc. may be selected. In yet another embodiment, the malignant tumor to which the present invention is applied is preferably a malignant tumor for which a known immune checkpoint inhibitor has already been approved, and such a malignant tumor is a malignant Melanoma, non-small cell lung cancer, renal cell carcinoma, Hodgkin's lymphoma, head and neck cancer, gastric cancer, pleural mesothelioma, colorectal cancer, esophageal cancer, urothelial cancer, solid tumor, Merkel cells It is selected from the group consisting of cancer, small cell lung cancer, hepatocellular carcinoma and breast cancer. In yet another aspect, malignant tumors can be selected from non-small cell lung cancer, colon cancer, uterine cancer, bladder cancer, head and neck cancer, or malignant melanoma because of their many gene mutations.

As used herein, the term "immune checkpoint inhibitor" means a molecule capable of inhibiting an immune checkpoint. Immune checkpoint inhibitors include, in particular, molecules that inhibit immune checkpoint functions of PD-1, PD-L1, CTLA-4, LAG-3, TIM-3, TIGIT, KIR, PD-L1/TGFβ trap, Specifically, anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-KIR antibody, PD-L1/TGFβ trap inhibition Fusion proteins, anti-PD-1/CTLA-4 dual antibodies, and the like. Among them, anti-PD-1 antibody, anti-PD-L1 antibody and anti-CTLA-4 antibody are preferred, and anti-PD-1 antibody is more preferred.

These antibodies as immune checkpoint inhibitors used in the present invention are capable of specifically binding to their corresponding antigens, and are human-derived antibodies, mouse-derived antibodies, rat-derived antibodies, rabbit-derived antibodies, or goat-derived antibodies. It may be derived from any antibody, and may be either a polyclonal antibody or a monoclonal antibody, and may be a complete form or a fragment such as an F(ab')2, Fab', Fab or Fv fragment. , chimerized antibodies, humanized antibodies or fully human antibodies. Among them, monoclonal antibodies are preferred, and humanized monoclonal antibodies and fully human monoclonal antibodies are more preferred.

These antibodies as immune checkpoint inhibitors used in the present invention inhibit immune checkpoints by binding PD-1, PD-L1, CTLA-4, LAG-3, TIM-3, TIGIT, KIR, etc. It can be prepared according to a method known in the technical field, using a region capable of as an antigen. For monoclonal antibodies, after determining the amino acid sequence or DNA sequence of the antibody, it can be modified by genetic recombination techniques to produce a genetically modified antibody. It can also be produced by a known method using Antibodies against PD-1, PD-L1, CTLA-4, LAG-3, TIM-3, TIGIT, KIR may also be obtained commercially.

As the anti-PD-1 antibody used in the present invention, a commercially available antibody can be used, particularly an antibody reported to be effective against malignant tumors as an immune checkpoint inhibitor, such as Nivolumab, pembrolizumab, spartalizumab, semiplimab, etc. are preferred, especially from the viewpoint of reducing the time and cost required for clinical application, which are already presently used as immune checkpoint inhibitors, e.g., malignant melanoma, non-small cell Nivolumab used in lung cancer, renal cell carcinoma, Hodgkin lymphoma, head and neck cancer, gastric cancer, malignant pleural mesothelioma, colorectal cancer, esophageal cancer, or e.g. malignant melanoma, non-small cell lung cancer, renal cell carcinoma Pembrolizumab, which has been used in , Hodgkin's lymphoma, head and neck cancer, urothelial cancer, and solid tumors, is preferred. A formulation containing nivolumab as an active ingredient is available from Ono Pharmaceutical Co., Ltd. as “Opdivo (registered trademark),” and a formulation containing pembrolizumab as an active ingredient is available from MSD Co., Ltd. as “Keytruda (registered trademark).” )”.

As the anti-PD-L1 antibody used in the present invention, a commercially available antibody can be used. As an inhibitor, e.g. avelumab, known for use in Merkel cell carcinoma, renal cell carcinoma, urothelial carcinoma, or atezolizumab, known for e.g. or durvalumab, which is known to be used for non-small cell lung cancer, extensive small cell lung cancer, for example. A formulation containing avelumab as an active ingredient is available from Merck BioPharma K.K. under the name BAVENCIO®. Trademark)” and a formulation with durvalumab as the active ingredient is available from AstraZeneca as “Imfinzi®”.

As the anti-CTLA-4 antibody used in the present invention, a commercially available antibody can be used, particularly an antibody reported to be effective against malignant tumors as an immune checkpoint inhibitor, such as Ibilimumab, tremelimumab and the like are preferred, and especially from the viewpoint of reducing the time and cost required for clinical application, currently already as immune checkpoint inhibitors, for example malignant melanoma, renal cell carcinoma, colorectal cancer, Known ibilimumab, which is used for non-small cell lung cancer and malignant pleural mesothelioma, is preferred. A formulation containing ibilimumab as an active ingredient is available as "Yervoy (registered trademark)" from Bristol-Myers Squibb K.K.

The malignant tumor therapeutic drug of the present invention can be administered systemically or locally, and can be administered orally or parenterally. Parenteral routes of administration include intramuscular, intraperitoneal, intrasternal, intravenous, subcutaneous, and the like. According to a specific embodiment, the therapeutic drug for malignant tumor of the present invention is preferably administered intraperitoneally or intravenously, especially by intravenous drip.

The therapeutic agent for malignant tumors of the present invention may be administered as an active ingredient, the immune checkpoint inhibitor itself, but in general, at least one pharmaceutically acceptable additive is added according to the route of administration. It is preferably administered in the form of a pharmaceutical composition containing, for example.

The dosage forms of the drug for treating malignant tumors of the present invention include, for oral administration, liquids, tablets (including orally disintegrating tablets, sublingual tablets, oral patch tablets, etc.), pills, capsules, powders, and granules. For parenteral administration, injections, infusions, external preparations, suppositories, inhalants, intranasal preparations and the like can be mentioned. Injections and infusions may be liquid formulations such as solutions, suspensions and emulsions, and may also include solid formulations such as freeze-dried formulations prepared as liquid formulations at the time of use.

The additives used in the drug for treating malignant tumors of the present invention are not particularly limited, but are tonicity agents, buffers, pH adjusters, and other additives (excipients, binders, disintegrants, , preservatives, lubricants, stabilizers, swelling agents, etc.), and can be appropriately selected and used according to the dosage form. In a specific embodiment, the drug for treating malignant tumors of the present invention can be made into an infusion, especially a liquid for intravenous infusion, using tonicity agents, buffers, pH adjusters and other additives. preferable.

The tonicity agent is not particularly limited, but is selected from the group consisting of sodium chloride, potassium chloride, sodium citrate, sucrose, glucose, mannitol, sorbitol, xylitol, arginine, cysteine, histidine and glycine. At least one tonicity agent and the like are included. A tonicity agent may be used alone or in combination of two or more.

Examples of buffering agents include, but are not limited to, histidine, histidine hydrochloride, phosphoric acid, citric acid, maleic acid, acetic acid, glacial acetic acid, succinic acid, tartaric acid, lactic acid, malic acid, boric acid, ascorbic acid, at least one buffer selected from the group consisting of glycine, glutamic acid, arginine, tris-(hydroxymethyl)-aminomethane (tris) and diethanolamine or salts thereof; A buffering agent may be used individually and may be used in combination of 2 or more types.

Examples of pH adjusters include, but are not limited to, bases such as sodium hydroxide, aqueous ammonia, potassium hydroxide, calcium hydroxide, magnesium hydroxide, sodium hydrogen carbonate, and sodium carbonate, as well as hydrochloric acid, citric acid, Acids such as succinic acid, acetic acid, glacial acetic acid, tartaric acid, carbon dioxide, lactic acid, sulfuric acid and phosphoric acid. The pH adjusters may be used alone or in combination of two or more.

Other additives include, but are not limited to, mannitol, sorbitol, sucrose, refined sucrose, trehalose, xylitol, glucose, lactose, glycerol, maltose, inositol, bovine serum albumin (BSA), dextran, PVA, hydroxyl Propylmethylcellulose (HPMC), polyethyleneimine, gelatin, polyvinylpyrrolidone (PVP), hydroxyethylcellulose (HEC), polyethylene glycol, ethylene glycol, dimethylsulfoxide (DMSO), dimethylformamide (DMF), proline, sodium L-serine glutamate, glutamic acid. Potassium, Alanine, Glycine, Lysine Hydrochloride, Sarcosine, γ-Aminobutyric Acid, Diethylenetriaminepentaacetic Acid, Polysorbate 20, Polysorbate 80, SDS, Polyoxyethylene Copolymer, Potassium Phosphate, Sodium Acetate, Ammonium Sulfate, Magnesium Sulfate, Sodium Sulfate, Trimethylamine at least one additive selected from the group consisting of N-oxides, betaine, zinc ions, copper ions, calcium ions, manganese ions, magnesium ions, CHAPS, sucrose monolaurate and 2-O-β-mannoglycerate etc.

The dose and frequency of administration of the drug for treating malignant tumors of the present invention are not particularly limited. Although it varies depending on the administration method, etc., it is determined by the general condition, responsiveness to treatment, degree of side effects, social factors, and the like. Specifically, the therapeutic drug for malignant tumor of the present invention is preferably 0.01 to 10000 mg, more preferably 0.1 to 5000 mg, more preferably 1 to 2000 mg as an immune checkpoint inhibitor per administration for adults. preferable. Any immune checkpoint inhibitor of the present invention, for example, every day, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, every week, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, multiple times, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 times every 6, 7, 8, 9 or 10 weeks administered more than once.

In addition, for immune checkpoint inhibitors that have already been clinically used against malignant tumors, it is preferable to administer them based on the known and approved dosage and administration with reference to their package inserts, but BNCT It can also be changed and used in consideration of the combined use with. Of course, as the malignant tumor therapeutic drug of the present invention, it is preferable that the package insert of the immune checkpoint inhibitor clearly states that it is used in combination with BNCT.

Specifically, when the immune checkpoint inhibitor is nivolumab, 1 to 1000 mg of nivolumab, preferably 10 to 700 mg, more preferably 80 to 480 mg of nivolumab is intravenously administered multiple times at intervals of 1 to 6 weeks to adults. Specifically, 240 mg can be administered as multiple doses at 2- to 3-week intervals, or 480 mg can be administered as multiple doses at 4-week intervals.

According to another aspect, when the immune checkpoint inhibitor is pembrolizumab, 1 to 1000 mg of pembrolizumab, preferably 10 to 500 mg, more preferably 200 mg of pembrolizumab is intravenously infused multiple times at 3-week intervals to adults. can be done.

According to another aspect, when the immune checkpoint inhibitor is avelumab, the dose of avelumab to adults is 1 to 100 mg/kg (body weight), preferably 5 to 50 mg/kg (body weight), more preferably 10 mg/kg. (body weight) can be administered intravenously multiple times at intervals of 1 to 3 weeks, preferably at intervals of 2 weeks.

According to another aspect, when the immune checkpoint inhibitor is atezolizumab, an adult receives 100-1500 mg, preferably 500-1500 mg, preferably 840-1200 mg of atezolizumab once every 2-4 weeks, preferably 2-4 weeks. It can be intravenously infused multiple times at 3-week intervals. Specifically, 1200 mg can be intravenously infused multiple times at 3-week intervals, or 840 mg can be intravenously infused multiple times at 1- to 2-week intervals. .

According to another aspect, when the immune checkpoint inhibitor is durvalumab, 0.1 to 5000 mg/kg (body weight), preferably 1 to 1500 mg/kg (body weight), more preferably 5 mg/kg (body weight), more preferably 5 mg/kg (body weight) of durvalumab is administered to an adult once. Up to 1300 mg/kg (body weight) can be intravenously infused multiple times at intervals of 2 to 4 weeks, preferably at intervals of 2 to 3 weeks, specifically, 10 mg/kg (body weight) once at intervals of 2 weeks. Multiple doses or multiple doses of 1500 mg/kg (body weight) can be intravenously infused at 4-week intervals.

According to another aspect, when the immune checkpoint inhibitor is ibilimumab, 0.05 to 100 mg/kg (body weight), preferably 0.1 to 50 mg/kg (body weight), more preferably 0.1 to 50 mg/kg (body weight) of ibilimumab is administered to an adult once. 0.5 to 5 mg/kg (body weight) can be intravenously infused multiple times at intervals of 1 to 8 weeks, preferably at intervals of 2 to 7 weeks, specifically, 3 mg/kg (body weight) once. Multiple intravenous infusions, for example 4 times, can be performed at 3-week intervals.

In the present invention, BNCT is preferably performed after starting administration of an immune checkpoint inhibitor. Specifically, after administering an immune checkpoint inhibitor once, twice or three times or more, administering a boron compound, 1 to 3 hours after that, neutron irradiation, and then further immune checkpoint inhibitor It is preferable to administer once, twice, or three times or more, and further preferably to continuously administer an immune checkpoint inhibitor.

The boron compound used for BNCT in the present invention is not particularly limited, and various compounds conventionally used in particle beam therapy can be used. Specifically, BPA, BSH, BSH Derivatives, and other known boron compounds for BNCT that further contain boron. Cluster molecules such as BSH and BSH derivatives are preferred because they contain many boron atoms in one molecule, and BPA is more preferred because it is a drug targeting malignant tumors focused on melanin biosynthesis. These boron compounds can be produced by known methods and are commercially available. A preparation containing BPA is available from Stella Pharma Co., Ltd. as "Stebronine (registered trademark)". Of course, according to the present invention, the package insert of the boron compound preferably clearly states that it is used in combination with an immune checkpoint inhibitor.

The boron compound used in the present invention is generally administered intravenously or intraarterially. In addition, administration by oral medicine, administration by intramuscular injection, transdermal administration, etc. can also be effectively used. .

The boron compound used in the present invention is administered in a concentration and amount that allows it to accumulate in tumor cells in a therapeutically effective amount. It is preferable to administer so that the final boron concentration in the tumor tissue area is 25 ppm or more. Further, it is more preferable that the ratio of the boron concentration in the tumor tissue to that in the normal tissue is 2 times or more. For example, in the case of BPA, such dosage is preferably 100-500 mg/kg, more preferably 250-500 mg/kg. It is preferable to administer such a dose by intravenous drip infusion over 1 to 3 hours at a rate of 100 to 200 mg/kg per hour, and neutron irradiation is preferably performed after administration or during continuous administration. Further, the boron compound can be additionally administered during neutron irradiation, and can be intravenously infused at a rate of 50 to 500 mg/kg, preferably 50 to 100 mg/kg per hour.

Neutron irradiation is preferably performed after sufficient time for the boron compound to reach the site to be treated and before the boron compound is reduced from the neutron-irradiated tumor site. It is more preferable to carry out after ~4 hours, more preferably after 1 to 3 hours. For neutron irradiation, using a nuclear reactor or an accelerator-type neutron generator, treatment such as neutron dose, irradiation time, tumor site boron concentration, blood boron concentration, pharmacokinetic results by boron compound-PET (BPA-PET, etc.) determine the conditions necessary for

Since BNCT is radiotherapy, it targets a limited area. BNCT treatment should preferably be performed as few times as possible. In principle, when the drug for treating malignant tumors of the present invention is used, it should be performed once during combination therapy with an immune checkpoint inhibitor. However, in consideration of the progress of the disease, the range of the irradiation site, etc., it may be considered to perform the treatment two or more times.

The BNCT immune combination therapy with the therapeutic agent for malignant tumors of the present invention can effectively treat patients who do not respond to immunotherapy, especially treatment with immune checkpoint inhibitors, and patients at advanced stage who have metastasis. For example, patients suffering from advanced-stage malignant melanoma can be expected to be particularly effective as described above.

In another embodiment of the invention, (i) anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-KIR antibody , an immune checkpoint inhibitor selected from the group consisting of a PD-L1/TGFβ trap inhibitory fusion protein and an anti-PD-1/CTLA-4 bispecific antibody, and (ii) boron for use in boron neutron capture therapy. A pharmaceutical combination for treating malignant tumors for patients who do not respond to the immune checkpoint inhibitor is provided, comprising the compound. The form of this combination medicine is not particularly limited, and (i) anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT an immune checkpoint inhibitor selected from the group consisting of antibodies, anti-KIR antibodies, PD-L1/TGFβ trap inhibitory fusion proteins and anti-PD-1/CTLA-4 bispecific antibodies, and (ii) boron neutron capture therapy. and the kit can include instructions describing the relationship between administration of the immune checkpoint inhibitor and administration of BNCT.

Unless otherwise contradicted, the above explanations about "therapeutic drugs for malignant tumors" shall apply to "combined drugs for treating malignant tumors" in the same way. shall apply equally to “malignant tumor treatment” above.

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these as long as it does not deviate from the gist and scope of application.

[Preparation of advanced melanoma model mice]
This animal experiment was properly conducted based on the approval of the Animal Care and Use Committee of National University Corporation Okayama University and National University Corporation Kyoto University. Mouse melanoma cells B16F10 were purchased from National University Corporation Tohoku University Institute of Aging and Cancer Medical Cell Resource Center/Cell Bank. Culture was performed using RPMI-1640+10% FBS (fetal bovine serum)+1% P/S (penicillin/streptomycin) using a 5% CO ₂ cell culture incubator. Cells were harvested using 0.25% trypsin under 70-90% confluent conditions to prepare a cell suspension of 1.0×10 ⁴ cells/μL. 5-week-old C57BL/6JJcl female mice (purchased from CLEA Japan, Inc.) were used, and 30 μL (3.0×10 ⁵ cells) were injected into the muscle of the right thigh and left flank of each mouse. 50 μL (5.0×10 ⁵ cells) of BL6F10 was implanted subcutaneously. It was confirmed that the produced advanced melanoma model mouse does not respond to treatment with an immune checkpoint inhibitor (anti-PD-1 antibody), as described later.

[Measurement of boron concentration]
A boron drug, borofarane ( ¹⁰ B) (trade name: steboronine, manufactured by Stella Chemifa Co., Ltd.) was subcutaneously administered to advanced melanoma model mice (n=4) at 500 mg/kg body weight. Boron concentration in each tissue was measured at 1, 2 and 3 hours after administration. Table 1 shows the results.

From Table 1, the boron concentration in the tumor site in the muscle of the right lower limb, which is the site to be irradiated, was 29.9 ppm, while the boron concentration in the normal muscle of the same site was 10 ppm. In addition, the boron concentration in the subcutaneous tumor site (planned shielding site) on the left side of the abdomen and the boron concentration in the normal skin of the right thigh, which is the planned irradiation site, were almost the same at 13.2 ppm and 13.1 ppm, respectively. . The melanoma cells used for the left flank subcutaneous tumor and the right thigh intramuscular tumor are the same cell line (B16-F10), but the boron concentration in the subcutaneous tumor area is 29 The low value of 13.2 ppm compared to 0.9 ppm is due to the fact that intramuscular tumor blood vessels are more likely to be formed and more likely to contribute to tumorigenesis. Neutron irradiation, which will be described later, was performed 2 hours after administration of the boron drug because the boron concentration in the right thigh was stable from 29.9 ppm to 31.8 ppm from 2 hours to 3 hours.

Example 1
Advanced melanoma model mice used in the test, control (neutron irradiation only) group (n = 9), anti-PD-1 antibody administration group (n = 9), BNCT treatment group (n = 9), and anti-PD-1 They were divided into antibody administration + BNCT treatment groups (n=9), and corresponding drug administration and neutron irradiation were performed as described below.

[Administration of immune checkpoint inhibitor]
On the 4th, 7th, 10th and 13th days after production, the mouse anti-PD-1 antibody (manufactured by Bio X cell, InVivoMab anti -mouse PD-1 (CD279), clone#RMP1-14) was administered intraperitoneally at 250 mg per mouse.

[Boron drug BPA administration]
Boron drug borofarane ( ¹⁰ B) (trade name: steboronine, manufactured by Stella Chemifa Co., Ltd.) was subcutaneously administered to advanced melanoma model mice at 500 mg/kg body weight. Two hours after administration, neutron irradiation was performed.

[Neutron irradiation]
On the 8th day after the model mouse was produced, neutron irradiation was performed at the trolley animal irradiation facility in Kyoto University's Institute for Integrated Radiation and Nuclear Science during operation of the 5 MW nuclear reactor. An outline of the irradiation method is shown in FIG. Neutron irradiation was performed for 12 minutes on the entire lower extremity (irradiated part) including the tumor in the right thigh muscle of the melanoma model mouse from a neutron source, and the left flank subcutaneous tumor was used as the tumor in the shielded part. High-concentration lithium fluoride was used for neutron shielding. That is, as shown in FIG. 1, a model mouse was placed on a ring-shaped shielding material made of high-concentration lithium fluoride, and neutron irradiation was performed from the shielding material side from a neutron source. As a result, neutrons were irradiated to the left thigh intramuscular tumor located where there was no shielding material, and neutrons did not reach the left abdominal tumor blocked by the shielding material. Experiments were performed on model mice under general anesthesia in order to minimize the effects of mouse movement. The average thermal neutron fluence in the irradiated area of the right lower extremity is 2.67E+12 (cm ^-2 ), and the average thermal neutron fluence within the shield of the left abdomen is 6.427E+11 (cm ^-2 ), which is about 1/4 of the irradiated area. Sufficient neutron shielding was confirmed.

[Measurement of tumor volume]
Right thigh intramuscular tumor volume (mm ³ ) and left flank subcutaneous tumor volume (mm ³ ) were measured on days 15, 20, 23 and 27 after neutron irradiation. Furthermore, the dose in each tissue (irradiated right thigh intramuscular tumor, shielded left flank subcutaneous tumor, irradiated right thigh normal muscle, irradiated right thigh normal skin) Determined by boron concentration in tissue.

FIG. 2 shows the results of measuring the tumor volume of the tumor in the right thigh muscle. From FIG. 2, 27 days after transplantation, the anti-PD-1 antibody administration + BNCT treatment group had the smallest tumor volume (mean 926 (mm ³ )). On the other hand, the BNCT-treated group showed the effect of BNCT at 1850 (mm ³ ). In the anti-PD-1 antibody-administered group, it was 3242 (mm ³ ). Twenty-three days after transplantation, the control group already exceeded 2200 (mm ³ ) and had the largest tumor volume compared to the other treatment groups.

FIG. 3 shows the results of measuring the left flank subcutaneous tumor volume. From FIG. 3, 27 days after transplantation, the tumor volume was the smallest in the anti-PD-1 antibody administration+BNCT treatment group (average 1291 (mm ³ )). On the other hand, the BNCT-treated group showed the effect of BNCT at 2015 (mm ³ ). The anti-PD-1 antibody-administered group was 4127 (mm ³ ). The control group already exceeded 5700 (mm ³ ) 23 days after transplantation, and had the largest tumor volume compared to the other treatment groups.

Table 2 shows the dose of each tissue converted to radiation.

These results show that this B16F10 transplanted melanoma model is resistant to anti-PD-1 antibodies, that is, it is an immunotherapy-resistant melanoma model that does not respond (does not respond) to anti-PD-1 antibody therapy. .

In the intramuscular tumor of the right thigh, the BNCT treatment group with BNCT alone has a certain degree of local tumor suppressive effect, but the effect is not so great, and the combined use of immunotherapy (administration of anti-PD-1 antibody + BNCT treatment group), the tumor volume was significantly reduced to less than half that of the BNCT treatment group at 27 days after transplantation.

In addition, the radiation dose to the intramuscular tumor site of the right thigh was extremely high at 24.8 Gy-eq, while the dose to the surrounding normal tissue was 5.9 Gy-eq for the muscle and 2.0 Gy-eq for the skin. eq is very small, and it can be seen that damage to surrounding normal tissues is suppressed.

Furthermore, from the results in the shielded area, it can be seen that the anti-PD-1 antibody administration + BNCT treatment group was able to significantly suppress tumor growth at distant sites. In other words, when irradiation was performed at a sufficient tumor dose at the site of direct irradiation, we succeeded in inducing a therapeutic effect at a distant site through immunotherapy and therapeutic effects at the site of direct irradiation.

When examining these results for the anti-PD-1 antibody administration group (single) and BNCT treatment group (single) at the shielded site, and the anti-PD-1 antibody administration + BNCT treatment group (combination), It showed an effect exceeding both. Therefore, it was found that anti-PD-1 antibody administration and BNCT treatment ultimately exhibited a synergistic effect when combined in both direct irradiation and shielded areas.

Example 2
Boron neutron capture therapy combined immunotherapy (B-NIT), which is a combination of boron neutron therapy and immunotherapy, was performed using the advanced stage melanoma model mouse prepared above. Model mice control (neutron irradiation only) group (n = 9), anti-PD-1 antibody administration group (n = 9), BNCT treatment group (n = 9), and anti-PD-1 antibody administration + BNCT treatment group (n =9), and administration of immune checkpoint inhibitors and boron agents and neutron irradiation were performed in the same manner as in Example 1.

[Tumor infiltrating lymphocytes (TIL) in the neutron irradiation area]
The ratio of CD8+/CD45+ tumor-infiltrating lymphocytes (TIL) in the neutron-irradiated tumor area of each group (n=3 each) 15 days after neutron irradiation was confirmed by flow cytometry and immunohistochemical staining. Specifically, a piece of melanoma tumor tissue was collected from the tumor site in the right femoral muscle of a mouse under general anesthesia, and a single cell suspension was prepared by a conventional method and used for flow cytometry. For flow cytometry, first stain with Zombie Aqua™ Fixable Viability Kit (BioLegend, San Diego, Calif., USA) to label dead cells and Fc Block (purified anti-mouse CD16/32 Pretreatment was performed with Clone 93; manufactured by Biolegend). Cells were then stained with antibodies and analyzed with an LSRFortessa™ flow cytometer (manufactured by BD Biosciences). As staining antibodies, FITC anti-mouse CD45 (clone 30-F11) and BV786 rat anti-mouse CD8a (clone 53-6.7) purchased from Biolegend were used and diluted according to the manufacturer's instructions. Data were analyzed using FlowJo Software (manufactured by BD Biosciences). The results are shown in FIG. As shown in FIG. 4, the CD8+/CD45+ tumor-infiltrating lymphocytes were 24.5% and 25.34% in the control group and the anti-PD-1 antibody-administered group, respectively, and there was no change. On the other hand, it increased to 49.2% in the BNCT treatment group, and further increased to 64.6% in the anti-PD-1 antibody administration + BNCT treatment group (Fig. 4). After fixing a sample collected from the tumor tissue with 4% paraformaldehyde, a frozen pathological section sample was prepared, and immunohistochemical staining of the tumor area was performed using an anti-CD8 antibody. Tissue pieces at multiple locations such as the tumor center and tumor margins were observed with a fluorescence microscope. FIG. 5 shows fluorescence photomicrograph data (×10) of the boundary region with the normal region around the tumor. In FIG. 5, 1 indicates nuclei (Hoechst staining: blue) and 2 indicates tumor-infiltrating lymphocytes (anti-CD8 antibody: red). From FIG. 5, tumor infiltrating lymphocytes (TIL) 2 could hardly be confirmed in the control group and the anti-PD-1 antibody administration group, but in the BNCT treatment group, the proportion of TIL2 increased significantly, and the anti-PD-1 antibody administration + BNCT treatment group , the proportion of TIL2 is further increased, supporting the results of the above flow cytometry. In general, since the blood flow is difficult to reach the center of the tumor, it is difficult for lymphocytes to reach it as well. be. However, as shown in FIG. 5, even in such a region, lymphocyte infiltration is low in the control group and the anti-PD-1 antibody-administered group, indicating that this model mouse is anti-PD-1 antibody-resistant. It is shown that. Even in such mice, the infiltration of lymphocytes attacking the tumor is significantly observed in the BNCT-treated group and the anti-PD-1 antibody-administered + BNCT-treated group. Based on the above, tumor tissue infiltrating lymphocytes (TIL), which are important in immunotherapy, were significantly observed in the tumor-normal boundary in the BNCT treatment group and the anti-PD-1 antibody administration + BNCT treatment group inside the tumor at the irradiation site. Therefore, it was considered to indicate a pathological condition of infiltration and aggregation around the tumor. This indicates that BNCT treatment or combination therapy with an immune checkpoint inhibitor and BNCT treatment induces a high degree of TIL, suggesting a very good effect of combined therapy. In addition, CD8a monoclonal antibody (4SM15) and eBioscience (manufactured by Thermo Fisher Scientific) were used as the primary antibody for immunohistochemical staining, and Alexa Flour 594 anti-mouse (manufactured by Thermo Fisher Scientific) was used as the secondary antibody. board.

[Tumor-infiltrating lymphocytes (TIL) in the shielded part]
In the same manner as the measurement of TIL in the neutron-irradiated area, the ratio of CD8 + / CD45 + tumor-infiltrating lymphocytes (TIL) in the shielded tumor area of each group (n = 3 each) was measured as described above [Tumor-infiltrating lymphocytes in the neutron-irradiated part Spheres (TIL)] were confirmed by flow cytometry and immunohistochemical staining. The results of flow cytometry are shown in FIG. 6, and the results of immunohistochemical staining are shown in FIG. As shown in FIG. 6, 14.4% in the control group, 14.1% in the anti-PD-1 antibody administration group, and 12.1% in the BNCT treatment group, and no change was observed between these groups. rice field. On the other hand, in the anti-PD-1 antibody administration + BNCT treatment group, the proportion of TIL increased greatly to 45.6%. For immunostaining, tissue sections at multiple sites such as the tumor center and tumor margins were observed with a fluorescence microscope. Figure 7 shows fluorescence micrograph data (x 10) of the boundary region between the tumor and the normal area around the tumor. : red). From FIG. 7, almost no TILs were confirmed in the control group, the anti-PD-1 antibody administration group and the BNCT treatment group, but many TILs were confirmed in the anti-PD-1 antibody administration + BNCT treatment group. I know it backs it up. From the above, in the anti-PD-1 antibody administration + BCNT treatment group, tumor tissue infiltrating lymphocytes (TIL), which are important in immunotherapy, are significantly observed at the tumor-normal border within the tumor at the shielded site. It was considered to indicate a pathological condition of infiltration and aggregation around the This indicates that the combined use of the immune checkpoint inhibitor and BNCT treatment produces a unique effect even in the shielded area that cannot be obtained with the immune checkpoint inhibitor alone or the BNCT treatment alone.

[HMGB1 (high mobility group box 1) in serum]
Among the molecules released from the inside of cells when cells are damaged and on the verge of death, groups of molecules that particularly affect immune reactions are called damage-associated molecular patterns (DAMPs). HMGB1 (high mobility group box 1) is one of the most important DAMPs induced by cell-killing treatments such as radiotherapy. It has been reported that HMGB1 binds to TLRs (Toll-like receptors) on dendritic cells, activates dendritic cells, leads to an increase in the ability to present tumor antigens, and leads to the activation of tumor-toxic T cells. there is In conjunction with the measurement of [tumor-infiltrating lymphocytes (TIL) in the neutron-irradiated part] and [tumor-infiltrating lymphocytes (TIL) in the shielded part], blood was collected separately from the same mouse, and HMGB1 in the mouse serum was measured. An ELISA kit (ARG81310, Arigo Biolaboratories Corp.) was used (n=3 for each group). 4.4 (ng/mL) in the control group and 6.5 (ng/mL) in the anti-PD-1 antibody administration group, showing no significant difference between these groups, but 9.5 in the BNCT treatment group. (ng/mL), and further increased to 14.1 (ng/mL) in the anti-PD-1 antibody administration + BNCT treatment group (Fig. 8). This increase in serum HMGB1 indicates that BNCT treatment or anti-PD-1 antibody administration+BNCT treatment induces immune induction via HMGB1.

Therefore, this blood (serum) HMGB1 level is considered to be a new biomarker in BNCT immune combination therapy (B-NIT).

[Percentage of effector memory T cells (Tem) in tumor infiltrating lymphocytes (TIL)]
We evaluated what types of T cells were increased among tumor tissue infiltrating lymphocytes (TILs). Cytotoxic T cells (CTLs), which play an important role in anti-tumor immune responses, attack and kill cancer cells by being activated. Such activated T cells (effector T cells) differentiate into T cells with various functions in the extracellular environment. Effector T cells are mostly dead after 1-2 weeks unless exposure to the same antigen is sustained. Some remaining effector T-cells exist as effector memory T-cells (Tem) capable of rapid immune response over a long period of time in the tumor rather than in secondary lymphoid tissues. Tem present within the tumor microenvironment circulates primarily in secondary lymphoid and tumor tissues and responds rapidly upon re-exposure to the same antigen.

All mice in each group (n = 6 each) were sacrificed 22 to 27 days after neutron irradiation, and the above [tumor-infiltrating lymphocytes (TIL) in the neutron-irradiated part] and [tumor-infiltrating lymphocytes (TIL) in the shielded part] TILs were sorted by flow cytometry as in . In the resulting TIL population, the Tem marker CD44+/CD62L-/CD8+ cells were counted by flow cytometry. Specifically, after cell washing with phosphate-buffered saline (PBS) containing 2% fetal calf serum (FCS), monoclonal antibodies specific for CD8, CD44, CD62L, and fixable viability dyes were used. Stained (PE-conjugated anti-CD8α (53-6.7) monoclonal antibody, Brilliant Violet 510 (BV510)-conjugated anti-CD44 (53-6.7) monoclonal antibody, PE-conjugated anti-CD62L (MEL-14) Monoclonal antibodies were purchased from BD Biosciences, Inc. eFluor 780-conjugate immobilizable dye was purchased from Thermo Fisher Scientific). After washing, the stained cells were analyzed using an LSRFortessa™ flow cytometer (manufactured by BD Biosciences) and FlowJo Software (manufactured by BD Biosciences). Staining antibodies were diluted according to the manufacturer's instructions. The results are shown in Figures 9A and 9B.

Tem (effector memory T cells) in the neutron-irradiated tumor showed a high Tem presence of 24.5% in the control group and 66.1% in the anti-PD-1 antibody administration + BNCT treatment group ( Figure 9A). In addition, Tem in the tumor of the shielded part was 82.7% in the anti-PD-1 antibody administration + BNCT treatment group compared to 43.0% in the control group, which also showed the presence of high Tem in the combined therapy (Fig. 9B).

In both the neutron-irradiated and shielded tumors, Tem was significantly increased in the anti-PD-1 antibody administration + BNCT treatment group compared to the control group, suggesting the long-term targeting of the tumor within the tumor. It can be seen that T cells (Tem) capable of immune response (tumor immune memory) are induced by treatment over a period of time.

Example 3
Boron neutron capture therapy combined immunotherapy (B-NIT), which is a combination of boron neutron therapy and immunotherapy, was performed using the developed advanced-stage melanoma model mice. The model mice were divided into a neutron-irradiated group and a neutron-non-irradiated group. Then, in the neutron irradiation group, the model mice were further controlled (neutron irradiation only) group (n = 9), anti-PD-1 antibody administration group (n = 9), BNCT treatment group (n = 9), and anti-PD- 1 antibody administration + BNCT treatment group (n=9), administration of immune checkpoint inhibitors and boron agents, and neutron irradiation were carried out in the same manner as in Example 1. In the neutron non-irradiation group, the model mice were further controlled (no neutron irradiation) group (n = 9), anti-PD-1 antibody administration group (n = 9), BPA administration group (n = 9), anti-PD-1 antibody The mice were divided into administration + BPA administration groups (n=9), and administration of immune checkpoint inhibitors and boron agents was carried out in the same manner as in Example 1.

[T cells in spleen tissue]
Immune cells are cells differentiated from hematopoietic stem cells present in the bone marrow. Tissues such as bone marrow where immune cells are produced and proliferate are called primary lymphoid tissues, and tissues such as lymph nodes and spleen where the produced immune cells are activated and immune reactions occur are called secondary lymphoid tissues. In this secondary lymphoid tissue, the spleen, immune cells recognize antigens at antigen receptors and become more potent T cells. Therefore, by looking at the number of T cells in the splenic tissue, it is possible to evaluate whether a systemic cell-mediated immune response is taking place.

Therefore, 15 days after the neutron irradiation group was irradiated with neutrons, CD8 + /CD45 + T cells in the spleen of each group (n = 3 each) were collected from the spleen in the same manner as TIL in Example 2, and single cell suspensions were performed. A turbid solution was prepared and adjusted and measured by flow cytometry. In the neutron irradiation group (HOT group), the control group had 5.4% and the anti-PD-1 antibody administration group had 7.1%, showing no change between these groups (Fig. 10A). On the other hand, it increased to 10.2% in the BNCT treatment group, and further increased to 14.4% in the anti-PD-1 antibody administration + BNCT treatment group (Fig. 10A). On the other hand, in the neutron non-irradiation group (COLD group), 4.8% in the neutron non-irradiation control group, 6.1% in the anti-PD-1 antibody administration group, 6.9% in the BPA administration group, and 6.9% in the anti-PD-1 antibody administration group The CD8+/CD45+ T cells in the spleen did not increase to 5.1% in the administration + BPA administration group (Fig. 10B). From the above, CD8 + / CD45 + T cells (activated T cells) in the spleen are increased in the BNCT treatment group subjected to neutron irradiation, and are further increased in the anti-PD-1 antibody administration + BNCT treatment group. Therefore, it is considered that a systemic cellular immune response occurs in the anti-PD-1 antibody administration+BNCT treatment group.

Other Embodiments Although the present invention has been described in detail above, it can be rephrased in other embodiments as follows. Therefore, unless there is a particular contradiction, the above explanations about the "therapeutic drug for malignant tumor" or "combination drug for treating malignant tumor" are also applied to each of the following embodiments.

Embodiment 1. A method of treating a malignant tumor comprising administering an immune checkpoint inhibitor to a patient, the method of treatment being combined with boron neutron capture therapy.

Embodiment 2. A method of treating a malignant tumor comprising administering to a patient an immune checkpoint inhibitor and a boron compound for use in boron neutron therapy, wherein the patient is unresponsive to treatment with the immune checkpoint inhibitor, Immune checkpoint inhibitor is anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-KIR antibody, PD-L1/TGFβ A therapeutic method selected from the group consisting of a trap-inhibiting fusion protein and an anti-PD-1/CTLA-4 bispecific antibody.

In Embodiment 1 or 2 above, it is preferable to administer the immune checkpoint inhibitor at least once, then administer the boron compound, and then irradiate with neutrons, since the effect of the combination is likely to be exhibited.

In the above embodiment 1 or 2, after administering an immune checkpoint inhibitor at least once, administering a boron compound, then irradiating with neutrons, and then administering an immune checkpoint inhibitor at least once is a combination It is preferable from the point that it is easy to exhibit an effect.

Embodiment 3. A medicament comprising an immune checkpoint inhibitor for use in treating malignant tumors in combination with boron neutron capture therapy.

Embodiment 4. (i) anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA-4 antibodies, anti-LAG-3 antibodies for use in the treatment of malignancies in patients who do not respond to treatment with immune checkpoint inhibitors an immune checkpoint inhibitor selected from the group consisting of an antibody, an anti-TIM-3 antibody, an anti-TIGIT antibody, an anti-KIR antibody, a PD-L1/TGF beta trap inhibitory fusion protein and an anti-PD-1/CTLA-4 bispecific antibody and (ii) a pharmaceutical combination comprising a boron compound for use in boron neutron capture therapy.

Embodiment 5. Use of an immune checkpoint inhibitor for the manufacture of a medicament for treating malignant tumors, wherein the medicament for treating malignant tumors is used in combination with boron neutron capture therapy.

Embodiment 6. for the manufacture of a combination medicament for the treatment of malignant tumors in patients who do not respond to treatment with said immune checkpoint inhibitor,
(i) anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-KIR antibody, PD-L1/TGFβ trap blocking fusion protein and an immune checkpoint inhibitor selected from the group consisting of anti-PD-1/CTLA-4 bispecific antibodies, and (ii) the use of boron compounds for use in boron neutron capture therapy.

In any of the above embodiments 1, 3 and 5, the immune checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, an anti-TIM-3 antibody, preferably selected from the group consisting of anti-TIGIT antibodies, anti-KIR antibodies, PD-L1/TGFβ-trap inhibitory fusion proteins and anti-PD-1/CTLA-4 bispecific antibodies, wherein the immune checkpoint inhibitor is a human Preferably, the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, or semiplimab, and the anti-PD-L1 antibody is preferably avelumab, atezolizumab, or durvalumab. , the anti-CTLA-4 antibody is preferably ibilimumab or tremelimumab, the boron compound used in boron neutron capture therapy is preferably borofarane ( ¹⁰ B), and patients who do not respond to immune checkpoint inhibitors Preferably, the subject and the immune checkpoint inhibitor is an immune checkpoint inhibitor to which the patient did not respond, and the malignant tumor is carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma, leukemia, glioma, It is preferably selected from the group consisting of malignant melanoma and lymphoma, the malignant tumor is more preferably malignant melanoma, and the malignant melanoma is more preferably advanced stage malignant melanoma. These preferred aspects apply in any of the first, third and fifth embodiments individually or in combination.

In Embodiments 2, 4 and 6 above, the malignant tumor is preferably selected from the group consisting of carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma, leukemia, glioma, malignant melanoma, and lymphoma. is more preferably malignant melanoma, and more preferably the malignant melanoma is advanced stage malignant melanoma.

1 nucleus 2 tumor infiltrating lymphocytes (TIL)

Claims (15)

1. A therapeutic drug for malignant tumors, which contains an immune checkpoint inhibitor and is used in combination with boron neutron capture therapy.
2. immune checkpoint inhibitor, anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-KIR antibody, PD-L1/TGFβ The therapeutic agent for malignant tumor according to claim 1, which is selected from the group consisting of a trap-inhibition fusion protein and an anti-PD-1/CTLA-4 bispecific antibody.
3. 3. The therapeutic drug for malignant tumor according to claim 2, wherein the immune checkpoint inhibitor is a humanized monoclonal antibody.
4. 4. The therapeutic drug for malignant tumor according to claim 2 or 3, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, or semiplimab.
5. 4. The therapeutic drug for malignant tumor according to claim 2 or 3, wherein the anti-PD-L1 antibody is avelumab, atezolizumab, or durvalumab.
6. 4. The therapeutic drug for malignant tumor according to claim 2 or 3, wherein the anti-CTLA-4 antibody is ibilimumab or tremelimumab.
7. The therapeutic drug for malignant tumors according to any one of claims 1 to 6, wherein the boron compound used in boron neutron capture therapy is borofaran ( ¹⁰ B).
8. The malignant tumor treatment according to any one of claims 1 to 7, wherein the subject is a patient who does not respond to an immune checkpoint inhibitor, and the immune checkpoint inhibitor is an immune checkpoint inhibitor to which the patient has not responded. medicine.
9. The therapeutic drug for malignant tumor according to any one of claims 1 to 8, wherein the malignant tumor is selected from the group consisting of carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma, leukemia, glioma, malignant melanoma, and lymphoma. .
10. 10. The therapeutic drug for malignant tumor according to claim 9, wherein the malignant tumor is malignant melanoma.
11. 11. The therapeutic drug for malignant tumor according to claim 10, wherein the malignant melanoma is advanced stage malignant melanoma.
12. (i) anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM-3 antibody, anti-TIGIT antibody, anti-KIR antibody, PD-L1/TGFβ trap blocking fusion protein and an immune checkpoint inhibitor selected from the group consisting of an anti-PD-1/CTLA-4 bispecific antibody, and (ii) said immune checkpoint inhibitor comprising a boron compound for use in boron neutron capture therapy. A combination drug for the treatment of malignancies in patients who do not respond to therapy.
13. 13. The combination pharmaceutical for treating malignant tumor according to claim 12, wherein the malignant tumor is selected from the group consisting of carcinoma, squamous cell carcinoma, adenocarcinoma, sarcoma, leukemia, glioma, malignant melanoma and lymphoma.
14. The combination pharmaceutical for treating malignant tumor according to claim 13, wherein the malignant tumor is malignant melanoma.
15. The combination pharmaceutical for treating malignant tumor according to claim 14, wherein the malignant melanoma is advanced stage malignant melanoma.

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