CANCER TREATMENT METHOD AND MEDICINE

Abstract

The present disclosure provides a composition, a combination product, a medical device and the like for treating or preventing cancer or a tumor or preventing the recurrence of the cancer or the tumor. The present disclosure provides a composition, a combination product and a medical device for treating or preventing cancer or a tumor or preventing the recurrence of the cancer or the tumor, each of which comprises an immune checkpoint inhibitor and a dendritic cell direct activator or means. In another aspect, the present disclosure provides: a novel cancer treatment method which comprises carrying out a treatment of cancer by employing a combination of a treatment by the administration of an immune checkpoint inhibitor and a treatment for improving the sensitivity to the immune checkpoint inhibitor and, therefore, can be used as an immunotherapy that can be expected to have an excellent therapeutic effect; and a medicine which can be used for the cancer treatment method.

Claims

1.-51. (canceled)

52. A method for treatment, prevention, or prevention of recurrence of cancer or tumor in a subject, comprising administering: a) an effective amount of an immune checkpoint inhibitor; and b) an effective amount of a direct dendritic cell activating agent or means to the subject at different times or at the same time.

53. The method of claim 52, wherein b) the direct dendritic cell activating agent or means comprises b1) an immunoadjuvant.

54. The method of claim 52, wherein the direct dendritic cell activating agent or means comprises a radiation therapy providing means and an immunoadjuvant.

55. The method of claim 52, wherein the immune checkpoint inhibitor comprises at least one agent selected from the group consisting of a PD-1 inhibitor, a PD-L1 inhibitor, and a CTLA-4 inhibitor.

56. The method of claim 52, wherein the method is performed on a subject who has not received therapy with an immune checkpoint inhibitor.

57. The method of claim 52, wherein the method is performed on a subject for whom therapy with an immune checkpoint inhibitor was determined to be ineffective.

58. The method of claim 52, wherein the method is performed on a subject who received therapy with an immune checkpoint inhibitor which was effective, but subsequently experienced progressive disease (PD).

59. The method of claim 52, wherein the cancer or tumor has an EGFR gene mutation.

60. The method of claim 52, wherein the method is performed on a subject who has received therapy with a tyrosine kinase inhibitor.

61. The method of claim 52, wherein the subject is a CD8.sup.+ T cell.sup.low subject.

62. The method of claim 52, wherein a regulatory T cell inhibitor is further administered.

63. The method of claim 62, wherein the regulatory T cell inhibitor comprises a COX-2 inhibitor.

64. A method for directly activating a dendritic cell in a subject, comprising administering an effective amount of an immunoadjuvant to the subject and/or using a radiation therapy providing means for the subject, wherein the subject has received an immune checkpoint inhibitor.

65. The method of claim 64, wherein the immunoadjuvant comprises a human Mycobacterium tuberculosis hot water extract or a portion thereof.

66. The method of claim 64, wherein the activation is determined by measuring an increase in an amount of expression of CD80/86 on a surface of a dendritic cell or an increase of CD8 positive T cells in the presence of the immunoadjuvant compared to in the absence of the immunoadjuvant.

67. The method of claim 65, wherein a radiation therapy is administered to the subject when the immunoadjuvant is administered.

68. A method for treatment, prevention, or prevention of recurrence of cancer or tumor of a subject with a specific regimen, the method comprising administering a combination of an effective amount of an immune checkpoint and an effective amount of a dendritic cell activating agent or means, wherein the regimen comprises: a) a step of administering an effective amount of an immune checkpoint inhibitor; and b) a step of administering an effective amount of a dendritic cell activating agent.

69. The method of claim 68, wherein the subject is a subject who has not received therapy with an immune checkpoint inhibitor or a subject for whom therapy with an immune checkpoint inhibitor alone is ineffective.

70. The method of claim 68, wherein the specific regimen further comprises: c) a step of performing radiation therapy.

71. The method of claim 68, wherein the dendritic cell activating agent or means comprises Extract Z that is administered by aspirating a liquid medicine into a 1 mL syringe and subcutaneously injecting 0.05 mL thereof twice a week.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0511] FIG. 1 is a block diagram showing the configuration of a system that utilizes a program in the present embodiment.

[0512] FIG. 2 is a flowchart showing the treatment procedure in the present embodiment.

[0513] FIG. 3 is a diagram describing the change over time in the therapeutic approach in Example 1.

[0514] FIG. 4 is a diagram describing the change over time in the therapeutic approach in Example 2.

[0515] FIG. 5 is a diagram describing the change over time in the tumor marker measurement value and the therapeutic approach in Example 3.

[0516] FIG. 6 is a diagram describing the change over time in the tumor marker measurement value and the therapeutic approach in Example 4.

[0517] FIG. 7 is a diagram showing enhancement of CXCL10 production by an immunoadjuvant in Example 13.

[0518] FIG. 8 is a diagram showing the effect of promoting infiltration of CD8.sup.+ T cells into tumor in Example 15.

[0519] FIG. 9 is a diagram showing PD-L1 expression in cultured Sq-1979 cells in Example 20.

[0520] FIG. 10 is a diagram describing the change over time in the therapeutic approach in Example 23.

[0521] FIG. 11 is a diagram describing the change over time in the tumor marker measurement value and the therapeutic approach in Example 23.

DESCRIPTION OF EMBODIMENTS

[0522] The present disclosure is described hereinafter while showing some of the best modes thereof. Throughout the entire specification, a singular expression should be understood as encompassing the concept thereof in the plural form, unless specifically noted otherwise. Thus, singular articles (e.g., “a”, “an”, “the”, and the like in the case of English) should also be understood as encompassing the concept thereof in the plural form, unless specifically noted otherwise. Further, the terms used herein should be understood as being used in the meaning that is commonly used in the art, unless specifically noted otherwise. Therefore, unless defined otherwise, all terminologies and scientific technical terms that are used herein have the same meaning as the general understanding of those skilled in the art to which the present disclosure pertains. In case of a contradiction, the present specification (including the definitions) takes precedence.

Definitions

[0523] The terms used herein are described hereinafter.

[0524] As used herein, an “immune checkpoint inhibitor” is a substance capable of disabling suppression of activation of a T cell by an immune checkpoint molecule by binding to an immune checkpoint molecule or a ligand thereof to inhibit immunosuppressive signaling. Specific examples include inhibitors against PD-1, PD-L1, PD-L2, CTLA-4, LAG-3, TIM-1, TIM-3, TIM-4, VISTA, BTLA, TIGIT, A2AR, 4-1BB, 4-1BBL, 2B4 (CD244), KIR family receptors, B7.1, B7.2, B7-H2, B7-H3, B7-H4, B7-H6, BATE, CD39, CD40, CD47, CD48, CD73, CD94/NKG2A, CD96, CD160, CD200, CD200R, CD274, butyrophilins, CEACAM1, CSF-1R, DcR3, EDO, Foxp1, GARP, GITR, gp49B, HHLA2, HVEM, ICOS, IDO, ILT-2, ILT-4, LAIR-1, MAFB, MICA./B, NKG2A/HLA-E, NR4A2, OCT-2, OX-40, PIR-B, Rara (retinoic acid receptor alpha), SIRP, TDO, TLR3, and TNFR. Preferred examples include inhibitors against PD-1, PD-L1, and CTLA-4. An anti-PD-1 antibody, which is one of PD-1 inhibitors, binds to PD-1 on a T cell to inhibit binding between PD-1 and PD-L1, thereby blocking suppressive signaling and maintaining activation of the T cell. An anti-PD-L1 antibody, which is one of PD-L1 inhibitors, binds to PD-L1 expressed on a cancer cell or an antigen-presenting cell to inhibit interaction with PD-1 on a T cell. As a result, suppressive signaling to the T cell is inhibited, and activation of the T cell is maintained. An anti-CTLA-4 antibody, which is one of CTLA-4 inhibitors, competes with a CD28 ligand on a dendritic cell and blocks a suppressive signal of an immune cell via CD28 to maintain activation of a T cell. Examples of an anti-PD-1 antibody include nivolumab, pembrolizumab, spartalizumab, and cemiplimab. Examples of an anti-PD-L1 antibody include atezolizumab, durvalumab, and avelumab. Examples of an anti-CTLA-4 antibody include ipilimumab and tremelimumab.

[0525] As used herein, an “immune checkpoint factor” is a factor that suppresses an autoimmune response in order to maintain homeostasis and also suppresses an excessive immune reaction. An “immune checkpoint factor” originally exists in order to suppress excessive activation of a T cell and not to attack itself. However, an immune checkpoint factor is utilized by a cancer cell to avoid an attack from the immune system and proliferate during the carcinogenesis process. Thus, inhibition of an immune checkpoint factor in a cancer cell enables a T cell to attack the cancer cell. Representative examples of an “immune checkpoint factor” include PD-1, PD-L1, PD-L2, CTLA-4, LAG-3, TIM-1, TIM-3, TIM-4, VISTA, BTLA, TIGIT, A2AR, 4-1BB, 4-1BBL, 2B4 (CD244), KIR family receptors, B7.1, B7.2, B7-H2, B7-H3, B7-H4, B7-H6, BATE, CD39, CD40, CD47, CD48, CD73, CD94/NKG2A, CD96, CD160, CD200, CD200R, CD274, butyrophilins, CEACAM1, CSF-1R, DcR3, EDO, Foxp1, GARP, GITR, gp49B, HHLA2, HVEM, ICOS, IDO, ILT-2, ILT-4, LAIR-1, MAFB, MICA./B, NKG2A/HLA-E, NR4A2, OCT-2, OX-40, PIR-B, Rara (retinoic acid receptor alpha), SIRP, TDO, TLR3, and TNFR.

[0526] As used herein, a “direct dendritic cell activating agent or means (device)” refers to an agent or means capable of directly (in other words, not via other molecules) activating a dendritic cell inside or outside the body of a subject. Representative examples of a “direct dendritic cell activating agent or means” include a radiation therapy providing means and an immunoadjuvant.

[0527] In the present disclosure, whether a certain substance “directly activates” a “dendritic cell” can be determined in the following manner: as exemplified in the Examples or the like, when a target substance or factor that is a candidate has higher expression of CD80/86 that is a cell membrane surface marker than that in the absence of the target substance or factor (e.g., saline)+the control antibody group, the target substance or factor that is a candidate is considered as directly activating the cell, and it can be determined that the target substance or factor falls under the “direct dendritic cell activating agent or means (device)”.

[0528] As used herein, a “radiation therapy providing means” refers to a method, an apparatus, an equipment, an agent, a device and the like for providing radiation therapy to a subject.

[0529] As used herein, a “medical device” refers to an object that is inserted or transplanted into a target or applied to the surface of a target for treatment, prevention, or prevention of recurrence. In addition to any device that is used for radiation therapy, general examples of the medical device include stents, fasteners, ports, catheters, scaffolds, grafts, and the like.

[0530] As used herein, an “EGFR gene mutation” refers to a mutation in the EGFR gene. “EGFR” means epidermal growth factor receptor and functions in proliferation and growth of a cell. When a mutation occurs in the EGFR gene, proliferation and growth of a cell are constantly activated, which causes canceration. An “EGFR gene mutation” is known to cause non-squamous cell carcinoma, which is one type of lung cancer.

[0531] As used herein, a “tyrosine kinase inhibitor (TKI)” refers to an agent that inhibits tyrosine kinase. A tyrosine kinase inhibitor has an action of suppressing cancer proliferation by blocking a signaling pathway involved in proliferation of a cancer cell. Representative examples of a “tyrosine kinase inhibitor” include afatinib, erlotinib, osimertinib (AZD9291), AZD3759, gefitinib, canertinib, lapatinib, cetuximab, matuzumab, zalutumumab, and panitumumab.

[0532] As used herein, an “immunoadjuvant” refers to any agent or factor that assists an immune reaction. Representative examples thereof can include a Mycobacterium tuberculosis extract such as a human Mycobacterium tuberculosis hot water extract or a portion thereof.

[0533] As used herein, “CXCL10” is an abbreviation of C-X-C motif chemokine ligand 10 and is also known as IP-10, interferon gamma-induced protein 10, or Small-inducible cytokine B10. CXCL10 is an 8.7 kDa protein that, in humans, is encoded by the CXCL10 gene, is a non-glycosylated protein, and consists of 77 amino acids. CXCL10 is a chemokine that is produced in response to treatment of a monocyte, an endothelial cell, or a fibroblast with IFNγ. IP-10 functions as a chemotaxis inducer cell expressing a G protein-coupled receptor, CXCR3 that has been found mainly in an activated T cell or NK cell. In addition, IP-10 is also known as CXCL10, C7, IFI10, INP10, IP-10, SCYB10, crg-2, gIP-10, mob-1, C-X-C motif chemokine ligand 10, C-X-C motif chemokine 10, or the like. IP-10 is also known by its IDs, NM_001565 (nucleic acid) or NP 001556 (protein).

[0534] As used herein, “enhancement of CXCL10 production” refers to increasing the production (or amount of substance) of CXCL10.

[0535] As used herein, “cancer or tumor that is CXCL10 positive” refers to cancer or tumor being CXCL10 positive.

[0536] As used herein, “CXCR3” has the same meaning as the meaning commonly used in the art, and is one of CXC chemokine receptor families, which are G protein-coupled receptors. In addition to G protein-coupled receptor 9 (GPR9) and CD183, CXCR3 may be referred to as CD182; CKR-L2; CMKAR3; IP10-R; Mig-R; MigR, or the like. Two mutants of CXCR3 are known. While CXCR3-A, one of the mutants, binds to CXCL9 (MIG), CXCL10 (IP-10), and CXCL11 (I-TAC), which are CXC chemokines, CXCR3-B can further bind to CXCL4 in addition to those chemokines. While examples of the nucleic acid ID include NM_001142797, NM_001504, and the like, examples of the protein ID can include NP_001136269, NP_001495, and the like.

[0537] As used herein, a “CD8.sup.+ T cell.sup.low” refers to a T cell positive for CD8 expression. As used herein, infiltration of CD8 positive T cells are evaluated in accordance with the cell count counted by confirming a slide using at least three different high magnification fields (40 power objective and 10 power eyepiece at the maximum). The number of cells stained as CD8 positive is recorded, and the case where the number of cells is 5 or less in three fields is defined as low infiltration, while the case where the number of cells is more than 5 is defined as high infiltration.

[0538] As used herein, a “human Mycobacterium tuberculosis hot water extract” is typically a substance produced from a human Mycobacterium tuberculosis, which is a mixture including polysaccharides with arabinose, mannose, and glucose as the primary ingredients. While anticancer effects due to human Mycobacterium tuberculosis hot water extracts have been studied for a long time, the detailed mechanism of action thereof is not necessarily elucidated. Further, the extract has not been used as a prophylactic drug. The extract can also comprise a trace amount of ingredients such as a protein, peptide, amino acid, nucleic acid, or lipid (glycolipid) when appropriate. Examples of a human Mycobacterium tuberculosis hot water extract can include Extract Z described herein or the like. Thus, the technology of the present disclosure can use Extract Z described in detail herein or any formulation manufactured using Extract Z as an active pharmaceutical ingredient.

[0539] The following is a representative manufacturing method of human Mycobacterium tuberculosis hot water extracts.

[0540] Human Mycobacterium tuberculosis is cultured for 3 to 7 weeks in a 37° C. thermostatic vessel. The film of microbial cells formed on a medium is then filtered out. The moist microbial cells with medium components removed by washing with water are used as the extracted raw material. The microbial cells are floated in a distilled water at an amount that is 15 to 40-fold of the wet weight thereof, and heated for 80 to 180 minutes at 90 to 120° C. for extraction. The residue of the microbial cells is removed with a sterilization filter, and the extracted solution is concentrated to 60% or less, and then acetone, trichloroacetate, ammonium sulfate, sulfosalicylic acid, or the like is added thereto so as to arrive at 0.5 to 3% (w/v), and stirred and left standing. The deposited precipitate is then centrifuged and removed, and the supernatant is subjected to running water dialysis. Inner fraction of dialysis fluid is subjected to vacuum concentration to a 1/20 to ¼ volume, and sodium chloride is added to the concentrate so as to arrive at 0.5 to 1% (w/v). 2 to 4-fold volume of ethanol is added and left standing, and then the precipitate is removed by centrifugation. After adding an additional 2 to 6-fold volume of ethanol and incubating, a precipitating polysaccharide is obtained by centrifugation or the like. Those skilled in the art can understand that each of the conditions described above can be appropriately changed to obtain the same product.

[0541] As used herein, “prophylaxis” or “prevention” is an act of administering an active ingredient in the present disclosure to an individual who has not developed the target disease, intended to for example prevent development of the disease.

[0542] As used herein, “treatment” or “therapy” is, for example, an act of administering an active ingredient of the present disclosure to an individual (subject, patient) diagnosed as having a developed disease by a physician or a similar practitioner, intended to, for example, alleviate the disease or condition, not increase carcinoma, or revert back to the state before the development of the disease. Even if the objective of administration is prevention of exacerbation of the disease or condition or prevention of increase in the carcinoma, the administration is a therapeutic act if administered to a patient.

[0543] As used herein, “radiation therapy” refers to a therapeutic method by irradiation of a radiation. Examples of a radiation include X-rays, gamma rays, electron beams, proton beams, heavy particle beams, and the like. For a radiation, one type of radiation may be used, or two or more types of radiation may be used. In one aspect, examples of the “radiation therapy providing means” of the present disclosure can include a radiosensitizer, a radiation irradiating device, a radioactive substance, and the like. In the present disclosure, the “radiation therapy providing means” is preferably configured to provide palliative irradiation. More preferably, the “radiation therapy providing means” is configured to irradiate to have an abscopal effect. In still another embodiment, the “radiation therapy providing means” is configured to irradiate a measurable lesion that is present other than a lesion that is a target.

[0544] As used herein, a “regulatory T cell inhibitor” refers to any agent, device, or the like that suppresses regulatory T cells. Whether regulatory T cells is suppressed can be determined by, for example, the following approach:

(1) Collecting CD4.sup.+CD25.sup.+ cells from the spleen or tumor from a mouse and co-culturing the cells with effector T cells collected/differentiated from the spleen of a mouse to confirm proliferation of the effector T cells. Performing treatment with an agent by administering the agent in vivo or adding the agent in vitro when culturing the CD4.sup.+CD25.sup.+ cells.
(2) Transplanting cancer cells into a mouse and administering the agent to the mouse. Collecting tumor and calculating the ratio of FoxP3.sup.+ cells in CD4.sup.+CD25.sup.+ cells infiltrating into the tumor.
(3) Transplanting cancer cells into a mouse and administering the agent to the mouse. Collecting tumor and confirming FoxP3 gene expression.

[0545] The target animal is not limited to mice and may be other animals (which can include humans). Examples of regulatory T cells inhibitor can include, but are not limited to, a COX-2 inhibitor and the like.

[0546] As used herein, a “COX-2 inhibitor” refers to an inhibitor of prostaglandin-endoperoxide synthase 2. Examples can include celecoxib, etodolac, meloxicam, nabumetone, zaltoprofen, lornoxicam, and the like.

[0547] The term “carrier” as used herein refers to a pharmaceutically acceptable substance, composition, or excipient such as, for example, a liquid or solid bulking agent, diluent, additive, solvent, or capsule forming agent, which is associated with or enables the transport or carriage of a target pharmaceutical compound from an organ or portion of the body to another organ or portion of the body. “Pharmaceutically acceptable” refers to being compatible with other raw materials in a formulation and being harmless to patients. Non-limiting examples of pharmaceutically acceptable carriers, carriers, and/or diluents can include sugars such as lactose, glucose, and sucrose, starch such as corn starch and potato starch, cellulose and derivatives thereof such as carboxymethylcellulose sodium, ethyl cellulose, and cellulose acetate, excipients such as powdered tragacanth, malt, gelatin, talc, cocoa powder, and suppository wax, oil such as peanut oil, cotton seed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil, glycols such as propylene glycol, polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol, esters such as ethyl oleate and ethyl laureate, buffering agents such as agar, magnesium hydroxide, and aluminum hydroxide, alginic acid, pyogenic substance-free water, isotonic saline, Ringer's solution, ethyl alcohol, phosphate buffer, and other nontoxic compatible substances used in a pharmaceutical formulation. A humectant, emulsifier, and lubricant such as sodium lauryl sulfate, magnesium stearate, or polyethylene oxide-polypropylene oxide copolymer, as well as a colorant, releasing agent, coating agent, sweetener, flavoring agent, fragrance, preservative, and antioxidant can also be included in a composition.

[0548] As used herein, “parenteral administration” refers to a dosage form for any route that is not oral administration. Any mode for administration in a mode and level that are effective for treating or preventing a disease intended for cancer treatment or prevention is employed. Examples of means of parenteral administration include administration through transdermal absorption or transmucosal absorption, as well as injection, infusion, and combinations thereof. For example, administration through transdermal absorption or transmucosal absorption exerts an effect by contacting a transdermally absorbed formulation such as a paste agent, adhesive formulation, or spray with the skin or mucous membrane so that a drug in the formulation migrates into the body through the skin or mucous membrane. Examples of administration via injection or infusion include intravenous, intradermal, subcutaneous, intramuscular, and enteral administration (intestinal infusion), which can also be administered as a bolus and/or sustained infusion. Injection or infusion can use a suspension, liquid agent, emulsion, or implanted agent in an oily or aqueous medium, comprising another formulation substance such as a suspending agent, stabilizer, and/or a dispersant. Enteral administration (intestinal infusion) can provide sustained drug delivery to the proximal small intestine by using a tube or portable infusion pump by percutaneous endoscopic gastrostomy. More preferably, the administration can be subcutaneous or intradermal administration. Parenteral administration (e.g., transdermal administration) can be performed with a tape/patch agent or powder, spray, ointment, paste, cream, lotion, gel, solution, or the like. A composition suitable for parenteral administration can comprise at least one type of a pharmaceutically acceptable aseptic isotonic aqueous or non-aqueous solution, dispersant, suspension, emulsion, implanted agent, or aseptic powder that can be reconstituted in an aseptic injection solution or dispersant immediately before use.

DESCRIPTION OF PREFERRED EMBODIMENTS

[0549] The preferred embodiments of the present disclosure are described hereinafter. It is understood that the embodiments provided hereinafter are provided to better facilitate the understanding of the present disclosure, and thus the scope of the present disclosure should not be limited by the following descriptions. Thus, it is apparent that those skilled in the art can refer to the descriptions herein to make appropriate modifications within the scope of the present disclosure. It is also understood that the following embodiments of the present disclosure can be used alone or in combination.

[0550] <Direct Dendritic Cell Activation>

[0551] In one aspect, the present disclosure provides various agents or means for directly activating a dendritic cell. In one embodiment, the present disclosure provides a composition or use for directly activating a dendritic cell which comprises an immunoadjuvant, and a method using the composition or use. In an exemplary embodiment, an immunoadjuvant comprises a Mycobacterium tuberculosis extract such as a human Mycobacterium tuberculosis hot water extract or a portion thereof.

[0552] In another embodiment, the present disclosure provides a composition or medical device or use for activating a dendritic cell which comprises a radiation therapy providing means, and a method using the composition or medical device or use. In this case, the radiation therapy providing means comprises at least one selected from the group consisting of a radiosensitizer, a radiation irradiating device, and a radioactive substance. For example, a radiation therapy providing means is configured to provide palliative irradiation. In a preferred embodiment, a radiation therapy providing means is configured to irradiate to have an abscopal effect. In a specific example, a radiation therapy providing means is configured to irradiate a measurable lesion that is present other than a lesion that is a target.

[0553] In the present disclosure, whether a dendritic cell is directly activated can be determined by confirming an increase in the presence of a target substance or factor compared to in the absence of the target substance or factor in an experiment comprising measuring an increase in the amount of expression of CD80/86 on the surface of the dendritic cell in the presence of an immunoadjuvant compared to in the absence of the immunoadjuvant.

[0554] <Combined Use of Immune Checkpoint Inhibition and Direct Dendritic Cell Activation>

[0555] The present disclosure is based on the discovery that combined use of an immune checkpoint inhibitor and a direct dendritic cell activating agent or means attains a therapeutic and preventive effect, which is specific and effective against neoplasm such as cancer.

[0556] The present disclosure generally provides a combination for treatment, prevention, or prevention of recurrence of cancer or tumor of a subject by combined use of an immune checkpoint inhibitor and a direct dendritic cell activating agent or means, a method for the treatment, prevention, or prevention of recurrence, and a combination of an immune checkpoint inhibitor and a direct dendritic cell activating agent or means for use in the treatment, prevention, or prevention of recurrence. The immune checkpoint inhibitor and the direct dendritic cell activating agent or means may be administered or provided at the same time or at different times. Any of administration of the immune checkpoint inhibitor and administration of the direct dendritic cell activating agent or means may precede.

[0557] In another aspect, the present disclosure provides a composition, use or a medical device, use, a therapeutic method, a preventive method, or a recurrence preventing method for treatment, prevention, or prevention of recurrence of cancer or tumor of a subject, using a direct dendritic cell activating agent or means. The direct dendritic cell activating agent or means is administered or used in combination with an immune checkpoint inhibitor.

[0558] In still another aspect, the present disclosure provides a composition, use or a medical device, use, a therapeutic method, a preventive method, or a recurrence preventing method for treatment, prevention, or prevention of recurrence of cancer or tumor of a subject, using an immune checkpoint inhibitor. The immune checkpoint inhibitor is administered or used in combination with a direct dendritic cell activating agent or means.

[0559] In one representative aspect, examples of a direct dendritic cell activating agent or means can include an immunoadjuvant, a radiation therapy providing means, a radioactive substance, a combination thereof, and the like.

[0560] The immune checkpoint inhibitor or the direct dendritic cell activating agent or means of the present disclosure can be provided in a dosage form of a pharmaceutical composition. In a specific embodiment, a pharmaceutical composition can comprise one or more types of compounds and at least one type of pharmaceutically acceptable carrier, wherein the one or more types of compounds can be converted into, for example, at least one type of compound of a Mycobacterium tuberculosis extract in a subject (in other words, the one or more types of compounds may be provided as a prodrug).

[0561] In one embodiment, examples of an immunoadjuvant that can be used in the present disclosure can include a human Mycobacterium tuberculosis hot water extract or a portion thereof, and the like.

[0562] In one embodiment, any means (such as a device) that provides an X-ray, a gamma ray, an electron beam, a proton beam, a heavy particle beam, or the like can be used as a radiation therapy providing means, wherein one type of radiation may be used, or two or more types of radiation may be used as a radiation. In one aspect, the radiation therapy providing means of the present disclosure may comprise a radiosensitizer, may be used with a radiation irradiating device, may be provided with a radioactive substance, or may comprise any combination thereof. In the present disclosure, a radiation therapy providing means is preferably configured to provide palliative irradiation. More preferably, a radiation therapy providing means is configured to irradiate to have an abscopal effect. Furthermore, in one embodiment, a radiation therapy providing means is configured to irradiate a measurable lesion that is present other than a lesion that is a target.

[0563] In the present disclosure, whether a dendritic cell is directly activated can be determined by confirming an increase in the presence of a target substance or factor compared to in the absence of the target substance or factor in an experiment comprising measuring an increase in the amount of expression of CD80/86 on the surface of the dendritic cell in the presence of an immunoadjuvant compared to in the absence of the immunoadjuvant.

[0564] In one preferred embodiment, an immune checkpoint inhibitor can be an inhibitor of a factor such as PD-1, PD-L1, or CTLA-4. An immune checkpoint inhibitor may be a combination of inhibitors of these factors. In one example, an immune checkpoint inhibitor may be a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, or the like. One agent may be an inhibitor of a plurality of factors, PD-1, PD-L1, and CTLA-4. One specific example can be, but is not limited to, nivolumab, pembrolizumab, atezolizumab, or the like.

[0565] In one embodiment, it is preferable, but is not limited to, that cancer expresses PD-L1. In another embodiment, cancer can include, but is not limited to, lung cancer. In a preferred embodiment, cancer or tumor can be, but is not limited to, cancer or tumor having an EGFR gene mutation. Thus, in one example, treatment, prevention, or prevention of recurrence can be for a subject who has received therapy with a tyrosine kinase inhibitor (e.g., EGFR-TKI). In one specific embodiment, therapy, prevention, or prevention of recurrence can be performed on a subject who has not received therapy with an immune checkpoint inhibitor. In another embodiment, treatment, prevention, or prevention of recurrence can be performed on a subject for whom therapy with an immune checkpoint inhibitor was determined to be ineffective. In one specific embodiment, treatment, prevention, or prevention of recurrence of the present disclosure can be performed on, but not limited to, a subject who received therapy with an immune checkpoint inhibitor which was effective, but subsequently experienced progressive disease (PD).

[0566] In one embodiment, a subject is a subject who is CXCL10 positive in the subject's tumor because a CXCR3 positive cell can easily invade. In another embodiment, the technology of the present disclosure is used for enhancement of CXCL10 production. In another embodiment, a subject is a CD8.sup.+ T cell.sup.low subject. Thus, the technology of the present disclosure can be used to promote infiltration of CD8.sup.+ T cells into tumor. In a specific embodiment, the present disclosure can be used for treatment, prevention, or prevention of recurrence of Tumor Mutational Burden.sup.high and CD8.sup.+ T cell.sup.low cancer that is inoperable or metastatic solid cancer for which there is no other therapeutic choice.

[0567] In an embodiment of the present disclosure, the present disclosure may be used with a regulatory T cell inhibitor. A COX-2 inhibitor can be used as a regulatory T cell inhibitor. Preferably, a COX-2 inhibitor can be used as a regulatory T cell inhibitor. Examples that can be used as a COX-2 inhibitor include, but are not limited to, celecoxib, other COX-2 inhibitors (e.g., etodolac, meloxicam, nabumetone, zaltoprofen, and lornoxicam), and the like.

[0568] (Medical Technology Such as Medicaments or Therapy)

[0569] A medicament for use in the technology for treatment, prevention, or prevention of recurrence of cancer of the present disclosure can be used by any approach that is known as a pharmaceutical product in the art.

[0570] In a specific embodiment, a pharmaceutical composition can comprise one or more types of compounds and at least one type of pharmaceutically acceptable carrier, wherein the one or more types of compounds can be converted into at least one type of Mycobacterium tuberculosis extract (i.e., prodrug) in a subject. A plurality of agents, when included, can be included in a single composition (combined agent) or in separate compositions. The agents can be formulated as a single composition using a known embodiment in the art, including those exemplified herein. A plurality of agents may be provided to achieve a therapeutic method (e.g., anticancer agent administration, radiation therapy, or the like) or with one or more other medicaments (e.g., surgery or an anticancer agent such as a chemotherapeutic agent) in addition to the immune checkpoint inhibitor and/or direct dendritic cell activating agent or means of the present disclosure. The immune checkpoint inhibitor and/or direct dendritic cell activating agent or means of the present disclosure can be provided or administered in combination with one or more other medicaments or therapeutic methods (e.g., surgery, chemotherapeutic agent, radiation therapy, or anticancer agent). In one embodiment, one or more other medicaments or therapeutic methods (e.g., surgery, chemotherapeutic agent, radiation therapy, or anticancer agent) may be administered after an appropriate period has elapsed from administration of the immune checkpoint inhibitor and/or direct dendritic cell activating agent or means of the present disclosure. When administered separately, two or more medicaments may be provided as a kit. Non-limiting examples of anticancer agents include chemotherapeutic agents such as antimetabolites and alkylating agents, growth inhibitors, cytotoxic agents, agents used in radiation therapy, antiangiogenesis agents, apoptosis agents, anti-tubulin agents, anticancer antibiotics, anti-microtubule drugs, tyrosine kinase inhibitors, proteasome inhibitors, anaplastic lymphoma kinase inhibitors, Janus kinase inhibitors, CDK inhibitors, MEK inhibitors, Raf kinase inhibitors, PARP inhibitors, antibody drugs, other molecularly targeted drugs, platinum formulations, immunotherapy such as dendritic cell therapy, gene therapy, other low molecule drugs, other agents for treating cancer, and the like.

[0571] The compositions disclosed herein that are suitable for oral administration can be in a dosage form of a capsule, cachet, pill, tablet, lozenge (generally using a fragrance base tragranth or acacia and sucrose), powder, granule, aqueous or non-aqueous liquid solution, aqueous or non-aqueous liquid suspension, oil-in-water emulsion, water-in-oil emulsion, elixir, syrup, troche (using inactive base such as gelatin, glycerin, sucrose, and/or acacia), and/or mouthwash, which each comprises a predetermined amount of at least one type of compound in the present disclosure.

[0572] A composition disclosed herein can be administered as a bolus, an electuary or paste.

[0573] The immune checkpoint inhibitor and/or direct dendritic cell activating agent or means of the present disclosure can be administered in any dosage form. Any dosage form, whether oral administration or parenteral administration, can be used, as long as the effect can be exerted. Preferably, parenteral administration is used.

[0574] A solid dosage form for oral administration (capsule, tablet, pill, sugar coated tablet, powder, granule, or the like) can be mixed with any of one or more types of pharmaceutically acceptable carrier such as sodium citrate or dicalcium phosphate, and/or a filler or bulking agent such as starch, lactose, sucrose, glucose, mannitol, and/or silisic acid, binding agent such as carboxymethyl cellulose, alginic acid salt, gelatin, polyvinyl pyrrolidone, sucrose, and/or acacia, a moisturizing agent such glycerol, a disintegrant such as agar, calcium carbonate, potato or tapioca starch, alginic acid, specific silicic acid salt, sodium carbonate, and sodium starch glycolate, a dissolution delaying agent such as paraffin, an absorption promotor such as a quaternary ammonium compound, humectant such as cetyl alcohol, glycerol monostearate, and polyethylene oxide-polypropylene oxide copolymer, an absorbent such as kaolin and bentonite clay, a lubricant such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixture thereof, and a colorant. For a capsule, tablet, and pill, a pharmaceutical composition can also comprise a buffering agent. A similar type of solid composition can also be used as a filler in a soft and hard filled gelatin capsule using an additive such as lactose and high molecular weight polyethylene glycol.

[0575] A liquid dosage form for oral administration can comprise a pharmaceutically acceptable emulsion, microemulsion, solution, suspension, syrup, and elixir. In addition to the active ingredient, a liquid dosage form can comprise an inactive diluent used in conventional art, such as water or other solvent, solubilizing agent, and emulsifying agent, e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil (especially cotton seed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol, sorbitan fatty acid ester, mixture thereof, and the like. Furthermore, a compound can be dissolved using cyclodextrin such as hydroxypropyl-β-cyclodextrin.

[0576] The component of the present disclosure can comprise an adjuvant such as a humectant, emulsifying and suspending agent, sweetener, flavoring agent, colorant, fragrance, or preservative. In addition to one or more types of compounds according to the present disclosure, a suspension can comprise a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan ester, microcrystalline cellulose, aluminum methhydroxide, bentonite, agar, tragacanth, or mixture thereof.

[0577] The combination disclosed herein can be a suppository for rectal or vaginal administration, which can be prepared by mixing one or more types of compounds according to the present disclosure with one or more types of suitable non-stimulatory additives or carriers including cocoa butter, polyethylene glycol, suppository wax, salicylate, or the like. The composition is a solid at room temperature, but a liquid at body temperature. Thus, the composition melts in the rectal or vaginal cavity and releases the compound of the present disclosure. A pharmaceutical composition suitable for vaginal administration can also comprise a pessary, tampon, cream, gel, paste, foam, or spray formulation comprising a carrier known to be suitable in conventional art.

[0578] The dosage form for topical or transdermal administration of the combination of the present disclosure can comprise powder, spray, ointment, paste, cream, lotion, gel, solution, patch, and inhalant. A pharmaceutical composition or pharmaceutical tablet can be mixed with a pharmaceutically acceptable carrier and a preservative, buffering agent, or high pressure gas that may be required under aseptic conditions.

[0579] An ointment, paste, cream, and gel can comprise, in addition to the combination of the present disclosure, an additive such as animal or vegetable fat, oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silicic acid, talc, zinc oxide, or mixture thereof.

[0580] Powder or spray can comprise, in addition to the pharmaceutical composition or pharmaceutical tablet of the present disclosure, an additive such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, or polyamide powder, or a mixture thereof. Furthermore, spray can comprise common high pressure gas such as chlorofluorohydrocarbon and volatile unsubstituted hydrocarbon such as butane and propane.

[0581] Ophthalmic formulations, optical ointment, powder, solution, and the like are also understood to be within the scope of the present disclosure.

[0582] A composition suitable for parenteral administration can comprise at least one type of pharmaceutically acceptable aseptic isotonic aqueous or non-aqueous solution, dispersant, suspension, emulsion, or aseptic powder that can be reconstituted in an aseptic injection solution or dispersant immediately before use.

[0583] The term “salt” as used herein includes acid and/or base salt formed with inorganic and/or organic acid and base. As used herein, the term “pharmaceutically acceptable salt” refers to a salt which is suitable for use in contact with tissue of a subject without excessive toxicity, stimulation, allergic reaction, and/or similar events with a reasonable balance of effect/risk ratio within the scope of a definitive medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in detail in Berge et al., J. Pharmaceutical Sciences (1977) 66: 1-19.

[0584] Pharmaceutically acceptable salts can be produced with an inorganic or organic acid. Non-limiting examples of suitable inorganic salts include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid. Non-limiting examples of suitable organic salts include acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, and malonic acid. Other non-limiting examples of suitable pharmaceutically acceptable salts include adipic acid salt, alginic acid salt, ascorbic acid salt, aspartic acid salt, benzenesulfonic acid salt, besilate, benzoic acid salt, bisulfuric acid salt, boric acid salt, butyric acid salt, camphoric acid salt, camphorsulfonic acid salt, citric acid salt, cyclopentanepropionic acid salt, digluconic acid salt, dodecylsulfuric acid salt, ethanesulfonic acid salt, formic acid salt, fumaric acid salt, glucoheptonic acid salt, glycerophosphoric acid salt, gluconic acid salt, hemisulfuric acid salt, heptanoic acid salt, hexanoic acid salt, hydroiodic acid salt, 2-hydroxy-ethanesulfonic acid salt, lactobionic acid salt, lactic acid salt, lauric acid, lauryl sulfate, malic acid salt, maleic acid salt, malonic acid salt, methanesulfonic acid salt, 2-naphthalenesulfonic acid salt, nicotinic acid salt, nitric acid salt, oleic acid salt, oxalic acid salt, palmitic acid salt, pamoic acid salt, pectinic acid salt, persulfuric acid salt, 3-phenylpropionic acid salt, phosphoric acid salt, picric acid salt, pivalic acid salt, propionic acid salt, stearic acid salt, succinic acid salt, sulfuric acid salt, tartaric acid salt, thiocyanic acid salt, p-toluenesulfonic acid salt, undecanoic acid salt, and valeric acid salt. In some embodiments, examples of organic acids that can produce a salt include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, lactic acid, trifluoroacetic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid.

[0585] A salt can be prepared at the time of separation and purification of a disclosed compound or separately by reacting the compound with a suitable base or acid. Non-limiting examples of pharmaceutically acceptable salts obtained from a base include alkali metals, alkali earth metals, ammonium, and N+(C1-4 alkyl)4 salts. Non-limiting examples of suitable alkali or alkali earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salt. Furthermore, non-limiting examples of suitable pharmaceutically acceptable salts optionally include nontoxic ammonium, quaternary ammonium, and amine cation formed using a counter ion such as a halide ion, hydroxide ion, carboxylic acid ion, sulfuric acid ion, phosphoric acid ion, nitric acid ion, lower alkyl sulfonic acid ion, and aryl sulfonic acid ion. Non-limiting examples of suitable organic bases that can produce a salt include primary amine, secondary amine, tertiary amine, substituted amine including naturally-occurring substituted amine, cyclic amine, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanol amine, and other basic ion exchange resins. In a specific embodiment, a pharmaceutically acceptable base addition salt can be selected from ammonium, potassium, sodium, calcium, and magnesium salt.

[0586] In an embodiment of the present disclosure, a target subject can be a patient in a state before onset of cancer, after a cancer treatment, an early stage of onset of cancer, or a precancerous condition. Alternatively, a target subject can be a healthy individual. If a healthy individual is a subject, the disclosure is performed as a preventive method.

[0587] Examples of cancer targeted in the present disclosure include, but are not limited to esophageal cancer, gastroesophageal junction cancer, renal cell cancer, lung cancer, digestive organ cancer, leukemia, lymphoma, myeloma, brain cancer, pancreatic cancer, endometrial cancer, prostate cancer, liver cancer, bladder cancer, gastroesophageal adenocarcinoma, chondrosarcoma, colorectal adenocarcinoma, colorectal cancer, breast cancer, renal cell cancer, ovarian cancer, head and neck cancer, melanoma, gastric adenocarcinoma, sarcoma, urogenital cancer, gynecological cancer, adrenocortical cancer, and the like. In a specific embodiment, cancer is lung cancer. In a specific embodiment, cancer is colorectal cancer. In a specific embodiment, cancer is colorectal adenocarcinoma. In a specific embodiment, cancer is melanoma. In a specific embodiment, cancer is breast cancer. In a specific embodiment, cancer is bladder cancer. In a specific embodiment, cancer is renal cell cancer. In a specific embodiment, cancer is pancreatic cancer. In a specific embodiment, cancer is endometrial cancer. In a specific embodiment, cancer can be unresectable. In a specific embodiment, cancer can be progressive. In a specific embodiment, cancer can be refractory. In a specific embodiment, cancer can be recurrent. In a specific embodiment, cancer can be metastatic. In various embodiments of the present disclosure, target cancer can include normal carcinoma, carcinoma with a relatively slow progression (e.g., cancer with low sensitivity to the immune system), oral squamous cell cancer, cervical cancer, MHC class I negative carcinoma on which CD8 positive T cells are generally less effective, immune checkpoint inhibitor resistant cancer, and the like. A cancer patient refers to a patient suffering from a “cancer” described above. In one embodiment, the target disease, disorder, or condition of the present disclosure comprises melanoma.

[0588] Although not wishing to be bound by any theory, a human T cell has a portion that can bind to a factor of an autologous cell in order not to attack a cell other than a T cell of an individual. The representative factor (molecule) of the portion is PD-1. Meanwhile, a general cell has a factor (molecule) such as PD-L1. For example, PD-L1 binds to PD-1, whereby a general cell escapes an attack from a T cell (immune cell). This mechanism is an immune checkpoint system as a system suppressing the activity of a T cell. Since a cancer cell itself is also an autologous cell, a cancer cell escapes an attack from a T cell (immune cell) by the immune checkpoint system.

[0589] Although not wishing to be bound by any theory, an immune checkpoint inhibitor (ICI) is, for example, an agent that inhibits the immune checkpoint system by inhibiting a stimulus to an immune checkpoint molecule (PD-1), which is an immune function inhibitory receptor present in a T cell that is an immune cell. Since an immune checkpoint inhibitor is an agent that inhibits a system suppressing the immunity, administration of the agent activates the immune function of a patient. In addition, inhibition of an immune checkpoint molecule in a cancer cell enables a T cell to attack the cancer cell.

[0590] Although not wishing to be bound by any theory, Extract Z is a suitable therapeutic agent that is used to increase sensitivity to an immune checkpoint inhibitor. Extract Z was developed as a therapeutic drug of tuberculosis, is a colorless and transparent subcutaneous injection solution, and comprises, as primary ingredients, a polysaccharide called Lipoarabinomannan extracted from human Mycobacterium tuberculosis, a nucleic acid, and a lipid.

[0591] Although not wishing to be bound by any theory, administration of Extract Z attains an effect of increasing sensitivity to an immune checkpoint inhibitor (ICI). Apart from administration of these therapeutic agents, radiation irradiation can also be expected to attain an effect of increasing sensitivity to an immune checkpoint inhibitor (ICI).

[0592] In one embodiment, therapy with an immune checkpoint inhibitor (first therapy) and therapy for increasing sensitivity to an immune checkpoint inhibitor (second therapy) as described above are combined to treat cancer of a patient. In other words, the method for cancer treatment of the present embodiment comprises a step of performing therapy with administration of an immune checkpoint inhibitor (first therapy) on a cancer patient and a step of performing therapy for increasing sensitivity to the immune checkpoint inhibitor (second therapy) on the cancer patient.

[0593] In one embodiment, nivolumab, pembrolizumab, or atezolizumab, which is an antibody against PD-1 of a T cell, is used as an immune checkpoint inhibitor in first therapy, thereby inhibiting an immune checkpoint of PD-1 of a T cell and PD-L1 of a cancer cell. Radiation irradiation, administration of Extract Z, or a combination thereof is performed as therapy in second therapy.

[0594] In one embodiment, first therapy and second therapy may be combined in an embodiment wherein these first therapy and second therapy are performed on a patient at the same time for a predetermined period. First therapy and second therapy may also be combined in an embodiment wherein either one of the first therapy and the second therapy is performed on a patient for a predetermined period, and the other therapy is then performed on the patient for a predetermined period. First therapy and second therapy may also be combined in an embodiment wherein either one of the first therapy and the second therapy is performed on a patient for a predetermined period, and both of them are then performed on the patient for a predetermined period.

[0595] First therapy and second therapy may also be combined in an embodiment wherein both of the first therapy and the second therapy are performed on a patient for a predetermined period, and only either one of them is then performed on the patient for a predetermined period. Furthermore, the embodiments as described above may be appropriately combined and periodically repeated.

[0596] In various embodiments described above, the timing at which therapy is changed is identified based on a predetermined change in the test result with respect to a patient (such as a medical image of the patient, a measurement result (measurement value) of a tumor marker of the patient, or a PET test result of the patient) as described below.

[0597] In one embodiment, since first therapy and second therapy are combined, it is possible to eliminate or reduce cancer tissue by the synergistic action of the effect of the first therapy and the effect of the second therapy.

[0598] Although not wishing to be bound by any theory, cancer tissue of a patient can be eliminated or reduced by enabling a T cell to exert its cytotoxic power on a cancer cell by the first therapy (administration of an immune checkpoint inhibitor) while increasing sensitivity to the immune checkpoint inhibitor by the second therapy (radiation irradiation, administration of Extract Z, or combined use of administration of Extract Z+radiation irradiation).

[0599] FIG. 1 is a block diagram showing the configuration of a system that utilizes a program in the present embodiment. FIG. 2 is a flowchart showing the therapy procedure in the present embodiment.

[0600] A system 1 shown in FIG. 1 comprises a control unit 11, a read unit 12, a storage unit 13, a display unit 14, an input unit 15, and a bus 16. The control unit 11 is connected with other hardware units constituting the system 1 via the bus 16.

[0601] The control unit 11 is configured by using a CPU (Central Processing Unit), an MPU (Micro Processing Unit), or the like and executes a program in the present embodiment. The read unit 12 reads out a program 3 in the present embodiment which is recorded in a portable recording medium 2 such as flexible discs from the portable recording medium. The storage unit 13 is composed of an SRAM (Static Random Access Memory), a DRAM (Dynamic Random Access Memory), or the like and stores information necessary during processing performed by the control unit 11 and the program 3 read out by the read unit 12.

[0602] The display unit 14 is composed of a liquid crystal display panel or the like and displays history information of a patient, a result of processing by the control unit 11 (type of treatment performed on a patient, period during which each treatment is performed) or the like. The input unit 15 is composed of a keyboard, a mouse, or the like and receives input information from a user.

[0603] An image obtaining apparatus 4 for obtaining a medical image (such as CT images, X images, or PET images) of a patient is connected to the system 1 so that the medical image of the patient is input to the system 1 (control unit 11) regularly or in response to a request from the system 1 (control unit 11). A tumor marker measurement instrument 5 for measuring a tumor marker (specifically, CEA: Carcinoembryonic Antigen) of a patient is also connected to the system 1 so that a measurement value of a tumor marker (CEA) of the patient is input to the system 1 (control unit 11) regularly or in response to a request from the system 1 (control unit 11).

[0604] Although the program 3 is read out from the portable recording medium 2 by using read unit 12 in the example described above, the program 3 may be stored in the storage unit 13 in advance. Alternatively, the program 3 may be obtained from an external server via a network not illustrated.

[0605] Next, the treatment procedure in the present embodiment is described while referring to the flowchart of FIG. 1.

[0606] The control unit 11 identifies a first period during which either one of therapy with an immune checkpoint inhibitor (first therapy) and therapy for increasing sensitivity to the immune checkpoint inhibitor (second therapy) is performed (step S1). In this step, it is also identified which of radiation irradiation, administration of Extract Z, and combined use of administration of Extract Z+radiation irradiation is performed as the second therapy. In this case, for example, the first period and the type of the second therapy are identified depending on an input from a user received via the input unit 15. The control unit 11 itself may identify the first period and the type of the second therapy based on the medical history, therapy history information in other cases of a patient, or the like. The identified first period and type of the second therapy are displayed on the display unit 14.

[0607] While continuing either one of the therapies, the control unit 11 obtains a medical image of the patient obtained at the image obtaining apparatus 4 (step S2).

[0608] The control unit 11 determines whether cancer tissue has reduced based on the obtained medical image (step S3). When cancer tissue has reduced (step S3: YES), the processing is returned to step S2.

[0609] Meanwhile, when cancer tissue has not reduced (step S3: NO), the control unit 11 obtains a measurement value of a tumor marker in the patient obtained by the tumor marker measurement instrument 5 (step S4).

[0610] The control unit 11 determines whether the obtained measurement value of the tumor marker has increased (step S5). If the measurement value has not increased (step S5: NO), the processing is returned to step S2.

[0611] Meanwhile, when the measurement value of the tumor marker has increased (step S5: YES), the control unit 11 identifies a second period during which the other of the therapy with an immune checkpoint inhibitor (first therapy) or the therapy for increasing sensitivity to the immune checkpoint inhibitor (second therapy) is performed (step S6), and the processing is then terminated. In this case, for example, the second period is identified depending on an input from a user received via the input unit 15. The control unit 11 itself may identify the second period based on the medical history, therapy history information in other cases of the patient, or the like. The identified second period is displayed on the display unit 14.

[0612] Although the period during which the other of the first therapy and the second therapy is performed is identified in step S6 in the embodiment described above, the period during which each of the first therapy and the second therapy is performed may be identified.

[0613] Although the period during which the other (or both) of the therapies is performed is identified when the measurement value of the tumor marker has increased in step S5 in the embodiment described above, the period during which the other (or both) of the therapies is performed may be identified when incorporation of glucose has increased based on the PET test result. It may be determined in step S5 whether the processing proceeds to the processing of step S6 (identifying the period during which the other (or both) of the therapies is performed) depending on both the variation in the measurement value of the tumor marker and the variation in incorporation of glucose based on the PET test result.

[0614] As used herein, “or” is used when “at least one or more” of the listed matters in the sentence can be employed. When explicitly described herein as “within the range of two values”, the range also includes the two values themselves.

[0615] Reference literatures such as scientific literatures, patents, and patent applications cited herein are incorporated herein by reference to the same extent that the entirety of each document is specifically described.

[0616] As described above, the present disclosure has been described while showing preferred embodiments to facilitate understanding. The present disclosure described hereinafter is based on the Examples. The above descriptions and the following Examples are not provided to limit the present disclosure, but for the sole purpose of exemplification. Thus, the scope of the present disclosure is not limited to the embodiments and Examples specifically described herein and is limited only by the scope of claims.

EXAMPLES

[0617] The Examples are described hereinafter. When necessary, animals used in the following Examples were handled in compliance with relevant ethical standards and guidelines, based on the Declaration of Helsinki. While the specific products described in the Examples were used, reagents can be substituted with an equivalent product from another manufacturer (Sigma-Aldrich, Wako Pure Chemical, Nacalai Tesque, R & D Systems, USCN Life Science INC, or the like).

Manufacturing Example: Extract Z

[0618] Extract Z used in this Example was manufactured in the following manner.

[0619] Human Mycobacterium tuberculosis strain Aoyama B which had been lyophilized and stored (−20° C.) was subjected to seed culture at 37±1° C. in a Sauton potato medium.sup.(1). The cultured bacteria were transferred to a production medium.sup.(2) and cultured (primary culture) for 5 to 7 weeks at 37±1° C. The resulting cells were washed with water for injection. To the cells, water for injection was then added in an amount 20-fold of the weight of the wet cells. The mixture was heated at 100° C. for 120 minutes to obtain an extract. The extract was filtered with a 0.45 μm-membrane filter and then concentrated under reduced pressure so that the saccharide content (in terms of D-arabinose by the phenol-sulfuric acid method) would be 4.0 to 6.0 mg/ml to obtain a concentrate. Subsequently, in order to remove proteins, 1 w/v % of sulfosalicylic acid was added to the concentrate. The mixture was left standing for 15 to 20 minutes at 10° C. or lower. Precipitates were then removed by centrifugation (10° C. or lower, 1,150×G, 10 minutes) to recover the supernatant. The protein concentration of the supernatant was 0.30 mg/ml or lower (Lowry method, in terms of tyrosine). The supernatant was further processed to remove sulfosalicylic acid until the concentration was at or below the detection limit (10 ppm or less, method using ferric chloride solution). The resultant solution was concentrated under reduced pressure so that the saccharide content would be 1.8 to 2.2 mg/ml, and the concentrate was combined with sodium chloride (0.9 w/v %) and cold ethanol at the same volume as the concentrate. The mixture was left standing for 40 hours or longer at 10° C. or lower, and then the precipitates (polysaccharide of high molecular weight region) were removed by centrifugation (10° C. or lower, 2,040×G, 10 minutes). Subsequently, the supernatant was combined with four times the amount of cold ethanol, and the mixture was left standing for 40 hours or longer at 10° C. or lower and centrifuged (10° C. or lower, 2,040×G, 10 minutes) to recover precipitates. The precipitates were dissolved in water for injection. After the saccharide content was adjusted to 1.8 to 2.2 mg/ml, the solution was filtered with a 0.45 μm membrane filter and sterilized with high pressure steam (121° C., 20 minutes) to prepare an Extract Z solution.

(1) Sauton-Potato Medium

[0620] Washed potato slices were soaked in a Sauton medium, sterilized for 15 minutes at 115° C., and then used as a Sauton-potato medium.

Sauton Medium

[0621] L-asparagine (monohydrate) 4.0 g

[0622] Citric acid (monohydrate) 2.0 g

[0623] Magnesium sulfate (heptahydrate) 0.5 g

[0624] Potassium monohydrogenphosphate (anhydride) 0.5 g

[0625] Ammonium iron citrate 0.05 g

[0626] Glycerol 60 ml

[0627] The above ingredients were dissolved in water to prepare a 1,000 ml solution. pH was adjusted to 7.0 to 7.3 by using a sodium hydroxide solution.

(2): Production Medium

[0628] L-asparagine (monohydrate) 4.0 g

[0629] Citric acid (monohydrate) 2.0 g

[0630] Magnesium sulfate (heptahydrate) 0.5 g

[0631] Potassium monohydrogenphosphate (anhydride) 0.5 g

[0632] Ammonium iron citrate 0.05 g

[0633] Glycerol 60 ml

[0634] The above ingredients were dissolved in water to prepare a 1,000 ml solution and sterilized with high pressure steam (121° C., 20 minutes). pH was adjusted to 7.0 to 7.3 by using a sodium hydroxide solution.

[0635] The physicochemical properties of the resulting Extract Z solution were as follows.

(1) Appearance:

[0636] Pale yellow clear liquid

(2) pH:

[0637] 4.50 to 5.30

(3) Protein Content:

[0638] 3.5 wt. % (as an amino acid) in a lyophilized product

(4) Nucleic Acid Content:

[0639] 0.1 wt. % in a lyophilized product

(5) Primary Constituent Monosaccharides of Polysaccharide:

[0640] Mannose 43.4 wt. %, arabinose 18.2 wt. %, and glucose 10.4 wt. % (hydrolyzed with 2N trifluoroacetic acid for two hours at 100° C., and then subjected to liquid chromatography using 2-cyanoacetamide fluorescent derivative (S. Honda, et al., Anal. Chem., 52, 1079 (1980)).

[0641] The Extract Z solution prepared by the method described in the Manufacturing Example described above can be appropriately diluted prior to use. In the following Examples, the Extract Z solution was diluted 1 to 50,000-fold and adjusted to a suitable concentration for use.

[0642] The manufacturers and catalogue numbers of the antibodies and staining reagents used in the following Examples are as follows.

CD80 (Miltenyi Biotec, catalogue number 130-102-372)
CD86 (Miltenyi Biotec, catalogue number 130-102-506)
CD11b (Miltenyi Biotec, catalogue number 130-113-811)
CD11c (Miltenyi Biotec, catalogue number 130-122-016)
CD45 (Miltenyi Biotec, catalogue number 130-119-130)
CD4 (Miltenyi Biotec, catalogue number 130-123-899)
CD8 (Life Technologies Corporation, catalogue number 25-0081-82)
TCRβ (BioLegend Incorporated, catalogue number 109220) NK (anti-CD49b antibody; Miltenyi Biotec, catalogue number 130-102-258)
MHC class II (Miltenyi Biotec, catalogue number 130-123-785)
PD-L1 (anti-CD274 antibody; Life Technologies Corporation, catalogue number 12-5982-81)
PI (Propidium Iodide Solution; dead cell marker; Miltenyi Biotec, catalogue number 130-093-233)

[0643] Examples of specific treatment for cancer patients are described below while referring to FIG. 3 (Example 1: example 1 that is immune checkpoint inhibitor+radiation irradiation alone), FIG. 4 (Example 2: example 2 that is immune checkpoint inhibitor+radiation irradiation alone), FIG. 5 (Example 3: example of an immune checkpoint inhibitor after administration of Extract Z), FIG. 6 (Example 4: example that is combined use of immune checkpoint inhibitor+administration of Extract Z+radiation irradiation), and FIGS. 10 and 11 (example that is combined use of immune checkpoint inhibitor+administration of Extract Z+radiation irradiation for a subject who received therapy with a tyrosine kinase inhibitor and therapy with an immune checkpoint inhibitor which were effective, but subsequently experienced progressive disease (PD)).

Example 1

[0644] This Example presents a case of a 69-year-old male diagnosed with lung squamous cell carcinoma without a driver gene mutation, with clinical TNM classification: cTxN3M1b and at stage: IVb.

[0645] From September, 2017 to November, 2018, nivolumab, which is one type of anti-PD-1 antibody that is an immune checkpoint inhibitor, was administered via intravenous drip injection at 240 mg per dose at an interval of 2 weeks. However, swelling in the cervical lymph node to which cancer had been metastasized progressed as shown in the CT images of FIG. 3. In December, 2018, the cervical lymph node was irradiated with a radiation of 30 Gy/10 fr. In addition, administration of nivolumab via intravenous drip injection at 240 mg per dose at an interval of 2 weeks has been continued from December, 2018 to the present (December, 2019). As a result, the swelling in the cervical lymph node was suppressed (April and October, 2019) and partially remitted, wherein the treatment has exerted the effect.

Example 2

[0646] This Example presents a case of a 60-year-old male diagnosed with lung adenocarcinoma without a driver gene mutation, with recurrent TNM classification: rTxN3M1b and at stage: IVb.

[0647] From November, 2017 to February, 2019, nivolumab was administered via intravenous drip injection at 240 mg per dose at an interval of 2 weeks. However, swelling in the cervical lymph node to which cancer had been metastasized progressed as shown in the CT images of FIG. 4. In March, 2019, the cervical lymph node was irradiated with a radiation of 30 Gy/10 fr. In addition, administration of nivolumab via intravenous drip injection at 240 mg per dose at an interval of 2 weeks has been continued from March, 2019 to the present (December, 2019). As a result, the swelling in the cervical lymph node was suppressed (May, 2019) and partially remitted, wherein the treatment has exerted the effect.

Example 3

[0648] This Example presents a case of a 67-year-old female diagnosed with EGFR gene mutation positive (exon 19 deletion) lung adenocarcinoma, with clinical TNM classification: cT1bN3M1b and at stage: IVb.

[0649] From Apr. 1, 2019, administration of Extract Z was started. Since the value of a tumor marker (CEA) increased from 57.0 to 277.0, administration of Extract Z was discontinued on Jun. 3, 2019, and administration of nivolumab via intravenous drip injection at 240 mg per dose at an interval of 2 weeks was started. Thereafter, the value of the tumor marker (CEA) drastically decreased from 277.0 to 63.2.

[0650] As shown in the CT images of FIG. 5, cancer tissue that had existed before the treatment was almost eliminated after the treatment. The synergistic action of administration of Extract Z and administration of nivolumab resulted in an excellent therapeutic effect.

Example 4

[0651] This Example presents a case of a 55-year-old female diagnosed with EGFR gene mutation positive (exon 19 deletion) lung adenocarcinoma, with clinical TNM classification: cT3bN2M1c and at stage: IVb.

[0652] From Jun. 20, 2019, pembrolizumab, which is one type of anti-PD-1 antibody that is an immune checkpoint inhibitor, started to be administered via intravenous drip injection at 200 mg per dose at an interval of 3 weeks. Since the value of a tumor marker (CEA) increased from 25.0 to 98.8, administration of Extract Z was started on Aug. 15, 2019 while maintaining the administration of pembrolizumab. In doing so, radiation irradiation at 30 Gy/10 fr was performed in combination with the administration of Extract Z. Thereafter, the measurement value of the tumor marker (CEA) drastically decreased from 98.8 to 41.5.

[0653] As shown in the X-ray images of FIG. 6, although the cancer tissue region once became larger on August 15 during the treatment (before administration of Extract Z), the region became much smaller after the treatment (administration of Extract Z+radiation irradiation) as compared to the region before the treatment. The synergistic action of administration of pembrolizumab and combined use of administration of Extract Z+radiation irradiation resulted in an excellent therapeutic effect.

Example 5: Application of Immunoadjuvant/Radiation Therapy for Directly Activating a Dendritic Cell

[0654] In this Example, application of an immunoadjuvant/radiation therapy for directly activating a dendritic cell was confirmed.

[0655] (Material and Method)

[0656] Immunoadjuvant=Extract Z

[0657] Radiation Therapy

Example 6: Example of Direct Activation of Dendritic Cells by Administration of an Immunoadjuvant

[0658] This Example shows direct activation of dendritic cells by administration of an immunoadjuvant.

[0659] 1 mg/kg of saline or Extract Z is subcutaneously administered to the right inguinal region of a C3H/HeN mouse once daily (20 mice for each group). 1×10.sup.6 oral squamous cell carcinoma Sq-1979 cells are subcutaneously infused on the ventral side on day 29 after starting the administration of saline or Extract Z.

[0660] On day 36 after starting the administration of saline or Extract Z, tumor and lymph nodes are collected, the collected samples of 5 mice are pooled, and measurement is performed using a flow cytometer.

[0661] (Antibody Used)

[0662] Each antibody against CD80, CD86, CD11b, and CD11c

[0663] (Result)

[0664] An increase in the amount of expression of CD80/86 on the surface of a dendritic cell was observed in the Extract Z administered group. This result suggests that administration of Extract Z activates a dendritic cell.

Example 7: Example of Direct Activation of a Dendritic Cell by Administration of Radiation Therapy

[0665] This Example shows direct activation of a dendritic cell by administration of radiation therapy.

[0666] Cancer cells are subcutaneously administered to mice to create mice with subcutaneous tumor. After subcutaneous transplantation of cancer cells, half the mice were subcutaneously and topically irradiated with a radiation. After the irradiation, subcutaneous tumor and lymph nodes are collected from the mice of the irradiation group and the mice of the non-irradiation group, and the amount of expression of CD80/86 on the surface of a dendritic cell is measured with a flow cytometer.

[0667] On day 36 after starting the irradiation (at the same timing for the non-irradiation group), tumor and lymph nodes are collected, the collected samples of 5 mice are pooled, and measurement is performed using a flow cytometer.

[0668] (Antibody Used)

[0669] Each antibody against CD80, CD86, CD11b, and CD11c

Example 8: Combined Use of a Plurality of Techniques of Directly Activating a Dendritic Cell

[0670] This Example shows combined use of a plurality of techniques of directly activating a dendritic cell.

[0671] Saline or Extract Z is administered to a group of patients with cervical cancer at a stage for which chemical radiation therapy is suitable. Administration of a platinum formulation (cisplatin or carboplatin) and radical radiation irradiation are performed at the same time as or at a different time from saline or Extract Z. Subsequently, immune parameters (CD11c, CD14, CD16, CD19, CD24, CD27, CD38, CD80, CD86, CD123, CD138, CCRS, CCR7, CXCR3, HLA-DR, CD3, CD4, CD8, CD45RA, CD56, CD69, CD159a, CTLA-4, NKp46, PD-1, IFNγ, TNFα, perforin, granzyme B, IL4, FoxP3, IL17A, Ki67, CXCL9, CXCL10, IL10, IL12p70), PFS (Progression-Free Survival), OS (Overall Survival), and ORR (Objective Response Rate) in the group of patients are measured.

Example 9: Combined Use of an Immune Checkpoint Inhibitor and an Immunoadjuvant

[0672] This Example shows combined use of an immune checkpoint inhibitor and an immunoadjuvant.

[0673] Saline or Extract Z is administered to a group of patients who have solid tumor, have no existing therapeutic choice, and are Tumor Mutational Burden.sup.high and CD8.sup.+ T Cell

[0674] An immune checkpoint inhibitor (e.g., pembrolizumab) is administered at the same time as or at a different time from saline or Extract Z. Subsequently, PFS (Progression-Free Survival), OS (Overall Survival), and ORR (Objective Response Rate) in the group of patients are measured.

Example 10: Another Example of Combined Use of an Immune Checkpoint Inhibitor and an Immunoadjuvant

[0675] This Example shows one example of combined use of an immune checkpoint inhibitor and an immunoadjuvant.

[0676] Extract Z is administered to a case with non-small cell lung cancer for which therapy with an immune checkpoint inhibitor alone was ineffective. Administration of an immune checkpoint inhibitor and palliative radiation therapy are performed at the same time as or at a different time from Extract Z. Subsequently, PFS (Progression-Free Survival), OS (Overall Survival), and ORR (Objective Response Rate) in the group of patients are measured.

Example 11: Combined Use of an Immune Checkpoint Inhibitor, an Immunoadjuvant, and Radiation Therapy

[0677] This Example shows an example of combined use of an immune checkpoint inhibitor, an immunoadjuvant, and radiation therapy.

[0678] Saline or Extract Z is administered to a group of patients who have non-small cell lung cancer, have no existing therapeutic choice, and EGFR mutation positive and EGFR-TKI intolerant/refractory. Administration of an immune checkpoint inhibitor and palliative radiation therapy are performed at the same time as or at a different time from saline or Extract Z. Subsequently, PFS (Progression-Free Survival), OS (Overall Survival), and ORR (Objective Response Rate) in the group of patients are measured.

Example 12: Combined Use of an Immune Checkpoint Inhibitor, an Immunoadjuvant, Radiation Therapy, and a COX-2 Inhibitor

[0679] Extract Z is administered to a case with non-small cell lung cancer for which therapy with an immune checkpoint inhibitor alone was ineffective. Administration of an immune checkpoint inhibitor, palliative radiation therapy, and administration of a regulatory T cell inhibitor are performed at the same time as or at a different time from Extract Z. Subsequently, PFS (Progression-Free Survival), OS (Overall Survival), and ORR (Objective Response Rate) in the group of patients are measured.

Example 13: Example of Enhancement of CXCL10 Production by an Immunoadjuvant

[0680] This Example demonstrates enhancement of CXCL10 production by the agent of the present disclosure.

[0681] Bone marrow-derived cells were collected from a C3H/HeJ mouse (male), and the collected bone marrow-derived cells were cultured at 4×10.sup.6/plate for 6 days (in the presence of 20 ng/mL GM-CSF and 20 ng/mL IL-4). The bone marrow-derived cells after culture were collected, seeded at 2×10.sup.5/well, and stimulated with 0.4 μg/mL, 0.8 μg/mL, 1.6 μg/mL, and 3.2 μg/mL of Extract Z. The culture supernatant was collected after 6 hours from the stimulation, and the concentration of CXCL10 was measured by ELISA method.

[0682] FIG. 7 shows the result. An increase in the concentration of CXCL10 in the culture supernatant was observed as the concentration of Extract Z used for stimulation was increased. Thus, it was found that Extract Z has an action of enhancing CXCL10 production.

Example 14: Example of Treatment of Cancer or Tumor that is CXCL10 Positive by an Immunoadjuvant

[0683] (CXCL10 Positive)

[0684] An immunoadjuvant is administered to a model subcutaneously transplanted with cancer cells to confirm the antitumor effect or life-prolonging effect. Tumor is collected from a subcutaneously transplanted model in which the antitumor effect or life-prolonging effect was observed, and the amount of expression of the CXCL10 gene in the tumor is studied to confirm that the amount of expression is high.

[0685] (CXCR3 Positive)

[0686] An immunoadjuvant is administered to a model subcutaneously transplanted with cancer cells to confirm the antitumor effect or life-prolonging effect. Tumor is collected from a subcutaneously transplanted model in which the antitumor effect or life-prolonging effect was observed, and the CXCR3 positive cells in the tumor are studied by a flow cytometer or immunostaining to confirm that the positive cell ratio is high.

Example 15: Example of the Effect of Promoting Infiltration of CD8.SUP.+ T Cells into Tumor

[0687] This Example shows the effect of the agent of the present disclosure to promote infiltration of CD8.sup.+ T cells into tumor.

[0688] 1 mg/kg of saline or Extract Z was subcutaneously administered to the right inguinal region of a C.sub.3H/HeN mouse once daily (40 mice for each group). 1×10.sup.6 oral squamous cell carcinoma Sq-1979 cells were subcutaneously infused on the ventral side on day 29 after starting the administration of saline or Extract Z. Tumor was collected after 7 days from introduction of Sq-1979 cells, the samples of 5 mice were pooled, and measurement was performed using a flow cytometer.

[0689] (Antibody and Reagent Used)

[0690] Antibody against CD45

[0691] Antibody against CD8

[0692] Antibody against TCRβ chain

[0693] PI (dead cell marker)

[0694] (Result)

[0695] FIG. 8 shows the result. Administration of Extract Z resulted in an increase in CD8.sup.+ T cells in tumor. Meanwhile, since the action of Extract Z is non-specific immunostimulatory, CD8.sup.+ T cells infiltrating into tumor include both cells having the antitumor effect and cells that express CTLA-4 and inhibit the antitumor effect.

Example 16: Example of Treatment or Prevention of Tumor Mutational Burden.SUP.high .and CD8.SUP.+ T Cell^{low .Inoperable or Metastatic Solid Cancer for which there is No Other Therapeutic Choice, Wherein the Treatment or Prevention Comprises an Immunoadjuvant}

[0696] This Example shows demonstration of treatment or prevention of Tumor Mutational Burden.sup.high and CD8.sup.+ T cell.sup.low inoperable or metastatic solid cancer for which there is no other therapeutic choice, wherein the treatment or prevention comprises an immunoadjuvant.

[0697] Saline or Extract Z is administered to a group of patients who have solid cancer, have no existing therapeutic choice, and are Tumor Mutational Burden.sup.high and CD8.sup.+ T Cell invasion.sup.low. An immune checkpoint inhibitor (e.g., pembrolizumab) is administered at the same time as or at a different time from saline or Extract Z. Subsequently, PFS (Progression-Free Survival), OS (Overall Survival), and ORR (Objective Response Rate) in the group of patients are measured.

Example 17: Regimen Treatment Example

[0698] This Example shows an example of a combined regimen of an immune checkpoint inhibitor, radiation therapy, and Extract Z.

[0699] (1) Immune checkpoint inhibitor: any of the following agents is used alone.

[0700] (a) Intravenous drip injection of 240 mg of nivolumab for every 2 weeks in principle

[0701] (b) Intravenous drip injection of pembrolizumab at 200 mg per dose for 30 minutes at an interval of 3 weeks, or intravenous drip injection of pembrolizumab at 400 mg per dose for 30 minutes at an interval of 6 weeks in principle

[0702] (c) Intravenous drip injection of atezolizumab at 1200 mg per dose for 30 minutes at an interval of 3 weeks in principle

[0703] (2) Radiation Therapy

[0704] Palliative irradiation other than brain metastasis, wherein irradiation is performed at 2 Gy×20 to 25 fractions or a dose equivalent dose: e.g., 3 Gy×10 fractions. The irradiation method of palliative irradiation depends on the judgement of a radiation oncologist of each facility.

[0705] (3) Extract Z

[0706] (a) Aspirating a liquid medicine into a 1 mL syringe and subcutaneously injecting a portion thereof (preferably about 0.05 mL) to the upper arm.

[0707] (b) Twice a week

[0708] (c) From the start of radiation therapy to the end of the radiation therapy (8 weeks at the maximum)

[0709] (4) Repeating Radiation Therapy and Extract Z

[0710] After 9 months from the start of the therapy, radiation therapy and Extract Z may be repeated in another site.

Example 18: Example Showing the Ratio of Tumor Infiltrating Cells as a Result of Administration of Extract Z

[0711] This Example shows analysis of the ratio of tumor infiltrating cells as a result of administration of the agent of the present disclosure.

[0712] 1 mg/kg of saline or Extract Z was repeatedly and subcutaneously administered to the right inguinal region of a C3H/HeN mouse once daily for 35 days (20 mice for each group). For Table 1, 1×10.sup.6 oral squamous cell carcinoma Sq-1979 cells were subcutaneously infused on the ventral side on day 29 after starting the administration of saline or Extract Z, and 200 μg/body of anti-PD-1 antibody was intraperitoneally infused on day 28, day 31, and day 34 after starting the administration of saline or Extract Z. For Table 2, 1×10.sup.6 oral squamous cell carcinoma Sq-1979 cells were subcutaneously infused on the ventral side on day 29 after starting the administration of saline or Extract Z, and 200 μg/body of anti-PD-1 antibody or Control antibody was intraperitoneally infused on day 28, day 31, and day 34 after starting the administration of saline or Extract Z.

[0713] On day 36 after starting the administration of saline or Extract Z, tumor and lymph nodes were collected, the collected samples of 5 mice were pooled, and measurement was performed using a flow cytometer.

[0714] (Antibody and Reagent Used)

[0715] Antibodies against CD45, CD4, CD8, and TCR (T cell receptor)

[0716] Antibodies against NK, MHC class II, and PD-L1

[0717] PI (dead cell marker)

[0718] (Result)

[0719] Table 1 and Table 2 show the results. While the percentage of CD8.sup.+ T cells in tumor infiltrating cells is 4.34% in the saline administered group, the percentage of CD8.sup.+ T cells is 10.43% in the Extract Z administered group (Table 1). In addition, while the group administered with an anti-PD-1 antibody alone shows 3.91%, the group administered with both Extract Z and an anti-PD-1 antibody shows 11.58% (Table 2). These results demonstrate that administration of Extract Z induces infiltration of CD8.sup.+ T cells into tumor.

TABLE-US-00001 TABLE 1 Mean (cell ratio [%]) SE CD8+T Saline 4.34 0.07 {close oversize bracket} ** Extract Z 10.43 0.50 CD4+T Saline 20.54 0.58 Extract Z 22.46 1.05 MHC class II Saline 6.71 0.53 Extract Z 5.64 0.68 NK Saline 39.79 1.35 Extract Z 38.07 0.50 PD-L1+/CD45+ Saline 68.62 1.36 Extract Z 65.04 1.71 PD-L1+/CD8+T Saline 96.79 0.18 Extract Z 98.35 0.11 n = 4, considering a pool of 5 mice as n = 1 ** p < 0.01 (Aspin-Welch t-test)

TABLE-US-00002 TABLE 2 Cell ratio (%) Cell type Sample Mean SE CD8+T/CD45+ Saline + anti-PD-1 3.91 0.31 {close oversize bracket} *** Extract Z + anti-PD-1 11.58 0.42 CD4+T/CD45+ Saline + anti-PD-1 29.93 2.98 Extract Z + anti-PD-1 26.58 2.72 MHC class II+/ Saline + anti-PD-1 4.81 1.08 CD45+ Extract Z + anti-PD-1 4.36 0.19 NK/CD45+ Saline + anti-PD-1 27.02 1.21 Extract Z + anti-PD-1 30.88 1.87 PD-L1+/CD45+ Saline + anti-PD-1 67.88 2.19 Extract Z + anti-PD-1 66.42 3.52 PD-L1+/CD8+T Saline + anti-PD-1 99.02 0.28 Extract Z + anti-PD-1 99.30 0.15 N = 4, considering a pool of 5 mice as N = 1 *** p < 0.001 (student's t-test)

Example 19: Example Showing the Ratio of Antigen-Presenting Cell Markers in a Lymph Node as a Result of Administration of Extract Z

[0720] This Example shows analysis of the ratio of antigen-presenting cell markers in a lymph node as a result of administration of the agent of the present disclosure.

[0721] 1 mg/kg of saline or Extract Z was repeatedly and subcutaneously administered to the right inguinal region of a C3H/HeN mouse once daily for 35 days (20 mice for each group). For Table 3, 1×10.sup.6 oral squamous cell carcinoma Sq-1979 cells were subcutaneously infused on the ventral side on day 29 after starting the administration of saline or Extract Z, and 200 μg/body of anti-PD-1 antibody was intraperitoneally infused on day 28, day 31, and day 34 after starting the administration of saline or Extract Z. For Table 4, 1×10.sup.6 oral squamous cell carcinoma Sq-1979 cells were subcutaneously infused on the ventral side on day 29 after starting the administration of saline or Extract Z, and 200 μg/body of anti-PD-1 antibody or Control antibody was intraperitoneally infused on day 28, day 31, and day 34 after starting the administration of saline or Extract Z.

[0722] On day 36 after starting the administration of saline or Extract Z, tumor and lymph nodes were collected, the collected samples of 5 mice were pooled, and measurement was performed using a flow cytometer.

[0723] (Antibody and Reagent Used)

[0724] Antibodies against CD45, CD80, CD86, CD11b, CD11c, MHC class II, and PD-L1

[0725] PI (dead cell marker)

[0726] (Result)

[0727] Tables 3 and 4 show the results. While the percentage of CD80.sup.+/MHC class II cells in antigen-presenting cell markers in lymph nodes is 6.09% in the saline administered group, the percentage of CD80.sup.+/MHC class II cells is 8.38% in the Extract Z administered group. In addition, while the group administered with an anti-PD-1 antibody alone shows 6.08%, the group administered with both Extract Z and an anti-PD-1 antibody is 6.89% (Table 4). These results demonstrate that administration of Extract Z enhances CD80% expression.

TABLE-US-00003 TABLE 3 Mean (cell ratio [%]) SE CD80+/MHC class II Saline 6.09 0.12 {close oversize bracket} * Extract Z 8.38 0.60 CD86+/MHC class II Saline 28.44 1.01 Extract Z 26.37 1.11 CD11b+CD11c−/ Saline 0.85 0.03 MHC class II Extract Z 1.12 0.03 CD11b+CD11c+/ Saline 1.28 0.06 MHC class II Extract Z 1.51 0.08 CD11b−CD11c+/ Saline 13.03 0.44 MHC class II Extract Z 9.22 0.53 n = 4, considering a pool of 5 mice as n = 1 * p < 0.05 (Aspin-Welch t-test)

TABLE-US-00004 TABLE 4 Cell ratio (%) Cell type Sample Mean SE CD80+/MHC class II Saline + anti-PD-1 6.08 0.27 Extract Z + anti-PD-1 6.89 0.22 CD86+/MHC class II Saline + anti-PD-1 25.90 0.82 Extract Z + anti-PD-1 24.65 1.78 CD11b+CD11c−/ Saline + anti-PD-1 1.18 0.11 MHC class II Extract Z + anti-PD-1 1.41 0.06 CD11b+CD11c+/ Saline + anti-PD-1 0.84 0.06 MHC class II Extract Z + anti-PD-1 0.86 0.04 CD11b−CD11c+/ Saline + anti-PD-1 10.51 0.50 MHC class II Extract Z + anti-PD-1 9.76 0.61 N = 4, considering a pool of 5 mice as N = 1

Example 20: Example Showing PD-L1 Expression in Cultured Sq-1979 Cells

[0728] This Example shows analysis of PD-L1 expression in cultured Sq-1979 cells by the agent of the present disclosure.

[0729] Cultured oral squamous cell carcinoma Sq-1979 cells were collected and stained with an anti-PD-L1 antibody and a Control antibody, and measurement was performed using a flow cytometer.

[0730] (Result)

[0731] FIG. 9 shows the result. When stained with an anti-PD-L1 antibody, 88.78% cells were positive, whereas when stained with a Control antibody, 1.57% cells were positive. Thus, it was demonstrated that Sq-1979 cells are PD-L1 positive cells.

Example 21: The Effect of Combined Use of Extract Z and an Immune Checkpoint Inhibitor

[0732] In this Example, Extract Z and an anti-CTLA-4 antibody are administered to a mouse tumor model to confirm that the antitumor effect or life-prolonging effect synergistically increases.

[0733] A splenocyte or lymph node is collected from a mouse, and an immune cell is isolated and Extract Z and an anti-CTLA-4 antibody are added thereto. Extract Z exhibits a IFN-γ production action (T cell activation action) by being added in vitro. Thus, it is confirmed whether the IFN-γ production synergistically increases by combined use with a CTLA-4 antibody.

Example 22: Example Showing that a CD8.SUP.+ T Cell Infiltrating into Tumor Expresses CTLA-4.SUP.+ by Administration of Extract Z

[0734] This Example confirms an increase in CTLA-4 expression in a tumor infiltrating CD8.sup.+ T cell which is found as a result of administration of Extract Z.

[0735] It is confirmed that administration of Extract Z not only promotes infiltration of overall CD8.sup.+ T cells but also promotes infiltration of CTLA-4.sup.+CD8.sup.+ T cells that inhibits antitumor effect.

Example 23: Example of Treatment with Extract Z, an Immune Checkpoint Inhibitor, and Radiation Irradiation

[0736] In this Example, the therapeutic effect of combined use of Extract Z, an immune checkpoint inhibitor, and radiation irradiation was confirmed.

[0737] This Example presents a case of a 57-year-old male diagnosed with EGFR gene mutation positive (exon 19 deletion) lung adenocarcinoma, with clinical TNM classification: cT2aN2M1c and at stage: IVb.

[0738] After administration of afatinib was started from October, 2015, recurrence having a T790M mutation was observed in 2016. Thus, administration of osimertinib was started from December, 2016. Since progression was observed in June, 2017, chemotherapy with cisplatin+pemetrexed was performed, followed by performing treatment with erlotinib+bevacizumab. However, progression was observed again. Therefore, administration of nivolumab via intravenous drip injection at 240 mg per dose at an interval of 2 weeks was started from September, 2017. As a result, primary foci were almost eliminated and metastasis to the lymph nodes was eliminated, but progression of primary tumor, multiple metastases in the lung, and metastasis to the right adrenal gland were observed on Nov. 6, 2020.

[0739] From Nov. 17, 2020 to Nov. 28, 2020, radiation irradiation at 30 Gy/10 fr and four times of administration of Extract Z were performed on the right adrenal gland metastatic site while continuing the administration of nivolumab.

[0740] As a result, the measurement value of a tumor marker (CEA) decreased from 35.3 to 22.7. Furthermore, it was found that the primary foci slightly reduced and some of the metastatic foci in the lung obviously reduced after the treatment (administration of Extract Z+radiation irradiation) as shown in the CT images of FIG. 10.

[0741] It is considered that the synergistic effect of administration of nivolumab and combined use of administration of Extract Z+radiation irradiation resulted in an abscopal effect. It was also demonstrated that an excellent therapeutic effect is attained in a later therapy-line patient, in which progression is observed although all standard therapies were performed.

[0742] (Note)

[0743] As described above, the present disclosure is exemplified by the use of its preferred embodiments. However, it is understood that the scope of the present disclosure should be interpreted solely based on the Claims. It is also understood that any patent, any patent application, and any references cited herein should be incorporated herein by reference in the same manner as the contents are specifically described herein. The present application claims priority to Japanese Patent Application No. 2019-238657 filed on Dec. 27, 2019 with the Japan Patent Office, and Japanese Patent Application No. 2020-171493 filed on Oct. 9, 2020 with the Japan Patent Office. It is understood that the entire content thereof is incorporated herein by reference.

INDUSTRIAL APPLICABILITY

[0744] The present disclosure provides a method of preventing and treating a disease such as cancer based on a conventionally unavailable mechanism.