Methods and systems for improving anti-cancer treatment

Abstract

This disclosure provides improved methods, compositions, agents, therapeutic regimens and systems for cancer treatment, including improving outcomes of chemotherapy and / or immunotherapy, and / or inducing autophagy in a subject through inhibition of extracellular diazepam-binding inhibitors (DBIs) in various ways. The composition comprises (a) an agent against extracellular human diazepam-binding inhibitors (DBIs) in an amount sufficient to inhibit extracellular human DBIs (an anti-DBI agent), and (b) one or more anticancer agents.

Description

[Technical Field]

[0001] Cross-references to related applications This application claims priority under European Patent Application No. 23168612.2, filed on 18 April 2023, the disclosure thereof being incorporated herein by reference in its entirety.

[0002] Technical field This disclosure relates to methods, compositions, agents, treatment regimens and systems for improving anticancer therapies such as chemotherapy and / or immunotherapy by inhibiting diazepam binding inhibitors (DBIs), such as extracellular DBIs. [Background technology]

[0003] background Diazepam-binding inhibitors (DBIs), also known as acyl-coenzyme A binding proteins (ACBPs), are proteins that are ubiquitously expressed in all tissues of the human body and can be released into the circulation. Plasma DBI concentrations increase with age and are correlated with body mass index, cardiovascular metabolic risk factors (e.g., high fasting blood glucose, systolic blood pressure, total free cholesterol, triglycerides, and subnormal high-density lipoprotein levels), signs of liver damage (e.g., high circulating transaminase concentrations), impaired renal function (e.g., high creatinine levels or decreased clearance), signs of systemic inflammation (e.g., high plasma concentrations of interleukin-1β or tumor necrosis factor), and impaired control of viral infections (e.g., HIV-1 and SARS-CoV-2).

[0004] Neutralization (i.e., inhibition) of extracellular DBI by monoclonal antibodies (mAbs) protects various organs (heart, liver, and lungs, etc.) from damage caused by various invasive factors, including toxic diets (e.g., high-fat diets, methionine / choline-deficient diets, etc.), drugs (e.g., anthracyclines, bleomycin, paracetamol, etc.), toxins (e.g., carbon tetrachloride, concanavalin A, etc.), and ischemia or other types of physical injury (e.g., bile duct ligation in the case of the liver). These organ-protective effects include reduced cell loss or cytoprotection, reduced inflammation, and inhibition of fibrosis. Mechanistically, they are related to the induction of autophagy. In fact, inhibition of autophagy by injection of high-dose 3-hydroxychloroquine or knockout of autophagy-related genes (e.g., Atg4b at systemic levels or Atg7 in specific cell types) eliminates the organ-protective effects of DBI neutralization.

[0005] Pharmacological treatments for cancer include chemotherapy, so-called targeted therapy, and immunotherapy. The long-term success of all drug-based anticancer therapies depends on their ability to induce an antitumor immune response. Therefore, chemotherapy is particularly efficient when it induces immunogenic cell death (ICD) of malignant cells. For example, mitoxantrone and oxaliplatin exemplify such ICD-inducing chemotherapy that elicits a potent anticancer immune response.

[0006] Most immunotherapies are designed to modulate, enhance, or induce the cancer immune dialogue by, for example, blocking inhibitory interactions between PD-1 and PD-L1, which constitute immune checkpoints. Therefore, immune checkpoint inhibitors (ICIs) are often drugs such as antibodies, antibody fragments, or small molecules that target PD-1 and PD-L1, for example.

[0007] Chemotherapy and immunotherapy are often combined as chemoimmunotherapy. In cancer patients, immunotherapies, including those targeting immune checkpoints such as PD-1 and PD-L1, frequently cause undesirable inflammatory and autoimmune side effects that need to be addressed with glucocorticoids (also known as corticosteroids or corticosteroid therapy). However, glucocorticoids are immunosuppressive, and their administration to cancer patients treated with immunotherapy (e.g., ICI treatment) is associated with poor outcomes because it also inhibits the antitumor effect of the ICI treatment. Furthermore, psychological stress or treatment-induced stress, as well as disease-related discomfort and pain, can lead to increased production of endogenous glucocorticoids through activation of the hypothalamic-pituitary-adrenal axis, thereby causing immunosuppression and impairing the effectiveness of anticancer treatment.

[0008] Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide and is associated with a range of established risk factors, including metabolic dysregulation (e.g., obesity and diabetes, which contribute to the development of metabolic dysfunction associated with steatohepatitis (MASH), as well as progression to advanced fibrosis and cirrhosis), repeated exposure to hepatotoxins (e.g., alcohol and aflatoxins), and infection with liver viruses (e.g., hepatitis B and C viruses). Despite an increasingly detailed understanding of the molecular and cellular pathways leading to HCC development, the options for successful treatment remain relatively limited. Curative attempts (e.g., transplantation, resection, and thermal ablation) are typically limited to early-stage HCC patients (BCLC-0 and BCLC-A), while treatments such as transarterial chemoembolization, transarterial radioembolization, and systemic interventions (tyrosine kinase inhibitors and immunotherapy) are usually reserved for intermediate (BCLC-B) and advanced (BCLC-C) stage patients.

[0009] Therefore, there remains a need to provide novel and improved anti-cancer chemotherapy, immunotherapy and / or chemoimmunotherapy, as well as methods for treating HCC, that overcome at least some of the above problems, at least to some extent. [Overview of the Initiative] [Means for solving the problem]

[0010] concise summary The applicants have surprisingly found that administering drugs that stimulate autophagy by inhibiting extracellular human diazepam binding inhibitors (DBIs) (anti-DBI agents) can improve the therapeutic effects of immunotherapy, chemotherapy, and chemoimmunotherapy.

[0011] The applicants have also surprisingly found that neutralization or inhibition of extracellular diazepam binding inhibitor (DBI), also known as acyl coenzyme A binding protein (ACBP), improves outcomes of immunotherapy, chemotherapy, and chemoimmunotherapy in various ways in a variety of tumor models, resulting in improved reduction of tumor growth, higher cure rates, and / or extended animal survival.

[0012] Neutralization / inhibition of extracellular DBI can be achieved in several ways, for example, by vaccination that induces neutralizing autoantibodies, or by administration of drugs such as monoclonal antibodies (mAbs) that bind to or modulate the activity of extracellular DBI.

[0013] Even more surprisingly, the applicants found that extracellular DBI inhibition enhances cancer immune surveillance. This was demonstrated by changes in the composition of tumor immune infiltrates due to an increased ratio of cytotoxic T cells to regulatory T cells, and by effects on T cell activation and exhaustion markers.

[0014] Even more surprisingly, the applicants found that extracellular DBI inhibition reverses the immunosuppressive effects of corticosteroid therapy, in that if the treatment further includes inhibition of extracellular DBI, a complete or partial recovery of any therapeutic efficacy of immunotherapy, chemotherapy, or chemoimmunotherapy is observed, to the extent that such efficacy was potentially reduced or eliminated as an outcome of co-administration of corticosteroid therapy. The applicants' data show that the inhibition of immune checkpoint inhibitor (ICI) treatment by corticosteroids is overcome in the presence of anti-DBI agents, i.e., anti-DBI agents protect ICI-activated T cells from inhibition. Mouse experiments showed that injection of corticosteroids increased levels of extracellular DBI. While not wishing to be bound by theory, the applicants believe that extracellular DBI has a direct or indirect inhibitory effect on T cells.

[0015] Mammalian subjects, such as mice treated with corticosteroids and anti-DBI agents, showed a better vaccine response against cancer cells killed by chemotherapy, demonstrating an enhanced ability of dead-cell vaccines to control tumor progression.

[0016] Disclosed herein are methods for improving the therapeutic effect of immunotherapy in a subject, comprising administering to the subject: (a) an agent that stimulates autophagy by inhibiting extracellular human diazepam binding inhibitor (DBI) (an anti-DBI agent), and (b) one or more immunotherapy agents, wherein the administration of the anti-DBI agent and one or more immunotherapy agents is sufficient to improve the therapeutic effect of immunotherapy compared to an equivalent method without the administration of the anti-DBI agent. Agents, compositions, and combinations for use in such a method are also disclosed. Accordingly, according to one aspect of the present invention, an agent that stimulates autophagy by inhibiting extracellular human diazepam binding inhibitor (DBI), or a composition containing such an agent, is provided for use in such a method. According to a further aspect of the present invention, one or more immunotherapy agents (or compositions or compositions containing such agents) are provided for use in such a method. According to yet another aspect of the present invention, a combination of an agent that stimulates autophagy by inhibiting extracellular human diazepam binding inhibitor (DBI) and one or more immunotherapy agents is provided for use in such a method.

[0017] Disclosed herein are methods for treating cancer in subjects requiring such treatment, comprising administering to a subject (a) an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) in an amount sufficient to inhibit the extracellular human DBI, and (b) one or more anticancer agents, wherein the administration of the anti-DBI agent and one or more anticancer agents is sufficient to treat the cancer in the subject and result in an enhanced treatment of the cancer compared to an equivalent method without the administration of the anti-DBI agent. Agents, compositions and combinations for use in such methods are also disclosed. Accordingly, according to one aspect of the present invention, an agent for extracellular human diazepam-binding inhibitor (DBI), or a composition comprising such an agent, is provided for use in such methods. According to a further aspect of the present invention, one or more immunotherapeutic agents (or compositions comprising such agents or compositions) are provided for use in such methods. According to yet further aspects of the present invention, a combination of an agent for extracellular human diazepam-binding inhibitor (DBI) and one or more immunotherapeutic agents is provided for use in such methods.

[0018] In some embodiments, the methods disclosed herein further include administering one or more corticosteroid therapeutic agents. In some embodiments, the methods disclosed herein reduce the immunosuppressive effect of one or more corticosteroid therapeutic agents in subjects undergoing immunotherapy or cancer treatment compared to an equivalent method without the administration of an anti-DBI agent.

[0019] Disclosed herein are methods for enhancing immune surveillance in a subject with cancer to enhance the therapeutic effect of anticancer therapy, the methods comprising administering to the subject (a) an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) in an amount sufficient to inhibit the extracellular human DBI, and (b) one or more anticancer agents, wherein the administration of the anti-DBI agent and one or more anticancer agents is sufficient to induce one or more immune surveillance biomarkers resulting in enhanced therapeutic effect of anticancer therapy compared to an equivalent method without administration of the anti-DBI agent. Agents, compositions and combinations for use in such a method are also disclosed. Accordingly, according to one aspect of the present invention, an agent for extracellular human diazepam-binding inhibitor (DBI), or a composition comprising such an agent, is provided for use in such a method. According to a further aspect of the present invention, one or more immunotherapeutic agents (or compositions comprising such agents or compositions) for use in such a method are provided. A further aspect of the present invention provides a combination of a drug for use in such a manner with one or more immunotherapeutic agents for an extracellular human diazepam binding inhibitor (DBI).

[0020] Disclosed herein are methods for reducing the immunosuppressive effect of corticosteroid therapy in a subject receiving immunotherapy, the methods comprising administering to the subject (a) an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) in an amount sufficient to inhibit the extracellular human DBI, and (b) one or more corticosteroid therapeutic agents, wherein the administration of the anti-DBI agent and one or more corticosteroid therapeutic agents is sufficient to reduce the immunosuppressive effect of corticosteroid therapy in a subject receiving immunotherapy compared to an equivalent method without the administration of the anti-DBI agent. In some embodiments, the methods disclosed herein further include the administration of one or more anticancer agents. Agents, compositions and combinations for use in such methods are also disclosed. Accordingly, according to one aspect of the present invention, an agent for extracellular human diazepam-binding inhibitor (DBI), or a composition comprising such an agent, is provided for use in such a method. According to a further aspect of the present invention, one or more corticosteroid therapeutic agents (or compositions comprising such agents or compositions) for use in such a method are provided. A further aspect of the present invention provides a combination of an agent for extracellular human diazepam binding inhibitors (DBIs) and one or more corticosteroid therapeutic agents for use in such a manner.

[0021] In some embodiments of the methods disclosed herein, the anti-DBI agent is an antibody or an aptamer.

[0022] In some embodiments of the methods disclosed herein, one or more anticancer agents include chemotherapeutic agents, immunotherapeutic agents, or both. In some embodiments, the chemotherapeutic agent includes immunogenic cell death (ICD) inducing activity. In some embodiments, the chemotherapeutic agent is alkylating agent, alkyl sulfonate, aziridine, ethyleneimine, methylmelamine, altoretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, trimethylolomelamine, acetogenin, camptothecin, bryostatin, calistatin, CC-1065, cryptophycin, dorastatin, duocalmycin, elcerovine, pancratistatin, sarcodictiin, spongstatin, nitrogen mustard, nitrosourea, antibiotic, dynemycin, bisphosphonate, esperamycin, neocarcinostatin chromophore, related chromoprotein enediyne antibiotic chromophore, acrasinomycin, actinomycin, ausramycin, azaserin, bleomycin, kakutinomycin, kara Bicin, Caminocycline, Cartinophylline, Chromomycin, Dactinomycin, Daunorubicin, Detrubicin, 6-Diazo-5-Oxo-L-Norleucine, Doxorubicin, Epirubicin, Esolubicin, Idarubicin, Marcelomycin, Mitomycin, Antimetabolites, Folic Acid Analogues, Purine Analogues, Pyrimidine Analogues, Androgens, Antiadrenal Drugs, Folic Acid Supplements, Aceglaton, Aldophosphamide Glycoside, Aminolevulinic Acid, Enyluracil, Amsacrin, Bestlovesil, Bisanthren, Edatrexate, Defofamine, Demecolsin, Diadiquan, Elformitin, Elptinium acetate, Epotilon, Etoglucid, Gallium nitrate, Hydroxyurea, Lentinan, Lonidamine (lonidainine), Mytansinoid, Mitoguazone, Mitoxantrone, Mopidamol, Nitraeline, Pentostatin, Fenamet, Pirarubicin, Rosoxantrone, Podophyllic acid, 2-Ethylhydrazide, Procarbazine,PSK polysaccharide complex, razoxane, rhizoxin, schizofran, spirogermanium, tenuazonic acid, triadiquan, 2,2',2"-trichlorotriethylamine, trichothecene, urethane, vindesine, dacarbazine, mannomustine, mitobronitol, mitractol, pipobronate, gasitosine, arabinoside, cyclophosphamide, thiotepa, toxoid, chlorambucil, gemcitabine, 6-thioguanine, mercaptopurine, methotrexate, platinum coordination complex, vinblastine, platinum, etoposide (VP-16), ifosfamide, mitoxantrone, vincristine, vinorelbine, novantrone, teniposide, edatrexate, daunomycin, aminopterin, xeloda, ibandronate, irinotecan, topoisomerase inhibitor RFS 2000, difluoromethylornithine (DMFO), retinoids, capecitabine, any of the aforementioned pharmaceutically acceptable salts, acids or derivatives, or any combination thereof. In some embodiments, the chemotherapeutic agent includes thiotepa, cyclophosphamide, busulfan, improsulfan, piposulfan, benzodopa, carboquan, methuredopa, bratacin, bratacinone, topotecan, adzeresin, karzeresin, bizeresin synthetic analog, cryptophycin 1, cryptophycin 8, KW-2189, CB1-TM1, chlorambucil, chlornafadin, colophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobembiti Fenestrine, prednimustine, trophosphamide, uracil mustard, carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine, engine antibiotics, calicheamicin, calicheamicin gamma II, calicheamicin omega II, dynemycin A, clodronate, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, deoxydoxorubicin, mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, potiphyllomycin, puromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zolubicin,Methotrexate, 5-fluorouracil (5-FU), denopterin, methotrexate, pteropterin, trimethrexate, fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxyuridine, carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, aminoglutethimide, mitotane, This includes trilostane, folinic acid, mytacin, ansamitocin, T-2 toxin, veraculin A, loridine A, angidin, paclitaxel, docetaxel, cisplatin, oxaliplatin, carboplatin, CPT-11, retinoic acid, sorafenib, olaparib, cetuximab, gefitinib, sulfasalazine, β-elemen, trigonelline, vorinostat, sunitinib, artesunate, any of the pharmaceutically acceptable salts, acids or derivatives of the above, or any combination thereof.

[0023] In some embodiments, one or more anticancer agents may be ferroptosis inducers. For example, agents that reduce the activity or expression of DBI as described herein (e.g., agents that reduce the activity of DBI as described herein, e.g., anti-DBI antibodies, or agents that reduce the expression of DBI as described herein, e.g., siRNA or thyroid hormone receptor agonists (e.g., resmethirome)) may be used in combination with ferroptosis inducers such as sorafenib, olaparib, cetuximab, gefitinib, sulfasalazine, β-elemen, trigonelline, vorinostat, sunitinib, or artesunate. In some examples, agents that reduce the activity or expression of DBI improve the activity of the ferroptosis inducer against target cancer cells and reduce the activity or expression of DBI compared to the activity of the ferroptosis inducer in the absence of the agent. In some cases, drugs that reduce DBI activity or expression show a synergistic improvement in the activity of ferroptosis inducers against target cancer cells compared to the activity of ferroptosis inducers in the absence of the drug, while reducing DBI activity or expression.

[0024] In some embodiments of the methods disclosed herein, one or more immunotherapeutic agents include activity against immune checkpoints. In some embodiments, the immune checkpoints include PD1 (programmed death 1), PDL1 (programmed death ligand 1), CTLA4 (cytotoxic T lymphocyte-associated protein 4), PDL2 (programmed death ligand 2), KIR (killer cell immunoglobulin-like receptor), B7-H3, B7-H4, BTLA (B and T lymphocyte attenuator), LAG3 (lymphocyte activator gene 3), TIM-3 (T cell immunoglobulin and This includes mucin domain-containing proteins, VISTA (V-domain Ig suppressor for T cell activation), ILT2 / LILRB1 (Ig-like transcript 2 / leukocyte Ig-like receptor 1), ILT3 / LILRB4 (Ig-like transcript 3 / leukocyte Ig-like receptor 4), ILT4 / LILRB2 (Ig-like transcript 4 / leukocyte Ig-like receptor 2), TIGIT (T cell immune receptor with Ig and ITIM domains), NKG2A, PVRIG, CBLB (Casitas b lineage lymphoma oncogene B), CISH (cytokine-inducible SH2-containing protein), or any combination thereof.

[0025] In some embodiments of the methods disclosed herein, one or more immunotherapeutic agents include anti-PD1 agents, anti-PD-L1 agents, anti-CTLA4 agents, anti-PD-L2 agents, anti-KIR agents, anti-B7-H3 agents, anti-B7-H4 agents, anti-BTLA agents, anti-LAG3 agents, anti-TIM-3 agents, anti-VISTA agents, anti-ILT2 / LILRB1 agents, anti-ILT3 / LILRB4 agents, anti-ILT4 / LILRB2 agents, anti-TIGIT agents, anti-NKG2A agents, anti-PVRIG agents, anti-CBLB agents, anti-CISH agents, or any combination thereof.In some embodiments, the immunotherapy agent is an anti-PD1 antibody, anti-PD-L1 antibody, anti-CTLA4 antibody, anti-PD-L2 antibody, anti-KIR antibody, anti-B7-H3 antibody, anti-B7-H4 antibody, anti-BTLA antibody, anti-LAG3 antibody, anti-TIM-3 antibody, anti-VISTA antibody, anti-ILT2 / LILRB1 antibody, anti-ILT3 / LILRB4 antibody, anti-ILT4 / LILRB2 antibody, anti-TIGIT antibody, anti-NKG2A antibody, anti-PVRIG antibody, anti-CBLB antibody, anti-CISH antibody, or ipilim. Mab, Tremelimumab, MK-1308, FPT155, PRS010, BMS-986249, BPI-002, CBT509, JS007, ONC392, TE1254, IBI310, BR02001, CG0161, KN044, PBI5D3H5, BCD145, ADU1604, AGEN1884, AGEN1181, CS1002, CP675206, Pembrolizumab, Nivolumab, Pidilizumab, AMP-224, BMS-9365 59, Semiprimab, PDR001, MDX-1105, MEDI4736, Atezolizumab, Avelumab, BMS-936559, Durvalumab, Rilurumab (IPH2102), IPH2101, MGA271, FPA150, IMP321 (Eftilagimod alfa), Relatrimab, MK-4280, AVA017, BI754111, ENUM006, GSK2831781, INCAGN2385, LAG3Ig, LAG525, REGN This includes 3767, Sym016, Sym022, Sym023, TSR033, TSR075, XmAb22841, LY3321367, MBG453, TSR-022, JNJ-61610588, MK-7684, PTZ-201, RG6058, COM902, IPH-2201, COM701, CA-327, LAG525, REGN3767, BI754111, teboterimab, FS118, MGC018, or any combination thereof. In some embodiments, the immunotherapy agent includes an anti-PD1 antibody, an anti-PD-L1 antibody, an anti-CTLA4 antibody, or any combination thereof.

[0026] In some embodiments of the methods disclosed herein, the one or more corticosteroid therapy agents are selected from cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, beclomethasone, and any combination thereof.

[0027] In some embodiments of the methods disclosed herein, the cancer comprises an immunosuppressive tumor microenvironment.

[0028] In some embodiments, the methods disclosed herein induce at least one immune surveillance biomarker including an increase in CD8 + , + , + , - , + , + , + , + , + , - , + , reg expression, a decrease in CD4 + expression, a decrease in T reg cells (CD4 + Foxp3 + ), a decrease in T reg cells (CD4 + Foxp3 + ), an increase in the ratio of CD8 + T cells to CD4 H + cells (CD4 + Foxp3 - ), an increase in T H + cells (CD4 + Foxp3 - ), a decrease in Lag3 + expression in CD8 + cells, a decrease in Lag3 + expression in CD4 + cells, a decrease in Foxp3 + expression in T reg ICOS + GITR + Lag3 - CD4 + cells, or a combination thereof. In some embodiments, inducing at least one immune surveillance biomarker comprises an increase in CD8 +This includes increased expression. In some embodiments, inducing at least one immunosurveillance biomarker is equivalent to CD4 without administration of an anti-DBI agent. + This includes a decrease in expression. In some embodiments, inducing at least one immunosurveillance biomarker is compared to an equivalent method without administration of an anti-DBI agent. reg cells (CD4 + Foxp3 + This includes a decrease in ). In some embodiments, inducing at least one immunosurveillance biomarker is compared to an equivalent method without administration of an anti-DBI agent, reg cells (CD4 + Foxp3 + ) for CD8 + This includes an increase in the proportion of T cells. In some embodiments, inducing at least one immunosurveillance biomarker is performed compared to an equivalent method without administration of an anti-DBI agent. H + cells (CD4 + Foxp3 - This includes an increased incidence of ). In some embodiments, inducing at least one immunosurveillance biomarker is compared to an equivalent method without administration of an anti-DBI agent. H + cells (CD4 + Foxp3 - ) Lag3 + This includes a decrease in the expression of CD8. In some embodiments, inducing at least one immunosurveillance biomarker is compared to an equivalent method without administration of an anti-DBI agent. + Lag3 in cells + This includes a decrease in the expression of CD4. In some embodiments, inducing at least one immunosurveillance biomarker is compared to an equivalent method without administration of an anti-DBI agent. + Foxp3 in cells + This includes a reduction in the expression of [specific marker]. In some embodiments, inducing at least one immunosurveillance biomarker is equivalent to T [another specific marker] compared to an equivalent method without administration of an anti-DBI agent. reg ICOS + GITR + Lag3 - CD4 +This includes a decrease in the number of cells.

[0029] In some embodiments, the methods disclosed herein are T H + cells (CD4 + Foxp3 - ) CD8 + The ratio of T cells is maintained compared to an equivalent method without administration of an anti-DBI agent. In some embodiments, the methods disclosed herein involve CD4 + CD8 against T cells + The ratio of T cells is maintained compared to an equivalent method without administration of an anti-DBI agent. In some embodiments, the methods disclosed herein are T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained compared to an equivalent method without the administration of anti-DBI agents.

[0030] In some embodiments, the methods disclosed herein further include inhibiting at least one cancer progression biomarker, which includes a reduction in the target tumor volume, a reduction in cancer cell proliferation, an increase in cancer cell death, a reduction in cancer growth, or a combination thereof, compared to an equivalent method without the administration of an anti-DBI agent. In some embodiments, inhibition of at least one cancer progression biomarker includes a reduction in tumor volume, and the reduction in tumor volume includes a reduction in tumor area or tumor volume. In some embodiments, the reduction in tumor volume includes a reduction in tumor volume compared to an equivalent method without the administration of an anti-DBI agent at an equivalent time point. In some embodiments, the reduction in tumor volume is up to approximately 60%. In some embodiments, the reduction in tumor volume includes a reduction in tumor area compared to an equivalent method without the administration of an anti-DBI agent at an equivalent time point. In some embodiments, inhibiting at least one cancer progression biomarker includes a reduction in cancer cell proliferation compared to an equivalent method without the administration of an anti-DBI agent at an equivalent time point. In some embodiments, inhibiting at least one cancer progression biomarker includes an increase in cancer cell death compared to an equivalent method without the administration of an anti-DBI agent at an equivalent time point. In some embodiments, inhibiting at least one cancer progression biomarker includes a reduction in cancer growth compared to an equivalent method without administration of an anti-DBI agent at an equivalent time point.

[0031] In some embodiments, the methods disclosed herein further include an increase in the survival rate of subjects compared to equivalent methods without administration of anti-DBI agents at equivalent time points. In some embodiments, the survival rate increases by up to approximately 50% compared to equivalent methods without administration of anti-DBI agents at equivalent time points.

[0032] In some embodiments, the methods disclosed herein further include an increased incidence of cancer-free occurrences compared to equivalent methods without administration of anti-DBI agents at equivalent time points. In some embodiments, the incidence of cancer-free occurrences is increased by up to 40% compared to equivalent methods without administration of anti-DBI agents at equivalent time points.

[0033] In some embodiments, the methods disclosed herein further include an increase in the time to death of the subject compared to an equivalent method without administration of an anti-DBI agent at an equivalent time point.

[0034] In some embodiments of the methods disclosed herein, the cancer is refractory cancer. In some embodiments, the cancer is resistant to immune checkpoint inhibitor therapy. In some embodiments, the cancer is selected from solid tumors, hematological tumors, skin cancers, tissue cancers, organ cancers, bone cancers, cartilage cancers, blood cancers, vascular cancers, primary cancers, metastatic cancers, bladder cancers, bone marrow cancers, brain cancers, breast cancers, colon cancers, esophageal cancers, gastrointestinal cancers, gingival cancers, kidney cancers, liver cancers, lung cancers, nasopharyngeal cancers, cervical cancers, ovarian cancers, prostate cancers, stomach cancers, testicular cancers, tongue cancers, and uterine cancers. In some embodiments, cancer is defined as cancer, lung cancer, non-small cell lung cancer, breast cancer, neoplastic cancer, undifferentiated carcinoma, giant cell carcinoma, spindle cell carcinoma, small cell carcinoma, papillary carcinoma, squamous cell carcinoma, lymphoepithelial carcinoma, basal cell carcinoma, matrix carcinoma, transitional cell carcinoma, papillary transitional cell carcinoma, adenocarcinoma, gastrinoma, cholangiocarcinoma, hepatocellular carcinoma, combination of hepatocellular carcinoma and cholangiocarcinoma, cord-like adenocarcinoma, adenoid cystic carcinoma, adenocarcinoma in adenomatous polyps, familial adenomatous polyposis, solid tumors, carcinoid tumors, bronchioloalveolar adenocarcinoma, papillary adenocarcinoma, chromophobe carcinoma, oxiphilic adenocarcinoma, basophilic carcinoma, clear cell adenocarcinoma, granular cell carcinoma, follicular adenocarcinoma, papillary adenocarcinoma, follicular adenocarcinoma, non-encapsulating sclerosing carcinoma.Carcinoma, adrenocortical carcinoma, endometrioid carcinoma, adnexal carcinoma, apocrine gland carcinoma, sebaceous gland carcinoma, ceruminous gland carcinoma, mucoepidermoid carcinoma, cystadenocarcinoma, papillary cystadenocarcinoma, papillary serous cystadenocarcinoma, mucinous cystadenocarcinoma, mucinous adenocarcinoma, signet ring cell carcinoma, invasive ductal carcinoma, medullary carcinoma, lobular carcinoma, inflammatory carcinoma, Paget's disease, breast cancer, acinar cell carcinoma, adenosquamous cell carcinoma, adenocarcinoma with squamous metaplasia, thymoma, ovarian stromal tumor, follicular cell tumor, granulosa cell tumor, roblastoma, Sertoli Cellular carcinoma, Leydig cell tumor, lipid cell tumor, accessory ganglion, extramammary accessory ganglion, pheochromocytoma, glomus angiosarcoma, melanoma, achromatic melanoma, superficial spreading melanoma, melanoma within a giant pigmented nevus, epithelioid cell melanoma, blue nevus, sarcoma, fibrosarcoma, fibrous histiocytoma, myxosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, fetal rhabdomyosarcoma, alveolar rhabdomyosarcoma, stromal sarcoma, mixed tumor, Müllerian mixed tumor, nephroblastoma, hepatoblastoma, Carcinosarcoma, mesenchymal tumor, Brenner tumor, phyllodes tumor, synovial sarcoma, mesothelioma, ovarian tumor, embryonal carcinoma, teratoma, ovarian goiter, choriocarcinoma, mesonephroma, angiosarcoma, hemangioendothelioma, Kaposi's sarcoma, hemangiopericytoma, lymphangiosarcoma, osteosarcoma, subcortical osteosarcoma, chondrosarcoma, chondroblastoma, mesenchymal chondrosarcoma, giant cell tumor of bone, Ewing's sarcoma, odontogenic tumor, ameloblastic odontogenic sarcoma, ameloblastoma, ameloblastic fibrosarcoma, pineal tumor, chordoma, glioma, ependymoma Astrocytoma, protoplasmic astrocytoma, fibrous astrocytoma, astroblastoma, glioblastoma, oligodendroglioma, oligodendroglioma, primitive neuroectodermal tumor, cerebellar sarcoma, gangliblastoma, neuroblastoma, retinoblastoma, olfactory neuron tumor, meningioma, neurofibrosarcoma, schwannoma, granulocyte tumor, lymphoma, Hodgkin's disease, Hodgkin's lymphoma, paragranuloma, small lymphocytic lymphoma, large cell diffuse lymphoma, follicular lymphoma, mycosis fungoides, and other specified non-Hodgkin's lymphomas.The cancer is selected from lymphoma, histiocytic proliferative disorder, multiple myeloma, mast cell sarcoma, intestinal immunoproliferative disorder, leukemia, lymphocytic leukemia, plasma cell leukemia, erythroleukemia, lymphosarcoma cell leukemia, myeloid leukemia, basophilic leukemia, eosinophilic leukemia, monocytic leukemia, mast cell leukemia, megakaryoblastic leukemia, myeloid sarcoma, and hairy cell leukemia. In some embodiments, the cancer is malignant cancer. In some embodiments, the cancer is selected from carcinoma, lung cancer, non-small cell lung cancer, and breast cancer. In some embodiments, the cancer is a solid tumor cancer. In some embodiments, the cancer is a fibrous tumor.

[0035] In some embodiments of the methods disclosed herein, the subject is a human being.

[0036] Disclosed herein are therapeutic regimens for treating cancer, comprising (a) an extracellular human diazepam-binding inhibitor (DBI) agent in an amount sufficient to inhibit the extracellular human DBI (anti-DBI agent), and (b) one or more anticancer agents, wherein the anti-DBI agent and the one or more anticancer agents are present in an amount sufficient to inhibit one or more cancer progression biomarkers in the subject at the time of administration to the subject, compared to an equivalent regimen without administration of the anti-DBI agent. Agents, compositions, and combinations for use in such regimens are also disclosed. Accordingly, according to one aspect of the present invention, an extracellular human diazepam-binding inhibitor (DBI) agent, or a composition comprising such an agent, is provided for use in such a manner. According to a further aspect of the present invention, one or more anticancer agents (or compositions comprising such agents or compositions) are provided for use in such regimens. According to yet further aspects of the present invention, a combination of an extracellular human diazepam-binding inhibitor (DBI) agent and one or more anticancer agents is provided for use in such regimens.

[0037] In some embodiments of the therapeutic regimens disclosed herein, the anti-DBI agent is an antibody or an aptamer.

[0038] In some embodiments of the therapeutic regimens disclosed herein, one or more anticancer agents include chemotherapeutic agents, immunotherapeutic agents, or combinations thereof. In some embodiments, the chemotherapeutic agent includes immunogenic cell death (ICD) induction activity. In some embodiments, the immunotherapeutic agent includes activity against immune checkpoints. In some embodiments, the immune checkpoints include PD1 (programmed death 1), PDL1 (programmed death ligand 1), CTLA4 (cytotoxic T lymphocyte-associated protein 4), PDL2 (programmed death ligand 2), KIR (killer cell immunoglobulin-like receptor), B7-H3, B7-H4, BTLA (B and T lymphocyte attenuator), LAG3 (lymphocyte activation gene 3), and TIM-3 (containing T cell immunoglobulin and mucin domains). This includes (3) VISTA (V-domain Ig suppressor for T cell activation), ILT2 / LILRB1 (Ig-like transcript 2 / leukocyte Ig-like receptor 1), ILT3 / LILRB4 (Ig-like transcript 3 / leukocyte Ig-like receptor 4), ILT4 / LILRB2 (Ig-like transcript 4 / leukocyte Ig-like receptor 2), TIGIT (T cell immune receptor with Ig and ITIM domains), NKG2A, PVRIG, CBLB, CISH, or any combination thereof.

[0039] In some embodiments of the therapeutic regimens disclosed herein, the immunotherapy agent includes an anti-PD1 agent, an anti-PD-L1 agent, an anti-CTLA4 agent, an anti-PD-L2 agent, an anti-KIR agent, an anti-B7-H3 agent, an anti-B7-H4 agent, an anti-BTLA agent, an anti-LAG3 agent, an anti-TIM-3 agent, an anti-VISTA agent, an anti-ILT2 / LILRB1 agent, an anti-ILT3 / LILRB4 agent, an anti-ILT4 / LILRB2 agent, an anti-TIGIT agent, an anti-NKG2A agent, an anti-PVRIG agent, an anti-CBLB agent, an anti-CISH agent, or any combination thereof. In some embodiments, the immunotherapy agent is ipilimumab, tremelimumab, MK-1308, FPT155, PRS010, BMS-986249, BPI-002, CBT509, JS007, ONC392, TE1254, IBI310, BR02001, CG0161, KN044, PBI5D3H5, BCD145, ADU1604, AGEN1 884, AGEN1181, CS1002, CP675206, Pembrolizumab, Nivolumab, Pidilizumab, AMP-224, BMS-936559, Semiprimab, PDR001, MDX-1105, MEDI4736, Atezolizumab, Avelumab, BMS-936559, Durvalumab, Rilurumab (IPH2102), IPH2101, MGA271, FPA150, IMP321 (Eftiragimod Alpha), Relatrimab, MK-4280, AVA017, BI754111, ENUM006, GSK2831781, INCAGN2385, LAG3Ig, LAG525, REGN3767, Sym016, Sym022, Sym023, TSR033, TSR075, XmAb22 This includes 841, LY3321367, MBG453, TSR-022, JNJ-61610588, MK-7684, PTZ-201, RG6058, COM902, IPH-2201, COM701, CA-327, LAG525, REGN3767, BI754111, tevoterimab, FS118, MGC018, or any combination thereof.

[0040] In some embodiments, the therapeutic regimens disclosed herein further comprise one or more corticosteroid therapeutic agents. In some embodiments, one or more corticosteroid therapeutic agents are selected from cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, beclomethasone, and any combination thereof.

[0041] In some embodiments of the treatment regimens disclosed herein, the anti-DBI agent and one or more anticancer agents are separate components administered together or sequentially. In some embodiments, the anti-DBI agent is administered at least about two weeks before one or more anticancer agents. In some embodiments, the anti-DBI agent and one or more anticancer agents are co-administered. In some embodiments, one or more corticosteroid therapies are co-administered with or sequentially with the anti-DBI agent and / or one or more anticancer agents. In some embodiments, any two or more of the anti-DBI agent, one or more anticancer agents, and one or more corticosteroid therapies are co-administered.

[0042] Disclosed herein are systems comprising one or more components, the one or more components comprising: (a) an extracellular human agent (anti-DBI agent) in an amount sufficient to inhibit extracellular diazepam binding inhibitors (DBIs) in a subject; (b) one or more anticancer agents, optionally including chemotherapeutic agents, immunotherapeutic agents, or combinations thereof; (c) one or more corticosteroid therapeutic agents; (d) or one or more of any combination of (a) to (c) individually. [Brief explanation of the drawing]

[0043] [Figure 1-1]Figures 1A-1D: Improvement of chemoimmunotherapy treatment for MCA205 cancer by autoimmunization against ACBP / DBI. Anti-ACBP / DBI autoimmunity was induced in C57BL / 6J mice by four weekly doses of Klh-ACBP / DBI vaccine or non-ACBP / DBI-specific control KLH. After two weeks of immunization recovery, mice were subcutaneously injected with MCA205 cells and treated according to the chemoimmunotherapy cycle described in Figure 1A, consisting of one dose of oxaliplatin (OXA; 10 mg / kg) and three consecutive doses of anti-PD1 monoclonal antibody (αPD1; 200 μg / dose), or injected with a corresponding vehicle and isotype mAb. Average (mean ± SEM) tumor growth curves are reported in Figure 1B, and overall survival is reported in Figure 1C. Statistical analysis was performed by linear mixed-effects modeling of tumor size for longitudinal analysis and by a 2×2 log-rank Mantel-Cox test (GraphPad Prism) for survival. The calculated p-values ​​are presented in the table shown in Figure 1D, formatted in bold font when below the threshold of 0.05. TF: No tumor. [Figure 1-2] Same as above. [Figure 1-3] Same as above.

[0044] [Figure 2-1]Figures 2A-2D: Improvement of chemoimmunotherapy treatment for MCA205 cancer by injection of ACBP / DBI neutralizing monoclonal antibody. C57Bl / 6J mice were subcutaneously injected with MCA205 cells and administered chemoimmunotherapy according to the cycle shown in Figure 2A, consisting of one dose of oxaliplatin (OXA; 10 mg / kg) and three consecutive doses of anti-PD1 monoclonal antibody (αPD1; 200 μg / dose), or were injected with a corresponding vehicle and isotype mAb. Half of the mice received repeated doses of anti-ACBP / DBI mAb (αDBI; 2.5 mg / kg) immediately before and for one week after chemotherapy. The tumor size reduction achieved by chemoimmunotherapy was further improved when combined with ACBP / DBI mAb neutralization, as shown in Figure 2B, and overall survival was extended accordingly, as shown in Figure 2C. The tumor growth and survival curves shown in Figures 2B and 2C are from one representative experiment, which was repeated twice to obtain similar results. Statistical analysis was performed on pooled data from two independent experiments using linear mixed-effects modeling for tumor size for longitudinal analysis and a 2×2 log-rank Mantel-Cox test (GraphPad Prism) for survival. The calculated p-values ​​are presented in the table shown in Figure 2D, formatted in bold font if they fall below the threshold of 0.05. TF: No tumor. [Figure 2-2] Same as above. [Figure 2-3] Same as above.

[0045] [Figure 3-1]Figures 3A-3M: Immune control of MCA205 cancer during improvement of chemoimmunotherapy treatment by injection of ACBP / DBI neutralizing monoclonal antibody. As shown in Figure 3A, T cell populations of the tumor microenvironment were analyzed by flow cytometry 10 days after chemotherapy. The number of specific cell populations relative to the total number of single cells analyzed is presented for global cytotoxic CD8+ T cells as shown in Figure 3B, and for CD4+ T cells, T lymphocytes, more specifically CD4+FoxP3+ regulatory T cells (Treg) as shown in Figure 3C, and for CD4+FoxP3- helper T cells (TH) as shown in Figure 3G. The increased ratios between the number of CD8+ T cells and CD4+ T cells, as shown in Figure 3D, the number of CD8+ T cells and Tregs, as shown in Figure 3F, and the number of CD8+ T cells and THs, as shown in Figure 3H, correlate with increased cancer-targeting cytotoxicity, while the high proportion of FoxP3+ cells within CD4+ T cells, as shown in Figure 3I, indicates a decreased immune response to cancer. The ratio of THs shown in Figure 3J to CD8+ T cells shown in Figure 3K, where cells expressing the surface marker LAG3, is an indication of exhaustion of helper cells and cytotoxic cells infiltrating the tumor. More precisely, unsupervised clustering highlights two target clusters in CD4+ T cell polarization when anti-DBI and chemoimmunotherapy are combined: a cluster of activated TH cells that are dominant (FoxP3-, ICOSint, GITR+, Lag3-) and a cluster of activated Treg cells that are suppressed (FoxP3-, GITR+, Lag3-), as shown in Figure 3L. Compensation, scaling, and gating strategies were performed using the omiq.ai platform. Data from three independent experiments are presented, population relative counts are sorted by ROUT tests (outlier threshold = 1%), and p-values ​​are calculated by one-way ANOVA with Sidak correction for multiple comparisons. [Figure 3-2] Same as above. [Figure 3-3] Same as above. [Figure 3-4] Same as above. [Figure 3-5] Same as above.

[0046] [Figure 4-1] Figures 4A-4B: Efficacy of combined treatment with ACBP / DBI-neutralizing monoclonal antibodies and chemotherapy in TC1 lung cancer. Viable TC1 cells stably expressing luciferase activity (TC1 Luc, 5 × 10⁵ per mouse) were intravenously (iv) injected into wild-type C57BL / 6J mice (minimum 7 mice per group). The mice were then treated as follows: vehicle + isotype control antibody (CTR); monoclonal antibody against DBI (αDBI; 2 mg, 5 mg / kg); monoclonal antibody against PD1 (αPD1; 10 mg / kg) and / or oxaliplatin (OXA; 5 mg / kg). Tumor size was monitored every 4 days by luciferase activity (shown in Figure 4A). Representative time-lapse images of 3 CTR mice, 3 αDBI-, 3 OXA+αPD1-, and 3 OXA+αDBI+αPD1- treated mice are shown in Figure 4B. The cross shape indicates that the animal has died. [Figure 4-2] Same as above.

[0047] [Figure 5-1] Figures 5A-5C: Improvement of chemoimmunotherapy treatment for TC1 lung cancer by injection of ACBP / DBI neutralizing monoclonal antibody. Tumor size of orthotopic TC1 lung cancer was quantified as the total flux of bioluminescent photons obtained. The mean (mean ± SEM) tumor growth curve is shown in Figure 5A, and the overall survival rate is shown in Figure 5B. Statistical significance was calculated using ANOVA type 2 (Wald test) for tumor growth curves, or a 2×2 log-rank Mantel-Cox test (GraphPad Prism) for survival. The calculated p-values ​​are presented in the table shown in Figure 5C, formatted in bold font if they are below the threshold of 0.05. TF: No tumor. [Figure 5-2] Same as above.

[0048] [Figure 6-1]Figures 6A-6B: Efficacy of combined treatment with ACBP / DBI-neutralizing monoclonal antibodies and chemotherapy in the context of corticosteroid therapy in TC1 lung cancer. Viable TC1 cells stably expressing luciferase activity (TC1 Luc, 5 × 10⁵ per mouse) were intravenously (iv) injected into wild-type C57BL / 6J mice. The mice were then treated as follows: vehicle (CTR); anti-DBI antibody (αDBI; 5 mg / kg) or its isotype; anti-PD1 (αPD1; 10 mg / kg) antibody or its isotype; and oxaliplatin (OXA; 5 mg / kg). Tumor size was monitored every 4 days by luciferase activity (shown in Figure 6A). Representative time-lapse images of three CTR mice, three αPD1+OXA- treated mice, three OXA+αPD1+αDBI- treated mice, three CORT+OXA+αPD1- treated mice, and CORT+OXA+αDBI+αPD1- treated mice are shown in Figure 6B. The cross shape indicates that the animal died. [Figure 6-2] Same as above.

[0049] [Figure 7-1] Figures 7A-7C: Improvement of chemoimmunotherapy treatment for TC1 lung cancer with ACBP / DBI neutralizing monoclonal antibody injection in the context of corticosteroid therapy. Tumor size of orthotopic TC1 lung cancer was quantified as the total flux of bioluminescent photons obtained. The mean (mean ± SEM) tumor growth curve is shown in Figure 7A, and overall survival is shown in Figure 7B. Statistical significance was calculated using ANOVA type 2 (Wald test) for tumor growth curves, or a 2×2 log-rank Mantel-Cox test (GraphPad Prism) for survival. The calculated p-values ​​are presented in the table shown in Figure 7C, formatted in bold font if they are below the threshold of 0.05. TF: No tumor. [Figure 7-2] Same as above.

[0050] [Figure 8-1]Figures 8A-8D: Improvement of immunotherapy treatment of E0771 tumors (breast cancer) by injection of ACBP / DBI neutralizing monoclonal antibody in the absence and presence of corticosteroid therapy. Live E0771 cells (5 × 10⁵ cells per mouse) were subcutaneously injected into the right lower abdomen of C57BL / 6J mice, adjacent to the mammary gland (near the orthotopic site). When the tumor became palpable (day 8), the mice were treated every two days with neutralizing anti-DBI monoclonal antibody (5 mg / kg) or its isotype. Simultaneously, free corticosterone (Sigma-Aldrich) was added to drinking water (0.1 mg / ml; water / ethanol 0.66%). As shown in Figure 8A, anti-PD1 monoclonal antibody (αPD1; 10 mg / kg) was injected on days 12, 15, 18, and 21. The mean (mean ± SEM) tumor growth curve is shown in Figure 8B, and the overall survival rate is shown in Figure 8C. Statistical analysis was performed by linear mixed-effects modeling of tumor size for longitudinal analysis and by a 2×2 log-rank Mantel-Cox test (GraphPad Prism) for survival. The calculated p-values ​​are presented in the table shown in Figure 8D, formatted in bold font when below the threshold of 0.05. TF: No tumor. [Figure 8-2] Same as above. [Figure 8-3] Same as above.

[0051] [Figure 9-1]Figures 9A - 9D: Improvement of anti - cancer immune response by injection of ACBP / DBI - neutralizing monoclonal antibody in relation to corticosteroid therapy. Two weeks before vaccination, C57BL / 6J mice (8 - week - old females) were pretreated every other day (i.p., in 200 μl of PBS) with neutralizing anti - DBI monoclonal antibody (αDBI; 5 mg / kg) or its isotype (IgG2A; 5 mg / kg) and free corticosterone (CORT) in drinking water (0.1 mg / ml; ethanol / water 0.66%). Wild - type MCA205 cells were treated with mitoxantrone (MTX; 4 μM) for 24 hours. A suspension of 2×106 cells was injected s.c. into the left flank of immunocompetent C57BL / 6J mice. PBS was injected as a negative control. One week later, viable MCA205 cancer cells (2×105 cells per mouse) were injected into the right flank of vaccinated mice. Corticosterone (CORT) and anti - ACBP / DBI were administered continuously throughout the experiment shown in Figure 9A. The mean (Average) (mean ± SEM) tumor growth curves are shown in Figure 9B and the overall survival rate is shown in Figure 9C. Statistical analysis was performed by linear mixed - effects modeling of tumor size for longitudinal analysis and by a 2×2 log - rank Mantel - Cox test (GraphPad Prism) for survival. The calculated p - values are presented in the table shown in Figure 9D and are formatted in bold font when they fall below the threshold of 0.05. TF: tumor - free. [Figure 9-2] Same as above. [Figure 9-3] Same as above.

[0052] [Figure 10]Figures 10A - 10D show the individual MCA205 tumor growth curves after chemoimmunotherapy treatment in mice autoimmunized against ACBP / DBI. For mean ± SEM, see Figure 1B. Figure 10A shows the tumor volume (mm3) after KLH - DBI administration compared to KLH alone. Figure 10B shows the tumor volume (mm3) after KLH + OXA + αPD1 administration compared to KLH alone. Figure 10C shows the tumor volume (mm3) after KLH - DBI + OXA + αPD1 administration compared to KLH - DBI. Figure 10D shows the tumor volume (mm3) after KLH - DBI + OXA + αPD1 administration compared to KLH + OXA + αPD1.

[0053] [Figure 11] Figures 11A - 11D show the individual MCA205 tumor growth curves after chemoimmunotherapy treatment and ACBP / DBI neutralization. For mean ± SEM, see Figure 2B. Figure 11A shows the tumor volume (mm3) after αDBI administration compared to control (CTR). Figure 11B shows the tumor volume (mm3) after OXA + αPD1 administration compared to CTR. Figure 11C shows the tumor volume (mm3) after αDBI + OXA + αPD1 administration compared to αDBI. Figure 11D shows the tumor volume (mm3) after αDBI + OXA + αPD1 administration compared to OXA + αPD1.

[0054] [Figure 12] Figures 12A - 12D show the individual TC1 tumor growth curves after chemoimmunotherapy treatment and ACBP / DBI neutralization. For mean ± SEM, see Figure 5A. Figure 12A shows the total flux (p / s) after αDBI administration compared to control (CTR). Figure 12B shows the total flux (p / s) after OXA + αPD1 administration compared to CTR. Figure 12C shows the total flux (p / s) after αDBI + OXA + αPD1 administration compared to αDBI. Figure 12D shows the total flux (p / s) after αDBI + OXA + αPD1 administration compared to OXA + αPD1.

[0055] [Figure 13]Figures 13A to 13D show individual TC1 tumor growth curves after chemoimmunotherapy and ACBP / DBI neutralization in the context of corticosteroid therapy. For mean ± SEM, see Figure 7A. Figure 13A shows the total flux (p / s) after αPD1 + OXA administration compared with the control (CTR). Figure 13B shows the total flux (p / s) after CORT + αPD1 + OXA administration compared with αPD1 + OXA. Figure 13C shows the total flux (p / s) after CORT + αPD1 + OXA + αDBI administration compared with CORT + αPD1 + OXA. Figure 13D shows the total flux (p / s) after CORT + αPD1 + OXA + αDBI administration compared with αPD1 + OXA + αDBI.

[0056] [Figure 14] Figures 14A to 14D show individual E0771 tumor growth curves after immunotherapy and ACBP / DBI neutralization in the context of corticosteroid therapy. See Figure 8B for mean ± SEM. Figure 14A shows tumor area (mm2) after αPD1 administration compared to the control (CTR). Figure 14B shows tumor area (mm2) after CORT + αPD1 administration compared to αPD1. Figure 14C shows tumor area (mm2) after CORT + αPD1 + αDBI administration compared to CORT + αPD1. Figure 14D shows tumor area (mm2) after CORT + αPD1 + αDBI administration compared to αPD1 + αDBI.

[0057] [Figure 15]Figures 15A to 15D show individual MCA205 tumor growth curves after vaccination and ACBP / DBI neutralization in the context of corticosteroid therapy. For mean ± SEM, see Figure 9B. Figure 15A shows tumor area (mm2) after mitoxantrone (MTX) administration compared to the control (CTR). Figure 15B shows tumor area (mm2) after CORT + MTX administration compared to MTX alone. Figure 15C shows tumor area (mm2) after CORT + MTX + αDBI administration compared to CORT + MTX. Figure 15D shows tumor area (mm2) after CORT + MTX + αDBI administration compared to MTX + αDBI.

[0058] [Figure 16-1]Figures 16A–16O: Excess DBI (ACBP) in HCC patients, based on analysis of data from The Tumor Cell Genomic Atlas (TCGA) dealing with hepatocellular carcinoma (HCC or LIHC). Figure 16A shows higher DBI mRNA expression in HCC tissue compared to normal tissue. Figures 16B and 16C show that DBI mRNA levels are elevated in HCC tissue compared to normal tissue, regardless of tumor grade and TNM stage in TCGA. Figure 16D shows elevated DBI mRNA levels detected in tumor tissue of HCC patients with AFP > 400 ng / mL compared to AFP ≤ 400 ng / mL. Figure 16E shows that high DBI expression is associated with poor prognosis in HCC patients. Figures 16F–16L show analysis of data from the AP-HP cohort. Figure 16F shows increased DBI plasma levels in HCC patients compared to controls. Figures 16G–16I show that elevated DBI plasma levels are associated with advanced BCLC stages, vascular invasion, and tumor metastasis. Figures 16J–16K show a positive correlation between DBI and AFP at both mRNA and protein levels. Figure 16L shows that plasma DBI levels are positively associated with the maximum tumor size in HCC patients. Figures 16M–16O show the tumor origin of DBI in mice with human HCC. Figure 16M is a schematic diagram of orthotopic liver transplantation of HUH-7 cells in nude mice. Figure 16N shows the detection of human DBI in mouse plasma after tumor transplantation by ELISA. Figure 16O shows that human DBI derived from HUH-7 cells in mouse plasma correlates with tumor size. [Figure 16-2] Same as above. [Figure 16-3] Same as above. [Figure 16-4] Same as above. [Figure 16-5] Same as above.

[0059] [Figure 17-1]Figures 17A–17M: Knockdown of DBI / Dbi attenuates proliferation, reduces clonal formation ability, and halts the cell cycle of hepatocellular carcinoma cells. Figure 17A shows DBI mRNA and protein expression profiles in different hepatocellular carcinoma cell lines. Figures 17B–17D show DBI / Dbi depletion efficiency validated by qRT-PCR in single-cell clones derived from different parental cell lines, including HUH-7, HEP-G2, and Hep55.1C. Figures 17E–17G show CCK-8 proliferation assays of DBI / Dbi knockdown cell lines and control cell lines derived from HUH-7, HEP-G2, and Hep55.1C. Figures 17H–17J are representative images and quantifications of colony formation assays, and Figures 17K–17M show cell fluorescence analysis of the cell cycle distribution of the aforementioned hepatocellular carcinoma cell lines. [Figure 17-2] Same as above. [Figure 17-3] Same as above. [Figure 17-4] Same as above. [Figure 17-5] Same as above.

[0060] [Figure 18-1]Figures 18A - 18Q: Intracellular and extracellular DBI inhibition in the orthotopic transplantation model shows blunted tumorigenesis. Figure 18A is a schematic diagram of an orthotopic HCC model involving intrahepatic injection of DBI - depleted clones (SH1, SH2, SH3) derived from Hep55.1C cells or parental control clones (NC). Figures 18B - 18E show log - rank tests for survival curves, tumor incidence, number of HCC nodules, and tumor weight in mice at the endpoint (n = 19 - 23 animals / group). Figure 18F is a schematic diagram of an orthotopic HCC model using Hep55.1C - Luc - derived cell clones (NC, SH1, SH2, SH3). Figure 18G shows the survival of mice treated as shown in Figure 18F. Figure 18H shows representative IVIS images of HCC lesions for different experimental conditions. Figure 18I shows quantification of the total intensity of tumor lesions from IVIS images. Figures 18J - 18K show the number of HCC nodules and tumor weight in mice at the endpoint (n = 18 - 19 mice / group). Figure 18L is a schematic diagram of an orthotopic HCC mouse model (Hep55.1C - Luc) treated with KLH / KLH - DBI. Figure 18M shows the survival of mice treated as shown in Figure 18L. Figures 18N - 18O show representative IVIS images and quantification of tumor burden by IVIS imaging. Figures 18P - 18Q show HCC nodules (codules) and tumor weight in mice at the endpoint (n = 15 - 21 mice / group). [Figure 18-2] The same as above. [Figure 18-3] The same as above. [Figure 18-4] The same as above. [Figure 18-5] The same as above. [Figure 18-6] The same as above. [Figure 18-7] The same as above. [Figure 18-8] The same as above. [Figure 18-9] The same as above. [Figure 18-10] The same as above. [Figure 18-11] The same as above. [Figure 18-12] The same as above.

[0061] [Figure 19-1] Figures 19A–19L: Intracellular and extracellular DBI inhibition delays Myc+Ctnnb1-driven hepatic tumorigenesis. Figure 19A is a schematic diagram of Myc / Ctnnb1-induced hepatic carcinogenesis in tamoxifen-inducible conditional DBI knockout mice (Dbi- / -) and control mice (Dbi+ / +). Figure 19E is a schematic diagram of Myc / Ctnnb1-induced hepatic carcinogenesis in Gabrg2 mutant (Gabrg2F77I / F77I) mice and control (Gabrg2WT) mice. Figure 19I is a schematic diagram of Myc / Ctnnb1-induced hepatic tumorigenesis in KLH / KLH-DBI (ACBP) immunized mice. Figures 19B, 19F, and 19J show the survival days of the mice. Figures 19C, 19G, and 19K show the maximum tumor size per mouse. Figures 19D, 19H, and 19L show the quantification of the number of tumors with different sizes (<2mm, 2–5mm, 5–10mm, and >10mm). [Figure 19-2] Same as above. [Figure 19-3] Same as above. [Figure 19-4] Same as above. [Figure 19-5] Same as above. [Figure 19-6] Same as above. [Figure 19-7] Same as above.

[0062] [Figure 20-1]Figures 20A–20X: Inhibition of intracellular and extracellular DBI impaired NASH-driven hepatocyte carcinogenesis. Figure 20A is a schematic diagram of Western diet (WD) + CCl4-induced hepatocyte carcinogenesis in tamoxifen-inducible conditional DBI knockout mice (Dbi- / -) and control mice (Dbi+ / +). Figure 20G is a schematic diagram of WD + CCl4-induced hepatocyte carcinogenesis in Gabrg2 mutant (Gabrg2F77I / F77I) mice and control (Gabrg2WT) mice. Figure 20M is a schematic diagram of WD + CCl4-induced hepatocyte carcinogenesis in KLH / KLH-ACBP-immunized mice. Figure 20S is a schematic diagram of HFD + DEN-induced hepatocyte carcinogenesis in KLH / KLH-DBI-immunized mice. Figures 20B, 20H, 20N, and 20T show representative immunoblots for detecting DBI proteins in the liver. GAPDH was used as a loading control. Figures 20C, 20I, 20O, and 20U show the quantification of Western blots. The optical density measurement ratio of DBI / GAPDH was normalized to the control group and is shown as (Nor.). Figures 20D, 20J, 20P, and 20V show plasma DBI levels. The values ​​were normalized to the control group and are shown as (Nor.). Figures 20E, 20K, 20Q, and 20W show the quantification of the maximum tumor size (qQuantification) for each treatment group. Figures 20F, 20L, 20R, and 20X show the quantification of the number of tumors with different sizes. n=5-27 per group. [Figure 20-2] Same as above. [Figure 20-3] Same as above. [Figure 20-4] Same as above. [Figure 20-5] Same as above. [Figure 20-6] Same as above. [Figure 20-7] Same as above. [Figure 20-8] Same as above. [Figure 20-9] Same as above. [Figure 20-10] Same as above. [Figure 20-11] Same as above. [Figure 20-12] Same as above.

[0063] [Figure 21-1] Figures 21A–21F: DBI neutralization enhances the sensitivity of HCC to immunotherapy. Figure 21A is a heterogeneous comparison of dysregulated molecular pathways between three mouse models and human liver disease. Figure 21B shows representative KEGG pathways in the three mouse datasets generated in this study. Figure 21C is a schematic diagram of anti-DBI (ACBP) + anti-PD1 combination therapy tested in orthotopic Hep55.1C HCC models. Figure 21D shows mouse survival days. Figures 21E–21F show tumor weight and number of tumor nodules per mouse in each treatment group (n=9 per group). [Figure 21-2] Same as above. [Figure 21-3] Same as above. [Figure 21-4] Same as above.

[0064] [Figure 22-1] Figures 22A–22E: Transcriptome signatures of DBI inhibition in NASH-driven HCC models. Figure 22A is a heatmap of gene expression profiles in three NASH-driven HCC mouse models. Figure 22B shows genes that are generally upregulated or downregulated in the three mouse models. Figure 22C is a flowchart summarizing the strategy for identifying gene set O1. Gene set O1 was defined as genes that are overexpressed in human HCC and downregulated by DBI inhibition in two or more mouse NASH models associated with poor prognosis in TCGA-LIHC. Figure 22D is a heatmap of gene set O1 in Figure 22C. Figure 22E shows the gene ontology (GO) and KEGG pathway analysis of gene set O1. Enriched GO terminology is shown for biological processes (BP), molecular functions (MF), and cellular components (CC). [Figure 22-2] Same as above. [Figure 22-3] Same as above. [Figure 22-4] Same as above. [Figure 22-5] Same as above.

[0065] [Figure 23-1] Figures 23A-23B: DBI inhibition regulates cell cycle-related genes. Figure 23A shows that in four NASH-driven HCC models, DBI inhibition downregulated genes that are positive regulators of the cell cycle (Ccnd1, Cdk4, Cdk6, Ccne1, or Pcna) and upregulated genes that inhibit cell cycle progression (Atr, Gadd45a, Gadd45b, Cdkn1a, or Cdkn2a). Figure 23B is a heatmap showing the expression profiles of cell cycle-related genes in HEP-G2-derived DBI knockdown cell lines. [Figure 23-2] Same as above.

[0066] [Figure 24-1] Figures 24A–24I: DBI inhibition reduced cell proliferation. Figures 24A–24E are representative images and quantifications of Ki67 IHC staining in liver sections from separate mouse models (n=3–10 samples / treatment group). T / NT represents tumor / non-tumor. Figure 24F is a schematic diagram of the strategy for generating primary Dbi+ / + and Dbi- / - HCC cells in a WD+CCl4-induced mouse model. Figure 24G shows representative images of primary Dbi+ / + and Dbi- / - HCC cells and quantifications of DBI (ACBP), Ki67, PCNA, CK19, AFP, and GPC3 immunofluorescence staining. Figure 24H shows representative images and quantifications of colony formation in primary Dbi+ / + and Dbi- / - HCC cells. Figure 24I shows the CCK-8 proliferation assay in primary HCC cells. [Figure 24-2] Same as above. [Figure 24-3] Same as above. [Figure 24-4] Same as above. [Figure 24-5] Same as above.

[0067] [Figure 25-1]Figures 25A-25C: DBI inhibition enhances ferroptosis sensitivity at the transcriptional level. Figure 25A shows qRT-PCR analysis illustrating the upregulation of ferroptosis-promoting genes and downregulation of ferroptosis-inhibiting genes by DBI inhibition. The results are shown as a heatmap (n=5-10 samples / group). Figure 25B shows quantification of Western blots indicating ferroptosis and autophagy-related proteins after DBI inhibition. Figure 25C shows the spatial transcriptome landscape of liver sections from WD / CCl4-induced HCC models using Dbi+ / + / Dbi- / - mice or KLH / KLH-DBI-immunized mice. Black rectangles indicate the target region for each sample. [Figure 25-2] Same as above. [Figure 25-3] Same as above.

[0068] [Figure 26-1] Figures 26A–26P: DBI neutralization increased susceptibility to ferroptosis induction in vitro and in vivo. Figures 26A–26D show CCK-8 survival assays of Dbi+ / + and Dbi- / - HCC cells treated with ferroptosis inducers including RSL3, IKE (imidazole ketone elastin), LA (linoleic acid), and LNA (linolenic acid). Figures 26E and 26K are schematic diagrams of anti-DBI+RSL3 or IKE combination therapy tested in orthotopic Hep55.1C-Luc HCC models. Figures 26F and 26L show mouse survival days. Figures 26G, 26H, 26M, and 26N are representative IVIS images of HCC lesions, as well as quantification of total intensity of HCC lesions from IVIS images at 25 / 23 days, respectively. Figures 26I, 26J, 26O, and 26P show the tumor weight and number of HCC nodules in mice at the human endpoint (n=7-9 animals / group). [Figure 26-2] Same as above. [Figure 26-3] Same as above. [Figure 26-4] Same as above. [Figure 26-5] Same as above. [Figure 26-6] Same as above. [Figure 26-7] Same as above. [Figure 26-8] Same as above. [Figure 26-9] Same as above. [Modes for carrying out the invention]

[0069] Detailed explanation Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to whom this disclosure relates. Any methods, systems, and materials similar or equivalent to those described herein may be used in the implementation of the embodiments described herein. The following definitions are provided to facilitate the understanding of certain terms frequently used herein and are not intended to limit the scope of this disclosure.

[0070] definition As used herein, the term “administer” means physically introducing a composition containing a therapeutic agent into a target using any of the various methods and delivery systems known to those skilled in the art. In some examples, the routes of administration of the drugs described herein include intravenous, intramuscular, subcutaneous, intraperitoneal, intraspinal, or other parenteral routes, for example, by injection or infusion. As used herein, the term “parenteral administration” means a mode of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intrafocal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as in vivo electroperforation. Non-injectable routes include topical, cutaneous, oral, or mucosal routes of administration, such as intranasal, intravaginal, rectal, sublingual, or topical. The drug may be administered, for example, once, multiple times, and / or over one or more extended periods.

[0071] As used herein, the term “drug” or its grammatical equivalent means a chemical or biological entity, such as an antibody, its antigen-binding moiety, its fragments, aptamers, compounds, small molecules, drugs, etc., or its active moiety, that can induce a biological effect against a biological target of interest, such as an anti-DBI agent, an anticancer agent, a corticosteroid therapy agent, etc.

[0072] As used herein, the term “antibody” refers to an immunoglobulin molecule and a molecule containing the immunologically active portion of an immunoglobulin molecule, i.e., an antigen-binding site that binds immunospecifically to an antigen. Therefore, the term antibody encompasses not only the entire antibody molecule but also antibody fragments, as well as variants (including derivatives) of antibodies and antibody fragments. In natural antibodies, two heavy chains are linked to each other by disulfide bonds, and each heavy chain is linked to a light chain by disulfide bonds. There are two types of light chains: lambda(1) and kappa(k). There are five major heavy chain classes (or isotypes) that determine the functional activity of antibody molecules: IgM, IgD, IgG, IgA, and IgE. Each chain contains a different sequence domain. The light chain contains two domains: a variable domain (VL) and a constant domain (CL). The heavy chain contains three (α, δ, γ) to five (μ, ε) domains, a variable domain (VH), and three to four constant domains (CH1, CH2, CH3, and CH4 are collectively referred to as CH). The variable regions of both the light (VL) and heavy (VH) chains determine the binding recognition and specificity to the antigen. The term antibody also includes heavy chain antibodies, such as camel antibodies, which contain only two heavy chains and lack the two light chains typically found in other mammalian antibodies. The antibodies described herein also include single-monomer variable antibody domains, such as the antigen-binding fragment of a heavy chain antibody, V. HHThis includes single-domain antibodies (sdAbs), also known as nanobodies, which are antibody fragments consisting of a light chain (CL) and a heavy chain (CH). The constant domains of the light chain (CL) and heavy chain (CH) confer important biological properties such as antibody chain association, secretion, transplacental migration, complement binding, and binding to the Fc receptor (FcR). The Fv fragment is the N-terminal portion of the Fab fragment of immunoglobulins and consists of a variable region of one light chain and one heavy chain. Antibody specificity lies in the structural complementarity between the antibody binding site and the antigenic determinant. The antibody binding site is mainly composed of residues derived from the hypervariable or complementarity-determining region (CDR). Occasionally, residues derived from the non-hypervariable region or framework region (FR) may be involved in the antibody binding site or may affect the entire domain structure, and therefore the binding site. The CDR refers to the amino acid sequence that together defines the binding affinity and specificity of the native Fv region of the native immunoglobulin binding site. The light and heavy chains of immunoglobulins each contain three CDRs, called L-CDR1, L-CDR2, L-CDR3, and H-CDR1, H-CDR2, and H-CDR3, respectively. Therefore, the antigen-binding site typically contains six CDRs, including sets of CDRs derived from the heavy chain V region and the light chain V region, respectively. The framework region (FR) refers to the amino acid sequence interposed between the CDRs. Residues in the antibody variable domain are traditionally numbered according to a system devised by Kabat et al. This system is described in Kabat et al., 1987, in Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, NIH, USA (hereinafter referred to as "Kabat et al."). Kabat residue notation does not necessarily directly correspond to the linear numbering of amino acid residues in the sequence number sequence. The actual linear amino acid sequence may contain fewer or additional amino acids than the strict Kabat numbering corresponding to the shortening or insertion of structural components, regardless of the framework of the basic variable domain structure or the complementarity-determining region (CDR). The correct Kabat numbering of residues can be determined for a given antibody by aligning homologous residues in the antibody sequence with the "standard" Kabat numbering sequence.The CDRs of the heavy chain variable domain are located at residues 31-35B (H-CDR1), 50-65 (H-CDR2), and 95-102 (H-CDR3) according to the Kabat numbering system. The CDRs of the light chain variable domain are located at residues 24-34 (L-CDR1), 50-56 (L-CDR2), and 89-97 (L-CDR3) according to the Kabat numbering system.

[0073] As used herein, the term “antigen” refers to any known or unknown substance that can be recognized by an antibody, including proteins, glycoproteins, and carbohydrates. In some embodiments, these antigens include biologically active proteins such as hormones, cytokines and their cell surface receptors, bacterial or parasitic cell membranes or their purified components, and viral antigens. In one example, the antigen expressed on the surface of the cell is an antigen that is difficult to purify or an antigen that loses the desired epitope upon biotinylation, e.g., the antigens mentioned above. In another example, the antigen is unknown, and the antigen is any material that provides a potential source of antigens. In some embodiments, the material is of animal origin, e.g., mammalian, plant, yeast, bacterial, or viral origin. The material may be cells or cell populations from which antibodies are desirable to be isolated, such as mammalian cells, immunomodulatory cells, lymphocytes, monocytes, polymorphonuclear leukocytes, T cells, cancer cells, tumor cells, yeast cells, bacterial cells, infectious agents, parasites, and plant cells. In some embodiments, the cells are tumor cells.

[0074] As used herein, the term “aptamer” refers to a class of molecules that represent an antibody substitute for molecular recognition. Aptamers are oligonucleotide sequences that have the ability to recognize substantially any class of target molecules with high affinity and specificity. Such ligands can be isolated by phylogenetic evolution of ligands through exponential enrichment (SELEX) of a random sequence library. A random sequence library can be obtained by combinatorial chemosynthesis of DNA. In this library, each member is a linear oligomer with a unique sequence that is ultimately chemically modified. Peptide aptamers consist of a structurally constrained antibody variable region presented by a platform protein such as Escherichia coli (E. coli) thioredoxin A, selected from the combinatorial library by two hybrid methods (Colas et al., 1996).

[0075] As used herein, the term “anticancer agent” refers to any substance used to treat hyperproliferative diseases such as cancer in mammals, for example, humans. Examples of anticancer agents are disclosed herein.

[0076] As used herein, the term “B cells” includes any B cells or derivatives thereof that produce antibodies, such as B lymphocytes, plasma cells, plasmablasts, activated B cells, or memory B cells. These cells may secrete antibodies and / or maintain antibodies on their surface. A population of B cells for use in this disclosure is any population suspected to contain at least one B cell capable of producing an antibody having the desired function. B cells for use in this disclosure may be obtained from a variety of sources. For example, B cells may be obtained from animals that have been immunized with an antigen or that have shown an immune response to an antigen as a result of a disease. Alternatively, B cells may be obtained from, for example, immunized naive animals (or animals that are not known to have been exposed to the antigen of interest or are not thought to have been exposed to the antigen of interest) that have not been previously exposed to the antigen of interest.

[0077] As used herein, the term “binding” refers to the association of two molecules by covalent, electrostatic, hydrophobic, ionic, and / or hydrogen bonding interactions, including, for example, salt and water bridge interactions. As used herein, in relation to the binding of an antibody to a given target molecule (e.g., an antigen or epitope), the term “binding” typically refers to about 10 -7 M or less, for example, about 10 -8 M or less, for example, about 10 -9 M or less, approximately 10 -10 M or less, or about 10 -11 M or less than K D This is a binding at an affinity corresponding to the given conditions.

[0078] As used herein, the term “cancer” has its general meaning in the art and includes, but is not limited to, solid tumors and hematological tumors. The term “cancer” includes diseases of the skin, tissues, organs, bones, cartilage, blood, and blood vessels. The term “cancer” further encompasses both primary cancers and metastatic cancers. Examples of cancers that can be treated by the methods and treatment regimens of this disclosure include, but are not limited to, cancer cells from the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestinal tract, gums, head, kidney, liver, lung, nasopharynx, neck, ovaries, prostate, skin, stomach, testes, tongue, or uterus. Furthermore, cancer can be, in specific terms, of the following histological types, but is not limited to these: neoplastic carcinoma, undifferentiated carcinoma, giant cell carcinoma and spindle cell carcinoma, small cell carcinoma, papillary carcinoma, squamous cell carcinoma, lymphoepithelial carcinoma, basal cell carcinoma, matrix carcinoma, transitional cell carcinoma, papillary transitional cell carcinoma, adenocarcinoma, gastrinoma, cholangiocarcinoma, hepatocellular carcinoma, combination of hepatocellular carcinoma and cholangiocarcinoma, cord-like adenocarcinoma, adenoid cystic carcinoma, adenocarcinoma in adenomatous polyps, familial adenomatous polyposis, solid tumors, carcinoid tumors, bronchioloalveolar adenocarcinoma, papillary adenocarcinoma, chromophobe carcinoma, oxiphilic adenocarcinoma, basophilic carcinoma, clear cell adenocarcinoma, granular cell carcinoma, follicular adenocarcinoma, papillary adenocarcinoma and follicular adenocarcinoma, non-encapsulating sclerosing carcinoma.Carcinoma, adrenocortical carcinoma, endometrioid carcinoma, adnexal carcinoma, apocrine gland carcinoma, ceruminous gland sebaceous gland carcinoma, mucoepidermoid carcinoma, cystadenocarcinoma, papillary cystadenocarcinoma, papillary serous cystadenocarcinoma, mucinous cystadenocarcinoma, mucinous adenocarcinoma, signet ring cell carcinoma, invasive ductal carcinoma, medullary carcinoma, lobular carcinoma, inflammatory carcinoma, Paget's disease of the buccal mammal, acinar cell carcinoma, adenosquamous cell carcinoma, adenocarcinoma with squamous metaplasia, thymoma, ovarian stromal tumor, follicular cell tumor, granulosa cell tumor, roblastoma, Sertoli cell Cancer, Leydig cell tumor, lipid cell tumor, accessory ganglion, extramammary accessory ganglion, pheochromocytoma, glomus angiosarcoma, melanoma, achromatic melanoma, superficial spreading melanoma, melanoma within a giant pigmented nevus, epithelioid cell melanoma, blue nevus, sarcoma, fibrosarcoma, fibrous histiocytoma, myxosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, fetal rhabdomyosarcoma, alveolar rhabdomyosarcoma, stromal sarcoma, mixed tumor, Müllerian duct mixed tumor, nephroblastoma, hepatoblastoma, cancer Sarcoma, mesenchymal tumor, Brenner tumor, phyllodes tumor, synovial sarcoma, mesothelioma, ovarian tumor, embryonal carcinoma, teratoma, ovarian goiter, choriocarcinoma, mesoneflomatoma, angiosarcoma, hemangioendothelioma, Kaposi's sarcoma, hemangiopericytoma, lymphangiosarcoma, osteosarcoma, subcortical osteosarcoma, chondrosarcoma, chondroblastoma, mesenchymal chondrosarcoma, giant cell tumor of bone, Ewing's sarcoma, odontogenic tumor, ameloblastic odontogenic sarcoma, ameloblastoma, ameloblastic fibrosarcoma, pineal tumor, chordoma, glioma, ependymoma, Astrocytoma, protoplasmic astrocytoma, fibrous astrocytoma, astroblastoma, glioblastoma, oligodendroglioma, oligodendroglioma, primitive neuroectodermal tumor, cerebellar sarcoma, gangliblastoma, neuroblastoma, retinoblastoma, olfactory neuron tumor, meningioma, neurofibrosarcoma, schwannoma, granulocyte tumor, lymphoma, Hodgkin's disease, Hodgkin's lymphoma, paragranuloma, small lymphocytic lymphoma, large cell diffuse lymphoma, follicular lymphoma, mycosis fungoides, and other specified non-Hodgkin lymphomas. The cancer is selected from specified non-Hodgkin's lymphoma, histiocytic proliferative disorder, multiple myeloma, mast cell sarcoma, intestinal immunoproliferative disorder, leukemia, lymphocytic leukemia, plasma cell leukemia, erythroleukemia, lymphosarcoma cell leukemia, myeloid leukemia, basophilic leukemia, eosinophilic leukemia, monocytic leukemia, mast cell leukemia, megakaryoblastic leukemia, myeloid sarcoma, and hairy cell leukemia. In some embodiments, the cancer is malignant cancer.

[0079] As used herein, the term “co-administration” can mean any of the following: (i) combining two or more drugs and administering them together at one time; (ii) administering a first drug and then a second drug a short time later (e.g., 1, 2, 5, 10, 15, 20, 30, and 45 minutes; and 1, 2, 4, 6, 8, 16, and 24 hours later); (iii) administering a drug to a subject who has already received long-term treatment with the first and / or second drug; or (iv) simultaneously administering two or more drugs by different routes of administration. For example, a subject with cancer may be administered a single dose of a mixture of the anticancer drug and the anti-DBI agent described herein. In another example, the subject may be administered the anticancer drug by intravenous infusion and the anti-DBI agent described herein parenterally. All of these co-administrations can be performed three times a week over a course of an anticancer therapy cycle, such as a chemotherapy cycle, for example, over a period of three weeks. Furthermore, the subject may be administered parenterally with the anti-DBI agent composition during a period preceding the administration of anticancer drugs (e.g., 1, 2, 7, and 14 days). In some embodiments, cancer patients may receive the anti-DBI agent composition daily during a standard course of chemotherapy. Thus, "co-administration" encompasses the simultaneous, separate, or sequential administration of two or more agents.

[0080] As used herein, the terms “conjugated,” “joined,” and “linked,” and their grammatical equivalents, refer to any manner in which two components are joined to one another, including, but not limited to, the creation of ligand-pharmaceutical fusion constructs by chemical (either by proximal association of one molecule with a second molecule, or by covalent or noncovalent association of one component with a second component), electrostatically, or by using all tools of molecular biology.

[0081] As used herein, the term “DBI” has its general meaning in the art and refers to a diazepam binding inhibitor, or acyl-CoA binding protein encoded by the DBI gene (gene ID: 1622). In some embodiments, DBI refers to extracellular DBI. The term is also known as EP;ACBP;ACBD1; and CCK-RP. An exemplary amino acid sequence of DBI is represented by the NCNI reference sequence NP_001073331.1 (SEQ ID NO: 1) (Acyl-CoA binding protein isoform 1). An exemplary human nucleic acid sequence is represented by the NCNI reference sequence NM_001079862.2 (SEQ ID NO: 2) (Acyl-CoA binding protein isoform 1).

[0082] Sequence ID 1-MSQAEFEKAA EEVRHLKTKP SDEEMLFIYG HYKQATVGDI NTERPGMLDF TGKAKWDAWN ELKGTSKEDA MKAYINKVEE LKKKYGI

[0083] SEQ ID NO: 2-GCTCGCCCGA GCAGGGTTGG GGCGAGTGGA CCGCGCCTCT AAAGGCGCTT GCCAGTGCAA TCTGGGCGAT CGCTTCCTGG TCCTCGCCTC CTCCGCTGTC TCCCTGGAGT TCTTGCAAGT CGGCCAGGAT GTCTCAGGCT GAGTTTGAGA AAGCTGCAGA GGAGGTTAGG CACCTTAAGA CCAAGCCATC GGATGAGGAG ATGCTGTTCA TCTATGGCCA CTACAAACAA GCAACTGTGG GCGACATAAA TACAGAACGG CCCGGGATGT TGGACTTCAC GGGCAAGGCC AAGTGGGATG CCTGGAATGA GCTGAAAGGG ACTTCCAAGG AAGATGCCAT GAAAGCTTAC ATCAACAAAG TAGAAGAGCT AAAGAAAAAA TACGGGATAT GAGAGACTGG ATTTGGTTAC TGTGCCATGT GTTTATCCTA AACTGAGACA ATGCCTTGTT TTTTTCTAAT ACCGTGGATG GTGGGAATTC GGGAAAATAA CCAGTTAAAC CAGCTACTCA AGGCTGCTCA CCATACGGCT CTAACAGATT AGGGGCTAAA ACGATTACTG ACTTTCCTTG AGTAGTTTTT ATCTGAAATC AATTAAAAGT GTATTTGTTA CTTTAAATAA CTTTAAAAAA AAAA

[0084] As used herein, the term “DBI activity” refers to any biological activity of DBI, including, among other things, inhibition of autophagy, induction of hypoglycemia, stimulation of food intake, stimulation of weight gain, reduction of fatty acid oxidation, upregulation of glucose transporters, upregulation of PPARGs, stimulation of glucose uptake, stimulation of glycolysis or lipogenesis, or any combination thereof.

[0085] As used herein, the terms “coding” or “encoding” refer to a specific sequence of nucleotides in a polynucleotide that serves as a template for the synthesis of other polymers and macromolecules in a biological process having either a defined sequence of nucleotides (e.g., rRNA, tRNA, and mRNA) or a defined sequence of amino acids, as well as the inherent properties of the biological properties that result therefrom. Thus, a gene, cDNA, or RNA codes for a protein if the transcription and translation of the mRNA corresponding to that gene produces a protein in a cell or other biological system. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is typically provided in a sequence listing, and the non-coding strand, which is used as a template for the transcription of the gene or cDNA, can be said to code for a protein or other product of that gene or cDNA.

[0086] As used herein, the terms “enhance,” “increase,” “expand,” and “improve,” and their grammatical equivalents, mean that, with respect to the level of any molecule (e.g., amino acid sequence, nucleic acid sequence, antibody, etc.), cell (e.g., B cell, T cell, tumor cell), and / or phenomenon (e.g., disease treatment), the amount of molecules, cells, and / or phenomena in the first sample (or first subject) is greater than that of the second sample (or second subject or control) by any amount statistically significant using any statistical analysis method accepted in the art, compared to the second sample (or second subject or control). For example, this could refer to a natural, synthetic or engineered compound, drug, or component that has a biological effect that enhances the effectiveness of an immune checkpoint, anti-cancer treatment, immunotherapy treatment, or any combination thereof.

[0087] As used herein, the terms “immune checkpoint inhibitor” or “ICI” include, for example, PD1 (programmed death 1), PDL1 (programmed death ligand 1), CTLA4 (cytotoxic T lymphocyte-associated protein 4), PDL2 (programmed death ligand 2), KIR (killer cell immunoglobulin-like receptor), B7-H3, B7-H4, BTLA (B and T lymphocyte attenuator), LAG3 (lymphocyte activation gene 3), TIM-3 (T cell immunoglobulin and mucin domain-containing 3), VISTA (T cell activation V-domain This refers to drugs that target and inhibit immune checkpoints, such as the main Ig suppressor, ILT2 / LILRB1 (Ig-like transcript 2 / leukocyte Ig-like receptor 1), ILT3 / LILRB4 (Ig-like transcript 3 / leukocyte Ig-like receptor 4), ILT4 / LILRB2 (Ig-like transcript 4 / leukocyte Ig-like receptor 2), TIGIT (T cell immune receptor with Ig and ITIM domains), NKG2A, PVRIG, and in some cases, drugs that target so-called endogenous checkpoint blockade, such as molecules with ubiquitin ligase activity, such as CBLB (Casitas b lineage lymphoma proto-gene B) and CISH (cytokine-induced SH2-containing protein). The inhibitors described herein include drugs such as antibodies or chemical molecules that target immune checkpoints, including PD-1, CTLA4, TIGIT, PD-L1, PDL2, KIR, B7-H3, B7-H4, BTLA, LAG3, TIM-3, VISTA, ILT2 / LILRB1, ILT3 / LILRB4, ILT4 / LILRB2, TIGIT, NKG2A, PVRIG, CBLB, CISH, or any combination thereof. The terms “Programmed Death-1” or “PD1” refer to an immunosuppressive receptor belonging to the CD28 family. PD1 is primarily expressed on previously activated T cells in vivo and binds to two ligands, PDL1 and PDL2. As used herein, the term “PD1” includes human PD-1 (hPD1), variants, isoforms, and species homologs of hPD1, as well as analogs having at least one common epitope with hPD1. The complete hPD1 sequence can be found under GenBank accession number U64863.A PD1 antagonist or anti-PD1 agent refers to any agent that blocks the inhibitory effect of PD1 on the immune system. For example, PD1 antagonists include agents or parts that directly block the binding of PD1 to its receptor, and agents or parts that have an allosteric effect on the activity of PD1. The term “programmed death ligand-1” or “PD-L1” refers to one of two cell surface glycoprotein ligands to PD1 (the other being PDL2) that, when bound to PD1, downregulate T cell activation and cytokine secretion. As used herein, the term “PDL1” includes human PDL1 (hPDL1), variants, isoforms, and species homologs of hPDL1, as well as analogs having at least one common epitope with hPDL1. The complete hPDL1 sequence can be found under GenBank accession number Q9NZQ7.

[0088] As used herein, the terms “immunogenic cell death” or “ICD” refer to a form of cell death induced by several cell proliferation inhibitors, including oxaliplatin, cyclophosphamide, and mitoxantrone (Galluzzi et al., Cancer Cell. 2015 Dec. 14;28(6):690-714) and anthracyclines, bortezomib, radiotherapy, and photodynamic therapy (PDT) (Garg et al., (2010) “Immunogenic cell death, DAMPs and anti-cancer therapeutics: an emerging amalgamation” Biochim Biophys Acta, 1805(1):53-71). Unlike normal apoptosis, which is mostly non-immunogenic or even tolerogenic, immunogenic apoptosis in cancer cells can induce an effective antitumor immune response through the activation of dendritic cells (DCs) and the resulting activation of specific T cell responses. ICD is characterized by the secretion of damage-associated molecular patterns (DAMPs).

[0089] The term "immunotherapy" refers to any treatment whose mechanism of action is, partially or primarily, to enhance the individual's immune response.

[0090] As used herein, the term “immunotherapy agent” refers to any molecule, compound, or drug that induces an active immune response. In some cases, immunotherapy agents activate the immune system. In some cases, immunotherapy agents are cancer-targeted immunotherapy agents. In some cases, immunotherapy agents induce an active immune response against cancer or its cells. In some cases, immunotherapy agents induce a general immune response. In some cases, immunotherapy agents induce an immune response against a specific protein, group of proteins, transcript or group of transcripts. For example, immunotherapy agents may include immune checkpoint inhibitors, chimeric antigen receptor CAR-T cells, CAR-NK cells, cytotoxic monoclonal antibodies, and vaccines. Examples of immunotherapy agents are disclosed herein.

[0091] As used herein, “inhibit,” “reduce,” “suppress,” “decrease,” “neutralize,” and their grammatical equivalents mean that, with respect to the level of any molecule (e.g., amino acid sequence, nucleic acid sequence, antibody, etc.), cell (e.g., B cell, T cell, tumor cell), and / or phenomenon (e.g., disease symptom), the amount of the molecule, cell, and / or phenomenon in the first sample (or first subject) relative to the second sample (or second subject or control) is less than in the second sample (or second subject or control) by any amount statistically significant using any statistical analysis method recognized in the art. For example, it may refer to a natural, synthetic or engineered compound, drug, or component that has a biological effect of inhibiting proteins such as extracellular DBI.

[0092] As used herein, terms such as “monoclonal antibody,” “monoclonal Ab,” “monoclonal antibody composition,” and “mAb” refer to preparations of antibody molecules with a single molecular composition. Monoclonal antibodies are obtained from a substantially homogeneous population of antibodies; that is, the individual antibodies in the population are identical except for any naturally occurring variations that may be present in small amounts. Examples of such compounds are disclosed herein.

[0093] As used herein, the term “neutralizing anti-DBI monoclonal antibody” refers to an antibody or monoclonal antibody that has specificity for DBI and inhibits, reduces, or completely neutralizes the activity of DBI (e.g., extracellular DBI). Whether an antibody is a neutralizing antibody can be determined by an in vitro assay such as one of those described in the Examples. Typically, the neutralizing antibodies of this disclosure inhibit the activity of extracellular DBI by at least 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100%. Examples of such antibodies are disclosed herein.

[0094] As used herein, the term “protein” refers to a polymer of amino acids of any length. The term also includes amino acid polymers modified, for example, by the formation of disulfide bonds, glycosylation, lipidation, phosphorylation, or conjugation with a labeling component.

[0095] As used herein, the term “refractory cancer” refers to cancer that does not respond to treatment. Some cancer cells have ways of defending themselves against chemotherapy drugs, immunotherapies, biological agents and / or radiation therapy. In such cases, the cancer is called refractory.

[0096] As used herein, the term “separate administrations” means administering two or more compounds / drugs to a subject simultaneously, substantially simultaneously, or sequentially, in any order from non-fixed-dose formulations. There may or may not be specific time intervals between the administrations of each compound.

[0097] As used herein, the term “sequential” administration means administering two or more compounds / drugs to a subject from unfixed (separate) dosage forms in separate actions. The administration actions may or may not be associated by a specified time interval. For example, administering compounds over a specified period, such as once every 14 to 21 days.

[0098] As used herein, the terms “specific” or “specificity” refer to the ability of an antibody to detectably bind to a target molecule (e.g., an epitope presented on an antigen) with relatively little detectable reactivity to other target molecules. Specificity can be determined relatively by binding or competitive binding assays, for example, using a Biacore instrument. Specificity may be indicated, for example, by an affinity / binding activity ratio of about 10:1, about 20:1, about 50:1, about 100:1, 10,000:1, or more, for binding to a specific antigen compared to nonspecific binding to other unrelated molecules.

[0099] As used herein, the terms “subject” and “patient” are interchangeable and refer to any subject, such as a human, to whom diagnosis, treatment, or therapy is desired or administered. Other subjects may include cattle, dogs, cats, guinea pigs, rabbits, rats, mice, horses, and the like. In some embodiments, the subject is a human.

[0100] Where used herein, the term “therapeutic dose” refers to a sufficient amount of one or more of the agents of this disclosure to achieve a therapeutic effect. However, it will be understood that the total daily dose of the compounds / agents and compositions of this disclosure is to be determined by the attending physician within the bounds of sound medical judgment. The specific therapeutic dose level for any particular subject will depend on a variety of factors, including the disorder being treated and its severity; the activity of the particular compound / agent used; the particular composition used, the subject’s age, weight, overall health, sex, and diet; the timing of administration, route of administration, and excretion rate of the particular compound / agent used; the duration of treatment; drugs used in combination with or concurrently with the particular compound / agent used; and similar factors well known in the medical field. For example, it is well within the art of the art to start with a dose of the compound / agent at a level lower than necessary to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. However, the daily dose of the product can vary over a wide range of 0.01 to 4,000 mg per adult per day. In some examples, compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250, 500, and 1000 mg of the active ingredient for symptomatic dose adjustment to the subject being treated. Pharmaceuticals may contain approximately 0.01 mg to approximately 1000 mg of the active ingredient. Effective doses of compounds / drugs, such as therapeutic doses, may be supplied at dosage levels ranging from 0.0002 mg / kg to approximately 50 mg / kg body weight / day, particularly approximately 0.001 mg / kg to 10 mg / kg body weight / day.

[0101] As used herein, the term “treatment regimen” refers to a pattern of treatment for a disease, e.g., a pattern of dosing used during treatment. A treatment regimen may include induction regimens and maintenance regimens. The term “induction regimen” or “induction period” refers to a treatment regimen (or part of a treatment regimen) used for the initial treatment of a disease. A common goal of an induction regimen is to deliver a high level of drug / medication to the subject during the initial period of the treatment regimen. An induction regimen may (partially or entirely) be a “loading regimen,” which may include administering a higher dose of drug / medication than the physician uses during the maintenance regimen, administering drug / medication more frequently than the physician uses during the maintenance regimen, or both. The term “maintenance regimen” or “maintenance period” refers to a treatment regimen (or part of a treatment regimen) used to maintain the subject during treatment for a disease, e.g., to keep the subject in remission for an extended period (months or years). Maintenance regimens may be continuous therapy (e.g., administering drugs / medications at regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent intervention, intervention upon relapse, or intervention upon meeting specific predetermined criteria (e.g., pain, disease onset, etc.)).

[0102] As used herein, the terms “treatment” or “to treat” include both defensive or preventive treatments, as well as curative or disease-modifying treatments, and include treatment of subjects at risk of or suspected of having the disease, and subjects diagnosed with the disease or medical condition, including the suppression of clinical relapses. Treatments may be administered to subjects with or who may eventually acquire a medical impairment to prevent, cure, delay the onset of, reduce the severity of, or improve one or more symptoms of the disease or relapsing disease, or to extend the survival of the subject beyond the expected survival in the absence of such treatment.

[0103] This specification discloses the use of agents that stimulate autophagy by inhibiting extracellular diazepam binding inhibitors (DBIs) (i.e., anti-DBI agents) in subjects such as specimens, organisms, or human subjects. Such use may be useful in the therapeutic treatment of cancer. The disclosures and embodiments described herein are at least in part based on compelling evidence of the therapeutic effects of various tumor models upon inhibition of extracellular DBI by various methods. Embodiments arise from novel findings that extracellular DBI inhibition increases cancer immune surveillance, inhibits cancer progression including inhibition of cancer cell proliferation, and leads to improved therapeutic outcomes, as demonstrated, for example, by an increased ratio of cytotoxic T cells to regulatory T cells, and changes in the composition of tumor immune infiltrates showing effects on T cell activation and exhaustion markers. Further embodiments of the disclosure are based on the surprising finding that extracellular DBI inhibition can reduce and / or reverse the immunosuppressive effects of corticosteroid therapy and restore the therapeutic effects of chemotherapy, immunotherapy, or chemoimmunotherapy lost due to co-administration of corticosteroid therapy agents. Therefore, the systems and compositions described herein, as well as the methods of use thereof, are useful for improving the therapeutic effect of immunotherapy, inducing and / or enhancing cancer immune surveillance, inhibiting cancer progression, improving treatment prognosis, enhancing the therapeutic effect of anticancer therapy, reducing the immunosuppressive effect of corticosteroid therapy, treating cancer, or combinations thereof.

[0104] Anti-DBI agents The systems, methods, and compositions described herein may include agents or their use that inhibit extracellular DBI activity. In some embodiments, the agent that inhibits extracellular DBI activity is an anti-DBI agent. In some embodiments, the agent that inhibits DBI activity inhibits extracellular DBI. In some embodiments, the agent that inhibits extracellular DBI activity inhibits extracellular DBI expression. In some embodiments, extracellular DBI may be mammalian DBI, such as extracellular human DBI. In some embodiments, autophagy is stimulated by inhibiting extracellular DBI activity. In some embodiments, the agent that stimulates autophagy by inhibiting extracellular DBI activity is an anti-DBI agent. In some embodiments, the agent that stimulates autophagy by inhibiting extracellular human DBI is an anti-DBI agent. In some embodiments, the anti-DBI agent described herein is a biological molecule or a chemical molecule. In some embodiments, the biological molecule is an antibody, its antigen-binding moiety, or an aptamer. In some embodiments, the chemical molecule is a small molecule, a drug, or a compound. In some embodiments, the anti-DBI agent described herein is an antibody, its antigen-binding moiety, or an aptamer for extracellular DBI.

[0105] In some embodiments, the antibodies or aptamers described herein are for a fragment consisting of an amino acid sequence ranging from amino acid residue at position 43 to amino acid residue at position 50 of SEQ ID NO: 1 (i.e., octapeptide or OP). In some embodiments, the agent that inhibits the activity of DBI (e.g., extracellular DBI) is an aptamer for extracellular DBI.

[0106] In some embodiments, the agent that inhibits the activity of DBI (e.g., extracellular DBI) is an antibody against extracellular DBI. In some embodiments, the antibody of this disclosure is a chimeric antibody, typically a chimeric mouse / human antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a human antibody. Fully human monoclonal antibodies can also be prepared by immunizing transgenic mice for most of the human immunoglobulin heavy and light chain loci. See, for example, U.S. Patents 5,591,669, 5,598,369, 5,545,806, 5,545,807, 6,150,584, and the references cited herein, the contents of which are incorporated herein by reference.

[0107] In some embodiments, the antibodies described herein are neutralizing antibodies. In some embodiments, the neutralizing antibodies in this disclosure do not mediate antibody-dependent cell-mediated cytotoxicity and therefore do not contain an Fc moiety that induces antibody-dependent cytotoxicity (ADCC). In some embodiments, the neutralizing antibodies do not contain an Fc domain that can substantially bind to the FcgRIIIA(CD16) polypeptide. In some embodiments, the neutralizing antibodies lack an Fc domain (e.g., lack a CH2 and / or CH3 domain) or contain an Fc domain of an IgG2 or IgG4 isotype. In some embodiments, the neutralizing antibodies consist of or include multispecific antibodies containing Fab, Fab', Fab'-SH, F(ab')2, Fv, diabodies, single-chain antibody fragments, or multiple different antibody fragments. In some embodiments, the neutralizing antibodies are not linked to a toxic moiety. In some embodiments, one or more amino acids selected from amino acid residues can be replaced with different amino acid residues so that the antibody alters the C2q binding and / or reduces or eliminates complement-dependent cytotoxicity (CDC). This approach is described in more detail in U.S. Patent No. 6,194,551 by ldusogie et al.

[0108] Any anti-DBI antibody that inhibits DBI activity (e.g., extracellular DBI) is suitable for use in the methods and compositions described herein. Such anti-DBI antibodies are commercially available, documented in the literature, and their sequences are known or can be derived.For example, antibodies that inhibit DBI activity (e.g., extracellular DBI) and are suitable for use as described herein include ab231910 (rabbit polyclonal, abcam); ab232760 (rabbit polyclonal, abcam); ab16871 (rabbit polyclonal, abcam); sc-30190 (rabbit polyclonal, Santa Cruz Biotechnology); FNab02256 (rabbit polyclonal, Wuhan Fine Biotech Co); PA5-89139 (rabbit polyclonal, Invitrogen); OTI4A8 (mouse monoclonal, OriGene); OTI6E12 (mouse monoclonal, OriGene); mAb 7A (mouse monoclonal, Fred Hutch Antibody Technology); Abcam (catalog number ab16871; RRID: AB_302557); Santa Cruz Biotechnology Cruz DBI human or mouse FL-87 monoclonal antibody (catalog number sc-30190; RRID: AB_2211046); Santa Cruz DBI human C-9 polyclonal antibody (catalog number sc-376853; RRID: AB_2722761); Abcam DBI mouse polyclonal antibody (catalog number ab231910); Fred Hutch Antibody Technology's DBI mouse 7a monoclonal antibody, Invitrogen's DBI polyclonal antibody (catalog numbers PA5-89139, PA5-79138, PA5-40659, PA5-102751, PA5-84066, PA5-76729 and PA5-92426); OriGene's DBI monoclonal antibody (catalog numbers CF813069, CF813070, CF813117, TA813069, TA8 The antibody has at least 80%, at least 85%, at least 90%, at least 95%, or 100% sequence identity to the polypeptide sequence of an antibody selected from the group consisting of 1370 and TA813117); a Proteintech DBI polyclonal antibody (catalog number 14490-1-AP); an Abnova DBI polyclonal antibody (catalog number H00001622-D01P); or more than one of the above.

[0109] The antibodies described herein may be commercially obtained or synthesized by any suitable method. For example, anti-DBI human monoclonal antibodies may be synthesized using peptides derived from full-length human ACBP and phage display techniques. In some embodiments, the antibodies described herein are mutant antibodies. In some embodiments, the antibodies described herein are selected based on preferred kinetic parameters such as specificity or affinity for human ACBP. The specificity of the antibodies described herein can be verified by Western blotting, immunofluorescence, and flow cytometry against human ACBP / DBI knockout cell lines. In some embodiments, the antibodies described herein react only with human ACBP and not with mouse ACBP. KD measurement may also be performed if KD is the equilibrium dissociation constant (the kd / ka ratio between the antibody and its antigen) and KD and affinity are inversely correlated. In some embodiments, the antibodies described herein have high affinity for human ACBP. In some embodiments, the antibodies described herein have preferred kinetic parameters, which indicate faster association and slower dissociation. For example, the sensorgram shapes of the antibodies described herein show a clear concentration-response relationship at lower concentrations.

[0110] In some embodiments, the agent that inhibits extracellular DBI expression is an expression inhibitor. In some embodiments, the gene expression inhibitor is an siRNA, an endonuclease, an antisense oligonucleotide, a thyroid hormone receptor agonist, or a ribozyme.

[0111] In some embodiments, the agents that inhibit DBI expression are thyroid hormone receptor agonists. As disclosed herein, administration of thyroid hormone receptor agonists can be used to indirectly reduce or inhibit the expression of circulating DBI. While not wishing to be bound by theory, the applicants have found that administration of thyroid hormone receptor agonists (e.g., thyroid hormone receptor β (THR-β) agonists) to subjects results in down-transcriptional regulation of Acbp / Dbi mRNA in the subjects. Thus, thyroid hormone receptor agonists can be used to treat diseases characterized by elevated DBI levels in subjects. Furthermore, the applicants have demonstrated that the addition of thyroid hormone receptor agonists described herein in combination with anticancer agents described herein synergistically improves the activity of the anticancer agents when treating cancer. Accordingly, disclosed herein are compositions for use in treating cancer in a subject, and methods for treating cancer in a subject by administering the compositions, wherein the composition comprises an effective amount of a thyroid hormone receptor agonist as described herein and an effective amount of an anticancer agent as described herein, wherein the effective amount of the thyroid hormone receptor agonist results in an improvement of the anticancer agent in treating the cancer. In some embodiments, the improvement is a decrease in the effective amount of the anticancer agent required to treat the cancer compared to the effective amount required to treat the cancer in the absence of an effective amount of the thyroid hormone receptor agonist. In some embodiments, the improvement is an increase in the activity of an effective amount of the anticancer agent in treating the cancer compared to the activity of the same effective amount of the anticancer agent in the absence of an effective amount of the thyroid hormone receptor agonist in treating the cancer. In some embodiments, the improvement includes enabling an anticancer agent that cannot treat the cancer in the absence of an effective amount of the agent that reduces DBI activity or expression to treat the cancer in the presence of an effective amount of the agent that reduces DBI activity or expression.

[0112] Examples of thyroid receptor agonists described herein that can be used in combination with anticancer agents to treat cancer include thyroid hormone receptor β(THR-β) agonists, such as resmethirome, sovethyrom, eprothyrom, (2R,4S)-4-(3-chlorophenyl)-2-[(4-{[4-hydroxy-3-(propan-2-yl)phenyl]methyl}-3,5-dimethylphenoxy)methyl]-1,3,2-lambda-5-dioxaphosfinan-2-one, and (4-(3-benzyl-4-hydroxybenzyl)-3,5-dimethylphenyl)oxy)methyl)phosphonic acid. In some embodiments, the thyroid hormone receptor agonist includes thyroid hormone receptor β(THR-β) agonists. In some embodiments, the thyroid hormone receptor agonist comprises resmethirome, sovethyrom, eprothyrom, (2R,4S)-4-(3-chlorophenyl)-2-[(4-{[4-hydroxy-3-(propan-2-yl)phenyl]methyl}-3,5-dimethylphenoxy)methyl]-1,3,2-lambda-5-dioxaphosfinan-2-one, or (4-(3-benzyl-4-hydroxybenzyl)-3,5-dimethylphenyl)oxy)methyl)phosphonic acid. In some embodiments, the thyroid hormone receptor agonist comprises resmethirome. In some embodiments, the thyroid hormone receptor agonist comprises sovethyrom. In some embodiments, the thyroid hormone receptor agonist comprises eprothyrom. In some embodiments, the thyroid hormone receptor agonist comprises (2R,4S)-4-(3-chlorophenyl)-2-[(4-{[4-hydroxy-3-(propan-2-yl)phenyl]methyl}-3,5-dimethylphenoxy)methyl]-1,3,2-lambda-5-dioxaphosfinan-2-one. In some embodiments, the thyroid hormone receptor agonist comprises (4-(3-benzyl-4-hydroxybenzyl)-3,5-dimethylphenyl)methyl)phosphonic acid.

[0113] In some embodiments, agents that inhibit the activity of DBI (e.g., extracellular DBI) consist of vaccine compositions suitable for inducing neutralizing autoantibodies against extracellular DBI when administered to a subject. For the purposes of this disclosure, the term “vaccine composition” is intended to mean a composition that can be administered to a human or animal to induce an immune response, which may result in the production of antibodies against extracellular DBI. Typically, a vaccine composition comprises at least one antigen derived from DBI. As used herein, the term “antigen” refers to a molecule that, when processed and presented by an MHC molecule, can be specifically bound by an antibody or a T cell receptor (TCR). As used herein, the term “antigen” also encompasses a T cell epitope. The antigen may further induce humoral and / or cellular immune responses that may be recognized by the immune system and / or result in the activation of B lymphocytes and / or T lymphocytes. The antigen may have one or more epitopes or antigenic sites (B epitopes and T epitopes). In some embodiments, the antigens of this disclosure consist of polypeptides.

[0114] In some embodiments, the antigen of the present disclosure comprises a polypeptide having at least 80% identity with the sequence or fragment (e.g., epitope) of SEQ ID NO: 1. In some embodiments, the polypeptide comprises (i) an amino acid sequence having at least 80% identity with SEQ ID NO: 1, or (ii) an amino acid sequence having at least 80% identity with an amino acid sequence in the range from amino acid residue at position 17 to amino acid residue at position 50 in SEQ ID NO: 1, or (iii) an amino acid sequence having at least 80% identity with an amino acid sequence in the range from amino acid residue at position 33 to amino acid residue at position 50 in SEQ ID NO: 1, or (iv) an amino acid sequence having at least 80% identity with an amino acid sequence in the range from amino acid residue at position 43 to amino acid residue at position 50 in SEQ ID NO: 1.

[0115] In some embodiments, polypeptides are conjugated to carrier proteins that are generally sufficiently exogenous to induce a strong immune response to the vaccine. Exemplary carrier proteins are inherently highly immunogenic. Both bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH) are commonly used as carriers in the development of conjugated vaccines when experimenting with animals, and are intended herein as carrier proteins. Proteins used in the preparation of therapeutic conjugated vaccines include, but are not limited to, numerous toxins of pathogenic bacteria and their toxoids. Numerous suitable carrier molecules include, but are not limited to, bacterial toxins or products, e.g., cholera toxin B-(CTB), diphtheria toxin, tetanus toxoid, and pertussis toxin and filamentous hemagglutinin, Shiga toxin, Pseudomonas aeruginosa exotoxin; lectins, e.g., lysine B subunit, abrin, and sweet pea lectin; subviral particles, e.g., retroviral nucleoprotein (retro NP), rabies ribonucleoprotein (rabies RNP), plant viruses (e.g., TMV, cowpee virus, and cauliflower mosaic virus), vesicular stomatitis virus-nucleocapsid protein (VSV-N), poxvirus vectors, and Semryki forest virus vectors; artificial vehicles, e.g., multiantigenic peptides (MAPs), microspheres; yeast virus-like particles (VLPs); malaria protein antigens; and other proteins and peptides, as well as any of the above modifications, derivatives, or analogues. Other useful carriers include those with the ability to enhance mucosal responses, such as the LTB family of bacterial toxins, retroviral nucleoproteins (retro-NPs), rabies ribonucleoproteins (rabies RNPs), vesicular stomatitis virus-nucleocapsid proteins (VSV-Ns), and recombinant poxvirus subunits.

[0116] anticancer drugs The systems, methods, and compositions described herein may include one or more anticancer agents and their use.

[0117] In some embodiments, one or more anticancer agents include small molecules, compounds, drugs, antibodies, their antigen-binding moieties or fragments thereof, aptamers, expression inhibitors, etc. In some embodiments, one or more anticancer agents include antibodies, their antigen-binding moieties or fragments thereof, aptamers, or expression inhibitors. In some embodiments, one or more anticancer agents include small molecules, compounds, drugs, or antibodies.

[0118] In some embodiments, one or more anticancer agents described herein include a chemotherapeutic agent, an immunotherapy agent, or both. In some embodiments, one or more anticancer agents include a chemotherapeutic agent. In some embodiments, one or more anticancer agents include a chemotherapeutic agent. In some embodiments, one or more anticancer agents include an immunotherapy agent. In some embodiments, one or more anticancer agents include a chemotherapeutic agent and an immunotherapy agent. In some embodiments, the anticancer agent includes the administration of radiotherapy to a subject having cancer.

[0119] In some embodiments, the chemotherapeutic agents described herein are cytotoxic compounds / agents used to treat cancer. In some embodiments, the chemotherapeutic agents are alkylating agents (e.g., thiotepa and cyclophosphamide); alkyl sulfonate esters (e.g., busulfan, improsulfan and pigosulfan); aziridine compounds (e.g., benzodopa, carboquan, meturedopa and uredopa); ethyleneimines; methylmelamine; altoretamine; triethylenemelamine; triethylenephosphoramide; triethylenethiophosphoramide; trimethylolmelamine; acetogenins (e.g., bratacin and bratacinone); and camptothecin. (Topotecan, etc.); Briostatin; Callistatin; CC-1065 (and its synthetic analogs, adozelesin, carzelesin, and bizelesin, etc.); Cryptophycin (Cryptophycin 1 and Cryptophycin 8, etc.); Dorastatin; Duocalmycin (and its synthetic analogs, KW-2189 and CB1-TM1, etc.); Eleutherobin; Pancratistatin; Sarcodictin; Spongestatin; Nitrogen mustards (chlorambucil, chlornafadin, colophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobenbitin, fenestrine, prednimastine, trophosphamide, and uracil mustard, etc.); nitrosoureas (carmustine, chlorozotosine, fotemustine, lomustine, nimustine, and ranimustine, etc.); antibiotics (e.g., engine antibiotics, i.e., calicheamicin, calicheamicin gamma II) and calicheamicin omega-II, etc.); dynemycin (dynemycin A, etc.); bisphosphonates (clodronate, etc.); esperamycin; neocartinostatin chromophore; related chromoprotein enediyne antibiotics chromophore; acrasinomycin; actinomycin; austramycin; azaserin; bleomycin; kakutinomycin; carabicin; kaminomycin; cartinophylline; chromomycin; dactinomycin; daunorubicin; detrubicin;6-Diazo-5-oxo-L-norleucine; Doxorubicin (morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin, etc.); Epirubicin; Esolubicin; Idarubicin; Marcelomycin; Mitomycin (mitomycin C, mycophenolic acid, nogaramycin, olibomycins, peplomycin, potiphyllomycin, puoromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin and Zolbicin, etc.); antimetabolites (methotrexate and 5-fluorouracil (5-FU), etc.); folate analogs (denopterin, methotrexate, pteropterin and trimethrexate, etc.); purine analogs (fludarabine, 6-mercaptopurine, thiamiprine and thioguanine, etc.); pyrimidine analogs (ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine and phloxuridine, etc.); androgens (carsterone, dromostanolone propionate) , epithiostanol, mepitiostane and testolactone, etc.); anti-adrenal drugs (aminoglutethimide, mitotane and trilostane, etc.); folic acid supplements (folic acid, etc.); acegraton; aldofsphamide glycoside; aminolevulinic acid; enyluracil; amsacrin; bestrabusil; bisanthren; edatraxate; defofamine; demecolsin; diazicon; elformitin; eriptinium acetate; epotilon; toggleside; gallium nitrate; hydroxyurea; lentinan; ronidynin; meitansinoids (meitansin and ansamito Syn, etc.); Mitoguazone; Mitoxantrone; Mopidammol; Nitraerine; Pentostatin; Fenamet; Pirarubicin; Rosoxantrone; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; PSK polysaccharide complex; Lazoxane; Rhizoxin; Schizofuran; Spirogermanium; Tenuazonic acid; Triadiquan; 2,2',2”-Trichlorotriethylamine; Trichothecene (T-2 toxin, Veraclin A, Loridine A and Angidin, etc.); Urethane; Vindesine; Dacarbazine; Mannomastine; Mitobronitol; Mitractol;Pipobroman; gasitosine; arabinoside; cyclophosphamide; thiotepa; taxoids (e.g., paclitaxel and docetaxel); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum complex compounds (cisplatin, oxaliplatin and carboplatin, etc.); vinblastine; platinum compounds; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (CPT-11, etc.); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids (retinoic acid, etc.); capecitabine; pharmaceutically acceptable salts; any of the above acids or derivatives; or any combination thereof. ;

[0120] In some embodiments, the chemotherapeutic agent is thiotepa, cyclophosphamide, busulfan, improsulfan, piposulfan, benzodopa, carboquan, methuredopa, bratacin, bratacinone, topotecan, adzeresin, carzeresin, bizeresin synthetic analog, cryptophycin 1, cryptophycin 8, KW-2189, CB1-TM1, chlorambucil, chlornafadin, colophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, noben Vitin, Fenesterine, Prednimustine, Trophosphamide, Uracil Mustard, Carmustine, Chlorozotocin, Fotemustine, Lomustine, Nimustine, Ranimustine, Endiyne antibiotics, Calicheamicin, Calicheamicin Gamma II, Calicheamicin Omega II, Dynemycin A, Clodronate, Morpholino-Doxorubicin, Cyanomorpholino-Doxorubicin, 2-Pyrrolino-Doxorubicin, Deoxydoxorubicin, Mitomycin C, Mycophenolic Acid, Nogaramycin, Olivomycin Syn, Peplomycin, Potiphyllomycin, Puromycin, Queramycin, Rhodolubicin, Streptonigrin, Streptozocin, Tubercidine, Ubenimex, Dinostatin, Zolubicin, Methotrexate, 5-Fluorouracil (5-FU), Denopterin, Methotrexate, Pteropterin, Trimethrexate, Fludarabine, 6-Mercaptopurine, Thiamipurine, Thioguanine, Ancitabine, Azacitidine, 6-Azauridine, Carmofur, Cytarabine, Dideoxyuridine, Doxyfluridi This includes enocitabine, floxyuridine, carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, aminoglutethimide, mitotane, trilostane, folinic acid, mytacin, ansamitosin, T-2 toxin, veraculin A, loridine A, angidin, paclitaxel, docetaxel, cisplatin, oxaliplatin, carboplatin, CPT-11, retinoic acid, any of the pharmaceutically acceptable salts, acids or derivatives of the above, or any combination thereof.

[0121] In some embodiments, the chemotherapeutic agent includes immunogenic cell death (ICD) induction activity. In some embodiments, the chemotherapeutic agent having ICD activity includes oxaliplatin, mitoxantrone, or both.

[0122] In some embodiments, the chemotherapeutic agent is a small molecule, an antibody, its antigen-binding moiety or fragment thereof, an aptamer, an expression inhibitor, a small molecule, a drug, etc. In some embodiments, the chemotherapeutic agent can be delivered to a target as the same or different molecules as one or more components described herein (e.g., immunotherapeutic agents, anti-DBI agents, corticosteroid therapeutic agents, or more than one of the following, or a combination thereof). In some embodiments, the chemotherapeutic agent can be conjugated to one or more components described herein. In some embodiments, the chemotherapeutic agent can be fused to one or more components described herein. In some embodiments, the chemotherapeutic agent can be covalently bonded to one or more components described herein. In some embodiments, the chemotherapeutic agent can be recombinantly synthesized on one or more components described herein.

[0123] In some embodiments, the immunotherapeutic agents described herein may utilize or induce polypeptides, cells, and / or factors of the innate and adaptive immune systems to recognize and treat cancer. In some embodiments, the immunotherapeutic agents described herein include agents having activity against immune checkpoints, activated effector cells, nonspecific immunostimulants (e.g., cytokines), engineered activated effector cells (e.g., chimeric antigen receptor T cells, chimeric antigen receptor NK cells), cell therapies using engineered or unengineered tumor-infiltrating lymphocytes, cancer antigen adjuvants, and the like.

[0124] In some embodiments, the immunotherapy agent includes a drug that exhibits activity against immune checkpoints.

[0125] In some embodiments, immune checkpoints include PD1 (programmed death 1), PDL1 (programmed death ligand 1), CTLA4 (cytotoxic T lymphocyte-associated protein 4), PDL2 (programmed death ligand 2), KIR (killer cell immunoglobulin-like receptor), B7-H3, B7-H4, BTLA (B and T lymphocyte attenuator), LAG3 (lymphocyte activation gene 3), TIM-3 (T cell immunoglobulin and This includes mucin domain-containing proteins, VISTA (V-domain Ig suppressor for T cell activation), ILT2 / LILRB1 (Ig-like transcript 2 / leukocyte Ig-like receptor 1), ILT3 / LILRB4 (Ig-like transcript 3 / leukocyte Ig-like receptor 4), ILT4 / LILRB2 (Ig-like transcript 4 / leukocyte Ig-like receptor 2), TIGIT (T cell immune receptor with Ig and ITIM domains), NKG2A, PVRIG, CBLB (Casitas b lineage lymphoma oncogene B), CISH (cytokine-inducible SH2-containing protein), or any combination thereof.

[0126] In some embodiments, the immunotherapy agent includes an anti-PD1 agent, an anti-PD-L1 agent, an anti-CTLA4 agent, an anti-PD-L2 agent, an anti-KIR agent, an anti-B7-H3 agent, an anti-B7-H4 agent, an anti-BTLA agent, an anti-LAG3 agent, an anti-TIM-3 agent, an anti-VISTA agent, an anti-ILT2 / LILRB1 agent, an anti-ILT3 / LILRB4 agent, an anti-ILT4 / LILRB2 agent, an anti-TIGIT agent, an anti-NKG2A agent, an anti-PVRIG agent, an anti-CBLB agent, an anti-CISH agent, or any combination thereof. In some embodiments, the immunotherapy agent includes an anti-PD1 agent. In some embodiments, the immunotherapy agent includes an anti-PD-L1 agent. In some embodiments, the immunotherapy agent includes an anti-CTLA4 agent. In some embodiments, the immunotherapy agent includes an anti-PD1 agent and an anti-PD-L1 agent. In some embodiments, the immunotherapy agent comprises an anti-PD1 agent and an anti-CTLA4 agent. In some embodiments, the immunotherapy agent comprises an anti-PD-L1 agent and an anti-CTLA4 agent. In some embodiments, the immunotherapy agent comprises an anti-PD1 agent, an anti-PD-L1 agent, and an anti-CTLA4 agent. In some embodiments, the immunotherapy agent is a small molecule, a biologic, an antibody, its antigen-binding moiety or fragment thereof, an aptamer, an expression inhibitor, a small molecule, a drug, etc.

[0127] In some embodiments, the immunotherapy agent is one or more antibodies, small molecules, or biologics against one or more immune checkpoints. In some embodiments, one or more antibodies against one or more immune checkpoints include anti-PD1 antibody, anti-PD-L1 antibody, anti-CTLA4 antibody, anti-PD-L2 antibody, anti-KIR antibody, anti-B7-H3 antibody, anti-B7-H4 antibody, anti-BTLA antibody, anti-LAG3 antibody, anti-TIM-3 antibody, anti-VISTA antibody, anti-ILT2 / LILRB1 antibody, anti-ILT3 / LILRB4 antibody, anti-ILT4 / LILRB2 antibody, anti-TIGIT antibody, anti-NKG2A antibody, anti-PVRIG antibody, anti-CBLB antibody, anti-CISH antibody, or any combination thereof. In some embodiments, the immunotherapy agent includes an anti-PD1 antibody. In some embodiments, the immunotherapy agent includes an anti-PD-L1 antibody. In some embodiments, the immunotherapy agent includes an anti-CTLA4 antibody. In some embodiments, the immunotherapy agent includes both an anti-PD1 antibody and an anti-PD-L1 antibody. In some embodiments, the immunotherapy agent comprises an anti-PD1 antibody and an anti-CTLA4 antibody. In some embodiments, the immunotherapy agent comprises an anti-PD-L1 antibody and an anti-CTLA4 antibody, or any combination thereof.

[0128] In some embodiments, one or more small molecules or biologics against one or more immune checkpoints include: ipilimumab, tremelimumab, MK-1308, FPT155, PRS010, BMS-986249, BPI-002, CBT509, JS007, ONC392, TE1254, IBI310, BR02001, CG0161, KN044, PBI5 D3H5, BCD145, ADU1604, AGEN1884, AGEN1181, CS1002, CP675206, Pembrolizumab, Nivolumab, Pidilizumab, AMP-224, BMS-936559, Semiprimab, PDR001, MDX-1105, MEDI4736, Atezolizumab, Avelumab, BMS-936559, Durvalumab, Rilurumab ( IPH2102), IPH2101, MGA271, FPA150, IMP321 (Eftilagimod Alpha), Relatrimab, MK-4280, AVA017, BI754111, ENUM006, GSK2831781, INCAGN2385, LAG3Ig, LAG525, REGN3767, Sym016, Sym022, Sym023, TSR033, TSR0 This includes 75, XmAb22841, LY3321367, MBG453, TSR-022, JNJ-61610588, MK-7684, PTZ-201, RG6058, COM902, IPH-2201, COM701, CA-327, LAG525, REGN3767, BI754111, tevoterimab, FS118, MGC018, or any combination thereof.

[0129] In some embodiments, the immunotherapy agent can be delivered to a target as the same or different molecules as one or more components described herein (e.g., an immunotherapy agent, an anti-DBI agent, a corticosteroid therapy agent, more than one of the following, or a combination thereof). In some embodiments, the immunotherapy agent can be conjugated to one or more components described herein. In some embodiments, the immunotherapy agent can be fused to one or more components described herein. In some embodiments, the immunotherapy agent can be covalently bonded to one or more components described herein. In some embodiments, the immunotherapy agent can be recombinantly synthesized on one or more components described herein.

[0130] Corticosteroid therapy The systems, methods, and compositions described herein may comprise one or more corticosteroid therapeutic agents and their use. The corticosteroid therapeutic agents described herein are anti-inflammatory corticoid activators used to treat inflammatory immune responses to diseases.

[0131] In some embodiments, one or more corticosteroid therapeutic agents described herein include cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, beclomethasone, or any combination thereof. In some embodiments, the corticosteroid therapeutic agent includes two or more of the above. In some embodiments, the corticosteroid therapeutic agent includes three or more of the above. In some embodiments, the corticosteroid therapeutic agent includes four or more of the above. In some embodiments, the corticosteroid therapeutic agent includes five or more of the above. In some embodiments, the corticosteroid therapeutic agent includes six or more of the above. In some embodiments, the corticosteroid therapeutic agent includes seven or more of the above. In some embodiments, the corticosteroid therapeutic agent includes eight or more of the above. In some embodiments, the corticosteroid therapy agent includes nine or more of the above. In some embodiments, the corticosteroid therapy agent includes ten or more of the above.

[0132] In some embodiments, one or more corticosteroid therapeutic agents include cortisol. In some embodiments, one or more corticosteroid therapeutic agents include cortisone. In some embodiments, one or more corticosteroid therapeutic agents include prednisone. In some embodiments, one or more corticosteroid therapeutic agents include prednisolone. In some embodiments, one or more corticosteroid therapeutic agents or one or more corticosteroid therapeutic portions include methylprednisolone. In some embodiments, one or more corticosteroid therapeutic agents include dexamethasone. In some embodiments, one or more corticosteroid therapeutic agents include betamethasone. In some embodiments, one or more corticosteroid therapeutic agents include triamcinolone. In some embodiments, one or more corticosteroid therapeutic agents include deflazacort. In some embodiments, one or more corticosteroid therapeutic agents include fludrocortisone acetate. In some embodiments, one or more corticosteroid therapeutic agents include deoxycorticosterone acetate. In some embodiments, one or more corticosteroid therapeutic agents include aldosterone. In some embodiments, one or more corticosteroid therapeutic agents include beclomethasone.

[0133] system The Disclosure further provides a system comprising one or more components, each comprising individually (a) an agent against an extracellular human diazepam binding inhibitor in an amount sufficient to inhibit extracellular DBI (an anti-DBI agent); (b) one or more anticancer agents, optionally including chemotherapeutic agents, immunotherapeutic agents, or any combination thereof; (c) one or more corticosteroid therapeutic agents; or one or more of any combination of (a) to (c). In some embodiments, the system comprises two or more components of (a) to (c). In some embodiments, the system comprises all of the components of (a) to (c).

[0134] In some embodiments, the system described herein includes an amount of an extracellular human diazepam binding inhibitor (anti-DBI agent) sufficient to inhibit extracellular DBI. In some embodiments, the system described herein includes one or more anticancer agents, which may optionally include chemotherapeutic agents, immunotherapeutic agents, or any combination thereof. In some embodiments, the system described herein includes one or more corticosteroid therapeutic agents.

[0135] composition This specification discloses compositions comprising: (a) one or more agents against extracellular human diazepam binding inhibitors (anti-DBI agents) in an amount sufficient to inhibit extracellular DBI in a subject; (b) one or more anticancer agents, optionally including chemotherapeutic agents, immunotherapeutic agents, or any combination thereof; (c) one or more corticosteroid therapeutic agents; or one or more of any combination of (a) to (c). In some embodiments, the composition comprises two or more components of (a) to (c). In some embodiments, the composition comprises all components of (a) to (c). In embodiments, the composition may be for use in the methods, regimens, and systems disclosed herein.

[0136] In some embodiments, the compositions described herein include an amount of an extracellular human diazepam binding inhibitor (anti-DBI agent) sufficient to inhibit extracellular DBI in a subject. In some embodiments, the compositions described herein include one or more anticancer agents, which may optionally include chemotherapeutic agents, immunotherapeutic agents, or a combination thereof. In some embodiments, the compositions described herein include one or more corticosteroid therapeutic agents.

[0137] Pharmaceutical composition: The systems, compositions, and components described herein, such as one or more agents, one or more anticancer agents, and / or one or more corticosteroid therapies, that inhibit the activity or expression of DBI (e.g., extracellular DBI), are administered to patients in the form of pharmaceutical compositions comprising a pharmaceutically acceptable carrier. Examples of pharmaceutically acceptable carriers that may be used in these compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, e.g., human serum albumin, buffers, e.g., phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts, or electrolytes, e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin. For use in administration to a subject, the compositions are formulated for administration to the subject. The compositions of this disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, orally, vaginally, or via an implanted reservoir. As used herein, the term "parent" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intrafocal, and intracranial injection or infusion techniques. The sterile injectable forms of the compositions of this disclosure may be aqueous or oily suspensions. These suspensions may be formulated according to techniques known in the art using appropriate dispersants or wetting and suspending agents. The sterile injection preparations may also be sterile injection solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile fixatives have been conventionally used as solvents or suspension media. For this purpose, any non-irritating fixative containing synthetic monoglycerides or diglycerides may be used.Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectable formulations, as are naturally pharmaceutically acceptable oils such as olive oil or castor oil, particularly their polyoxyethylated versions. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose or similar dispersants commonly used in formulations of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tween®, Span®, and other emulsifiers or bioavailability enhancers commonly used in the manufacture of pharmaceutically acceptable solids, liquids, or other dosage forms, may also be used for formulation purposes. The compositions of this disclosure may be administered orally in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions, or solutions. For oral tablets, commonly used carriers include lactose and corn starch. Typically, lubricants such as magnesium stearate are also added. For oral administration in capsule form, a useful diluent is, for example, lactose. When an aqueous suspension is required for oral use, an agent that inhibits the activity or expression of extracellular DBI is combined with an emulsifier and a suspending agent. Specific sweeteners, flavorings, or colorants may be added, if desired. Alternatively, the compositions of this disclosure may be administered in the form of suppositories for rectal administration. These can be prepared by mixing one or more agents or one or more portions with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore melts in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol. The compositions of this disclosure may also be administered topically, particularly when the target of treatment includes areas or organs that are easily accessible by topical application (including diseases of the eyes, skin, or lower intestine). Suitable topical formulations are readily prepared for each of these areas or organs. For topical application, the compositions may be formulated into a suitable ointment containing the active agent suspended or dissolved in one or more carriers.Carriers for topical administration of the compounds / agents / parts of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying waxes, and water. Alternatively, the compositions may be formulated into suitable lotions or creams containing the active agent suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester waxes, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water. Topical application to the lower bowel can be done with rectal suppositories (see above) or suitable enema formulations. Patches may also be used. The compositions of this disclosure may also be administered by nasal aerosol or inhalation. Such compositions may be prepared according to well-known techniques in the field of pharmaceutical formulations and may be prepared as a solution in physiological saline using benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and / or other conventional solubilizers or dispersants.

[0138] In some embodiments, one or more agents that inhibit the activity or expression of DBI (e.g., extracellular DBI) of the Disclosure are administered directly to the target or isolated organ using injection, a pump device and / or any machine (e.g., a bypass machine). In some embodiments, an isolated organ suitable for transplantation is perfused with a preservation solution containing an effective amount of the agent that inhibits the activity or expression of extracellular DBI. As used herein, the terms “preservation solution” or “organ preservation solution” refer to an aqueous solution having a pH of 6.5 to 7.5, comprising salts, preferably chlorides, sulfates, sodium, calcium, magnesium, and potassium; sugars, mannitol, raffinose, sucrose, glucose, fructose, lactobionic acid (which is water-resistant), or gluconic acid; antioxidants, e.g., glutathione; activators, e.g., xanthine oxidase inhibitors, e.g., allopurinol, lactate; amino acids, e.g., histidine, glutamic acid (or glutamate), tryptophan; and optionally, colloids, e.g., hydroxyethyl starch, polyethylene glycol, or dextran. In some embodiments, an apparatus for preserving organs is used, comprising an organ container filled with a preservation solution, and further comprising one or more means for injecting one or more drugs (e.g., drugs that inhibit the activity or expression of extracellular DBI) into the organ container.

[0139] Treatment method This specification discloses, among other disclosures, methods for improving the effectiveness of immunotherapy, inducing immune surveillance, inhibiting cancer progression, enhancing cancer immune surveillance, inhibiting cancer progression, improving treatment outcomes, enhancing the therapeutic effect of anticancer therapy, reducing the immunosuppressive effect of corticosteroid therapy, treating cancer, or any combination thereof, in subjects having or requiring cancer. Agents, compositions, and combinations for use in such methods are also disclosed. In some embodiments, subjects having cancer are treated by the described methods for inducing immune surveillance, inhibiting cancer progression, improving treatment outcomes, enhancing the therapeutic effect of anticancer therapy, reducing the immunosuppressive effect of corticosteroid therapy, or a combination thereof. In some embodiments, the methods described herein are compared to equivalent methods without the administration of an anti-DBI agent. In some embodiments, the absence of an anti-DBI agent means administration of an anticancer agent alone or administration of a corticosteroid therapy agent alone. In some embodiments, the absence of an anti-DBI agent means administration of an anticancer agent together with a corticosteroid therapy agent.

[0140] In some embodiments, the methods described herein include administering an amount sufficient to inhibit extracellular human DBI, an agent against an extracellular human diazepam binding inhibitor (an anti-DBI agent), or a composition comprising such an agent; one or more anticancer agents or compositions comprising such agents; and / or one or more corticosteroid therapeutic agents or compositions comprising such agents, or any combination thereof. In some embodiments, one or more anticancer agents include chemotherapeutic agents, immunotherapeutic agents, or both.

[0141] In some embodiments, methods for improving the therapeutic effect of immunotherapy in a subject, as well as agents, compositions, and combinations for use in such methods, are disclosed herein.

[0142] In some embodiments, a method for improving the therapeutic effect of immunotherapy in a subject includes administering to the subject (a) an agent that stimulates autophagy by inhibiting extracellular human diazepam binding inhibitor (DBI) (an anti-DBI agent) and (b) one or more immunotherapy agents. In some embodiments, the method includes administering to the subject (a) an agent that stimulates autophagy by inhibiting extracellular human diazepam binding inhibitor (DBI) (an anti-DBI agent), (b) one or more immunotherapy agents, and (c) one or more chemotherapeutic agents. In some embodiments, the method includes administering to the subject (a) an agent that stimulates autophagy by inhibiting extracellular human diazepam binding inhibitor (DBI) (an anti-DBI agent), (b) one or more immunotherapy agents, (c) one or more chemotherapeutic agents, and (d) one or more corticosteroid therapies, or any combination thereof. In some embodiments, the administration of an anti-DBI agent and one or more immunotherapeutic agents is sufficient to improve the therapeutic effect of immunotherapy compared to an equivalent method without the administration of an anti-DBI agent.

[0143] In some embodiments, methods for enhancing immune surveillance in subjects with cancer, as well as agents, compositions, and combinations for use in such methods, are disclosed herein.

[0144] In some embodiments, methods for enhancing immune surveillance enhance the therapeutic effect of anti-cancer therapy.

[0145] In some embodiments, a method for enhancing immune surveillance includes administering an amount of an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) sufficient to inhibit extracellular human DBI. In some embodiments, a method for enhancing immune surveillance includes administering (a) an amount of an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) sufficient to inhibit extracellular human DBI, and (b) one or more anticancer agents. In some embodiments, a method for enhancing immune surveillance includes administering (a) an amount of an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) sufficient to inhibit extracellular human DBI, and (b) one or more corticosteroid therapies. In some embodiments, a method for enhancing immune surveillance includes administering (a) an extracellular human diazepam binding inhibitor (DBI) agent (anti-DBI agent) in an amount sufficient to inhibit extracellular human DBI, (b) one or more anticancer agents, and (c) one or more corticosteroid therapies, or any combination thereof.

[0146] In some embodiments, administration of an anti-DBI agent, one or more anticancer agents, and / or one or more corticosteroid therapies is sufficient to induce one or more immune surveillance biomarkers. In some embodiments, the method of enhancing immune surveillance results in an enhanced therapeutic effect of anticancer therapy compared to an equivalent method without administration of an anti-DBI agent.

[0147] In some embodiments, the method described herein induces at least one immunosurveillance biomarker in a sample or subject, and the incidence of at least one immunosurveillance biomarker is compared to the incidence of the immunosurveillance biomarker in the absence of administration of an anti-DBI agent, CD8 + Increased T cell expression; CD4 + Decreased T cell expression; T reg cells (CD4 + Foxp3 + ) decrease; T reg cells (CD4 + Foxp3 + ) for CD8 +Increase in the ratio of T cells; T H + cells (CD4 + Foxp3 - ); increase in T H + cells (CD4 + Foxp3 - ) in Lag3 + decrease in expression; CD8 + cells in Lag3 + decrease in expression; CD4 + decrease in Foxp3 + expression in T cells; T reg ICOS + GITR + Lag3 - CD4 + decrease in T cells; or combinations thereof. In some embodiments, the methods described herein induce at least 1, 2, 3, 4, 5, 6, 7, 8 or more immune surveillance biomarkers.

[0148] In some embodiments, the methods described herein reduce the immunosuppressive effect of corticosteroid therapy compared to equivalent methods without administration of the anti-DBI agents described herein. In some embodiments, methods of reducing the immunosuppressive effect of corticosteroid therapy enhance the therapeutic effect of anti-cancer therapy. In some embodiments, the methods of enhancing immune surveillance described herein induce one or more immune surveillance biomarkers compared to equivalent methods without administration of the anti-DBI agents described herein, thereby reducing the immunosuppressive effect of corticosteroid therapy in the administered subject. Without being bound by theory, it is believed that administration of the anti-DBI agents described herein together with anti-cancer agents and corticosteroid therapy agents induces an anti-inflammatory immune response. In some embodiments, such responses promote the antigen response and the cytotoxic T cell environment.

[0149] In some embodiments, provided herein is a method of reducing the immunosuppressive effect of corticosteroid therapy in a subject suffering from cancer, the method comprising administering an agent (anti-DBI agent) against extracellular human diazepam-binding inhibitor (DBI) in an amount sufficient to inhibit extracellular human DBI. In some embodiments, the method of reducing the immunosuppressive effect of corticosteroid therapy comprises (a) an agent (anti-DBI agent) against extracellular human diazepam-binding inhibitor (DBI) in an amount sufficient to inhibit extracellular human DBI, and (b) one or more anti-cancer agents. In some embodiments, the method of reducing the immunosuppressive effect of corticosteroid therapy comprises (a) an agent (anti-DBI agent) against extracellular human diazepam-binding inhibitor (DBI) in an amount sufficient to inhibit extracellular human DBI, and (b) one or more corticosteroid therapy agents. In some embodiments, the method of reducing the immunosuppressive effect of corticosteroid therapy comprises (a) an agent (anti-DBI agent) against extracellular human diazepam-binding inhibitor (DBI) in an amount sufficient to inhibit extracellular human DBI, and (b) one or more anti-cancer agents, and (c) one or more corticosteroid therapy agents, or any combination thereof.

[0150] In some embodiments, the reduction in immunosuppressive effect described herein is, compared to an equivalent method without administration of an anti-DBI agent, (a) an increase in CD8 + T cell expression; (b) a decrease in CD4 + T cell expression; (c) a decrease in T reg cells (CD4 + Foxp3 + ); (d) an increase in the ratio of CD8 reg T cells to T + cells (CD4 + Foxp3 + ); (e) an increase in T H + cells (CD4 + Foxp3 - ); (f) an increase in Lag3 H in T + + cells (CD4 - Foxp3 +Decreased expression of (g)CD8 + Lag3 in T cells + Decreased expression of (h)CD4 + Foxp3 in T cells + Decreased expression of (i)T reg ICOS + GITR + Lag3 - CD4 + (a) a decrease in T cells; (j) a decrease in tumor volume; (k) a decrease in cancer cell proliferation; (l) an increase in cancer cell death; (m) a decrease in cancer growth; (n) an increase in survival rate; (o) an increase in cancer-free occurrence rate; (p) an increase in time to death; or any one or more combinations of (a) to (p) in the sample.

[0151] In some embodiments, the reduction in immunosuppressive effect described herein includes any one of (a) to (p). In some embodiments, the reduction in immunosuppressive effect described herein includes any two or more of (a) to (p). In some embodiments, the reduction in immunosuppressive effect described herein includes any three or more of (a) to (p). In some embodiments, the reduction in immunosuppressive effect described herein includes any four or more of (a) to (p). In some embodiments, the reduction in immunosuppressive effect described herein includes any five or more of (a) to (p). In some embodiments, the reduction in immunosuppressive effect described herein includes any six or more of (a) to (p). In some embodiments, the reduction in immunosuppressive effect described herein includes any seven or more of (a) to (p). In some embodiments, the reduction in immunosuppressive effect described herein includes any eight or more of (a) to (p). In some embodiments, the reduction of immunosuppressive effects described herein includes any nine or more of (a) to (p). In some embodiments, the reduction of immunosuppressive effects described herein includes any ten or more of (a) to (p). In some embodiments, the reduction of immunosuppressive effects described herein includes any eleven or more of (a) to (p). In some embodiments, the reduction of immunosuppressive effects described herein includes any twelve or more of (a) to (p). In some embodiments, the reduction of immunosuppressive effects described herein includes any thirteen or more of (a) to (p). In some embodiments, the reduction of immunosuppressive effects described herein includes any fourteen or more of (a) to (p). In some embodiments, the reduction of immunosuppressive effects described herein includes any fifteen or more of (a) to (p). In some embodiments, the reduction of immunosuppressive effects described herein includes sixteen of (a) to (p).

[0152] In some embodiments, methods for inhibiting the progression of cancer in cancer patients, as well as agents, compositions, and combinations for use in such methods, are disclosed herein. In some embodiments, the cancer patients are undergoing chemotherapy.

[0153] In some embodiments, a method to inhibit cancer progression includes administering to a cancer patient an amount of an extracellular human diazepam binding inhibitor (anti-DBI agent) sufficient to inhibit extracellular human DBI. In some embodiments, a method to inhibit cancer progression includes administering to a cancer patient (a) an amount of an extracellular human diazepam binding inhibitor (anti-DBI agent) sufficient to inhibit extracellular human DBI, and (b) 1 or more immunotherapy agents. In some embodiments, a method to inhibit cancer progression includes administering to a cancer patient (a) an amount of an extracellular human diazepam binding inhibitor (anti-DBI agent) sufficient to inhibit extracellular human DBI, and (b) 1 or more corticosteroid therapy agents. In some embodiments, a method to inhibit cancer progression includes administering to a cancer patient (a) an amount of an extracellular human diazepam binding inhibitor (anti-DBI agent) sufficient to inhibit extracellular human DBI, (b) 1 or more anticancer agents, and (c) 1 or more corticosteroid therapy agents, or any combination thereof. In some embodiments, administration of an anti-DBI agent, one or more anticancer agents, and / or one or more corticosteroid therapies is sufficient to inhibit one or more cancer progression biomarkers compared to an equivalent method without administration of an anti-DBI agent.

[0154] In some embodiments, the methods described herein inhibit at least one cancer progression biomarker in a subject or sample, and compared to the expression of at least one cancer progression biomarker in an equivalent manner without administration of the anti-DBI agent described herein, the at least one cancer progression biomarker includes a decrease in tumor volume; a decrease in cancer cell proliferation; an increase in cancer cell death; a decrease in cancer growth; or a combination thereof. In some embodiments, the methods described herein induce at least one, two, three or more cancer progression biomarkers.

[0155] In some embodiments, methods for inhibiting cancer progression inhibit at least one cancer progression biomarker in a subject, thereby improving the treatment outcome in that subject. In some embodiments, methods for improving the treatment outcome in a subject are disclosed herein.

[0156] In some embodiments, a method to improve the treatment outcome of a subject includes administering an amount of an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) sufficient to inhibit extracellular human DBI. In some embodiments, a method to improve the treatment outcome of a subject includes administering (a) an amount of an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) sufficient to inhibit extracellular human DBI, and (b) one or more anticancer agents. In some embodiments, a method to improve the treatment outcome of a subject includes administering (a) an amount of an extracellular human diazepam-binding inhibitor (DBI) agent (anti-DBI agent) sufficient to inhibit extracellular human DBI, and (b) one or more corticosteroid therapies. In some embodiments, a method for improving the treatment outcome of a subject includes administering (a) an extracellular human diazepam binding inhibitor (DBI) agent (anti-DBI agent) in an amount sufficient to inhibit extracellular human DBI, (b) one or more anticancer agents, and (c) one or more corticosteroid therapies, or any combination thereof.

[0157] In some embodiments, improvement in therapeutic outcome in a subject or in vitro sample includes a reduction in tumor volume; a reduction in cancer cell proliferation; an increase in cancer cell death; a reduction in cancer growth; or a combination thereof, compared to the expression of at least one cancer progression biomarker in an equivalent manner without administration of an anti-DBI agent as described herein.

[0158] In some embodiments, methods for treating cancer in subjects requiring treatment, as well as agents, compositions, and combinations for use in such methods, are disclosed herein. In some embodiments, the method involves administering to a subject an amount of an agent against an extracellular human diazepam binding inhibitor (anti-DBI agent) sufficient to inhibit extracellular human DBI.

[0159] In some embodiments, a method for treating cancer comprises administering to a subject (a) an agent against extracellular human diazepam binding inhibitors (anti-DBI agents) in an amount sufficient to inhibit extracellular human DBI, and (b) one or more anticancer agents. In some embodiments, a method for treating cancer comprises administering to a subject (a) an agent against extracellular human diazepam binding inhibitors (anti-DBI agents) in an amount sufficient to inhibit extracellular human DBI, and (b) one or more anticancer agents. In some embodiments, a method for treating cancer comprises administering to a subject (a) an agent against extracellular human diazepam binding inhibitors (anti-DBI agents) in an amount sufficient to inhibit extracellular human DBI, and (b) one or more corticosteroid therapeutic agents. In some embodiments, a method for treating cancer involves administering to a subject (a) an agent against extracellular human diazepam binding inhibitors (anti-DBI agent) in an amount sufficient to inhibit extracellular human DBI, (b) one or more anticancer agents, and (c) one or more corticosteroid therapies, or any combination thereof. In some embodiments, the administration of the anti-DBI agent, one or more anticancer agents, and / or one or more corticosteroid therapies constitutes a therapeutic dose sufficient to treat cancer.

[0160] In some embodiments, cancer treatment was compared with an equivalent method without the administration of an anti-DBI agent, in which CD8 was present in the sample. + Increased T cell expression; CD4 + Decreased T cell expression; T reg cells (CD4 + Foxp3 + ) decrease; T reg cells (CD4 + Foxp3 + ) for CD8 + Increased ratio of T cells; T H + cells (CD4 + Foxp3 - ) increase; T H + cells (CD4 + Foxp3 - ) Lag3 + Decreased expression of CD8 + Lag3 in T cells + Decreased expression of CD4 + Foxp3 in T cells + Decreased expression; T reg ICOS + GITR + Lag3 - CD4 + This can be measured by: a decrease in T cells; a decrease in tumor volume; a decrease in cancer cell proliferation; an increase in cancer cell death; a decrease in cancer growth; an increase in survival rates; a rate of cancer-free occurrences; an increase in time to death; or a combination thereof.

[0161] In some embodiments, a method for treating cancer in a subject requiring treatment of cancer is disclosed herein, comprising administering to the subject (a) an agent against extracellular human diazepam binding inhibitors (anti-DBI agent) in an amount sufficient to inhibit extracellular DBI, (b) one or more anticancer agents, optionally including chemotherapeutic agents, immunotherapeutic agents, or combinations thereof, and (c) one or more corticosteroid therapies, or any combination of (a) to (c), wherein the administration of the anti-DBI agent and one or more anticancer agents is sufficient to treat the cancer compared to an equivalent method without the anti-DBI agent. In some embodiments, the method for treating cancer comprises administering two or more of (a) to (c) to the subject. In some embodiments, the method for treating cancer comprises administering all of (a) to (c) to the subject.

[0162] In some embodiments, the methods described herein compare CD8+ T cell expression in a sample or subject with CD8+ T cell expression in the absence of administration of the anti-DBI agent described herein. + It induces an increase in T cell expression. In some embodiments, CD8 + T cell expression increases by a maximum of approximately 10%. In some embodiments, CD8 + T cell expression increases by only about 9%. In some embodiments, CD8 + T cell expression increases by only about 8%. In some embodiments, CD8 + T cell expression increases by only about 7%. In some embodiments, CD8 + T cell expression increases by only about 6%. In some embodiments, CD8 + T cell expression increases by only about 5%. In some embodiments, CD8 + T cell expression increases by only about 4%. In some embodiments, CD8 + T cell expression increases by only about 3%. In some embodiments, CD8 + T cell expression increases by only about 2%. In some embodiments, CD8 + T cell expression increases by only about 1%. In some embodiments, CD8 +T cell expression increases by only about 0.5%. In some embodiments, CD8 + T cell expression increases by only about 0.1%.

[0163] In some embodiments, the methods described herein refer to the CD4 in the absence of administration of the anti-DBI agent described herein. + Compared to T cell expression, CD4 in the sample or subject + It induces a decrease in T cell expression. In some embodiments, CD4 + T cell expression decreases by only about 5% at most. In some embodiments, CD4 + T cell expression decreases by only about 4%. In some embodiments, CD4 + T cell expression decreases by only about 3%. In some embodiments, CD4 + T cell expression decreases by only about 2%. In some embodiments, CD4 + T cell expression decreases by only about 1%. In some embodiments, CD4 + T cell expression decreases by only about 0.5%. In some embodiments, CD4 + T cell expression decreases by only about 0.1%.

[0164] In some embodiments, the methods described herein provide a T in a sample or subject compared to an equivalent method without administration of the anti-DBI agent described herein. reg cells (CD4 + Foxp3 + This induces a decrease in T. In some embodiments, reg cells (CD4 + Foxp3 + ) Expression is reduced by only about 1% at most. In some embodiments, T reg cells (CD4 + Foxp3 + ) Expression decreases by only about 0.9%. In some embodiments, T reg cells (CD4 + Foxp3 + ) Expression decreases by only about 0.8%. In some embodiments, T reg cells (CD4 + Foxp3+ ) Expression is reduced by only about 0.7%. In some embodiments, T reg cells (CD4 + Foxp3 + ) Expression is reduced by only about 0.6%. In some embodiments, T reg cells (CD4 + Foxp3 + ) Expression is reduced by only about 0.5%. In some embodiments, T reg cells (CD4 + Foxp3 + ) Expression decreases by only about 0.4%. In some embodiments, T reg cells (CD4 + Foxp3 + ) Expression decreases by only about 0.3%. In some embodiments, T reg cells (CD4 + Foxp3 + ) Expression is reduced by only about 0.2%. In some embodiments, T reg cells (CD4 + Foxp3 + Expression decreases by only about 0.1%.

[0165] In some embodiments, the methods described herein provide a T in a sample or subject compared to an equivalent method without administration of the anti-DBI agent described herein. reg cells (CD4 + Foxp3 + ) for CD8 + It induces an increase in the proportion of T cells. In some embodiments, CD8 + T cells vs. T reg cells (CD4 + Foxp3 + The ratio of ) increases to a maximum of approximately 150:1. In some embodiments, CD8 + T cells vs. T reg cells (CD4 + Foxp3 + The ratio of ) increases to approximately 125:1. In some embodiments, CD8 + T cells vs. T reg cells (CD4 + Foxp3 +The ratio of ) increases to approximately 100:1. In some embodiments, CD8 + T cells vs. T reg cells (CD4 + Foxp3 + The ratio of ) increases to approximately 75:1. In some embodiments, CD8 + T cells vs. T reg cells (CD4 + Foxp3 + The ratio of ) increases to approximately 50:1. In some embodiments, CD8 + T cells vs. T reg cells (CD4 + Foxp3 + The ratio of ) increases to approximately 25:1.

[0166] In some embodiments, the methods described herein are equivalent to the methods described herein in the absence of administration of the anti-DBI agent described herein. H + cells (CD4 + Foxp3 - Compared to the amount of T in the sample or subject, H + cells (CD4 + Foxp3 - This induces an increase in T. In some embodiments, H + cells (CD4 + Foxp3 - ) Cells increase by up to approximately 1%. In some embodiments, T H + cells (CD4 + Foxp3 - ) The number of cells increases by only about 0.9%. In some embodiments, T H + cells (CD4 + Foxp3 - ) The number of cells increases by only about 0.8%. In some embodiments, T H + cells (CD4 + Foxp3 - ) The number of cells increases by only about 0.7%. In some embodiments, T H + cells (CD4 + Foxp3 - ) The number of cells increases by only about 0.6%. In some embodiments, T H + cells (CD4 + Foxp3- ) The number of cells increases by only about 0.5%. In some embodiments, T H + cells (CD4 + Foxp3 - ) The number of cells increases by only about 0.4%. In some embodiments, T H + cells (CD4 + Foxp3 - ) The number of cells increases by only about 0.3%. In some embodiments, T H + cells (CD4 + Foxp3 - ) The number of cells increases by only about 0.2%. In some embodiments, T H + cells (CD4 + Foxp3 - The number of cells increases by only about 0.1%.

[0167] In some embodiments, the methods described herein provide a T in a sample or subject compared to equivalent methods without the administration of the anti-DBI agents described herein and the agents, compositions, and combinations for use in such a manner. H + cells (CD4 + Foxp3 - ) Lag3 + It induces a decrease in the expression of T. In some embodiments, H + cells (CD4 + Foxp3 - ) Lag3 + The expression of is reduced by only about 10% at most. In some embodiments, T H + cells (CD4 + Foxp3 - ) Lag3 + The expression of is reduced by only about 9%. In some embodiments, T H + cells (CD4 + Foxp3 - ) Lag3 + The expression of is reduced by only about 8%. In some embodiments, T H + cells (CD4 + Foxp3 - ) Lag3 + The expression of is reduced by only about 7%. In some embodiments, TH + cells (CD4 + Foxp3 - ) Lag3 + The expression of is reduced by only about 6%. In some embodiments, T H + cells (CD4 + Foxp3 - ) Lag3 + The expression of is reduced by only about 5%. In some embodiments, T H + cells (CD4 + Foxp3 - ) Lag3 + Expression is reduced by only about 4%. In some embodiments, T H + cells (CD4 + Foxp3 - ) Lag3 + The expression of is reduced by only about 3%. In some embodiments, T H + cells (CD4 + Foxp3 - ) Lag3 + The expression of is reduced by only about 2%. In some embodiments, T H + cells (CD4 + Foxp3 - ) Lag3 + The expression of [the substance] decreases by only about 1%.

[0168] In some embodiments, the methods described herein provide a CD8 in a sample or subject compared to equivalent methods without the administration of the anti-DBI agents described herein and the agents, compositions, and combinations for use in such a method. + Lag3 in T cells + It induces a decrease in the expression of CD8. In some embodiments, + Lag3 in T cells + The expression of CD8 is reduced by only about 10% at most. In some embodiments, CD8 + Lag3 in T cells + The expression of CD8 decreases by only about 9%. In some embodiments, CD8 + Lag3 in T cells +The expression of CD8 decreases by only about 8%. In some embodiments, CD8 + Lag3 in T cells + The expression of CD8 decreases by only about 7%. In some embodiments, CD8 + Lag3 in T cells + The expression of CD8 decreases by only about 6%. In some embodiments, CD8 + Lag3 in T cells + The expression of CD8 decreases by only about 5%. In some embodiments, CD8 + Lag3 in T cells + The expression is reduced by only about 4%. In some embodiments, CD8 + Lag3 in T cells + The expression of CD8 decreases by only about 3%. In some embodiments, CD8 + Lag3 in T cells + The expression of CD8 decreases by only about 2%. In some embodiments, CD8 + Lag3 in T cells + The expression of [the substance] decreases by only about 1%.

[0169] In some embodiments, the methods described herein provide a CD4 level in a sample or subject compared to equivalent methods in the absence of anti-DBI agents described herein. + Foxp3 in T cells + It induces a decrease in the expression of CD4. + Foxp3 in T cells + The expression is reduced by only about 20% at most. In some embodiments, CD4 + Foxp3 in T cells + The expression is reduced by only about 19%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 18%. In some embodiments, CD4 + Foxp3 in T cells + The expression of CD4 decreases by only about 17%. In some embodiments, CD4 + Foxp3 in T cells +The expression is reduced by only about 16%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 15%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 14%. In some embodiments, CD4 + Foxp3 in T cells + The expression is reduced by only about 13%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 12%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 11%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 10%. In some embodiments, CD4 + Foxp3 in T cells + The expression is reduced by only about 9%. In some embodiments, CD4 + Foxp3 in T cells + The expression is reduced by only about 8%. In some embodiments, CD4 + Foxp3 in T cells + The expression of CD4 decreases by only about 7%. In some embodiments, CD4 + Foxp3 in T cells + The expression of CD4 decreases by only about 6%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 5%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 4%. In some embodiments, CD4 + Foxp3 in T cells + The expression of CD4 decreases by only about 3%. In some embodiments, CD4 + Foxp3 in T cells + Expression is reduced by only about 2%. In some embodiments, CD4 +Foxp3 in T cells + The expression of [the substance] decreases by only about 1%.

[0170] In some embodiments, the methods described herein provide a T in a sample or subject compared to an equivalent method without administration of the anti-DBI agent described herein. reg ICOS + GITR + Lag3 - CD4 + It induces a decrease in T cells. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 30% at most. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 29%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 28%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 27%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 26%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 25%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 24%. In some embodiments, ICOS + GITR + Lag3 - CD4 +T cells are reduced by only about 23%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 22%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 21%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 20%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 19%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 18%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 17%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 16%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 15%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 14%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 13%. In some embodiments, ICOS + GITR+ Lag3 - CD4 + T cells are reduced by only about 12%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 11%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 10%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells decrease by only about 9%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells decrease by only about 8%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells decrease by only about 7%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells decrease by only about 6%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells are reduced by only about 5%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells decrease by only about 4%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells decrease by only about 3%. In some embodiments, ICOS + GITR + Lag3 - CD4 +T cells decrease by only about 2%. In some embodiments, ICOS + GITR + Lag3 - CD4 + T cells decrease by only about 1%.

[0171] In some embodiments, the methods described herein result in a higher CD8 in a sample or subject compared to equivalent methods without the administration of the anti-DBI agent described herein. + T cells vs. T H + cells (CD4 + Foxp3 - ) maintain the ratio. In some embodiments, CD8 + T cells vs. T H + cells (CD4 + Foxp3 - The ratio of ) is maintained within approximately 3%. In some embodiments, CD8 + T cells vs. T H + cells (CD4 + Foxp3 - The ratio of ) is maintained within approximately 2%. In some embodiments, CD8 + T cells vs. T H + cells (CD4 + Foxp3 - The ratio of ) is maintained within approximately 1%. In some embodiments, CD8 + T cells vs. T H + cells (CD4 + Foxp3 - The ratio of ) is maintained within approximately 0.5%. In some embodiments, CD8 + T cells vs. T H + cells (CD4 + Foxp3 - The ratio of ) will be kept within approximately 0.1%.

[0172] In some embodiments, the methods described herein result in a higher CD4 concentration in a sample or subject compared to equivalent methods without the administration of anti-DBI agents described herein. + CD8 against T cells + Maintain the ratio of T cells. In some embodiments, CD8 +T cell vs. CD4 + The proportion of T cells is maintained within approximately 1%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.9%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.8%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.7%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.6%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.5%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.4%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.3%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.2%. In some embodiments, CD8 + T cell vs. CD4 + The proportion of T cells is maintained within approximately 0.1%.

[0173] In some embodiments, the methods described herein provide a higher T in a sample or subject compared to an equivalent method without administration of an anti-DBI agent. H ICOS int GITR + Lag3 - PD1 - CD4 + Maintain T cell development. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 +The T cell development rate is maintained within approximately 1%. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.9%. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.8%. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.7%. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.6%. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.5%. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.4%. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.3%. In some embodiments, T H ICOS int GITR+ Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.2%. In some embodiments, T H ICOS int GITR + Lag3 - PD1 - CD4 + The T cell development rate is maintained within approximately 0.1%.

[0174] In some embodiments, the tumor volume is reduced by approximately 60%. In some embodiments, the tumor volume is reduced by approximately 50%. In some embodiments, the tumor volume is reduced by approximately 40%. In some embodiments, the tumor volume is reduced by approximately 30%. In some embodiments, the tumor volume is reduced by approximately 20%. In some embodiments, the tumor volume is reduced by approximately 10%. In some embodiments, the tumor volume is reduced by approximately 5%.

[0175] In some embodiments, the reduction in tumor volume induces a reduction in tumor area compared to an equivalent method without administration of the anti-DBI agent described herein.

[0176] In some embodiments, the methods described herein induce a reduction in cancer cell proliferation in a sample or subject compared to an equivalent method at an equivalent time point without administration of the anti-DBI agent described herein.

[0177] In some embodiments, the methods described herein induce increased cancer cell death in a sample or subject compared to equivalent methods without the administration of the anti-DBI agent described herein.

[0178] In some embodiments, the methods described herein induce a reduction in cancer growth in a subject or sample compared to an equivalent method without administration of the anti-DBI agent described herein.

[0179] Treatment regimen This disclosure also provides therapeutic regimens for use as anticancer therapies, as well as agents, compositions, and combinations for use in such regimens. In some embodiments, the therapeutic regimen comprises (a) an agent against extracellular human diazepam binding inhibitors (anti-DBI agent) in an amount sufficient to inhibit extracellular human DBI, and (b) one or more anticancer agents. In some embodiments, the anti-DBI agent and one or more anticancer agents are present in the therapeutic regimen in an amount sufficient to induce one or more immunosurveillance biomarkers in the subject at the time of administration to the subject. In some embodiments, the therapeutic regimen causes an enhancement of the therapeutic effect of the anticancer therapy compared to an equivalent regimen without the administration of an anti-DBI agent. In some embodiments, the absence of administration of an anti-DBI agent is administration of an anticancer agent alone or administration of a corticosteroid therapy agent alone. In some embodiments, the absence of administration of an anti-DBI agent is administration of an anticancer agent together with corticosteroid therapy.

[0180] This disclosure further provides therapeutic regimens for treating cancer, as well as agents, compositions, and combinations for use in such regimens. In some embodiments, the therapeutic regimen comprises (a) an agent against extracellular human diazepam binding inhibitors (anti-DBI agent) in an amount sufficient to inhibit extracellular human DBI, and (b) one or more anticancer agents. In some embodiments, the anti-DBI agent and the preceding one or more anticancer agents are present in the therapeutic regimen in an amount sufficient to inhibit one or more cancer progression biomarkers in the subject at the time of administration to the subject, compared to an equivalent regimen without administration of the anti-DBI agent. In some embodiments, the absence of administration of the anti-DBI agent is administration of an anticancer agent alone or administration of a corticosteroid therapy agent alone. In some embodiments, the absence of administration of the anti-DBI agent is administration of an anticancer agent together with a corticosteroid therapy agent.

[0181] In some embodiments, the anti-DBI agent and one or more anticancer agents are separate components administered together. In some embodiments, the anti-DBI agent and one or more anticancer agents are separate components administered sequentially.

[0182] In some embodiments, the anti-DBI agent is administered at least about four weeks before one or more anticancer drugs. In some embodiments, the anti-DBI agent is administered at least about three weeks before one or more anticancer drugs. In some embodiments, the anti-DBI agent is administered at least about two weeks before one or more anticancer drugs. In some embodiments, the anti-DBI agent is administered at least about one week before one or more anticancer drugs.

[0183] In some embodiments, an anti-DBI agent and one or more anticancer agents are co-administered.

[0184] In some embodiments, one or more corticosteroid therapeutic agents are co-administered with an anti-DBI agent and one or more anticancer agents. In some embodiments, one or more corticosteroid therapeutic agents are co-administered with an anti-DBI agent. In some embodiments, one or more corticosteroid therapeutic agents are co-administered with one or more anticancer agents. In some embodiments, one or more corticosteroid therapeutic agents are co-administered with an anti-DBI agent and one or more anticancer agents. In some embodiments, one or more corticosteroid therapeutic agents are co-administered with an anti-DBI agent. In some embodiments, one or more corticosteroid therapeutic agents are co-administered with one or more anticancer agents.

[0185] In some embodiments, one or more corticosteroid therapeutic agents are administered sequentially together with an anti-DBI agent and one or more anticancer agents. In some embodiments, one or more corticosteroid therapeutic agents are administered sequentially together with an anti-DBI agent. In some embodiments, one or more corticosteroid therapeutic agents are administered sequentially together with one or more anticancer agents. In some embodiments, one or more corticosteroid therapeutic agents are administered sequentially together with an anti-DBI agent and one or more anticancer agents. In some embodiments, one or more corticosteroid therapeutic agents are administered sequentially together with an anti-DBI agent. In some embodiments, one or more corticosteroid therapeutic agents are administered sequentially together with one or more anticancer agents.

[0186] In some embodiments, any two or more of the following are co-administered: an anti-DBI agent, one or more anticancer agents, and one or more corticosteroid therapies.

[0187] cancer In some embodiments, cancer is characterized by a suppressive tumor microenvironment. In some embodiments, the cancers described herein utilize effective immune recognition and other mechanisms to evade elimination by the immune system. In some embodiments, cancers such as immunocompetent tumors generate a suppressive microenvironment that downregulates the immune response. In some embodiments, the suppressive tumor microenvironment includes the expression of immunosuppressive cells and factors (regulatory T cells, cancer-associated fibroblasts, myeloid inhibitory cells, and other cell types, etc.), hypoxia, low pH, and exhibits other endogenous and dynamic immunosuppressive properties. For example, the tumor microenvironment may be inherently immunosuppressive to protect cancer cells from immune surveillance, but it can also be dynamically adapted to rapid tumor growth and progression, and to counter stress and injury conditions such as chemotherapy.

[0188] In some embodiments of this disclosure, the cancer is refractory cancer. In some embodiments, the cancer does not respond to treatment. In some embodiments, cancer cells have a way of defending themselves against chemotherapeutic agents. In some embodiments, cancer cells have a way of defending themselves against immunotherapeutic agents. In some embodiments, cancer cells have a way of defending themselves against biological agents. In some embodiments, cancer cells have a way of defending themselves against radiotherapy.

[0189] In some embodiments, the cancer is resistant to immune checkpoint inhibitor therapy. In some embodiments, the cancer is selected from solid tumors, hematological tumors, skin cancers, tissue cancers, organ cancers, bone cancers, cartilage cancers, blood cancers, vascular cancers, primary cancers, metastatic cancers, bladder cancers, bone marrow cancers, brain cancers, breast cancers, colon cancers, esophageal cancers, gastrointestinal cancers, gingival cancers, kidney cancers, liver cancers, lung cancers, nasopharyngeal cancers, cervical cancers, ovarian cancers, prostate cancers, stomach cancers, testicular cancers, tongue cancers, and uterine cancers. In some embodiments, the cancer is a solid tumor cancer. In some embodiments, the cancer is a fibrous cancer.

[0190] In some embodiments, cancer is defined as cancer, lung cancer, non-small cell lung cancer, breast cancer, neoplastic cancer, undifferentiated carcinoma, giant cell carcinoma, spindle cell carcinoma, small cell carcinoma, papillary carcinoma, squamous cell carcinoma, lymphoepithelial carcinoma, basal cell carcinoma, matrix carcinoma, transitional cell carcinoma, papillary transitional cell carcinoma, adenocarcinoma, gastrinoma, cholangiocarcinoma, hepatocellular carcinoma, combination of hepatocellular carcinoma and cholangiocarcinoma, cord-like adenocarcinoma, adenoid cystic carcinoma, adenocarcinoma in adenomatous polyps, familial adenomatous polyposis, solid tumors, carcinoid tumors, bronchioloalveolar adenocarcinoma, papillary adenocarcinoma, chromophobe carcinoma, oxiphilic adenocarcinoma, basophilic carcinoma, clear cell adenocarcinoma, granular cell carcinoma, follicular adenocarcinoma, papillary adenocarcinoma, follicular adenocarcinoma, non-encapsulating sclerosing carcinoma.Carcinoma, adrenocortical carcinoma, endometrioid carcinoma, adnexal carcinoma, apocrine gland carcinoma, sebaceous gland carcinoma, ceruminous gland carcinoma, mucoepidermoid carcinoma, cystadenocarcinoma, papillary cystadenocarcinoma, papillary serous cystadenocarcinoma, mucinous cystadenocarcinoma, mucinous adenocarcinoma, signet ring cell carcinoma, invasive ductal carcinoma, medullary carcinoma, lobular carcinoma, inflammatory carcinoma, Paget's disease, breast cancer, acinar cell carcinoma, adenosquamous cell carcinoma, adenocarcinoma with squamous metaplasia, thymoma, ovarian stromal tumor, follicular cell tumor, granulosa cell tumor, roblastoma, Sertoli Cellular carcinoma, Leydig cell tumor, lipid cell tumor, accessory ganglion, extramammary accessory ganglion, pheochromocytoma, glomus angiosarcoma, melanoma, achromatic melanoma, superficial spreading melanoma, melanoma within a giant pigmented nevus, epithelioid cell melanoma, blue nevus, sarcoma, fibrosarcoma, fibrous histiocytoma, myxosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, fetal rhabdomyosarcoma, alveolar rhabdomyosarcoma, stromal sarcoma, mixed tumor, Müllerian mixed tumor, nephroblastoma, hepatoblastoma, Carcinosarcoma, mesenchymal tumor, Brenner tumor, phyllodes tumor, synovial sarcoma, mesothelioma, ovarian tumor, embryonal carcinoma, teratoma, ovarian goiter, choriocarcinoma, mesonephroma, angiosarcoma, hemangioendothelioma, Kaposi's sarcoma, hemangiopericytoma, lymphangiosarcoma, osteosarcoma, subcortical osteosarcoma, chondrosarcoma, chondroblastoma, mesenchymal chondrosarcoma, giant cell tumor of bone, Ewing's sarcoma, odontogenic tumor, ameloblastic odontogenic sarcoma, ameloblastoma, ameloblastic fibrosarcoma, pineal tumor, chordoma, glioma, ependymoma Astrocytoma, protoplasmic astrocytoma, fibrous astrocytoma, astroblastoma, glioblastoma, oligodendroglioma, oligodendroglioma, primitive neuroectodermal tumor, cerebellar sarcoma, gangliblastoma, neuroblastoma, retinoblastoma, olfactory neuron tumor, meningioma, neurofibrosarcoma, schwannoma, granulocyte tumor, lymphoma, Hodgkin's disease, Hodgkin's lymphoma, paragranuloma, small lymphocytic lymphoma, large cell diffuse lymphoma, follicular lymphoma, mycosis fungoides, and other specified non-Hodgkin lymphomas. The cancer is selected from specified non-Hodgkin's lymphoma, histiocytic proliferative disorder, multiple myeloma, mast cell sarcoma, intestinal immunoproliferative disorder, leukemia, lymphocytic leukemia, plasma cell leukemia, erythroleukemia, lymphosarcoma cell leukemia, myeloid leukemia, basophilic leukemia, eosinophilic leukemia, monocytic leukemia, mast cell leukemia, megakaryoblastic leukemia, myeloid sarcoma, and hairy cell leukemia. In some embodiments, the cancer is malignant cancer.

[0191] In some embodiments, the cancer is selected from carcinoma, lung cancer, non-small cell lung cancer, and breast cancer. In some embodiments, the cancer is carcinoma. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is non-small cell lung cancer. In some embodiments, the cancer is breast cancer.

[0192] In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a fibrous tumor. In some embodiments, the subject is a human.

[0193] While preferred embodiments of the Disclosure have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided only as examples. Without departing from the Disclosure, those skilled in the art will conceive of numerous variations, modifications, and substitutions. It should be understood that various alternative forms to the embodiments of the Disclosure described herein may be used when carrying out the Disclosure. The following claims define the scope of the Disclosure, and the methods and structures within these claims, as well as their equivalents, are intended to be encompassed thereby. [Examples]

[0194] Examples Example 1: The role of DBI in immunotherapy and corticosteroid therapy material and method Cell culture and treatment Both syngeneic cancer cell lines were maintained in Dulbecco's Modified Eagle Medium (fibrosarcoma cell line MCA205, lung cancer TC-1) or Roswell Park Memorial Institute Medium 1640 (mammary cancer E0771), supplemented with HEPES and 10% fetal bovine serum, and subcultured three times a week.

[0195] Mouse experiment C57BL / 6J mice were housed under temperature-controlled conditions and given free access to food and water. The experiment was conducted under FELASA guidelines and approved by the local ethics committee (project numbers #24410;#31018;2020_021_24771,2021_009_29509,2021_010_29520).

[0196] Subcutaneous tumor model MCA205 mice were lightly anesthetized with isoflurane and 0.3 × 10⁻⁶ mice were subjected to a 0.3 × 10⁻⁶ feeding. 6 Individual MCA205 cells were subcutaneously injected. The treatment consisted of oxaliplatin (Sigma, 10 mg / kg) for moderate tumors with a size of 50 mm. 3 When the tumor size reached a certain point, it was administered, followed by three doses of neutralizing anti-PD1 monoclonal antibody (BioXCell, clone 29F.1.A12, 200 μg per mouse, 8, 12, and 16 days after chemotherapy). From two days before chemotherapy to one day before immunotherapy, six injections of neutralizing anti-DBI monoclonal antibody (2.5 mg / kg) or its isotype (IgG2A, 2.5 mg / kg) were administered, if indicated, to combine this therapy or its corresponding control (vehicle + anti-PD1 isotype IgG2A). Tumor size was monitored three times weekly until one of the following endpoints was reached: tumor size > 1500 mm 3 Ulcer formation or weight loss >20%.

[0197] To establish E0771-symbolic solid tumors, 5 × 10⁶ wild-type E0771 cells were placed adjacent to the mammary gland (near the orthotopic site) in the lower right abdomen of C57BL / 6J mice (8-week-old females). 5The individual molecules were injected subcutaneously (sc). Approximately 7 days after injection, tumor growth was monitored with a digital caliper, and the mice were assigned to different groups. From day 8, the mice were treated every two days with neutralizing anti-DBI monoclonal antibody (5 mg / kg) or its isotype (IgG2A, 5 mg / kg). Simultaneously, free corticosterone (Sigma-Aldrich) was added to the drinking water (0.1 mg / ml; water / ethanol 0.66%). Anti-PD1 monoclonal antibody (αPD1; 10 mg / kg) was injected on days 12, 15, 18, and 21. Tumor growth was routinely monitored with a digital caliper.

[0198] To establish a TC-1-orthotopic TC1 model, wild-type TC1 Luc cells (5 × 10⁶ cells in 100 μL of PBS) were used. 5 The antibody was intravenously injected into wild-type C57BL / 6J mice. Tumor development and growth were monitored by in vivo photonic imaging of luciferase activity in tumor cells. Approximately 7 days after injection, tumor development in the lungs was detected with a 4-minute exposure time, and mice were randomized according to quantified tumor size and assigned to different groups for treatment as described below. Anti-DBI or isotype control antibody was administered intraperitoneally (ip) at a dose of 2.5 mg / kg body weight on day 9, again on day 10, and then every other day until day 18. Oxaliplatin or its vehicle (PBS) was administered via ip injection at a dose of 5 mg / kg body weight on day 11, and PD1 monoclonal antibody or its isotype was administered via ip injection at doses on days 19, 23, and 27. Anti-DBI or its isotype control was injected one day before each anti-PD1 injection and maintained on a weekly basis.

[0199] To acquire bioluminescence images, mice were administered a 3 mg dose of luciferase substrate (Beetle Luciferin potassium salt, Promega) via intravenous injection. Eight minutes after luciferin administration (in the case of the TC1 model), photons were acquired using the IVIS LUMINA III bioluminescence in vivo imaging system (PerkinElmer, Waltham, Massachusetts, USA). In vivo imaging was performed every 4-5 days with exposure times starting at 4 minutes, and then gradually reduced to 3 minutes, 2 minutes, and 1 minute when photon saturation occurred. Tumor-bearing mice that showed photon saturation at 1 minute of exposure were euthanized.

[0200] Antitumor cholinergic assay Two weeks prior to vaccination (-14 days), C57BL / 6J mice (8-week-old females) were pre-treated every two days with neutralizing anti-DBI monoclonal antibody (5 mg / kg) or its isotype (IgG2A, 5 mg / kg) (ip, in 200 μl of PBS), and free corticosterone (Sigma-Aldrich) was added to drinking water (0.1 mg / ml; ethanol / water 0.66%). Wild-type MCA205 cells were treated with MTX (4 μM) for 24 hours. Then, the supernatant and detached cells were collected, centrifuged, and the pellet was further washed with ice-cold PBS. 2 × 10⁶ cells in 100 μl of PBS 6 The cells were sc-injected into the left flank of immune C57BL / 6J mice (-7 days). PBS was injected as a negative control. After one week, all mice were confirmed to be tumor-free in the vaccinated flank, and 2 × 10⁶ cells (MCA205 cells) were found per mouse. 5 Cells were injected into the right flank of vaccinated mice (Day 0). Corticosterone and a neutralizing anti-DBI monoclonal antibody were administered continuously throughout the experiment. Tumor growth was monitored regularly over the following weeks.

[0201] Induction of autoimmunity against ACBP / DBI As previously described, anti-ACBP / DBI autoimmunity was induced in young adult mice (6-8 weeks old) using either a KLH-ACBP / DBI vaccine or a non-ACBP / DBI specific control KLH (77649, Thermo Fisher Scientific) along with the mineral adjuvant Montanide ISA 51 VG (Seppic) at a once-weekly dose for four weeks [Montegut L. et al., Immunization of mice with the self-peptide ACBP coupled to keyhole limpet hemocyanin, STAR Protoc, 2022, 3(1), p.101095.]. The induction of autoimmunity was confirmed by immunoblotting of RecDBI detected in mouse plasma. After two weeks of immunization recovery, 0.3 × 10⁴ cells were bled into the right flank of the mice. 6 Individual MCA205 cells were subcutaneously injected, and the treatment was continued as described above.

[0202] Immune infiltration in the MCA205 tumor microenvironment Ten days after chemotherapy, MCA205 tumors from three independent experiments were dissected and homogenized into single-cell suspensions by mechanical and enzymatic disruption according to the manufacturer's instructions (tumor dissection kit, 130-096-730, Miltenyi Biotec). Cells were stained with Live-Dead Fixable Yellow Dye (Thermo Fisher Scientific), then Fc receptors were blocked with unbound anti-mouse CD16 / CD32 antibody (BD BioSciences; clone 2.4G2), and fluorophore-conjugated antibodies were added for detection of surface markers (CD45, CD3, CD4, CD8a, ICOS, GITR, LAG3, PD1, TIGIT, and VISTA). Cells were fixed and permeabilized before staining with anti-FoxP3 (eBioscience FoxP3 / Transcription Factor staining buffer, Thermo Fisher Scientific). Fluorescence data were acquired using BD LSRFortessa X20 with BD FACS Diva software. Compensation, scaling, gating, and data analysis were performed on the omiq.ai platform. CD4 + and CD8 + Unsupervised clustering (FlowSOM algorithm) was performed on one experimental set to define a specific subpopulation of T cells. The gating strategy was inferred from the markers expressed by the target cluster, which were differentially present between the OXA+ anti-PD1 group and the anti-DBI+OXA+ anti-PD1 group (FoxP3). - ICOS int GITR + , Lag3 - activation T H FoxP3 + GITR + , Lag3 - activation T reg ), which was applied to three independent copies for statistical analysis.

[0203] Statistical analysis. For tumor growth experiments, longitudinal analysis was performed using linear mixed-effects modeling of tumor size. A type II ANOVA (Wald test) was used to calculate p-values ​​by testing together that both tumor growth gradients and intercepts were the same between the treatment groups of interest, and the p-values ​​were corrected for multiple comparisons using Holm's method. Survival curves were compared 2x2 using the log-rank Mantel-Cox test in GraphPad Prism (v9) software. For comparisons of immune cell infiltration, data from population relative counts were sorted using the ROUT test (outlier threshold = 1%), and statistical significance was tested using one-way ANOVA with Sidak correction for multiple comparisons.

[0204] result Improvement of outcomes in chemoimmunotherapy using autoantibodies that neutralize ACBP / DBI As a first approach to investigate the effects of ACBP / DBI on chemoimmunotherapy, 6-week-old C57BL / 6J mice were immunized with the immunogenic carrier protein keyhole limpet hemocyanin (KLH) alone (as a control) or with KLH conjugated to ACBP / DBI with the adjuvant montanaid. Four injections of the KLH-ACBP / DBI conjugate over consecutive weeks are known to induce a strong autoantibody response against ACBP / DBI, resulting in neutralization of ACBP / DBI. [Montegut, L. et al., Immunization of mice with the self-peptide ACBP coupled to keyhole limpet hemocyanin, STAR Protoc, 2022, 3(1), p.101095.] Two weeks after the completion of the vaccination protocol, MC205 dermatofibrosarcoma cells (syngeneic with C57BL / 6J) were subcutaneously inoculated. When the tumor becomes palpable (surface area 25-50 mm) 2Around 8 days post-inoculation, the mice received one cycle of intraperitoneal (ip) chemotherapy with oxaliplatin (OXA) (OXA or PBS as a solvent control), followed by three cycles of immunotherapy consisting of ip administration of PD-1-specific monoclonal antibodies (anti-PD1 antibodies or isotype-matched control antibodies) on 16, 20, and 24 days post-inoculation (Figure 1A). Tumor growth was monitored using calipers (Figure 1B and Figures 10A-10D), and mouse survival was recorded (Figure 1C). Notably, tumors transplanted into KLH-ACBP / DBI-vaccinated mice responded more efficiently to OXA + anti-PD1 chemoimmunotherapy than tumors transplanted into KLH-vaccinated control animals. This improved therapeutic effect was statistically significant at the levels of tumor growth and animal survival (Figures 1A-1D). In summary, these results indicate that autoantibodies that neutralize ACBP / DBI may improve outcomes in chemoimmunotherapy.

[0205] Improvement of chemoimmunotherapy outcomes with anti-ACBP / DBI mAbs In the following experiment, MC205 cells were sc-administered to 8-week-old C57BL / 6J mice. The mice received regular anti-ACBP / DBI mAbs (or IgG2A isotype control antibodies as a control) starting 6 days post-inoculation, and also received one dose of oxaliplatin (OXA) chemotherapy via intravenous injection (ip) on 8 days post-inoculation, followed by three cycles of immunotherapy autoantibodies (i.e., anti-PD1 mAbs) on 16, 20, and 24 days post-inoculation (Figure 2A). Tumors transplanted into anti-ACBP / DBI mAb-treated mice responded more efficiently to OXA + anti-PD1 chemoimmunotherapy than tumors in control mice, with 11 out of 23 anti-ACBP / DBI mAb-treated mice receiving chemoimmunotherapy showing a significant reduction in tumor growth with complete regression of the cancer, compared to only 2 out of 23 control mice receiving chemoimmunotherapy alone. This also resulted in a significant increase in animal survival (Figures 2B-2D and 11A-11D). Therefore, administration of mAbs specific to ACBP / DBI improves the effectiveness of chemoimmunotherapy.

[0206] In another experiment following a similar treatment schedule, mice were sacrificed 18 days post-inoculation (Figure 3A), and the composition of tumor immunoinfiltrates was determined by immunofluorescence staining and subsequent cytofluorometry analysis (Figures 3B-3M). Anti-ACBP / DBI mAbs were found to affect CD4 + T cells (Figure 3C), especially CD4 + Foxp3 - T helper cells (T) are defined as cells (Figure 3G). H It amplified some of the effects of chemoimmunotherapy (OXA + anti-PD1) on cancer immunoinvasions such as infiltration by ) and CD4. Anti-ACBP / DBI mAb amplified some of the effects of CD4. + Foxp3 + Regulatory T cells (T) are defined as cells. reg ) is better than CD8 + Improved the ratio of T cells (Figure 3F), CD4 + T cells within T cells reg The frequency of cells was reduced (Figure 3I). Anti-ACBP / DBI mAbs were found in T cells expressing the fatigue marker LAG3. H and CD8 + The proportion of T cells was reduced (Figures 3J and 3K). Furthermore, T H Cells and T reg Shifts in the activation / exhaustion markers ICOS and GITR were observed between cells (Figures 3L and 3M). All of these effects of anti-ACBP / DBI mAbs were detected only in the context of chemoimmunotherapy and not in its absence (Figures 3B-3M). In summary, these results suggest that anti-ACBP / DBI mAbs improve treatment-induced immune surveillance.

[0207] Next, we confirmed the positive effect of anti-ACBP / DBI mAbs on chemoimmunotherapy in another model, namely non-small cell lung cancer (NSCLC). When luciferase-expressing TC1 NSCLC cells were intravenously (iv) injected into syngeneic immunocompetent C57BL / 6J mice, orthotopic lung cancer was formed, which could be monitored by bioluminescence imaging. [Liu, P. et al., Crizotinib-induced immunogenic cell death in non-small cell lung cancer, Nature Communications, 2019, 10(1).] From day 9 after TC1 cell inoculation, NSCLC-carrying mice were regularly injected via ip (intravenous injection) with anti-ACBP / DBI mAbs (or isotype control antibodies), then treated with OXA (once via ip) on day 11 after inoculation, and treated with anti-PD1 mAbs (intravenous injection on days 19, 23, and 27 after TC1 inoculation) (Figure 4A), and tumor progression was determined by bioluminescence imaging (Figure 4B). Anti-ACBP / DBI mAbs improved tumor control achieved by chemoimmunotherapy (Figures 5A and 12A-12D). These results support the cancer treatment improvement effect of ACBP / DBI neutralization.

[0208] Improvement of chemoimmunotherapy outcomes by ACBP / DBI neutralization in association with corticosteroid therapy In the next experiment, five days after injection of TC1 cells, animals were treated with corticosterone (CORT) or, as needed, vehicle only (0.66% ethanol continuously present in drinking water from day 5 to day 9) and / or anti-ACBP / DBI mAb (or isotype control mAb). Furthermore, animals received one dose of oxaliplatin chemotherapy (ip administration on day 8 after TC1 injection, which was 3 days earlier than in the previous experiment), followed by PD-1 targeted immunotherapy (ip injections on days 16, 20, and 24 after TC1 injection), which was also 3 days earlier than in the previous experiment (compare Figure 5A and Figure 6A). Notably, corticosteroid therapy strongly interfered with the effect of this chemoimmunotherapy, but this was restored by injection of anti-ACBP / DBI antibody (Figures 6B, 7A, and 13A-13D). These data indicate that ACBP / DBI neutralization can reverse the immunosuppressive effects of corticosteroid therapy, which can interfere with the anti-cancer effects of chemoimmunotherapy.

[0209] Improvement of immunotherapy outcomes by ACBP / DBI neutralization in the absence and presence of corticosteroid therapy. Syngeneic E0771 mammary cancer cells were subcutaneously (sc) inoculated into C57BL / 6J mice. Six days later, treatment with corticosterone (administered in drinking water with 0.66% ethanol as a control using CORT or vehicle only) and anti-ACBP / DBI mAb (or isotype control mAb, both administered as intraperitoneal injections three times a week) was initiated, and tumor growth was monitored for several weeks. On days 12, 15, 18, and 21, the animals received intraperitoneal (ip) injections of PD-1 specific mAb (or its isotype control) (Figure 8A). This treatment typically reduces tumor growth [Zheng, X. et al., Increased vessel perfusion predicts the efficacy of immune checkpoint blockade, J Clin Invest, 2018, 128(5), pp.2104-2115.], but this effect was lost in CORT-treated animals unless they received anti-ACBP / DBI mAb (Figure 8B). The efficacy of immunotherapy with anti-PD1 mAbs, its loss with corticosteroid therapy, and its recovery with anti-ACBP / DBI mAbs were detectable at the levels of tumor growth and animal survival (Figures 8B and 14A-14D). Furthermore, anti-ACBP / DBI mAbs significantly improved the immunotherapy effect of αPD-1 at the animal survival level, even in the absence of corticosteroid therapy (Figures 8C and 8D). These findings suggest that ACBP / DBI neutralization can improve immunotherapy outcomes both in the absence and in the presence of corticosteroid therapy.

[0210] Improvement of anti-cancer immune response by ACBP / DBI neutralization in association with corticosteroid therapy C57BL / 6J mice were treated with the glucocorticoid corticosterone (CORT) (or vehicle only as a control, administered in 0.66% ethanol (in drinking water)) and anti-ACBP / DBI monoclonal antibody 7A (mAb, or its isotype control IgG2a antibody), and the antibody was administered by intraperitoneal (ip) injection three times a week as shown in the scheme (Figure 9A). One week after the start of treatment, the animals were subcutaneously (sc) injected with mitoxantrone (MTX)-treated MCA205 fibrosarcoma cells, and it is known that MTX induces ICD, and that MTX-treated cancer cells induce a protective anti-cancer immune response that suppresses, delays, or reduces tumor growth that may develop in naive mice after injection of live, untreated MCA205 fibrosarcoma cells. [Panaretakis T. et al., Mechanisms of pre-apoptotic calreticulin exposure in immunogenic cell death, EMNO J, 2009, 28(5), pp.578-590.] Therefore, when injected into the contralateral flank one week after vaccination with MTX-treated MCA205 cells, the growth of live MCA205 fibrosarcoma cells was suppressed, and the survival time of the animals was longer than when naive mice were inoculated with similar live cells. Treatment with CORT reduced this vaccination effect, but this was largely restored by anti-ACBP / DBI mAbs (Figure 9B and Figures 15A-15D). In conclusion, ACBP / DBI neutralization appears to be able to reverse the immunosuppressive effect of corticosteroid therapy on the anti-cancer immune response.

[0211] Example 2: The role of DBI in hepatocellular carcinogenesis The significance of DBI in malignant diseases has not been sufficiently studied, and there is no knowledge about the role of DBI in HCC. However, surprisingly, the applicants have found the important significance of DBI in hepatocellular carcinogenesis and tumor progression by using guidelines for methods of inhibiting DBI (e.g., by knockout, receptor mutation, or antibody-mediated neutralization). Notably, high intratumoral DBI mRNA expression and high levels of circulating DBI protein are associated with features of poor prognosis in HCC patients, supporting the clinical significance of these findings.

[0212] To investigate its role in hepatocellular carcinogenesis in mice, the applicant inhibited DBI using three methods: (i) inducible systemic knockout of DBI, (ii) point mutations in the DBI receptor (GABRG2), or (iii) induction of autoantibodies that neutralize DBI. The applicant found that DBI plays a major pro-oncogenic role in HCC induced by intrahepatic transplantation of HCC cell lines, transgenic co-expression of two proto-oncogenes Myc and Ctnnb1, and chronic challenge with a Western diet together with either CCl4 or diethylenenitrosamine. Inhibition of DBI nearly normalized HCC-related gene expression and thus reduced oncogenic changes in cell cycle-related, immunosuppressive, and ferroptosis-regulating genes. Functional experiments confirmed that DBI inhibition reduced the proliferation of normal and malignant hepatocytes, increased the HCC response to PD-1 blockade, and sensitized HCC to therapeutic induction of ferroptosis. While we do not wish to be constrained by theory, DBI constitutes a viable target involved in the pathogenesis of HCC.

[0213] Increased DBI levels in human HCC Nine different publicly available gene expression datasets, including the Cancer Genome Atlas (TCGA), consistently show that HCC tumors contain higher DBI mRNA levels than normal adjacent liver tissue (Figure 16A). According to TCGA, this DBI elevation arose independently of the tumor / lymph node / metastasis (TNM) staging system, Child-Pugh (CP) grade, tumor differentiation, vascular invasion, and levels of fibrosis and inflammation (Figures 16B and 16C). However, DBI mRNA was particularly abundant in patients with α-fetoprotein (AFP) protein levels exceeding 400 ng / mL (Figure 16D), and DBI mRNA levels above the median were associated with reduced overall survival in TCGA (Figure 16E).

[0214] Using ELISA, DBI protein levels were measured in the plasma of 146 HCC patients belonging to different BCLC stages and in 58 plasma samples from chronic liver disease patients without a history of HCC collected at the University Hospital (AP-HP, Bobigny, France). ELISA-detectable plasma DBI levels (ng / mL) were found to be higher in HCC patients compared to the group without current or previous HCC (Figure 16F) and correlated with poor prognostic features such as advanced BCLC stage (Figure 16G), vascular invasion (Figure 16H), and extrahepatic metastasis (Figure 16I). Furthermore, DBI mRNA correlated with AFP mRNA in liver biopsy (Figure 16J), reflecting the correlation between DBI protein levels and AFP protein levels in plasma (Figure 16K). Additionally, DBI plasma concentration correlated with tumor size, as detected by CT scans (Figure 16L). Overall, these results support the idea that advanced HCC is associated with high DBI levels.

[0215] Importantly, when human HCC cells were orthotopically inoculated into the livers of immunodeficient mice (Figure 16M), plasma concentrations of human DBI (but not mouse DBI), as measured by species-specific ELISA (which distinguishes between human and mouse DBI), gradually increased over time (Figure 16N) and correlated with tumor size (Figure 16O). These findings suggest that elevated circulating DBI may be directly attributable to HCC. While we do not wish to be bound by theory, overall, these findings suggest that HCC progression is associated with increased DBI expression by malignant tissue and elevated DBI plasma levels.

[0216] Therefore, the potential pathogenic role of DBI in HCC was investigated using appropriate preclinical models. Intracellular and extracellular DBI contribute to the pathogenesis of transplantable HCC, proto-oncogene-induced HCC, and oncogenic-induced HCC. Among a collection of different human HCC cell lines, HUH-7 cells exhibited the highest DBI mRNA and protein levels (Figure 17A). Knockdown of DBI using three different shRNAs yielded HUH-7 clones with less potential for proliferation and clonality, and a reduced proportion of cells in the S phase of the cell cycle. Similar results were obtained for human hepatoblastoma HEP-G2 cells and mouse HCC Hep55.1C cells subjected to DBI and DBI knockdown, respectively (Figures 17B-17M).

[0217] Compared to the parent cell line, three Hep55.1C clones subjected to DBI depletion (by three different shRNAs) were relatively less pathogenic when orthotopically inoculated into the livers of immunocompetent C57BL / 6 mice (Figure 18A), as indicated by reduced recipient mouse mortality (Figure 18B), a decrease in the number of mice developing macroscopic HCC (Figure 18C), and a decrease in the number and total weight of HCC nodules at the endpoint (Figures 18D and 18E). Continuous monitoring of tumor growth using luciferase-transduced Hep55.1C clones confirmed that DBI-depleted cells did not develop HCC (Figures 18F-18K). Notably, neutralizing the extracellular DBI protein by appropriate autovaccination protocols (i.e., inducing autoantibodies against DBI by inoculating DBI conjugated with immunogen keyhole limpet hemocyanin (KLH) along with an adjuvant) also delayed orthotopic growth of parental (DBI-expressing) luciferase-transformed Hep55.1C tumors compared to animals immunized with KLH alone as a control (Figures 18L-18Q).

[0218] Next, we studied a model of proto-oncogene-induced hepatocellular carcinogenesis in which two plasmids encoding Myc and Ctnnb1 were simultaneously transfected into hepatocytes by hydrodynamic injection. f / f ) is ubiquitously excised in mice (genotype: UBCcre / ERT2::DBI) using tamoxifen-inducible Cre recombinase (UBCcre / ERT2). f / f , Dbi + / + , contrast: DBI without CRE f / f , Dbi - / - ) was rather resistant to Myc / Ctnnb1-induced hepatocarcinogenesis (Figures 19A-19D). Furthermore, the γ2 subunit (Gabrg2) of the GABAA receptor was homozygously mutated (F77I) (genotype: Gabrg2 F77I / F77IMice that lost their interaction with DBI were relatively resistant to Myc / Ctnnb1-induced HCC (Figures 19E-19H). Finally, immunization with KLH-DBI conferred partial protection against Myc / Ctnnb1-mediated hepatocellular carcinogenesis (Figures 19I-19L).

[0219] Since the potential involvement of DBI in diet- and toxin-induced HCC is unclear, we investigated a hepatocarcinogenesis model combining a Western diet (WD) with weekly ip administration of the hepatotoxic substance CCl4. Immunization with Dbi (Figures 20A-20F), homozygous Gabrg2 mutation (Figures 20G-20L), and KLH-Dbi (Figures 20M-20R) mitigated the characteristic findings of WD+CCl4-induced liver injury (non-alcoholic steatohepatitis [NASH], lobular inflammation, ballooning degeneration with Mallory-Denk bodies, and fibrosis), reduced hepatic Dbi mRNA and protein expression and circulating DBi concentration, and inhibited the development of HCC. Therefore, all DBI inhibition protocols reduced the size and number of WD / CCl4-induced tumor lesions occurring in the liver (Figures 20E, 20F, 20K, 20L, 20Q, 20R). When the disease was induced by a combination of HFD and the carcinogen diethylnitrosamine (DEN), similar anti-NASH, anti-fibrotic, and tumor-suppressing effects were observed with KLH-Dbi vaccination (compared to a control vaccinated with KLH alone) (Figures 20S-20X).

[0220] Overall, these results indicate that hepatocarcinogenesis is promoted by DBI. This effect of DBI is at least partially mediated by the extracellular pool of DBI acting on GABAA receptors.

[0221] DBI neutralization improves immune surveillance. To elucidate the mechanism by which DBI promotes liver carcinogenesis, we will examine three of the above models, namely (i) WD + CCl4-treated DBI. + / + Mouse vs. Dbi - / -Liver bulk RNA sequencing was performed on mice, (ii) WD+CCl4 treated KLH-only versus KLH-DBI vaccinated mice, and (iii) HFD+DEN treated KLH-only versus KLH-DBI vaccinated animals. The effects of unrestricted DBI activity were then compared to 25 publicly available datasets detailing deregulation of molecular pathways in different liver diseases. In Figure 21A, the normalized enrichment score (NES) for each KEGG pathway was row-normalized and presented as a Z-score. Euclidean distances were calculated for row and column clustering analysis. Unsupervised hierarchical clustering showed that many molecular pathways downregulated by DBI inhibition were upregulated in multiple liver diseases, regardless of etiology (alcoholic, inflammatory, metabolic, or toxic), (or conversely, molecular pathways upregulated by DBI inhibition were downregulated in those liver diseases), with the exception of hepatitis B, C, and D virus infections, which did not belong to the same cluster as DBI inhibition (Figure 21A).

[0222] In three RNA-seq datasets reflecting DBI inhibition, several gene ontology items suggestive of immunosuppression were downregulated, corresponding to signatures specific to regulatory T cells in lymph nodes, inflammatory Th17 cells, and polarized Th2 cells (Figure 21B). Based on this prediction, orthotopic Hep55.1C HCC was highly responsive to combination treatment with a neutralizing mAb + PD-1 blocking antibody against DBI. This combination was significantly more efficient at reducing tumor growth than either of the two monotherapies (DBI alone or PD-1 blocking alone), and therefore significantly extended survival (Figures 21C-21F). Thus, DBI inhibition can enhance the sensitivity of HCC to immunotherapy.

[0223] DBI neutralization reduces HCC proliferation. Cell cycle-related genes were strongly downregulated in all three RNA-seq datasets reflecting DBI inhibition (Figure 21B). Bioinformatics analysis was performed using i) WD+CCl4 treated DBI + / + vs DBI- / - We identified genes that were commonly upregulated or downregulated in the livers of mice treated with (ii) WD+CCl4, or (iii) WD+DEN, and KLH-only versus KLH-ACBP / Dbi vaccine (Figures 22A-22B). We also identified genes that are overexpressed in human HCC and downregulated by DBI inhibition, which is associated with poor prognosis (Figures 22C and 22D). Many of these DBI-dependent, disease-related genes were involved in cell cycle progression (especially mitosis) (Figure 22E). Quantitative analysis by qRT-PCR confirmed that BDI inhibition downregulated genes necessary for cell cycle progression (Ccnd1, Ccne1, Cdk4, Cdk6, Pcna, etc.) and upregulated genes that inhibit the cell cycle (Atr, Gadd45a, Gadd45b, Cdkn1a, Cdkn2a, etc.) (Figure 23A). Similarly, knockdown of DBI in HEP-G2 cell lines induced transcriptional signs of cell cycle disruption, consistent with a decrease in the oncogenic and proliferative potential of such DBI-depleted cells (Figure 23B).

[0224] Prompted by these results, we evaluated cell proliferation in mouse livers subjected to oncogenic tumorigenesis using immunohistochemical detection of Ki67 (Figures 24A–24E) and PCNA. Notably, DBI inhibition by DBI KO, Gabrg2 mutation, or KLH- / Dbi vaccination resulted in reduced proliferation in both tumor lesions (where detectable) and non-malignant liver parenchyma, regardless of the exact oncogenic stimulus (CCl4 or DEN) (Figures 24A–24E). Neutralization of DBI with monoclonal antibodies (mAbs) inhibited hepatocyte proliferation both after sham surgery and after partial hepatectomy (inducing liver regeneration (Figure 24E)), suggesting a general proliferative effect of DBI in normal hepatocytes.

[0225] Finally, HCC cells were subjected to carcinogenesis caused by WD+CCl4 in Dbi + / + Mouse vs. Dbi - / - Isolated from mice (Figure 24F). Dbi - / -HCC cells showed relatively low expression of proliferation markers (Ki67, PCNA) and malignancy-associated markers (AFP, CK19, GPC3) (Figure 24G), weakened clonal potential (Figure 24H), and reduced in vitro proliferation (Figure 24I). These findings indicate that DBI is necessary for full-scale HCC proliferation.

[0226] While we do not wish to be bound by theory, DBI has a significant proliferative effect on normal and transformed hepatocytes.

[0227] DBI neutralization increases susceptibility to ferroptosis induction. The aforementioned bioinformatics analysis also identified changes in various pathways of cellular self-consumption. Thus, DBI inhibition was accompanied by a decrease in transcripts associated with apoptosis and necroptosis, but an increase in the expression of genes associated with ferroptosis (see Figure 21B). Furthermore, DBI inhibition enhanced the expression of autophagy-related genes and the proferroptotic effect of autophagy.

[0228] Validation by qRT-PCR confirmed i) WD+CCl4 treated Dbi - / - vs DBI + / + (ii) WD + CCl4 treatment Gabrg2 F77I / F77I vs Gabrg2 WT Upregulation of ferroptosis-driving genes and downregulation of ferroptosis-suppressing genes were observed in the liver of mice, (iii) WD+CCl4-treated, and (iv) WD+DEN-treated KLH-Dbi vs. KLH-only-vaccinated mice (Figure 25A). Furthermore, immunoblotting confirmed increased protein expression of ferroptosis effectors (e.g., ACSL4, KEAP1, NCOA4, and POR) and decreased expression of ferroptosis suppressors (ACSL3, BMAL1, GPX4, SQSTM1 / p62, etc.) after DBI inhibition (Figure 25B). Such a shift from ferroptosis suppression to execution was observed in WD+CCl4-treated DBI mice. - / - Mouse vs. Dbi + / +HCC clones derived from mouse liver were also observed at both mRNA and protein levels. Spatial transcriptomics or spatially resolved mass spectrometry metabolomics combined with hematoxylin-eosin staining allowed for differentiation between non-malignant and malignant regions of the liver (Figure 25C). Notably, inhibition of DBI by DBI knockout or KLH-Dbi vaccination revealed higher expression of ferroptosis effectors in seemingly (still) non-malignant tissue compared to the DBI-uninhibited control group (Figure 25C). Therefore, changes in ferroptosis-related gene expression levels observed throughout the liver are detected in both isolated HCC cells and non-malignant tissue.

[0229] Gene / protein expression data indicated that HCC cells were particularly vulnerable to loss of viability in response to a combination of Dbi knockout and the addition of pharmacological ferroptosis inducers including RSL3, IKE (imidazole ketone elastin), LA (linoleic acid), and LNA (linolenic acid) (Figures 26A-26D). The groups treated with ferroptosis inducers were compared with each Dbi knockout. + / + and Dbi / - The data was normalized compared to the vehicle (Veh) treatment control group.

[0230] Furthermore, orthotopic Hep55.1C cancer responded more efficiently to treatment with a combination of anti-DBI mAb plus ferroptosis induction (using RSL3 or IKE) than to monotherapy with either anti-DBI mAb or ferroptosis inducer (Figures 26E-26P). This combination was more effective in reducing tumor burden than either of the two monotherapies (anti-DBI alone or RSL3 / IKE alone) and therefore significantly extended survival (Figures 26E-26P).

[0231] While we do not wish to be bound by theory, inhibition of DBI sensitizes HCC to therapeutic intervention with ferroptosis-inducing agents.

[0232] In this disclosure, the applicants have surprisingly found that DBI contributes to the onset and progression of HCC induced by a variety of quite different methods, including (i) intrahepatic inoculation of HCC cells, (ii) proto-oncogene induction transformation, and (iii) long-term challenge with the hepatotoxin CCl4 or (iv) mutagenic DEN in association with a Western diet.

[0233] The applicants outlined genetic methods to inhibit the DBI system (i.e., knockdown or knockout of DBI and mutations in its receptor GABRG2), as well as immunological approaches (i.e., neutralizing antibodies against DBI), and found that all of these strategies mitigated hepatocarcinogenesis. This preclinical evidence supporting the HCC-promoting effect of DBI is supported by clinical correlations indicating that aggressive HCC is associated with upregulation of DBI mRNA and increased circulating ACBP / DBI protein concentrations.

[0234] Inhibition of DBI consistently reduced signs of hepatitis leading to excessive macrophage infiltration and fibrosis, but also reduced the expression of genes associated with immunosuppressive T cell subtypes (T regs , T H 2 and T H 17, etc.). Therefore, inhibition of extracellular DBI by monoclonal antibodies sensitized HCC to PD-1 blockade immunotherapy.

[0235] DBI inhibition also resulted in the upregulation of 13 proferoptosis mRNAs and the downregulation of several more antiferoptosis mRNAs. This effect, which could be verified at the protein level, was accompanied by increased sensitivity of HCC cells to pharmacological induction of ferroptosis both in vitro and in vivo.

[0001] While we do not wish to be bound by theory, neutralization of DBI reduces or delays the onset of HCC in various preclinical models. Furthermore, inhibition of DBI inhibits proliferation and sensitizes established HCC to PD-1 targeted immunotherapy and pharmacological induction of ferroptosis.

[0236] material and method HCC patient cohort Between March 2013 and May 2021, a total of 260 plasma samples were collected at Avicenne Tertiary University Hospital (Bovigny, France) from 146 patients with HCC and 58 patients with chronic liver disease without HCC. Two or more plasma samples were collected from 46 patients. Plasma samples were divided into two groups: (i) plasma collected at the time of HCC diagnosis, treatment day, or radiographic evaluation indicating active HCC (n=195), and (ii) plasma collected from patients with chronic liver disease without HCC or after HCC treatment without active tumors at the time of imaging (n=65). Whole blood (5 mL) was collected using EDTA tubes. Blood samples were centrifuged at 2000 × g for 10 minutes at room temperature and immediately stored at -80°C. All patients signed informed consent for sample collection, and the study was approved by the ethics committee.

[0237] Patient, tumor characteristics, and treatment type were obtained from medical records. Patient baseline characteristics were collected prior to treatment (including age, sex, etiology of liver disease, and presence of cirrhosis). Cirrhosis was defined by histology or by a combination of clinicobiological data, ultrasound, and liver stiffness measurement. HCC was diagnosed histologically or using non-invasive criteria via imaging techniques (magnetic resonance imaging [MRI] and / or triphasic computed tomography [CT]) in accordance with European Association for the Study of the Liver (EASL) guidelines. Tumor characteristics at imaging (tumor size and number, major vessel invasion, and metastasis), Barcelona Clinic Liver Cancer Staging System (BCLC), alpha-fetoprotein (AFP), and Child-Pugh (CP) scores were also collected. Data on the type of treatment (ablation, embolization, and / or systemic treatment) and radiological response, assessed by mRECIST criteria, were also recorded at 4 and 12 weeks post-regional and systemic treatments, respectively.

[0238] Preparation of stable cell lines, cell culture, and cell assays Stable DBI knockdown hepatocellular carcinoma cell lines were developed. Hepatocellular carcinoma cells were grown under the following conditions: HEP-G2 (EMEM + 10% FBS + 1% sodium pyruvate + 1% HEPES), HUH-7 (DMEM, 10% FBS), Hep55.1C (DMEM, 10% FBS), and Hep55.1C cells expressing firefly luciferase (Hep55.1C-luc, DMEM, 10% FBS, 0.1% blastosidine). These cancer cells were grown in 6 welldishes to a concentration density of 60% to 70%, and then transfected with 5 μg / mL polyblenn (5 μL to 8 μL) thoroughly mixed in 1 mL of medium and 25 μL to 35 μL of lentiviral shRNA targeting DBI (SH1, SH2, SH3) or a negative control (NC). After 24–48 hours, the culture medium was replaced with fresh medium, and the cells were maintained for another 24 hours. Transduced cells were selected using puromycin (10 μg / mL). Single-cell clones were isolated by single-cell FACS sorting in 96-well plates and gradually grown in 24-well, 12-well, and 6-well plates. All cell clones from the 6-well plates were replicated; one was stocked at -80°C for storage, and the other was collected for qRT-PCR to detect DBI knockdown efficiency. For further assays, the clone with the highest knockdown efficiency was selected.

[0239] Growth assay. The CCK-8 assay was performed using a commercially available kit. For the colony formation assay, HEPG-2 (NC, SH1, SH2, and SH3, 1000 cells / well), HUH-7 (NC, SH1, SH2, and SH3, 1000 cells / well), and Hep55.1C (NC, SH1, SH2, and SH3, 500 cells / well) cancer cells were seeded in 6-well plates in 2 mL of medium containing 10% FBS. The medium was changed every 3 days for 7–14 days until the control (NC) well approached confluence. After gentle washing twice with PBS, the colonies were fixed with 4% paraformaldehyde for 30 minutes and stained with 0.2% crystal violet at room temperature for 30 minutes. The number of colonies (colony > 0.3 mm) was counted using isoftware. All experiments were performed in triplicates.

[0240] Cell cycle analysis. Following the instructions of the commercially available kit, the aforementioned transfected cells (25 × 10) were used. 4 Cells were plated in a 6-well plate for 24 hours. The cell cycle was then synchronized for 24 hours in a medium containing 0.1% FBS, followed by 24 hours of incubation in a medium containing 10% FBS. Cells were harvested and fixed using ice-cold 70% ethanol. The cell cycle distribution was then determined by flow cytometry.

[0241] animal All mice used in this study had a C57BL / 6 background. Five mice / cage were housed in a 12-hour light / dark cycle under specific pathogen-free conditions. Mice were acclimated to their new housing facility for at least one week before use. All mouse experiments were conducted according to protocols approved by the local animal experiment ethics committee. Liver tissue and plasma samples were collected for further analysis. Liver tissue was freshly frozen in liquid nitrogen and stored at -80°C, fixed with 4% paraformaldehyde, or embedded in optimal cutting temperature compound (OCT).

[0242] In vitro inhibition of DBI Four strategies were developed to inhibit the expression or function of DBI: (i) Tamoxifen-induced ACBP / DBI knockout. Mice with tamoxifen-induced ACBP / DBI knockout were subjected to ACBP / DBI fl / fl Mice were bred with B6.Cg-Tg(UBC-Cre / ERT2)1Ejb / 1J mice, followed by tamoxifen injections (ip 75 mg / kg / day, 5 consecutive days). Control mice were ACBP / DBI, which does not contain Cre. fl / fl The subjects were mice. Tamoxifen was dissolved in corn oil (90%) + ethanol (10%) at a concentration of 20 mg / mL, divided into aliquots, and stored at -20°C. (ii) Constitutive Gabrg2 F77I / F77I Point mutation. Gabrg2 containing point mutation F77I. tm1Wul / J mice were compared to control mice with unmutated wild-type mice. (iii) Induction of ACBP / DBI-specific autoantibodies. As described above, KLH-DBI was generated by conjugating KLH with recombinant DBI in a molar ratio of 1:30. KLH alone was used as a control. Next, Montanide (and KLH / KLH-DBI) were mixed in a 1:1 volume ratio. Mice were inoculated via ip with the above mixture on day 0 (30 μg), day 7 (30 μg), day 14 (30 μg), and day 21 (10 μg of KLH-DBI) to induce anti-DBI autoantibodies. Immunoblots of recombinant DBI were incubated with plasma from KLH / KLH-DBI-immunized mice to detect anti-ACDBIBP antibodies. (iv) Monoclonal anti-DBI antibodies. Mice were inoculated via ip 3-4 times per week with an anti-DBI mAb (a-DBI, 5 mg / kg) or its control isotype (5 mg / kg) to neutralize DBI.

[0243] NASH-driven HCC model This study tested three main types of HCC mouse models: (i) Diet + Toxin Induction mouse models, each based on either a Western diet (WD) or a high-fat diet (HFD) combined with carbon tetrachloride (CCl4) or diethylnitrosamine (DEN). In most experiments, mice received a Western diet (WD, i.e., high-fat, high-fructose, and high-cholesterol diet), high-sugar water (23.1 g / L D-fructose + 18.9 g / L D-glucose), and weekly final doses of 2 μL / g body weight of carbon tetrachloride (CCl4, 1:10 dilution in corn oil) via ip injection. The high-sugar water was added to a sterile 450 mL sipper sack bag and replaced weekly. Food was also added weekly. Male mice were used in this HCC model. Three DBI inhibition strategies (DBI knockout, Gabrg2) were used. F77I / F77I Mutations and KLH-ACBP vaccination were combined with this mouse model. Mice were sacrificed after 27–33 weeks based on liver tumor detection by medical ultrasound (also known as osmosis). At necropsy, the number and size of tumors were determined by counting the number of visible tumors and measuring their size with calipers. Mouse body weight was monitored weekly. Data from the WD+CCl4 mouse model were confirmed using an HFD-DEN-induced HCC mouse model (combined with KLH-DBI). For this purpose, male C57BL / 6 mice (15 dpp) were treated with a single dose of diethylnitrosamine (DEN, N0258) dissolved in saline at a dose of 25 mg / kg body weight by intraperitoneal injection. Two weeks later, simultaneously with weaning, the mice were initiated with a vaccination protocol, and HFD consisted of 60% fat calories. Fresh diets were given every 2–3 days, and body weight was recorded monthly. Mice were sacrificed after 36 weeks, and their livers were removed to measure the number and size of tumors. Vaccination was performed using KLH / KLH-DBI. Specifically, DEN-injected mice (28 dpp) were randomized into two groups for the vaccination protocol.

[0244] Proto-oncogene-induced HCC Hydrodynamic transfection of the oncogene (Myc+Ctnnb1). A mixture of the transposase-encoding vector (SB100, 1.5 μg), the pT3-EF1a-Myc plasmid (7.5 μg), and the pT3-N90-Ctnnb1 plasmid (7.5 μg) was freshly prepared in 1 mL of 1 × PBS in a ratio of 1:5:5. The final solution was sterile filtered through a 0.22 μm filter and placed in a 37°C water bath before use. A volume equal to 10% of the mouse body weight (1 mL / 10 g) was injected into the tail vein of female mice (7 weeks old) under high pressure within 8–10 seconds using a 3 mL syringe with a 30 G × 1 / 2 needle. The four DBI inhibition strategies described above were tested in combination with this HCC mouse model, and tumorigenesis was monitored by ultrasound. Survival time was monitored, and mice were sacrificed at a humane endpoint defined by one of the following: mouse grimace scale score, significant abdominal distension due to tumor load, a decrease of more than 20% from initial body weight, or a maximum tumor diameter of 20 mm based on ultrasound. Tumor number and size were measured and recorded at necropsy.

[0245] Orthotopic transplantation of HCC cells Mouse HCC cell lines (Hep55.1C and Hep55.1C-derived cell lines such as Hep55.1C-Luc or Hep55.1C / Hep55.1C-Luc-NC, -SH1, -SH2 and -SH3, etc.), 30 × 10 in FBS-free DMEM 4 (50 μL / mouse) and human HCC cells (HUH-7, 200 × 10 in DMEM without FBS) 4 Luciferase-labeled Hep55.1C-Luc was transplanted into mouse livers (50 μL / mouse) to induce orthotopic liver tumors in female C57BL / 6J mice or male nude mice, respectively. Tumor growth in mouse models transplanted with luciferase-labeled Hep55.1C-Luc was monitored by the IVIS bioluminescence in vivo imaging system, and by ultrasound unless otherwise noted. Mice were monitored for survival and symptoms (e.g., skin jaundice). Mice were sacrificed at the above humane endpoints.

[0246] partial liver resection Nine-week-old male mice were subjected to 70% partial hepatectomy (Ph). Briefly, the mice were administered buprenorphine for analgesia and anesthetized by inhalation of isoflurane (2%). The midline abdominal skin and muscle were then incised to expose the abdominal cavity, followed by the removal of the left and midline hepatic lobes. First, a 4-0 silk suture was placed at the base of the left lateral lobe, a knot was tied, and the knotted lobe was cut just above the suture. The same procedure was performed on the middle lobe. Finally, the peritoneum was closed with a 5-0 suture, and the skin was closed with a wound clip. Pseudocontrol mice were subjected to the same surgical procedure without removal of the hepatic lobes. The mice were randomly divided into four groups (pseudo+ isotype, pseudo+ anti-ACBP, Ph+ isotype, and Ph+ anti-ACBP), with 9-10 mice per group. Anti-Acbp mAb (ip 2.5 mg / kg) or its isotype (ip 2.5 mg / kg) was administered 4 hours and 1 hour prior to surgery, with three additional doses given before sacrificial. Seven days after partial hepatectomy, the animals were sacrificial, the remaining liver was recovered, and processed for further analysis.

[0247] Isolation of primary HCC cells Six-well plates were coated with 2% FBS / PBS + 0.1% laminin. HCC tumors were treated with a NASH-driven mouse model (WD + D-fructose + water D-glucose + CCl4 in a DBI-based solution). + / + Mouse or DBI - / -The tumor tissue was isolated (from mice) and washed twice with ice-cold PBS. The tumor tissue was transferred to a 10 cm plate, completely chopped with a razor blade, and then transferred to a 15 mL Falcon tube. 10 mL of 2 mg / mL collagenase dispase solution was added, and the cells were incubated at 37°C for 30 minutes with rotation. After filtering through a 100 μm mesh into a 50 mL Falcon tube, the tissue was washed with 10 mL of 2% FBS / PBS. The sample was then sequentially filtered through 70 μm and 40 μm strainers to a volume of 15 mL and centrifuged again (room temperature (RT), 1000 rpm, 2 min). The cells were then resuspended in 5 mL of 1 × erythrocyte (RBC) lysis buffer, incubated on ice for 10 minutes, then 5 mL of 2% FBS / PBS was added and spun (room temperature, 1000 rpm, 2 min). The washing process was repeated twice, and the cells were resuspended in culture medium (500 mL DMEM + 10% FBS + 1 × ITS + 20 μl EGF (1 μg / μl) + 20 μl IGFII (0.2 μg / μl)) before being seeded into 6-well plates.

[0248] Multi-omics analysis Bulk RNA sequencing. Mouse liver tissue samples were homogenized in QIAzol lysis reagent. Total RNA was extracted and purified using the miRNeasy Mini Kit. Total RNA was subjected to RNA sequencing using a sequencing instrument. RNA sequencing data was obtained in Fasta file format. Reads were mapped to the mouse genome assembly (GRCm39, mm10) using HISAT2, followed by read counting using HTSeq counts. Differential expression analysis was estimated using the DESeq2 R-package. The count matrix was transformed and normalized using Dispersion Stabilization Transformation (VST) to be performed in DESeq2. Gene set enrichment analysis (GSEA)-based KEGG pathway, as well as gene ontology (GO) including biological processes (BP), molecular functions (MF), and cellular components (CC), was performed using differential expression data with web-based tools for functional enrichment analysis. Data was visualized using software packages.

[0249] Spatial transcriptome. Tissue samples were first screened for RNA quality to ensure a DV200 score above 50%, and spatial gene expression slides and reagent kits were used. 5-micrometer thick tissue sections were cut from FFPE tissue blocks and mounted on slides. After the sections underwent the initial deparaffinization, imaging, and permeabilization steps, they were carefully hybridized with probes. The successfully hybridized probe pairs were then ligated to facilitate junction sealing and finally released from the tissue in instrumentation and captured on FFPE Visium spatial gene expression slides placed within a fiducial frame.

[0250] Each capture region contained approximately 5000 gene expression spots, including sequencing primers, a 16-nucleotide (nt) spatial barcode, a 12-nt unique molecular identifier (UMI), and a 30-nt poly(dT) sequence (to capture the ligation product). These spots provided a resolution of approximately 5–10 cells. The ligation product was extended by adding the UMI, partial read 1, and spatial barcode to ultimately obtain a spatially barcoded product for subsequent library preparation. The cycle count was carefully confirmed using qRT-PCR, and the ligated, spatially barcoded product was indexed by Sample Index PCR. Library sequencing was performed using a sequencing instrument with an SP flow cell (100 cycles), and FASTQ files were processed to generate .cloup files. tSNE and spatial plots were performed and plotted using Loupe Browser (10× Genomics).

[0251] Spatial metabolomics. Details include sample preparation, H&E staining, mass spectrometry imaging (MSI) acquisition, and data analysis. (i) Sample preparation for MSI. Liver tissue was collected from mice, rinsed with PBS, placed in an embedding cassette containing a foam pad, and then gently frozen in a liquid nitrogen steam bath. The cassette was stored at -80°C. Tissue sections were prepared using a cryotome HM 500 O(Micro). The tissue was heated from -80°C to -15°C in 20 minutes. The tissue pieces were fixed on a Peltier platform with milliQ water and rapidly frozen for 10 minutes. The tissue sections were cut to a thickness of 20 μm and placed on labeled microscope slides. Alternatively, three slides were dedicated to mass spectrometry imaging (MSI) positive polarity mode, MSI negative mode, and hematoxylin and eosin (H&E) staining. MSI mass spectrometry imaging (MSI) sections were frozen in a slide mailer at -80°C and H&E staining was performed immediately after cutting. (ii) H&E staining. Sections on microscope slides were successively immersed in 100%, 70%, and 50% ethanol (EtOH) for 2 minutes each, followed by immersion in tap water for 2 minutes. The tissue was then stained four times with hematoxylin for 4.5 minutes each, and rinsed with water for 4 minutes. A second staining was performed with eosin for 2.2 minutes, followed again by rinsing with water for 1.5 minutes. Three final baths were applied to the sections for 1 minute each, using 50%, 70%, and 100% EtOH, respectively. Finally, the coverslips were quickly rinsed with xylene before mounting with Entellan. (iii) MSI acquisition. Full scans (50 m / z to 1200 m / z) were acquired in sensitivity mode using synapt XS waters with a DESI XS source. Three slices were acquired in positive polarity acquisition mode, and the other three were used in negative polarity acquisition mode. The tissue sections obtained in negative polarity were sprayed with 2 μl / min methanol / water buffer (96 / 4, 1 mmol ammonium acetate) containing leucine-enkephalin (250 ng / ml) using the rock spray method. The capillary voltage was set to 0.70 V, the sampling cone to 40 V, and the ion source temperature to 150 °C.For the negative polarity acquisition mode, the buffer solution at 2 μl / min consisted of methanol / water (82 / 18) containing 0.1% formic acid and 250 ng / ml leucine-enkephalin. The capillary voltage was set to 0.40 V, the sampling cone to 40 V, and the ion source temperature to 150 °C. The DESI source was 50 μm. 2 The tissue was scanned with a spatial resolution of / pixel, and the scan time was 0.153 seconds. Acquisition parameters were set in HDI v1.6 and acquired with MassLynx v4.2. (iv) Data analysis. Raw data was processed by HDI v1.6. A target list was constructed based on the top 1000 strongest ion signals from one replica. Then, a separate target list of special interest was added to this data-driven constructed list. The result is an identical list of m / z values ​​searched across all pixels (corresponding to mass spectra) of the entire set of MSI files. The HDI raw output was exported to a text file and imported into R software for further data processing. Data processing in R included raw data cleaning (image trimming, exclusion of replicas with acquisition errors) and normalization by total ion count (TIC). A smoothing function was applied to the MSI files to limit artifact signal variation between pixels. Next, artifact metabolites were removed: metabolites were excluded from subsequent analysis if the mean of the target tissue region (tissue ROI based on K-means pixel cluttering) was greater than half of the MSI file and less than the mean of the glass ROI. Then, all pixel values ​​corresponding to the tissue replicas were collected and analyzed by central and unscaled principal component analysis (PCA). Next, the PCA values ​​were clustered using the K-means method, and each pixel was assigned a color code according to its cluster. Then, images of the replicated tissue sections were reconstructed from this color code assignment using a common ROI for the entire set of files. Finally, each ion signal was averaged by the ROI, and the values ​​were calculated using a heatmap.

[0252] Combination therapy Anti-DBI + anti-PD-1. Hep55.1C cells (3 x 10⁶ cells in DMEM without FBS) 5 50 μL / mouse was inoculated into the livers of 7-week-old female C57BL / 6J mice. Ten days after tumor onset, the mice were randomly divided into four groups of 9-10 mice each (isotype 1 + isotype 2, isotype 1 + anti-PD1, anti-ACBP + isotype 2, and anti-ACBP + anti-PD1). Treatment with anti-DBI mAb (ip 5 mg / kg) or isotype 1 (ip 5 mg / kg) was then initiated and administered three times / week. Treatment with anti-PD1 / isotype 2 (ip 200 μg / mouse) was initiated one week after six doses of anti-ACBP / isotype 1. Mice were sacrificed at the humane endpoint described above. In Figures 21 and 26, Iso represents isotype, and DBI / αPD1 represents anti-DBI / anti-PD1, respectively.

[0253] Anti-DBI + ferroptosis inducer. Hep55.1C-Luc cells (3 x 10⁶ cells in DMEM without FBS) 5 50 μL of luciferin (15 mg / mL / mouse) was inoculated into the livers of 7-week-old female C57BL / 6J mice. Seven days after tumor initiation, tumors were evaluated by in vivo bioluminescence imaging using the IVIS system (200 μL of 15 mg / mL luciferin / mouse ip injection 5 minutes prior to detection). Mice were divided into four groups based on bioluminescence imaging (isotype + vehicle, isotype + RSL3 / IKE, anti-DBI + vehicle, anti-DBI + RSL3 / IKE). Treatment with anti-DBI / isotype (ip 5 mg / kg) was then initiated on a regular dosing schedule of 8 days out of 10, with 2 days of dosing and 1 day of rest. RSL3 and IKE were administered at 50 mg / kg four times over 2 consecutive weeks, starting on days 21 / 19 after tumor initiation. Tumor growth was monitored for 4 weeks. Mice were sacrificed at the above endpoints.

[0254] Histology and immunohistochemistry of the liver Liver tissue in the cassette was fixed with 4% paraformaldehyde for 24 hours and then transferred to 70% ethanol. After dehydration, the cassette was embedded in paraffin. For histological evaluation, liver sections were stained with hematoxylin and eosin (H&E) and Sirius Red. Slides were scanned and imaged using a slide scanner. The NAFLD activity score (NAS) was assessed by a specialist pathologist according to the NASH-CRN scoring system, without knowing the treatment received. Fibrosis stage was assessed by Sirius Red staining and quantified as % Sirius Red + area using QuPath software. Proliferation score was assessed by Ki67 and PCNA IHC staining of liver sections. Image analysis was performed using QuPath software as positive area %.

[0255] Immunofluorescence Primary HCC cells (DBI + / + and DBI - / - The cells were seeded at 2000 cells / well using four replicas. After 24 hours, the cells were fixed in 4% PFA / PBS (100 μL / well) at room temperature (RT) for 20 minutes. The fixed plates were washed twice with PBS (200 μL / well), and permeabilization and blocking were performed with 0.1% Triton® X-100 + 5% BSA + 10% FBS at RT for 60 minutes (100 μL / well). The primary antibody was diluted to the appropriate concentration in 1% BSA (100 μL / well) and incubated overnight at 4°C. After two PBS washes (200 μL / well), the secondary antibody (1:250) and DAPI (1:5000) in 1% BSA (50 μL / well) were subsequently added to the cells, and incubated at RT in the dark for 1 hour. The plates were washed twice with PBS, followed by the addition of PBS (100 μL / well). Image acquisition and analysis were performed using software.

[0256] Enzyme-linked immunosorbent assay Whole blood was collected from mice for enzyme-linked immunosorbent assay (ELISA). Plasma was isolated by centrifugation at 5000 × rpm at 4°C for 10 minutes. The level of DBI in the plasma was assayed by ELISA. High-binding ELISA plates were coated overnight at 4°C with 100 μL of anti-DBI antibody (human: MBS768488, mouse: ab231910). The plates were washed twice with 100 μL of washing buffer (PBS + 0.05% Tween® 20) and blocked with 200 μL of blocking buffer (PBS + 1% BSA + 0.05% Tween® 20) at room temperature for 2 hours. After washing, 100 μL of standard or diluted plasma sample (human: 1 / 50, mouse: 1 / 20) was added to the plate and incubated at room temperature for 2 hours. The plates were washed three times with washing buffer, followed by incubation with 1 μg / μl anti-DBI detection antibody (100 μL / well, human: LS-C299614, mouse: MBS2005521) at room temperature for 1 hour. After three wash buffer cycles, 100 μl of diluted avidin-HRP (human: 1 / 5000, mouse: 1 / 1000) was added to each well and incubated at room temperature for 30 minutes. The plates were washed three times, 100 μl of substrate was added, and incubated in the dark for 30 minutes. Then, 50 μl of stop solution was added, and the absorbance at 450 nm was read using a microplate reader.

[0257] Quantitative real-time PCR Liver tissue (approximately 25 mg) was homogenized in a lysis reagent. Total RNA was extracted and purified. Total RNA (1000 ng) was reverse transcribed to cDNA. Quantitative real-time PCR (qRT-PCR) detection was performed.

[0258] Immunoblotting Liver tissue was homogenized in a protein lysis buffer (20 mM Tris buffer pH 7.4 + 150 mM NaCl + 1% Triton® X-100 + 10 mM EDTA + protease inhibitor cocktail) using a homogenizer. Protein extracts were centrifuged at 12000 g (4°C) for 15 minutes, and the supernatant was collected. Protein concentration was determined by a BCA protein assay kit, and the samples were then boiled in Laemmli buffer at 100°C for 10 minutes. Total protein (25 μg) from each sample was loaded onto 4-12% NuPAGE Bis-Tris gels and transferred to PDVF membranes. To simultaneously detect different antigens within the same experiment, the membranes were horizontally sliced ​​into several sections based on the molecular weight of the target proteins. The membrane was blocked at RT for 1 hour with 5% (w / v) BSA in TBST (TBS containing 0.1% Tween®-20), and incubated overnight at 4°C with a suitable antibody in 5% (w / v) BSA in TBST. Detection was performed by incubation at RT for 1 hour with a suitable horseradish peroxidase (HRP) conjugate secondary antibody. The blot was visualized using ECL detection reagent. Optical density analysis was performed using software. GAPDH served as a loading control. The optical density ratio of target protein / GAPDH was normalized relative to the control group.

[0259] Bioinformatics analysis For differential expression of the liver disease (DBI) in cancer patients, RNA-seq data was downloaded from the TCGA database, extracted in TPM format, and presented as log2(value + 1). Differential expression of the liver disease (DBI) in HCC patients (presented as log2(value)) was further validated by analyzing data from HCC-related GEO datasets. In HCC patient plasma analysis, continuous variables are presented as median and interquartile range, and categorical variables as number and percentage. Correlations between continuous variables were assessed using Spearman's rank correlation analysis. Survival curves were plotted using Kaplan-Meier (KM) analysis. Liver disease-related GEO datasets were downloaded from the GEO database via the GEO query package. Data were normalized. When multiple probe values ​​were encountered for the same gene, only the probe with the highest signal value was retained. GSEA-based KEGG pathway analysis was performed using online tools for functional enrichment analysis in various biological contexts. The comprehensive transcriptome dysregulation similarities between the aforementioned liver disease GEO datasets and three NASH-driven mouse RNA-seq datasets (WD+CCl4_ACBP / DBI knockout, WD+CCl4_KLH-ACBP, and HFD+DEN_KLH-ACBP) in this study were analyzed by row and column clustering analysis (Euclidean distance) of normalized enrichment scores (NES). Statistical analysis and data visualization were performed.

[0260] statistical analysis Unless otherwise specified, data are presented as mean ± SEM. Normality was assessed by normality tests. Normally distributed data were analyzed using t-tests, one-way ANOVA, or two-way ANOVA. Non-normally distributed data were analyzed using Welch's t-test (two groups), Mann-Whitney U-test (two groups), or Kruskal-Wallis test (multiple groups), followed by Dunn's post-hoc test. Weight curves were analyzed using two-way ANOVA performed via the Tum-Growth online tool (https: / / kroemerlab.shinyapps.io / TumGrowth / ). A log-rank test was used for Kaplan-Meier (KM) survival analysis. Correlations were assessed using Spearman's rank correlation analysis. Unless otherwise specified, statistical analyses were performed using GraphPad Prism 8. [Table 1]

Claims

1. A composition for use in the treatment of cancer in the subject, (a) an amount of extracellular human diazepam binding inhibitor (DBI) sufficient to inhibit extracellular human DBI (anti-DBI agent), (b) One or more anticancer drugs Includes, A composition in which, when the anti-DBI agent and the one or more anticancer agents are administered to the subject in a sufficient therapeutic dose, the treatment of the cancer is enhanced compared to the administration of the one or more anticancer agents in a therapeutic dose without the anti-DBI agent.

2. The composition for use according to claim 1, further comprising one or more corticosteroid therapeutic agents.

3. The composition for use according to claim 2, for use in reducing the immunosuppressive effect of the one or more corticosteroid therapies in subjects undergoing immunotherapy or cancer treatment, compared to the level of immunosuppression with the administration of the one or more anticancer agents and one or more corticosteroid therapies without the anti-DBI agent.

4. The composition for use according to any one of claims 1 to 3, wherein the anti-DBI agent is an antibody or an aptamer.

5. The composition for use according to any one of claims 1 to 4, wherein the one or more anticancer agents comprises a chemotherapeutic agent, an immunotherapy agent, or both.

6. The composition for use according to claim 5, wherein the chemotherapeutic agent comprises immunogenic cell death (ICD) inducing activity.

7. The aforementioned chemotherapeutic agents include alkylating agents, ferroptosis inducers, alkyl sulfonates, aziridines, ethyleneimines, methylmelamine, altoretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, trimethylolmelamine, acetogenins, camptothecin, bryostatin, calistatin, CC-1065, cryptophycin, dorastatin, duocalmycin, elcerovine, pancratistatin, sarcodictiin, spongistin, nitrogen mustard, nitrosourea, antibiotics, dynemycin, bisphosphonates, esperamycin, neocartinostatin chromophore, related chromoprotein enediyne antibiotic chromophore, acrasinomycin, actinomycin, ausramycin, azaserin, bleomycin, kakutinomycin, carabicin, kaminomycin, cartinophylline, chromomycin, dactinomycin, daunorubicin, detrubicin, 6-Diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin, antimetabolites, folic acid analogs, purine analogs, pyrimidine analogs, androgens, anti-adrenal drugs, folic acid supplements, acegraton, aldofsphamide glycoside, aminolevulinic acid, enyluracil, amsacrin, Bestrabusil, Bisanthren, edatrexate, defofamine, demecolsin, diaziquan, elf Olumitin, eriptinium acetate, epotilon, etoglucid, gallium nitrate, hydroxyurea, lentinan, ronidamine, mytansinoid, mitoglucon, mitoxantrone, mopidammole, nitraerine, pentostatin, fenamet, pirarubicin, losoxantrone, podophyllic acid, 2-ethylhydrazide, procarbazine, PSK polysaccharide complex, razoxane, rhizoxin, schizofran, spirogermanium, tenuazonic acid, triadiquan, 2,2',2"-Trichlorotriethylamine, Trichothecene, Urethane, Vindesine, Dacarbazine, Mannomustine, Mitobronitol, Mitractol, Pipobroman, Gacitosine, Arabinoside, Cyclophosphamide, Thiotepa, Toxoid, Chlorambucil, Gemcitabine, 6-Thiogunine, Mercaptopurine, Methotrexate, Platinum-Coordinating Complex, Vinblastine, Platinum, Etoposide (VP-16), Ifosfamide, Mitoxantrone, Vincristine, Vinorelbine, Novantrone, Teniposide, Edatrexate, Daunomycin, Aminopterin, Xeloda, Ibandronate, Irinotecan, Topoisomerase Inhibitor RFS 2000, a composition for use according to claim 5, comprising difluoromethylornithine (DMFO), retinoids, capecitabine, sorafenib, olaparib, cetuximab, gefitinib, sulfasalazine, β-elemen, trigonelline, vorinostat, sunitinib, artesunate, any of the aforementioned pharmaceutically acceptable salts, acids or derivatives, or any combination thereof.

8. The aforementioned chemotherapeutic agents include thiotepa, cyclophosphamide, busulfan, improsulfan, piposulfan, benzodopa, carboquan, methuredopa, bratacin, bratacinone, topotecan, adzeresin, karzeresin, bizeresin synthetic analog, cryptophycin 1, cryptophycin 8, KW-2189, CB1-TM1, chlorambucil, chlornafadin, colophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobenbitin, fenestrine, Prednimustine, trophosphamide, uracil mustard, carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine, engine antibiotics, calicheamicin, calicheamicin gamma II, calicheamicin omega II, dynemycin A, clodronate, morpholino doxorubicin, cyanomorpholino doxorubicin, 2-pyrrolino doxorubicin, deoxydoxorubicin, mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, potifu Iromycin, puromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zolubicin, methotrexate, 5-fluorouracil (5-FU), denopterin, methotrexate, pteropterin, trimethrexate, fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxyuridine A composition for use according to claim 5, comprising: carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testactone, aminoglutethimide, mitotane, trilostane, folinic acid, mytacin, ansamitosine, T-2 toxin, veraculin A, loridine A, angidin, paclitaxel, docetaxel, cisplatin, oxaliplatin, carboplatin, CPT-11, retinoic acid, any of the pharmaceutically acceptable salts, acids or derivatives mentioned above, or any combination thereof.

9. The composition for use according to claim 5, wherein the one or more immunotherapeutic agents include activity against immune checkpoints.

10. The aforementioned immune checkpoints include PD1 (programmed death 1), PDL1 (programmed death ligand 1), CTLA4 (cytotoxic T lymphocyte-associated protein 4), PDL2 (programmed death ligand 2), KIR (killer cell immunoglobulin-like receptor), B7-H3, B7-H4, BTLA (B and T lymphocyte attenuator), LAG3 (lymphocyte activator gene 3), and TIM-3 (T cell immunoglobulin and mucin). The composition for use according to claim 9, comprising Main Content 3), VISTA (V-domain Ig suppressor for T cell activation), ILT2 / LILRB1 (Ig-like transcript 2 / leukocyte Ig-like receptor 1), ILT3 / LILRB4 (Ig-like transcript 3 / leukocyte Ig-like receptor 4), ILT4 / LILRB2 (Ig-like transcript 4 / leukocyte Ig-like receptor 2), TIGIT (T cell immune receptor having Ig and ITIM domains), NKG2A, PVRIG, CBLB (Casitas b lineage lymphoma oncogene B), CISH (cytokine-inducible SH2-containing protein), or any combination thereof.

11. The composition for use according to claim 5, wherein the one or more immunotherapeutic agents include an anti-PD1 agent, an anti-PD-L1 agent, an anti-CTLA4 agent, an anti-PD-L2 agent, an anti-KIR agent, an anti-B7-H3 agent, an anti-B7-H4 agent, an anti-BTLA agent, an anti-LAG3 agent, an anti-TIM-3 agent, an anti-VISTA agent, an anti-ILT2 / LILB1 agent, an anti-ILT3 / LILB4 agent, an anti-ILT4 / LILB2 agent, an anti-TIGIT agent, an anti-NKG2A agent, an anti-PVRIG agent, an anti-CBLB agent, an anti-CISH agent, or any combination thereof.

12. The immunotherapeutic agent may be anti-PD1 antibody, anti-PD-L1 antibody, anti-CTLA4 antibody, anti-PD-L2 antibody, anti-KIR antibody, anti-B7-H3 antibody, anti-B7-H4 antibody, anti-BTLA antibody, anti-LAG3 antibody, anti-TIM-3 antibody, anti-VISTA antibody, anti- ILT2 / LILRB1 antibody, anti-ILT3 / LILRB4 antibody, anti-ILT4 / LILRB2 antibody, anti-TIGIT antibody, anti-NKG2A antibody, anti-PVRIG antibody, anti-CBLB antibody, anti-CISH antibody, ipilimumab, tremelimumab, M K-1308, FPT155, PRS010, BMS-986249, BPI-002, CBT509, JS007, ONC392, TE1254, IBI310, BR02001, CG0161, KN044, PBI5D3H5, BCD145, ADU1604, AGEN1884, AGEN1181, CS1002, CP675206, Pembrolizumab, Nivolumab, Pidilizumab, AMP-224, BMS-936559, Semiprimab, PDR0 01, MDX-1105, MEDI4736, Atezolizumab, Avelumab, BMS-936559, Durvalumab, Rilurumab (IPH2102), IPH2101, MGA271, FPA150, IMP321 (Eftiragimod alfa), Relatrimab, MK-4280, AVA017, BI754111, ENUM006, GSK2831781, INCAGN2385, LAG3Ig, LAG525, REGN3767, Sym016, Sym0 A composition for use according to claim 5, comprising 22, Sym023, TSR033, TSR075, XmAb22841, LY3321367, MBG453, TSR-022, JNJ-61610588, MK-7684, PTZ-201, RG6058, COM902, IPH-2201, COM701, CA-327, LAG525, REGN3767, BI754111, teboterimab, FS118, MGC018, or any combination thereof.

13. The composition for use according to claim 5, wherein the immunotherapy agent comprises an anti-PD1 antibody, an anti-PD-L1 antibody, an anti-CTLA4 antibody, or any combination thereof.

14. A composition for use according to any one of claims 2 to 13, wherein the one or more corticosteroid therapeutic agents are selected from cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, beclomethasone, and any combination thereof.

15. The composition for use according to any one of claims 1 to 14, wherein the cancer comprises an inhibitory tumor microenvironment.

16. The administration of the composition results in the induction of at least one immune surveillance biomarker, including an increase in CD8 expression, a decrease in CD4 expression, a decrease in T cells (CD4 Foxp3), a decrease in T cells (CD4 Foxp3), an increase in the ratio of CD8 T cells to T cells (CD4 Foxp3), an increase in T cells (CD4 Foxp3), a decrease in Lag3 expression in T cells (CD4 Foxp3), a decrease in Lag3 expression in CD8 cells, a decrease in Foxp3 expression in CD4 cells, a decrease in T ICOSGITRLag3 CD4 cells, or a combination thereof, compared to an equivalent method without administration of the anti-DBI agent. A composition for use according to any one of claims 1 to 15. ＋ increase in expression, CD4 ＋ decrease in expression, T ｒｅｇ cells (CD4 ＋ Foxp3 ＋ ), decrease in T ｒｅｇ cells (CD4 ＋ Foxp3 ＋ ), increase in the ratio of CD8 ＋ T cells to T Ｈ + cells (CD4 ＋ Foxp3 － ), increase in T Ｈ + cells (CD4 ＋ Foxp3 － ), decrease in Lag3 ＋ expression in T ＋ cells (CD4 ＋ Foxp3 ＋ ), decrease in Lag3 ＋ expression in CD8 ｒｅｇ cells, decrease in Foxp3 ＋ expression in CD4 ＋ cells, decrease in T － ICOSGITRLag3 ＋ CD4 cells, or a combination thereof.

17. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent compared to CD8 ＋ A composition for use according to claim 16, comprising an increase in expression.

18. Inducing at least one of the aforementioned immune surveillance biomarkers is equivalent to CD4 without the administration of the anti-DBI agent. ＋ A composition for use according to claim 16, comprising a reduction in expression.

19. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to T ｒｅｇ cells (CD4 ＋ Foxp3 ＋ The composition for use according to claim 16, which includes a reduction of ).

20. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to T ｒｅｇ cells (CD4 ＋ Foxp3 ＋ ) CD8 ＋ A composition for use according to claim 16, comprising an increase in the proportion of T cells.

21. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to T Ｈ + cells (CD4 ＋ Foxp3 － The composition for use according to claim 16, comprising an increased incidence rate of ).

22. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to T Ｈ + cells (CD4 ＋ Foxp3 － ) Lag3 ＋ A composition for use according to claim 16, comprising a reduction in the expression of [the specified substance].

23. Inducing at least one immune surveillance biomarker is more effective than an equivalent method without the administration of the anti-DBI agent, compared to CD8 ＋ Lag3 in cells ＋ A composition for use according to claim 16, comprising a reduction in the expression of [the specified substance].

24. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to CD4 ＋ Foxp3 in cells ＋ A composition for use according to claim 16, comprising a reduction in the expression of [the specified substance].

25. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to T ｒｅｇ ICOS ＋ GITR ＋ Lag3 － CD4 ＋ A composition for use according to claim 16, comprising a reduction in cells.

26. T Ｈ + cells (CD4 ＋ Foxp3 － ) CD8 against cells ＋ The composition for use according to any one of claims 16 to 25, wherein the T cell ratio is maintained when the composition is administered compared to an equivalent method without administration of the anti-DBI agent.

27. CD4 ＋ CD8 against T cells ＋ The composition for use according to any one of claims 16 to 25, wherein the T cell ratio is maintained when the composition is administered compared to an equivalent method without administration of the anti-DBI agent.

28. T Ｈ ICOS ｉｎｔ GITR ＋ Lag3 － PD1 － CD4 ＋ The composition for use according to any one of claims 16 to 25, wherein the T cell generation rate is maintained when the composition is administered compared to an equivalent method without administration of the anti-DBI agent.

29. The composition for use according to any one of claims 1 to 28, wherein administration of the composition results in inhibition of at least one cancer progression biomarker, including a reduction in the target tumor volume, a reduction in cancer cell proliferation, an increase in cancer cell death, a reduction in cancer growth, or a combination thereof, compared to an equivalent method without administration of the anti-DBI agent.

30. The composition for use according to claim 29, wherein the inhibition of the at least one cancer progression biomarker comprises a reduction in tumor volume, and the reduction in tumor volume comprises a reduction in tumor area or tumor volume.

31. The composition for use according to claim 30, wherein the reduction in tumor volume includes a reduction in tumor volume compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

32. The composition for use according to claim 31, wherein the tumor volume is reduced by up to approximately 60%.

33. The composition for use according to claim 30, wherein the reduction in tumor volume includes a reduction in tumor area compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

34. The composition for use according to claim 29, wherein inhibiting the at least one cancer progression biomarker comprises reducing the proliferation of cancer cells compared to an equivalent method without administration of the anti-DBI agent at an equivalent time point.

35. The composition for use according to claim 29, wherein inhibiting the at least one cancer progression biomarker includes an increase in cancer cell death compared to an equivalent method without administration of the anti-DBI agent at an equivalent time point.

36. The composition for use according to claim 29, wherein inhibiting the at least one cancer progression biomarker comprises a reduction in cancer growth compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

37. The composition for use according to any one of claims 51 to 36, wherein administration of the composition results in an increased survival rate of the subject compared to the absence of administration of the anti-DBI agent at the same time point.

38. The composition for use according to claim 37, wherein the survival rate is increased by up to approximately 50% compared to an equivalent method without administration of the anti-DBI agent at an equivalent time point.

39. A composition for use according to any one of claims 1 to 38, wherein administration of the composition results in an increased incidence of cancer-free occurrences in the subject compared to the absence of administration of the anti-DBI agent at the same time point.

40. The composition for use according to claim 39, wherein the incidence of the cancer-free occurrence is increased by up to 40% compared to an equivalent method without administration of the anti-DBI agent at an equivalent time point.

41. The composition for use according to any one of claims 1 to 40, wherein the cancer is an intractable cancer.

42. The composition for use according to any one of claims 1 to 41, wherein the cancer is resistant to immune checkpoint inhibitor therapy.

43. The composition for use according to any one of claims 1 to 42, wherein the cancer is selected from solid tumors, hematological tumors, skin cancers, tissue cancers, organ cancers, bone cancers, cartilage cancers, blood cancers, vascular cancers, primary cancers, metastatic cancers, bladder cancers, bone marrow cancers, brain cancers, breast cancers, colon cancers, esophageal cancers, gastrointestinal cancers, gingival cancers, kidney cancers, liver cancers, lung cancers, nasopharyngeal cancers, cervical cancers, ovarian cancers, prostate cancers, stomach cancers, testicular cancers, tongue cancers, and uterine cancers.

44. The aforementioned cancers include: cancer, lung cancer, non-small cell lung cancer, breast cancer, neoplastic cancer, undifferentiated carcinoma, giant cell carcinoma, spindle cell carcinoma, small cell carcinoma, papillary carcinoma, squamous cell carcinoma, lymphoepithelial carcinoma, basal cell carcinoma, matrix carcinoma, transitional cell carcinoma, papillary transitional cell carcinoma, adenocarcinoma, gastrinoma, cholangiocarcinoma, hepatocellular carcinoma, combination of hepatocellular carcinoma and cholangiocarcinoma, cord-like adenocarcinoma, adenoid cystic carcinoma, adenocarcinoma in adenomatous polyps, familial adenomatous polyposis, solid tumors, carcinoid tumors, bronchioloalveolar adenocarcinoma, papillary adenocarcinoma, chromophobe carcinoma, oxiphilic adenocarcinoma, basophilic carcinoma, clear cell adenocarcinoma, granular cell carcinoma, follicular adenocarcinoma, papillary adenocarcinoma, follicular adenocarcinoma, non-encapsulating sclerosing carcinomaCarcinoma, adrenocortical carcinoma, endometrioid carcinoma, adnexal carcinoma, apocrine gland carcinoma, sebaceous gland carcinoma, ceruminous gland carcinoma, mucoepidermoid carcinoma, cystadenocarcinoma, papillary cystadenocarcinoma, papillary serous cystadenocarcinoma, mucinous cystadenocarcinoma, mucinous adenocarcinoma, signet ring cell carcinoma, invasive ductal carcinoma, medullary carcinoma, lobular carcinoma, inflammatory carcinoma, Paget's disease, breast cancer, acinar cell carcinoma, adenosquamous cell carcinoma, adenocarcinoma with squamous metaplasia, thymoma, ovarian stromal tumor, follicular cell tumor, granulosa cell tumor, lobulostomy, Sertoli cell carcinoma, Leydig cell tumor, lipid cell tumor, accessory ganglion, extramammary accessory ganglion, pheochromocytoma, globulinoma Mussian angiosarcoma, melanoma, achromatic melanoma, superficial spreading melanoma, melanoma within a giant pigmented nevus, epithelioid cell melanoma, blue nevus, sarcoma, fibrosarcoma, fibrous histiocytoma, myxosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, fetal rhabdomyosarcoma, alveolar rhabdomyosarcoma, stromal sarcoma, mixed tumor, Müllerian mixed tumor, nephroblastoma, hepatoblastoma, carcinosarcoma, mesenchymal tumor, Brenner tumor, phyllodes tumor, synovial sarcoma, mesothelioma, ovarian progenitor tumor, fetal carcinoma, teratoma, ovarian goiter, choriocarcinoma, mesonephroma, angiosarcoma, hemangioendothelioma, Kaposi's sarcoma, hemangiopericytoma, lymphangiosarcoma, bone Sarcoma, subcortical osteosarcoma, chondrosarcoma, chondroblastoma, mesenchymal chondrosarcoma, giant cell tumor of bone, Ewing's sarcoma, odontogenic tumor, ameloblastic odontogenic sarcoma, ameloblastoma, ameloblastic fibrosarcoma, pineal tumor, chordoma, glioma, ependymoma, astrocytoma, protoplasmic astrocytoma, fibrous astrocytoma, astroblastoma, glioblastoma, oligodendroglioma, oligodendroglioma, primitive neuroectodermal tumor, cerebellar sarcoma, ganglioma, neuroblastoma, retinoblastoma, olfactory neuron tumor, meningioma, neurofibrosarcoma, schwannoma, granulocyte tumor, lymphoma, Hodgkin's disease, Hodgkin's lymphoma A composition for use according to any one of claims 1 to 42, selected from tumors, paragranulomas, small lymphocytic lymphomas, large diffuse lymphomas, follicular lymphomas, mycosis fungoides, other specified non-Hodgkin lymphomas, histiocytic proliferative disorders, multiple myelomas, mast cell sarcomas, small intestinal immunoproliferative disorders, leukemias, lymphocytic leukemias, plasma cell leukemias, erythroleukemias, lymphosarcoma cell leukemias, myeloid leukemias, basophilic leukemias, eosinophilic leukemias, monocytic leukemias, mast cell leukemias, megakaryoblastic leukemias, myeloid sarcomas, and hairy cell leukemias.

45. The composition for use according to any one of claims 1 to 44, wherein the cancer is malignant cancer.

46. The composition for use according to any one of claims 1 to 45, wherein the cancer is selected from carcinoma, lung cancer, non-small cell lung cancer, and breast cancer.

47. The composition for use according to any one of claims 1 to 46, wherein the cancer is a solid tumor carcinoma.

48. The composition for use according to any one of claims 1 to 46, wherein the cancer is a fibrous tumor.

49. The composition for use according to any one of claims 1 to 48, wherein the subject is a human.

50. A method for treating cancer in a subject requiring treatment for cancer, wherein the method involves the subject, (a) an amount of extracellular human diazepam binding inhibitor (DBI) sufficient to inhibit extracellular human DBI (anti-DBI agent), (b) One or more anticancer drugs This includes administering, A method wherein the administration of the anti-DBI agent and the one or more anticancer agents is in a therapeutic dose sufficient to treat cancer, and results in an enhanced treatment of the cancer in the subject compared to an equivalent method without the administration of the anti-DBI agent.

51. A method for improving the therapeutic effect of immunotherapy in a subject, wherein the method applies to the subject, (a) Drugs that stimulate autophagy by inhibiting extracellular human diazepam binding inhibitors (DBIs) (anti-DBI agents), (b) One or more immunotherapeutic agents This includes administering, A method wherein the administration of the anti-DBI agent and the one or more immunotherapy agents is sufficient to improve the therapeutic effect of immunotherapy compared to an equivalent method without the administration of the anti-DBI agent.

52. The method according to claim 50 or 51, further comprising administering one or more corticosteroid therapeutic agents.

53. The method according to claim 52, wherein the method reduces the immunosuppressive effect of the one or more corticosteroid therapies in a subject receiving immunotherapy or cancer treatment, compared to an equivalent method without the administration of the anti-DBI agent.

54. A method for enhancing immune surveillance in a subject with cancer in order to enhance the therapeutic effect of anticancer therapy, wherein the method involves the subject, (a) an amount of extracellular human diazepam binding inhibitor (DBI) sufficient to inhibit extracellular human DBI (anti-DBI agent), (b) One or more anticancer drugs This includes administering, A method wherein the administration of the anti-DBI agent and the one or more anticancer agents is sufficient to induce one or more immune surveillance biomarkers that result in an enhancement of the therapeutic effect of the anticancer therapy compared to an equivalent method without the administration of the anti-DBI agent.

55. A method for reducing the immunosuppressive effect of corticosteroid therapy in a subject receiving immunotherapy, wherein the method involves the subject, (a) an amount of extracellular human diazepam binding inhibitor (DBI) sufficient to inhibit extracellular human DBI (anti-DBI agent), (b) One or more corticosteroid therapy agents This includes administering, A method wherein the administration of the anti-DBI agent and the one or more corticosteroid therapeutic agents is sufficient to reduce the immunosuppressive effect of the corticosteroid therapy in the subject receiving immunotherapy, compared to an equivalent method without the administration of the anti-DBI agent.

56. The method according to claim 55, further comprising administering one or more anticancer drugs.

57. The method according to any one of claims 50 to 56, wherein the anti-DBI agent is an antibody or an aptamer.

58. The method according to any one of claims 51 to 54, 56, or 57, wherein the one or more anticancer agents include a chemotherapeutic agent, an immunotherapy agent, or both.

59. The method according to claim 58, wherein the chemotherapeutic agent contains immunogenic cell death (ICD) inducing activity.

60. The aforementioned chemotherapeutic agents include alkylating agents, ferroptosis inducers, alkyl sulfonates, aziridines, ethyleneimines, methylmelamine, altoretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, trimethylolmelamine, acetogenins, camptothecin, bryostatin, calistatin, CC-1065, cryptophycin, dorastatin, duocalmycin, elcerovine, pancratistatin, sarcodictiin, spongistin, nitrogen mustard, nitrosourea, antibiotics, dynemycin, bisphosphonates, esperamycin, neocartinostatin chromophore, related chromoprotein enediyne antibiotic chromophore, acrasinomycin, actinomycin, ausramycin, azaserin, bleomycin, kakutinomycin, carabicin, kaminomycin, cartinophylline, chromomycin, dactinomycin, daunorubicin, detrubicin, 6-Diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin, antimetabolites, folic acid analogs, purine analogs, pyrimidine analogs, androgens, anti-adrenal drugs, folic acid supplements, acegraton, aldofsphamide glycoside, aminolevulinic acid, enyluracil, amsacrin, Bestrabusil, Bisanthren, edatrexate, defofamine, demecolsin, diaziquan, elf Olumitin, eriptinium acetate, epotilon, etoglucid, gallium nitrate, hydroxyurea, lentinan, ronidamine, mytansinoid, mitoglucon, mitoxantrone, mopidammole, nitraerine, pentostatin, fenamet, pirarubicin, losoxantrone, podophyllic acid, 2-ethylhydrazide, procarbazine, PSK polysaccharide complex, razoxane, rhizoxin, schizofran, spirogermanium, tenuazonic acid, triadiquan, 2,2',2"-Trichlorotriethylamine, Trichothecene, Urethane, Vindesine, Dacarbazine, Mannomustine, Mitobronitol, Mitractol, Pipobroman, Gacitosine, Arabinoside, Cyclophosphamide, Thiotepa, Toxoid, Chlorambucil, Gemcitabine, 6-Thiogunine, Mercaptopurine, Methotrexate, Platinum-Coordinating Complex, Vinblastine, Platinum, Etoposide (VP-16), Ifosfamide, Mitoxantrone, Vincristine, Vinorelbine, Novantrone, Teniposide, Edatrexate, Daunomycin, Aminopterin, Xeloda, Ibandronate, Irinotecan, Topoisomerase Inhibitor RFS The method according to claim 58, comprising 2000, difluoromethylornithine (DMFO), retinoids, capecitabine, sorafenib, olaparib, cetuximab, gefitinib, sulfasalazine, β-elemen, trigonelline, vorinostat, sunitinib, artesunate, any pharmaceutically acceptable salt, acid or derivative thereof, or any combination thereof.

61. The aforementioned chemotherapeutic agents include thiotepa, cyclophosphamide, busulfan, improsulfan, piposulfan, benzodopa, carboquan, methuredopa, bratacin, bratacinone, topotecan, adzeresin, karzeresin, bizeresin synthetic analog, cryptophycin 1, cryptophycin 8, KW-2189, CB1-TM1, chlorambucil, chlornafadin, colophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobenbitin, and fenesterine. Prednimustine, trophosphamide, uracil mustard, carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine, engine-based antibiotics, calicheamicin, calicheamicin gamma II, calicheamicin omega II, dynemycin A, clodronate, morpholino doxorubicin, cyanomorpholino doxorubicin, 2-pyrrolino doxorubicin, deoxydoxorubicin, mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, Potiphyllomycin, puromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zolubicin, methotrexate, 5-fluorouracil (5-FU), denopterin, methotrexate, pteropterin, trimethrexate, fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, flox The method according to claim 58, comprising siuridine, carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, aminoglutethimide, mitotane, trilostane, folinic acid, mytacin, ansamitosin, T-2 toxin, veraculin A, loridine A, angidin, paclitaxel, docetaxel, cisplatin, oxaliplatin, carboplatin, CPT-11, retinoic acid, any of the pharmaceutically acceptable salts, acids or derivatives described above, or any combination thereof.

62. The method according to claim 58, wherein the one or more immunotherapeutic agents include activity against immune checkpoints.

63. The aforementioned immune checkpoints include PD1 (programmed death 1), PDL1 (programmed death ligand 1), CTLA4 (cytotoxic T lymphocyte-associated protein 4), PDL2 (programmed death ligand 2), KIR (killer cell immunoglobulin-like receptor), B7-H3, B7-H4, BTLA (B and T lymphocyte attenuator), LAG3 (lymphocyte activator gene 3), and TIM-3 (T cell immunoglobulin and mucin). The method according to claim 62, comprising: Main component 3), VISTA (V-domain Ig suppressor for T cell activation), ILT2 / LILB1 (Ig-like transcript 2 / leukocyte Ig-like receptor 1), ILT3 / LILB4 (Ig-like transcript 3 / leukocyte Ig-like receptor 4), ILT4 / LILB2 (Ig-like transcript 4 / leukocyte Ig-like receptor 2), TIGIT (T cell immune receptor having Ig and ITIM domains), NKG2A, PVRIG, CBLB (Casitas b lineage lymphoma oncogene B), CISH (cytokine-inducible SH2-containing protein), or any combination thereof.

64. The method according to any one of claims 58 to 63, wherein the one or more immunotherapeutic agents include an anti-PD1 agent, an anti-PD-L1 agent, an anti-CTLA4 agent, an anti-PD-L2 agent, an anti-KIR agent, an anti-B7-H3 agent, an anti-B7-H4 agent, an anti-BTLA agent, an anti-LAG3 agent, an anti-TIM-3 agent, an anti-VISTA agent, an anti-ILT2 / LILB1 agent, an anti-ILT3 / LILB4 agent, an anti-ILT4 / LILB2 agent, an anti-TIGIT agent, an anti-NKG2A agent, an anti-PVRIG agent, an anti-CBLB agent, an anti-CISH agent, or any combination thereof.

65. The immunotherapeutic agent may be anti-PD1 antibody, anti-PD-L1 antibody, anti-CTLA4 antibody, anti-PD-L2 antibody, anti-KIR antibody, anti-B7-H3 antibody, anti-B7-H4 antibody, anti-BTLA antibody, anti-LAG3 antibody, anti-TIM-3 antibody, anti-VISTA antibody, anti- ILT2 / LILRB1 antibody, anti-ILT3 / LILRB4 antibody, anti-ILT4 / LILRB2 antibody, anti-TIGIT antibody, anti-NKG2A antibody, anti-PVRIG antibody, anti-CBLB antibody, anti-CISH antibody, ipilimumab, tremelimumab, MK -1308, FPT155, PRS010, BMS-986249, BPI-002, CBT509, JS007, ONC392, TE1254, IBI310, BR02001, CG0161, KN044, PBI5D3H5, BCD145, ADU1604, AGEN1884, AGEN1181, CS1002, CP675206, Pembrolizumab, Nivolumab, Pidilizumab, AMP-224, BMS-936559, Semiprimab, PDR001 MDX-1105, MEDI4736, Atezolizumab, Avelumab, BMS-936559, Durvalumab, Rilurumab (IPH2102), IPH2101, MGA271, FPA150, IMP321 (Eftiragimod alfa), Relatrimab, MK-4280, AVA017, BI754111, ENUM006, GSK2831781, INCAGN2385, LAG3Ig, LAG525, REGN3767, Sym016, Sym022, The method according to any one of claims 58 to 64, comprising Sym023, TSR033, TSR075, XmAb22841, LY3321367, MBG453, TSR-022, JNJ-61610588, MK-7684, PTZ-201, RG6058, COM902, IPH-2201, COM701, CA-327, LAG525, REGN3767, BI754111, tevoterimab, FS118, MGC018, or any combination thereof.

66. The method according to any one of claims 58 to 64, wherein the immunotherapy agent comprises an anti-PD1 antibody, an anti-PD-L1 antibody, an anti-CTLA4 antibody, or any combination thereof.

67. The method according to any one of claims 52 to 66, wherein the one or more corticosteroid therapeutic agents are selected from cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, beclomethasone, and any combination thereof.

68. The method according to any one of claims 51 to 67, wherein the cancer includes an inhibitory tumor microenvironment.

69. The method, compared to an equivalent method without the administration of the anti-DBI agent, shows that the CD8 in the sample ＋ Increased expression, CD4 ＋ Decreased expression, T ｒｅｇ cells (CD4 ＋ Foxp3 ＋ ) decrease, T ｒｅｇ cells (CD4 ＋ Foxp3 ＋ ) CD8 ＋ Increase in the ratio of T cells, T Ｈ + cells (CD4 ＋ Foxp3 － ) increase, T Ｈ + cells (CD4 ＋ Foxp3 － ) Lag3 ＋ Decreased expression of CD8 ＋ Lag3 in cells ＋ Decreased expression, CD4 ＋ Foxp3 in cells ＋ Decreased expression of T ｒｅｇ ICOS ＋ GITR ＋ Lag3 － CD4 ＋ The method according to any one of claims 50 to 68, further comprising inducing at least one immunosurveillance biomarker, which includes a decrease in cells or a combination thereof.

70. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent compared to CD8 ＋ The method according to claim 69, including an increase in expression.

71. Inducing at least one of the aforementioned immune surveillance biomarkers is equivalent to CD4 without the administration of the anti-DBI agent. ＋ The method according to claim 69, including a reduction in expression.

72. Inducing said at least one immune surveillance biomarker comprises a decrease in T ｒｅｇ cells (CD4 ＋ Foxp3 ＋ ) as compared to an equivalent method without administration of said anti-DBI agent, according to the method of claim 69.

73. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to T ｒｅｇ cells (CD4 ＋ Foxp3 ＋ ) CD8 ＋ The method according to claim 69, comprising an increase in the proportion of T cells.

74. Inducing said at least one immune surveillance biomarker comprises an increase in the incidence of T Ｈ + cells (CD4 ＋ Foxp3 － ), the method according to claim 69.

75. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to T Ｈ + cells (CD4 ＋ Foxp3 － ) Lag3 ＋ The method according to claim 69, which includes a reduction in the expression of [the substance].

76. Inducing at least one immune surveillance biomarker is more effective than an equivalent method without the administration of the anti-DBI agent, compared to CD8 ＋ Lag3 in cells ＋ The method according to claim 69, which includes a reduction in the expression of [the substance].

77. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to CD4 ＋ Foxp3 in cells ＋ The method according to claim 69, which includes a reduction in the expression of [the substance].

78. Inducing at least one of the aforementioned immune surveillance biomarkers is more effective than an equivalent method without the administration of the anti-DBI agent, compared to T ｒｅｇ ICOS ＋ GITR ＋ Lag3 － CD4 ＋ The method according to claim 69, comprising a reduction in cells.

79. The above method, T Ｈ + cells (CD4 ＋ Foxp3 － ) CD8 against cells ＋ The method according to any one of claims 69 to 78, wherein the ratio of T cells is maintained compared to an equivalent method without the administration of the anti-DBI agent.

80. The above method, CD4 ＋ CD8 against T cells ＋ The method according to any one of claims 69 to 78, wherein the ratio of T cells is maintained compared to an equivalent method without the administration of the anti-DBI agent.

81. The above method, T Ｈ ICOS ｉｎｔ GITR ＋ Lag3 － PD1 － CD4 ＋ The method according to any one of claims 69 to 78, which maintains the T cell incidence rate compared to an equivalent method without administration of the anti-DBI agent.

82. The method according to any one of claims 51 to 81, further comprising inhibiting at least one cancer progression biomarker, including a reduction in the target tumor volume, a reduction in cancer cell proliferation, an increase in cancer cell death, a reduction in cancer growth, or a combination thereof, compared to an equivalent method without the administration of the anti-DBI agent.

83. The method according to claim 82, wherein the inhibition of the at least one cancer progression biomarker includes a reduction in tumor volume, and the reduction in tumor volume includes a reduction in tumor area or tumor volume.

84. The method according to claim 83, wherein the reduction in tumor volume includes a reduction in tumor volume compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

85. The method according to claim 84, wherein the tumor volume is reduced by up to approximately 60%.

86. The method according to claim 83, wherein the reduction in tumor volume includes a reduction in tumor area compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

87. The method according to claim 82, wherein inhibiting the at least one cancer progression biomarker comprises reducing the proliferation of cancer cells compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

88. The method according to claim 82, wherein inhibiting the at least one cancer progression biomarker includes an increase in cancer cell death compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

89. The method according to claim 82, wherein inhibiting the at least one cancer progression biomarker includes a reduction in cancer growth compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

90. The method according to any one of claims 50 to 89, wherein the method further comprises an increase in the incidence of survival of the subject compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

91. The method according to claim 90, wherein the survival rate is increased by up to approximately 50% compared to an equivalent method without administration of the anti-DBI agent at the same time point.

92. The method according to any one of claims 51 to 89, further comprising an increase in the incidence of cancer-free occurrences in the subject compared to an equivalent method without administration of the anti-DBI agent at equivalent time points.

93. The method according to claim 92, wherein the incidence of the cancer-free occurrence is increased by up to 40% compared to an equivalent method without administration of the anti-DBI agent at the same time point.

94. The method according to any one of claims 50 to 93, wherein the method further comprises an increase in the time to death of the subject compared to an equivalent method without administration of the anti-DBI agent at an equivalent time point.

95. The method according to any one of claims 51 to 94, wherein the cancer is an intractable cancer.

96. The method according to any one of claims 51 to 95, wherein the cancer is resistant to immune checkpoint inhibitor therapy.

97. The method according to any one of claims 51 to 96, wherein the cancer is selected from solid tumors, hematological tumors, skin cancers, tissue cancers, organ cancers, bone cancers, cartilage cancers, blood cancers, vascular cancers, primary cancers, metastatic cancers, bladder cancers, bone marrow cancers, brain cancers, breast cancers, colon cancers, esophageal cancers, gastrointestinal cancers, gingival cancers, kidney cancers, liver cancers, lung cancers, nasopharyngeal cancers, cervical cancers, ovarian cancers, prostate cancers, stomach cancers, testicular cancers, tongue cancers, and uterine cancers.

98. The aforementioned cancers include cancer, lung cancer, non-small cell lung cancer, breast cancer, neoplastic cancer, undifferentiated carcinoma, giant cell carcinoma, spindle cell carcinoma, small cell carcinoma, papillary carcinoma, squamous cell carcinoma, lymphoepithelial carcinoma, basal cell carcinoma, matrix carcinoma, transitional cell carcinoma, papillary transitional cell carcinoma, adenocarcinoma, gastrinoma, cholangiocarcinoma, hepatocellular carcinoma, combination of hepatocellular carcinoma and cholangiocarcinoma, cord-like adenocarcinoma, adenoid cystic carcinoma, adenocarcinoma within adenomatous polyps, familial adenomatous polyposis, solid tumors, carcinoid tumors, bronchioloalveolar adenocarcinoma, papillary adenocarcinoma, chromophobe carcinoma, oxiphilic adenocarcinoma, basophilic carcinoma, clear cell adenocarcinoma, granular cell carcinoma, follicular adenocarcinoma, papillary adenocarcinoma, follicular adenocarcinoma, and unencapsulated adenocarcinoma. Sclerosing carcinoma, adrenocortical carcinoma, endometrioid carcinoma, adnexal carcinoma, apocrine gland carcinoma, sebaceous gland carcinoma, ceruminous gland carcinoma, mucoepidermoid carcinoma, cystadenocarcinoma, papillary cystadenocarcinoma, papillary serous cystadenocarcinoma, mucinous cystadenocarcinoma, mucinous adenocarcinoma, signet ring cell carcinoma, invasive ductal carcinoma, medullary carcinoma, lobular carcinoma, inflammatory carcinoma, Paget's disease, breast cancer, acinar cell carcinoma, adenosquamous cell carcinoma, adenocarcinoma with squamous metaplasia, thymoma, ovarian stromal tumor, follicular cell tumor, granulosa cell tumor, lobulostomy, Sertoli cell carcinoma, Leydig cell tumor, lipid cell tumor, accessory ganglion, extramammary accessory ganglion, pheochromocytoma, glomus angiosarcoma, melanoma, achromatic melanoma, superficial Melanoma, melanoma within a giant pigmented nevus, epithelioid cell melanoma, blue nevus, sarcoma, fibrosarcoma, fibrous histiocytoma, myxosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, fetal rhabdomyosarcoma, alveolar rhabdomyosarcoma, stromal sarcoma, mixed tumor, Müllerian mixed tumor, nephroblastoma, hepatoblastoma, carcinosarcoma, mesenchymal tumor, Brenner tumor, phyllodes tumor, synovial sarcoma, mesothelioma, ovarian progenitor tumor, fetal carcinoma, teratoma, ovarian goiter, choriocarcinoma, mesonephroma, angiosarcoma, hemangioendothelioma, Kaposi's sarcoma, hemangiopericytoma, lymphangiosarcoma, osteosarcoma, subcortical osteosarcoma, chondrosarcoma, chondroblastoma, mesenchymal chondrosarcoma, giant cell tumor of bone Ewing's sarcoma, odontogenic tumor, ameloblastic odontogenic sarcoma, ameloblastoma, ameloblastic fibrosarcoma, pineal tumor, chordoma, glioma, ependymoma, astrocytoma, protoplasmic astrocytoma, fibrous astrocytoma, astroblastoma, glioblastoma, oligodendroglioma, oligodendroglioma, primitive neuroectodermal tumor, cerebellar sarcoma, gangliblastoma, neuroblastoma, retinoblastoma, olfactory neuron tumor, meningioma, neurofibrosarcoma, schwannoma, granulocyte tumor, lymphoma, Hodgkin's disease, Hodgkin's lymphoma, paragranuloma, small lymphocytic lymphoma, large cell diffuse lymphoma, follicular lymphoma, mycosis fungoides,The method according to any one of claims 51 to 96, selected from other specified non-Hodgkin lymphoma, histiocytic proliferative disorder, multiple myeloma, mast cell sarcoma, intestinal immunoproliferative disorder, leukemia, lymphocytic leukemia, plasma cell leukemia, erythroleukemia, lymphosarcoma cell leukemia, myeloid leukemia, basophilic leukemia, eosinophilic leukemia, monocytic leukemia, mast cell leukemia, megakaryoblastic leukemia, myeloid sarcoma, and hairy cell leukemia.

99. The method according to any one of claims 51 to 98, wherein the cancer is malignant cancer.

100. The method according to any one of claims 51 to 99, wherein the cancer is selected from carcinoma, lung cancer, non-small cell lung cancer, and breast cancer.

101. The method according to any one of claims 51 to 100, wherein the cancer is a solid tumor carcinoma.

102. The method according to any one of claims 51 to 100, wherein the cancer is a fibrous tumor.

103. The method according to any one of claims 51 to 102, wherein the subject is a human.

104. An anti-DBI agent for use in the method according to any one of claims 50 to 103.

105. An immunotherapeutic agent for use in the method according to any one of claims 50, 52, 53, or 57 to 103.

106. A composition comprising an immunotherapy agent for use in the method according to any one of claims 50, 52, 53, or 57 to 103.

107. An anticancer agent for use in the method according to any one of claims 51 to 103.

108. A composition comprising an anticancer agent for use in the method according to any one of claims 51 to 103.

109. A corticosteroid therapeutic agent for use in the method according to any one of claims 52, 53, or 55 to 103.

110. A composition comprising a corticosteroid therapeutic agent for use in the method according to any one of claims 52, 53, or 55 to 103.

111. A combination of an anti-DBI agent and an immunotherapy agent for use in the method according to any one of claims 50, 52, 53, or 57-103.

112. A combination of an anti-DBI agent and an anticancer agent for use in the method according to any one of claims 51 to 103.

113. A combination of an anti-DBI agent and a corticosteroid therapy agent for use in the method according to any one of claims 52, 53, or 55 to 103.

114. A treatment regimen for treating cancer, (a) an amount of extracellular human diazepam binding inhibitor (DBI) sufficient to inhibit extracellular human DBI (anti-DBI agent), (b) One or more anticancer drugs Includes, A treatment regime in which the anti-DBI agent and the one or more anticancer agents are present in the treatment regime in an amount sufficient to inhibit one or more cancer progression biomarkers in the subject at the time of administration to the subject, compared to an equivalent regime in which the anti-DBI agent is not administered.

115. The therapeutic regimen according to claim 114, wherein the anti-DBI agent is an antibody or an aptamer.

116. The treatment regimen according to claim 114 or 115, wherein the one or more anticancer agents include a chemotherapeutic agent, an immunotherapy agent, or any combination thereof.

117. The therapeutic regimen according to claim 116, wherein the chemotherapeutic agent comprises immunogenic cell death (ICD) inducing activity.

118. The treatment regimen according to claim 116, wherein the immunotherapy agent comprises activity against immune checkpoints.

119. The aforementioned immune checkpoints include PD1 (programmed death 1), PDL1 (programmed death ligand 1), CTLA4 (cytotoxic T lymphocyte-associated protein 4), PDL2 (programmed death ligand 2), KIR (killer cell immunoglobulin-like receptor), B7-H3, B7-H4, BTLA (B and T lymphocyte atenuator), LAG3 (lymphocyte activating gene 3), TIM-3 (containing T cell immunoglobulin and mucin domain 3), and VISTA (T cell). A therapeutic regimen according to claim 118, comprising: activating V-domain Ig suppressor), ILT2 / LILRB1 (Ig-like transcript 2 / leukocyte Ig-like receptor 1), ILT3 / LILRB4 (Ig-like transcript 3 / leukocyte Ig-like receptor 4), ILT4 / LILRB2 (Ig-like transcript 4 / leukocyte Ig-like receptor 2), TIGIT (T-cell immune receptor having Ig and ITIM domains), NKG2A, PVRIG, CBLB, CISH, or any combination thereof.

120. The treatment regimen according to any one of claims 116 to 119, wherein the immunotherapy agent comprises an anti-PD1 agent, an anti-PD-L1 agent, an anti-CTLA4 agent, an anti-PD-L2 agent, an anti-KIR agent, an anti-B7-H3 agent, an anti-B7-H4 agent, an anti-BTLA agent, an anti-LAG3 agent, an anti-TIM-3 agent, an anti-VISTA agent, an anti-ILT2 / LILRB1 agent, an anti-ILT3 / LILRB4 agent, an anti-ILT4 / LILRB2 agent, an anti-TIGIT agent, an anti-NKG2A agent, an anti-PVRIG agent, an anti-CBLB agent, an anti-CISH agent, or any combination thereof.

121. The aforementioned immunotherapy agents are ipilimumab, tremelimumab, MK-1308, FPT155, PRS010, BMS-986249, BPI-002, CBT509, JS007, ONC392, TE1254, IBI310, BR02001, CG0161, KN044, PBI5D3H5, BCD145, ADU1604, AGEN1884, AGEN1181, C S1002, CP675206, Pembrolizumab, Nivolumab, Pidilizumab, AMP-224, BMS-936559, Semiprimab, PDR001, MDX-1105, MEDI4736, Atezolizumab, Avelumab, BMS-936559, Durvalumab, Rilurumab (IPH2102), IPH2101, MGA271, FPA150, IMP32 1 (Eftilagimod alfa), Relatrimab, MK-4280, AVA017, BI754111, ENUM006, GSK2831781, INCAGN2385, LAG3Ig, LAG525, REGN3767, Sym016, Sym022, Sym023, TSR033, TSR075, XmAb22841, LY3321367, MBG453, TS A therapeutic regimen according to any one of claims 116 to 120, comprising R-022, JNJ-61610588, MK-7684, PTZ-201, RG6058, COM902, IPH-2201, COM701, CA-327, LAG525, REGN3767, BI754111, tevoterimab, FS118, MGC018, or any combination thereof.

122. A therapeutic regimen according to any one of claims 114 to 121, further comprising one or more corticosteroid therapeutic agents.

123. The treatment regimen according to claim 122, wherein the one or more corticosteroid therapeutic agents are selected from cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, beclomethasone, and any combination thereof.

124. The treatment regimen according to any one of claims 114 to 123, wherein the anti-DBI agent and the one or more anticancer agents are separate components administered together or sequentially.

125. The treatment regimen according to any one of claims 114 to 123, wherein the anti-DBI agent is administered at least about two weeks before the one or more anticancer agents.

126. The treatment regimen according to any one of claims 114 to 123, wherein the anti-DBI agent and the one or more anticancer agents are administered together.

127. The treatment regimen according to claim 122 or 123, wherein the one or more corticosteroid therapeutic agents are co-administered with the anti-DBI agent and / or the one or more anticancer agents, or administered sequentially.

128. The treatment regimen according to claim 122 or 123, wherein any two or more of the anti-DBI agent, the one or more anticancer agents, and the one or more corticosteroid therapeutic agents are co-administered.

129. An anti-DBI agent for use in a therapeutic regimen according to any one of claims 114 to 128.

130. A composition comprising an anti-DBI agent for use in a therapeutic regimen according to any one of claims 114 to 128.

131. An anticancer agent for use in a therapeutic regimen according to any one of claims 114 to 128.

132. A composition comprising an anticancer agent for use in a therapeutic regimen according to any one of claims 114 to 128.

133. A corticosteroid therapeutic agent for use in a therapeutic regimen according to any one of claims 122 to 128.

134. A composition comprising a corticosteroid therapeutic agent for use in a therapeutic regimen according to any one of claims 122 to 128.

135. A combination of an anti-DBI agent and an anticancer agent for use in a therapeutic regimen according to any one of claims 114 to 128.

136. A system comprising one or more components, wherein the one or more components are: (a) an amount of extracellular human DBI-anti-DBI agent sufficient to inhibit extracellular diazepam binding inhibitors (DBIs) in the subject; (b) One or more anticancer agents, which optionally include a chemotherapeutic agent, an immunotherapy agent, or a combination thereof; (c) One or more corticosteroid therapeutic agents; (d) or any combination of (a) to (c) A system that includes one or more of these individually.

137. A composition for use in the treatment of hepatocellular carcinoma (HCC) in a subject, wherein the composition comprises an agent that reduces human diazepam-binding inhibitor (DBI) activity or expression in an amount sufficient to treat the HCC when administered to the subject.

138. The composition for use according to claim 137, wherein the amount is sufficient to treat the symptoms of HCC.

139. The composition for use according to claim 137 or 138, wherein the agent reduces DBI activity compared to the amount of activity obtained by administering a composition lacking the agent that reduces DBI activity.

140. The composition for use according to claim 137, wherein the agent comprises a DBI-binding polypeptide.

141. The composition for use according to claim 140, wherein the DBI-binding polypeptide is an anti-DBI antibody.

142. The composition for use according to claim 141, wherein the anti-DBI antibody is a monoclonal antibody.

143. The composition for use according to claim 141, wherein the anti-DBI antibody is a polyclonal antibody.

144. The composition for use according to claim 140, wherein the DBI-binding polypeptide is an anti-DBI antibody fragment.

145. The composition for use according to claim 140, wherein the DBI-conjugated polypeptide is an antibody fragment comprising a single-chain Fv, a Fab' fragment, or a nanobody.

146. The composition for use according to claim 137, wherein the agent comprises a polypeptide antigen that induces the production of a neutralizing anti-DBI antibody.

147. The composition for use according to claim 137 or 138, wherein the drug reduces DBI expression compared to the expression level before administration.

148. The composition for use according to claim 147, wherein the drug comprises a thyroid hormone receptor agonist.

149. The composition for use according to claim 148, wherein the thyroid receptor agonist comprises a thyroid hormone receptor β (THR-β) agonist.

150. The composition for use according to claim 148, wherein the thyroid receptor agonist comprises resmethylome, sovethyrom, eprothyrom, (2R,4S)-4-(3-chlorophenyl)-2-[(4-{[4-hydroxy-3-(propan-2-yl)phenyl]methyl}-3,5-dimethylphenoxy)methyl]-1,3,2-lambda-5-dioxaphosfinan-2-one, or (4-(3-benzyl-4-hydroxybenzyl)-3,5-dimethylphenyl)oxy)methyl)phosphonic acid.

151. The composition for use according to claim 148, wherein the thyroid hormone receptor agonist comprises resmethylome.

152. The composition for use according to claim 147, wherein the agent is siRNA, an endonuclease, an antisense oligonucleotide, or a ribosome.

153. The composition for use according to claim 147, wherein the drug is the siRNA.

154. A method for treating liver cancer in a subject, the method comprising administering to the subject a composition comprising a drug that reduces the activity or expression of human diazepam binding inhibitor (DBI), wherein the administration is sufficient to treat the liver cancer in the subject.

155. The method according to claim 154, wherein the liver disease is hepatocellular carcinoma (HCC).

156. The method according to claim 155, wherein the HCC is induced by non-alcoholic steatohepatitis (NASH).

157. The method according to any one of claims 154 to 156, wherein the drug reduces DBI activity compared to the activity level before administration.

158. The method according to claim 157, wherein the drug comprises a DBI-binding polypeptide.

159. The method according to claim 158, wherein the DBI-binding polypeptide is an anti-DBI antibody.

160. The method according to claim 159, wherein the anti-DBI antibody is a monoclonal antibody.

161. The method according to claim 159, wherein the anti-DBI antibody is a polyclonal antibody.

162. The method according to claim 158, wherein the DBI-binding polypeptide is an anti-DBI antibody fragment.

163. The method according to claim 158, wherein the DBI-conjugated polypeptide is a single-chain Fv, a Fab' fragment, or an antibody fragment comprising a nanobody.

164. The method according to claim 157, wherein the agent comprises a polypeptide antigen that induces the production of a neutralizing anti-DBI antibody.

165. The method according to any one of claims 154 to 156, wherein the drug reduces DBI expression compared to the expression level before administration.

166. The method according to claim 165, wherein the drug comprises a thyroid hormone receptor agonist.

167. The method according to claim 166, wherein the thyroid receptor agonist comprises a thyroid hormone receptor β (THR-β) agonist.

168. The method according to claim 166, wherein the thyroid receptor agonist comprises resmethylome, sovethyrom, eprothyrom, (2R,4S)-4-(3-chlorophenyl)-2-[(4-{[4-hydroxy-3-(propan-2-yl)phenyl]methyl}-3,5-dimethylphenoxy)methyl]-1,3,2-lambda-5-dioxaphosfinan-2-one, or (4-(3-benzyl-4-hydroxybenzyl)-3,5-dimethylphenyl)oxy)methyl)phosphonic acid.

169. The method according to claim 166, wherein the thyroid hormone receptor agonist comprises resmethylome.

170. The method according to claim 165, wherein the agent is siRNA, an endonuclease, an antisense oligonucleotide, or a ribosome.

171. The method according to claim 165, wherein the drug is an siRNA that inhibits DBI expression.

172. A method for treating hepatocellular carcinoma (HCC) in a subject, the method comprising administering to the subject a composition comprising a drug that reduces the activity or expression of human diazepam binding inhibitor (DBI), wherein the administration is sufficient to treat the HCC.

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