Pharmaceutical composition and combination formulation for immunotherapy of cancer, pharmaceutical use, and treatment method

Abstract

Provided are a pharmaceutical composition and combination formulation for immunotherapy of cancer, pharmaceutical use, and a treatment method. By means of the combined use of different active pharmaceutical ingredients, namely a TLR agonist, a tumor antigen, and an antibody for treating cancer, wherein each of the active pharmaceutical ingredients has a different anti-tumor immune activation effect, a synergistic effect is achieved. Also provided is a combination therapy of a tumor antigen-releasing therapy with the use of a TLR agonist and an antibody for treating cancer. By means of the anti-tumor immune activation effect of radiotherapy-induced tumor antigen release combined with the TLR agonist and the antibody, a synergistic effect is achieved. Compared with therapies relying on a single drug species, the combined use of different active pharmaceutical ingredients or the combination therapy can improve the efficacy, and can reduce the dose, thereby lowering the likelihood of toxicity and adverse events, and reducing the possibility of acquired resistance to immunotherapy.

Description

A pharmaceutical composition, combination formulation, pharmaceutical use, and treatment method for immunotherapy of cancer.

[0001] Cross-references to related applications

[0002] This application claims priority to Chinese Patent Application No. CN 202510124980.4, filed on January 26, 2025, entitled "A pharmaceutical composition, combination formulation, and pharmaceutical use and method of treatment for immunotherapy of cancer," and Chinese Patent Application No. CN 202510915752.9, filed on July 3, 2025, entitled "A pharmaceutical composition, combination formulation, and pharmaceutical use and method of treatment for immunotherapy of cancer," the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] This application belongs to the field of oncology technology, and more specifically, relates to a pharmaceutical composition, combination formulation, pharmaceutical use, and treatment method for immunotherapy of cancer. Background Technology

[0004] Immunotherapy and targeted therapy have become important tools in clinical cancer treatment. Immunotherapy works by activating or enhancing the body's own immune system, enabling it to recognize and attack tumor cells, thereby achieving a therapeutic effect. Current clinical approaches to cancer immunotherapy rely on the systemic administration of high doses of one or two biological immunomodulators. These drugs aim to restore pre-set conditions of tumor-induced immunosuppression while stimulating adaptive immune responses against tumor-specific antigens (TSA) and tumor-associated antigens (TAA).

[0005] Toll-like receptors (TLRs) are present on many cells of the immune system and participate in innate immune responses. The TLR receptor family includes six transmembrane TLRs (TLR-1, 2, 4, 5, 6, and 10) and four endosomal TLRs (TLR-3, 7, 8, and 9). Each PAMP (pathogen-associated molecular pattern) is recognized by a different TLR: lipopolysaccharide (TLR4), lipopeptide (TLR2 and TLR6 or TLR1), flagellin (TLR5), single-stranded RNA (TLR7 / 8), double-stranded RNA (TLR3), and DNA containing a CpG motif (TLR9).

[0006] Several TLR agonists for cancer treatment are currently under development, including TLR9 agonists such as CpG oligodeoxynucleotide (CpG ODN); TLR7 / 8 agonists such as retsimod (R848), BDB001, BDB018, and NKTR-262; TLR8 agonists such as VTX-2337 (Motolimod); TLR7 agonists such as imiquimod, RO7119929, SHR2150, TQ-A3334, and LHC165; TLR5 agonists such as CBLB502 (also known as Entolimod); TLR4 agonists such as GLA-SE (glucopyranosyl lipid A stable emulsion) and GSK1795091; and TLR3 agonists such as rintatolimod and poly-ICLC. Clinical trials have already investigated the ability of various TLR agonists to promote anti-tumor immunity. Antitumor responses are largely attributed to their ability to stimulate APCs (such as dendritic cells (DCs)) and thereby activate tumor-specific T cell responses.

[0007] CpG ODN is a synthetically produced deoxyoligonucleotide (ODN) containing unmethylated cytosine-phosphate-guanine dinucleotide (CpG). Upon recognition by the individual's immune system, CpG ODN exhibits strong immunostimulatory activity, stimulating both innate and adaptive immune responses. CpG ODN can activate plasmacytoid dendritic cells (pDCs), causing them to secrete type I interferon (IFN), and can also promote the expression of co-stimulatory molecules such as CD80 and CD86 on DCs and tumors expressing TLR9. This can lead to the secretion of various cytokines / chemokines and activate natural killer (NK) cells, TH1 cells, and cytotoxic T lymphocytes (CTLs). Encouraging results have been achieved in phase I and II clinical trials using CpG ODN as an adjuvant for tumor vaccines or in combination therapy with chemotherapy for cancer treatment.

[0008] Targeted therapy involves intervening in specific molecules or signaling pathways on tumor cells by targeting their molecular targets, using drugs or other treatments to directly interfere with or block the growth and spread of tumor cells, thereby killing or inhibiting them. Targeted therapy primarily utilizes over 100 antibodies approved in recent years. Generally, therapeutic antibodies kill tumor cells through three mechanisms: (1) Direct antibody action, which involves blocking or activating ligand / receptor signaling activity, inducing apoptosis, and delivering drugs or cytotoxic agents. Antibody receptor activation can directly kill tumor cells. Antibodies can also mediate tumor cell killing through receptor antagonistic activity. (2) Immune-mediated cell killing mechanisms, including complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), T cell function regulation, etc. Immune-mediated tumor cell killing can be accomplished through the following mechanisms: induction of phagocytosis, activation of complement, antibody-dependent cell-mediated cytotoxicity, targeting of tumors by genetically modified T cells via single-chain variable fragments (scFv), activation of T cells through antibody-mediated antigen cross-presentation by dendritic cells, and inhibition of T cell inhibitory receptors. (3) The specific effects of antibodies on the tumor vascular system and matrix induce ablation of vascular and stromal cells by capturing vascular receptor antagonists or ligands.

[0009] Therapeutic monoclonal antibody drugs have advanced the research and development of anticancer drugs, and significant benefits have been obtained from their use. However, some issues still need further research and resolution, such as antibody immunogenicity; a significant proportion of patients do not respond to treatment or become refractory; and more than half of the antibodies fail to achieve long-term effective inhibition and killing of tumor cells. Furthermore, the use of high systemic doses of monoclonal antibodies is accompanied by a high frequency of serious adverse events.

[0010] To address the limitations of the aforementioned therapies in cancer treatment, targeted therapy and immunotherapy have begun to evolve towards combination therapy, and more combination therapy regimens are expected to emerge in the future to further improve treatment efficacy. Some companies have conducted beneficial explorations in combination therapy with cancer immunotherapy. For example, the I / II clinical trial ILLUMINATE-204 (Clinicaltrials.gov, NCT02644967), involving the intratumoral TLR9 agonist IMO-2125 (Tilsotolimod) combined with the CTLA4 antibody ipilimumab for the treatment of PD-1 inhibitor-refractory advanced melanoma, showed that this combination therapy was generally well-tolerated and effective in PD-1 inhibitor-refractory advanced melanoma, including durable responses (Cara L. Haymaker1, Robert H. Andtbacka, Douglas B. Johnson, etc., 2020 Virtual ESMO Congress). However, the pivotal registrational phase 3 clinical trial ILLUMINATE-301 (Clinicaltrials.gov, NCT03445533), in combination with the CTLA4 antibody ipilimumab for the treatment of PD-1 refractory advanced melanoma, failed to meet the primary endpoint of ORR.

[0011] In summary, new approaches to cancer immunotherapy are still needed in this field to improve cancer treatment outcomes. Summary of the Invention

[0012] While current cancer immunotherapy methods offer benefits to a limited number of patients, their adherence and applicability are limited by acquired resistance and a high rate of adverse immune events. Furthermore, while concurrent administration of multiple antibodies significantly enhances antitumor efficacy, it can also trigger a range of adverse reactions, leading to poor tolerability and making treatment unsustainable.

[0013] To address the aforementioned problems in the prior art, this application provides a pharmaceutical composition, combination formulation, and its pharmaceutical uses and treatment methods for cancer immunotherapy, overcoming many limitations of current cancer immunotherapy. This application combines pharmaceutical active ingredients with different anti-tumor immune-activating effects, resulting in a synergistic effect. This application also provides a combination therapy of releasing tumor antigens with the use of TLR agonists and antibodies, such as the synergistic effect of releasing tumor antigens through radiotherapy combined with the anti-tumor immune-activating effect of TLR agonists. By combining different pharmaceutical active ingredients or combination therapies, this application can improve efficacy compared to therapies relying on a single drug, and can reduce dosage, thereby reducing the likelihood of toxicity and adverse events, and reducing the possibility of acquired immune resistance.

[0014] In a first aspect, this application provides a pharmaceutical composition comprising:

[0015] (a) One, two, or more TLR agonists;

[0016] (b) Tumor antigens;

[0017] (c) One, two, or more antibodies for treating cancer;

[0018] (d) Pharmaceutically acceptable carriers or excipients.

[0019] In some embodiments, the pharmaceutical composition does not contain other active ingredients.

[0020] In a second aspect, this application provides a combination formulation comprising:

[0021] (a) One, two, or more TLR agonists;

[0022] (b) Tumor antigens;

[0023] (c) One, two, or more antibodies for treating cancer.

[0024] In some embodiments, the combination formulation does not contain other active ingredients.

[0025] In some embodiments, when (a) there are two or more TLR agonists in the combination formulation, each TLR agonist is formulated as a separate pharmaceutical composition; or at least two TLR agonists are formulated together as a pharmaceutical composition [e.g., when there are three TLR agonists in (a), two are formulated together as a pharmaceutical composition and the other is formulated as a separate pharmaceutical composition]; or all TLR agonists are formulated as a single pharmaceutical composition.

[0026] In some embodiments, when (c) comprises two or more antibodies for treating cancer, each antibody is formulated as a separate pharmaceutical composition; or at least two antibodies are formulated together as a pharmaceutical composition [e.g., when (c) comprises three antibodies, two are formulated together as a pharmaceutical composition and the other is formulated as a separate pharmaceutical composition]; or all antibodies are formulated as a single pharmaceutical composition.

[0027] In some embodiments, in the combination formulation, (a), (b), and (c) are each formulated as separate pharmaceutical compositions; or in the combination formulation, (a) and (b) are formulated together as a pharmaceutical composition, and (c) is formulated as a separate pharmaceutical composition; or (a), (b), and a portion thereof are formulated together as a pharmaceutical composition, and the remainder thereof is formulated as a separate pharmaceutical composition.

[0028] In some embodiments, the pharmaceutical composition contains a pharmaceutically acceptable carrier or excipient.

[0029] The pharmaceutical compositions described in the first aspect and the combination formulations described in the second aspect are further described in detail below.

[0030] In some embodiments, the TLR agonist is selected from TLR1 agonists, TLR2 agonists, TLR3 agonists, TLR4 agonists, TLR5 agonists, TLR6 agonists, TLR7 agonists, TLR8 agonists, TLR7 / 8 dual agonists, TLR9 agonists, and TLR10 agonists. Preferably, the TLR agonist is selected from TLR3 agonists, TLR7 agonists, TLR8 agonists, TLR7 / 8 dual agonists, and TLR9 agonists.

[0031] In some embodiments, the TLR agonist is selected from CpG ODN, R848, BDB001, Poly I:C, BDB018, NKTR-262, VTX-2337 (Motolimod), Imiquimod, RO7119929, SHR2150, TQ-A3334, LHC165, CBLB502 (also known as Entolimod), GLA-SE (glucopyranosyl lipid astable emulsion), GSK1795091, Rintatolimod, poly-ICLC, and pharmaceutically acceptable salts thereof. The CpG ODN includes types A, B, and C.

[0032] In some embodiments, the TLR9 agonist is CpG ODN or a pharmaceutically acceptable salt thereof, including type A, type B, and type C. Preferably, the TLR9 agonist is type C CpG ODN or a pharmaceutically acceptable salt thereof.

[0033] In some embodiments, the CpG ODN includes chemical modifications. In some embodiments, the CpG ODN includes modification with one or more phosphate ester groups. In some embodiments, the modification with one or more phosphate ester groups is a thiophosphate bond. In some embodiments, the phosphate backbone of the CpG ODN is fully modified with thiophosphate.

[0034] In some embodiments, the CpG ODN is CpG HP007, CpG HP021, CpG HP026, CpG 684, SD-101, G10 CpG ODN, CpG 1018, CpG 7909, IMO-2125, and pharmaceutically acceptable salts thereof.

[0035] In some embodiments, the TLR9 agonist is a pharmaceutically acceptable salt of CpG HP007. In some embodiments, the TLR7 / 8 dual agonist is R848 or a pharmaceutically acceptable salt thereof. The TLR3 agonist is Poly I:C or a pharmaceutically acceptable salt thereof.

[0036] In some embodiments, (a) is a TLR9 agonist; or a TLR7 / 8 agonist; or a TLR3 agonist; or a combination of a dual TLR7 / 8 agonist and a TLR9 agonist; or a combination of a TLR7 / 8 agonist and a TLR3 agonist; or a combination of a TLR3 agonist and a TLR9 agonist; or a combination of a TLR7 / 8 agonist, a TLR3 agonist, and a TLR9 agonist; preferably, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof.

[0037] In some embodiments, the tumor antigen is whole-tumor lysate (WTL), neoantigen, genetically engineered protein, polypeptide, nucleic acid, DNA, etc.

[0038] In some embodiments, the tumor antigen is a whole tumor lysate, and the tumor is colorectal cancer, melanoma, breast cancer, lung cancer, gastric cancer, head and neck tumors, prostate cancer, liver cancer, nasopharyngeal carcinoma, esophageal cancer, cervical cancer, etc.

[0039] In some embodiments, the tumor antigen is an allogeneic tumor antigen or a xenogeneic tumor antigen.

[0040] In some embodiments, the whole tumor lysate is prepared by resuspending whole tumor cells in sterile PBS to form a cell suspension, repeatedly freezing and thawing for lysis, centrifuging, and collecting the supernatant by filtration. The concentration of the cell suspension is 1×10⁻⁶. 6 -1×10 8 / mL, preferably 1×10 7 -5×10 7 / mL. The repeated freeze-thaw conditions are 2-10 freeze-thaw cycles at -80℃ and 37℃, preferably 4-8. The centrifugation conditions are 3-20 min at 500-5000 rpm, preferably 5-15 min at 1000-3000 rpm.

[0041] In some embodiments, the whole tumor lysate is prepared by resuspending whole tumor cells in sterile PBS to prepare lysates with a cell count of 4 × 10⁻⁶ cells per milliliter. 7 Cell suspensions were divided into two 1.5 mL centrifuge tubes, each containing 0.5 mL of the lysis buffer. The tubes were subjected to six freeze-thaw cycles at -80°C and 37°C (freezing for 15 min, thawing for 5 min, with shaking to mix in between). Lysis was determined by counting cells using phenol blue staining. The cells were centrifuged at 2,000 rpm for 10 min, and the supernatant was collected. All supernatants were combined and filtered through a 0.22 μm filter. Total protein concentration was determined using the BCA method. The remaining tumor cell lysis mixture was stored at -80°C.

[0042] In some embodiments, the tumor antigen is a neotumor. The screening method for neotumor antigens includes: extracting tumor cell DNA and RNA, performing whole-exome sequencing and RNA sequencing, sequence alignment, variant detection, and gene data annotation; (In silicon peptide construction and HLA-peptide binding predictions) constructing mutation-related peptides using sequencing data and computer simulation methods (In silicon), predicting the affinity of the constructed peptides for HLA binding, selecting high-affinity peptides based on affinity strength, and listing them; further screening using RNA sequencing data based on the list, reordering peptides according to HLA affinity and gene expression level, prioritizing peptides with high HLA affinity and high gene expression levels, which are more likely to initiate an anti-tumor immune response; synthesizing antigen peptides according to the predicted peptide sequences; (In vitro pMHC binding assay for peptides) verifying the binding ability of candidate peptides to MHC through in vitro experiments. Peptide-driven refolding experiments are used to detect the ability of peptides to bind to their corresponding MHC / HLA molecules in vitro, helping to determine which peptides can effectively bind to specific MHC / HLA molecules. In vitro validation experiments of pMHC binding are typically performed in peptide selection to ultimately confirm the selection of antigenic peptides. In vivo immunogenicity testing verifies whether potential antigens can elicit an effective immune response in the real immune system, ensuring immunoprotective effects in practical applications.

[0043] In some embodiments, the cancer-treating antibody targets PD-1, CTLA-4, TIGIT, PD-L1, PD-L2, OX40, 4-1BB, ICOS, LAG3, TIM3, IDO, HER2, CD20, CD22, CD27, CD40, CD40L, GITR, EGFR, CD2, CD19, CD33, CD37, CD38, CD44, CD45, CD47, CD52, CD56, CD70, CD79, 5T4, AGS-5, AGS-16, angiopoietin 2, B7.1, and B7. 2. B7DC, B7H1, B7H2, B7H3, BT-062, BTLA, CAIX, carcinoembryonic antigen, Cripto, ED-B, ErbB1, ErbB2, ErbB3, ErbB4, EGFL7, EpCAM, EphA2, EphA3, EphB2, FAP, fibronectin, folate receptor, ganglioside GM3, GD2, glucocorticoid-induced tumor necrosis factor receptor GITR, gp100, gpA33, GPNMB, ICOS, IGF1R, integrin αν, integrin ανβ, KIR, Lewis Y, mesothelin, c-MET, MN carbonic anhydrase IX, MUC1, MUC16, cohesin-4, NKGD2, NOTCH, OX40L, PSCA, PSMA, RANKL, ROR1, ROR2, SLC44A4, STING, multiligand proteoglycan-1, TACI, TAG-72, tendinin, TIM3, TRAILR1, TRAILR2, VEGFR-1, VEGFR-2, VEGFR-3, SSTR, SSTR2, CXCR4, NTR, GRPR. Preferably, the antibody for treating cancer targets one, two, or more (e.g., three) of PD-1, CTLA-4, TIGIT, OX40, and CD40. More preferably, the antibody for treating cancer or a combination thereof targets one, two, or more (e.g., three) of PD-1, CTLA-4, TIGIT, and CD40.

[0044] In some embodiments, the antibody used to treat cancer is a monoclonal antibody, a bispecific antibody, a trispecific antibody, an antibody-drug conjugate (ADC), or a radionuclide conjugate (RDC).

[0045] In some embodiments, (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody.

[0046] In some embodiments, the anti-PD-1 antibody is a monoclonal antibody selected from Keytruda, Opdivo, cimiprimab, toripalimab, sintilimab, camrelizumab, patelilimab, sugelilimab, penapralimab, cepalimumab, and slulilimab; the anti-CTLA4 antibody is a monoclonal antibody selected from ipilimumab and tremelimumab; and the anti-TIGIT antibody is a monoclonal antibody selected from domvanalimab, belrestotug, and tiragol. The following are listed: umab, ospermumab, JS-006, vibostolimab, Lancastotug, Renvistobart, Etigilimab, HB-0030, PM-1021, AB-308, COM-902, M6223, ASP-8374, BAT-6005, Tamgiliblimab; the anti-CD40 antibody is a monoclonal antibody selected from SEA-CD40, Sotigalimab, BSI-038, and Anti-CD40. ChiLob7 / 4 (BioNTech SE), 2141-V11, CDX-1140, YH-003, Selicrelumab, Mitazalimab, ILB-2101, ABBV-927, LVGN-7409; the anti-OX40 antibody is a monoclonal antibody selected from BGB-A445, INBRX-106, YH-002, INCAGN-1949, MEDI-6469, GEN-1055, BAT-6026, IBI-101, ABBV-368, MEDI-0562, HFB-3010.

[0047] In some embodiments, the bispecific antibody is selected from: epalolimbab / tovoretalimab (CTLA4 inhibitor / PD1 inhibitor), canducilin / tovoretalimab (CTLA4 inhibitor / PD1 inhibitor), Volrustomig (CTLA4 inhibitor / PD1 inhibitor), Danvilostomig (CTLA4 inhibitor / PD1 inhibitor), Lorigerlimab (CTLA4 inhibitor / PD1 inhibitor), Vudalimab (CTLA4 inhibitor / PD1 inhibitor), SHS-010 (CTLA4 inhibitor / PD1 inhibitor), REMD-511 (CTLA4 inhibitor / PD1 inhibitor), YH-008 (CD40 inhibitor / PD1 inhibitor), FS-120 (4-1BB agonist / OX40 agonist), YH-006 (CTLA4 inhibitor / OX40 agonist), ATOR-1015 (CTLA4 inhibitor / OX40 agonist), APVO-603 (4-1BB agonist / OX40 agonist). The following are selected as specific antibodies: CD40 agonist, GEN-1042 (4-1BB agonist / CD40 agonist), IMB-071703 (4-1BB agonist / CD40 agonist), SM2256 (4-1BB agonist / CD40 agonist), Rilvegostomig (PD1 inhibitor / TIGIT inhibitor), ZG-005 (PD1 inhibitor / TIGIT inhibitor), REMD-532 (PD1 inhibitor / TIGIT inhibitor), IBI-321 (PD1 inhibitor / TIGIT inhibitor), BC008-1A (PD1 inhibitor / TIGIT inhibitor), ZGGS15 (LAG3 inhibitor / TIGIT inhibitor); and trispecific antibodies selected from GBD209 (CTLA4 inhibitor / PD1 inhibitor / TIGIT inhibitor), FG-T903 (CD112R antagonist / PD1 inhibitor / TIGIT inhibitor), and HC010 (hcbiopharma) (CTLA4 inhibitor / PD1 inhibitor / VEGF inhibitor).

[0048] In some embodiments, the ADC drug is selected from: Mylotarg, Besponsa, Kadcyla, Polivy, Lumoxiti, Enhertu, Adcetris, Padcev, Tivdak, Blenrep, Trodelvy, Akalux, Zynlonta, Adrenaline, Luconsartuzumab, and Elahere.

[0049] In some embodiments, the radionuclide conjugate drug RDC is selected from: Pluvicto and Lutathera.

[0050] In some embodiments, the pharmaceutical composition or combination formulation comprises:

[0051] (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof;

[0052] (b) is a tumor antigen;

[0053] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody. The antibody is preferably a monoclonal antibody.

[0054] In some embodiments, the pharmaceutical composition or combination formulation comprises:

[0055] (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0056] (b) Tumor antigens;

[0057] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; wherein the antibody is preferably a monoclonal antibody.

[0058] In some embodiments, the pharmaceutical composition or combination formulation comprises:

[0059] (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0060] (b) Tumor antigens;

[0061] (c) Anti-CD40 antibody. The antibody is preferably a monoclonal antibody.

[0062] In some embodiments, the pharmaceutical composition or combination formulation comprises:

[0063] (a) R848 or a pharmaceutically acceptable salt thereof;

[0064] (b) Tumor antigens;

[0065] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; wherein the antibody is preferably a monoclonal antibody.

[0066] In some embodiments, the pharmaceutical composition or combination formulation comprises:

[0067] (a) Poly I:C or a pharmaceutically acceptable salt thereof;

[0068] (b) Tumor antigens;

[0069] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; wherein the antibody is preferably a monoclonal antibody.

[0070] In some embodiments, the pharmaceutical composition or combination formulation comprises:

[0071] (a) CpG HP007 or a pharmaceutically acceptable salt thereof, and R848 or a pharmaceutically acceptable salt thereof;

[0072] (b) Tumor antigens;

[0073] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; wherein the antibody is preferably a monoclonal antibody.

[0074] In some embodiments, the mass ratio of (a):(b):(c) in the pharmaceutical composition or formulation is (1-1000):(10-1000):(1-6000), preferably (2-500):(100-500):(2-3000), more preferably (20-500):(200-500):(10-3000). Most preferably (30-200):(300-400):(20-1500).

[0075] In one set of embodiments, the tumor antigen in the pharmaceutical composition or combination formulation is 10-1000 parts, such as 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000 parts, preferably 100-500 parts, more preferably 200-500 parts, and most preferably 300-400 parts; a TLR9 agonist (such as CpG) When present, the amount is 1-1000 parts, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1000 parts, preferably 2-500 parts, more preferably 20-500 parts, and most preferably 100- 200 parts; when a TLR7 / 8 dual agonist (such as R848 or a pharmaceutically acceptable salt thereof) is present, it is 1-200 parts, such as 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200 parts, preferably 2-100 parts, more preferably 20-100 parts, and most preferably 30-50 parts; a TLR3 agonist (such as Poly... When I:C or its pharmaceutically acceptable salts are present, the amount is 1-200 parts, such as 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200 parts, preferably 2-100 parts, more preferably 20-100 parts, most preferably 30-50 parts; when anti-CTLA4 antibody is present, it is 1-2000 parts, such as 1, 2 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000 portions, preferably 2- 1000 parts, more preferably 10-1000 parts, most preferably 300-500 parts; when anti-CD40 antibody is present, it is 1-500 parts, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500 parts, preferably 2-200 parts, more preferably 10-200 parts, most preferably 20-50 parts; anti-PD When antibodies are present, they are available in quantities ranging from 1 to 2000, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700.

[0076] 1800, 1900, 2000 parts, preferably 2-1000 parts, more preferably 10-1000 parts, and most preferably 300-500 parts; when anti-OX40 antibody is present, it is 1-500 parts, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500 parts, preferably 2-200 parts, more preferably 10-200 parts, and most preferably 20-50 parts; anti-TIG When IT antibodies are present, the quantity is 1-1000 parts, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 300, 400, 500, 600, 700, 800, 900, 1000 parts, preferably 2-500 parts, more preferably 10-500 parts, and most preferably 100-200 parts.

[0077] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 1-1000 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 1-200 parts of R848 or a pharmaceutically acceptable salt thereof; or 1-200 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 1-1000 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 1-200 parts of R848 or a pharmaceutically acceptable salt thereof; or 1-200 parts of R848 or a pharmaceutically acceptable salt thereof, and 1-200 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 1-1000 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 1-200 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 1-1000 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 1-200 parts of R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or its pharmaceutically acceptable salt, 1-200 parts;

[0078] (b) Tumor antigens are 10-1000 samples;

[0079] (c) 1-2000 doses of anti-CTLA4 antibody; or 1-500 doses of anti-CD40 antibody; or 1-2000 doses of anti-PD-1 antibody and 1-2000 doses of anti-CTLA4 antibody; or 1-500 doses of anti-CD40 antibody and 1-2000 doses of anti-CTLA4 antibody; or 1-2000 doses of anti-PD-1 antibody, 1-2000 doses of anti-CTLA4 antibody and 1-1000 doses of anti-TIGIT antibody; or 1-2000 doses of anti-PD1 antibody, 1-2000 doses of anti-CTLA4 antibody and 1-500 doses of anti-CD40 antibody; or 1-500 doses of anti-OX40 antibody, 1-2000 doses of anti-CTLA4 antibody and 1-1000 doses of anti-TIGIT antibody.

[0080] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 2-500 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 2-100 parts of R848 or a pharmaceutically acceptable salt thereof; or 2-100 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 2-500 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 2-100 parts of R848 or a pharmaceutically acceptable salt thereof; or 2-100 parts of R848 or a pharmaceutically acceptable salt thereof, and 2-100 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 2-500 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 2-100 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 2-500 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 2-100 parts of R848 or a pharmaceutically acceptable salt thereof, and 2-100 parts of Poly I:C or a pharmaceutically acceptable salt thereof.

[0081] (b) Tumor antigens are 100-500 samples;

[0082] (c) 2-1000 doses of anti-CTLA4 antibody; or 2-200 doses of anti-CD40 antibody; or 2-1000 doses of anti-PD-1 antibody and 2-1000 doses of anti-CTLA4 antibody; or 2-200 doses of anti-CD40 antibody and 2-1000 doses of anti-CTLA4 antibody; or 2-1000 doses of anti-PD-1 antibody, 2-1000 doses of anti-CTLA4 antibody and 2-500 doses of anti-TIGIT antibody; or 2-1000 doses of anti-PD1 antibody, 2-1000 doses of anti-CTLA4 antibody and 2-200 doses of anti-CD40 antibody; or 2-200 doses of anti-OX40 antibody, 2-1000 doses of anti-CTLA4 antibody and 2-500 doses of anti-TIGIT antibody.

[0083] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 20-500 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 20-100 parts of R848 or a pharmaceutically acceptable salt thereof; or 20-100 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 20-500 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 20-100 parts of R848 or a pharmaceutically acceptable salt thereof; or 20-100 parts of R848 or a pharmaceutically acceptable salt thereof, and 20-100 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 20-500 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 20-100 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 20-500 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 20-100 parts of R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof. I:C or its pharmaceutically acceptable salt, 20-100 parts;

[0084] (b) Tumor antigens: 200-500 samples;

[0085] (c) 10-1000 doses of anti-CTLA4 antibody; or 10-200 doses of anti-CD40 antibody; or 10-1000 doses of anti-PD-1 antibody and 10-1000 doses of anti-CTLA4 antibody; or 10-200 doses of anti-CD40 antibody and 10-1000 doses of anti-CTLA4 antibody; or 10-1000 doses of anti-PD-1 antibody, 10-1000 doses of anti-CTLA4 antibody and 10-500 doses of anti-TIGIT antibody; or 10-1000 doses of anti-PD1 antibody, 10-1000 doses of anti-CTLA4 antibody and 10-200 doses of anti-CD40 antibody; or 10-200 doses of anti-OX40 antibody, 10-1000 doses of anti-CTLA4 antibody and 10-500 doses of anti-TIGIT antibody.

[0086] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 100-200 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 30-50 parts of R848 or a pharmaceutically acceptable salt thereof; or 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100-200 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of R848 or a pharmaceutically acceptable salt thereof; or 30-50 parts of R848 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100-200 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100-200 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof. 30-50 parts of I:C or its pharmaceutically acceptable salt;

[0087] (b) The tumor antigen is 300-400 samples;

[0088] (c) 300-500 doses of anti-CTLA4 antibody; or 20-50 doses of anti-CD40 antibody; or 300-500 doses of anti-PD-1 antibody and 300-500 doses of anti-CTLA4 antibody; or 20-50 doses of anti-CD40 antibody and 300-500 doses of anti-CTLA4 antibody; or 300-500 doses of anti-PD-1 antibody, 300-500 doses of anti-CTLA4 antibody and 100-200 doses of anti-TIGIT antibody; or 300-500 doses of anti-PD1 antibody, 300-500 doses of anti-CTLA4 antibody and 20-50 doses of anti-CD40 antibody; or 20-50 doses of anti-OX40 antibody, 300-500 doses of anti-CTLA4 antibody and 100-200 doses of anti-TIGIT antibody.

[0089] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 25, 100, or 300 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 30 parts of R848 or a pharmaceutically acceptable salt thereof; or 30 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 25, 100, or 300 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30 parts of R848 or a pharmaceutically acceptable salt thereof; or 30 parts of R848 or a pharmaceutically acceptable salt thereof, and 30 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 25, 100, or 300 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 25, 100, or 300 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30 parts of R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or 25, 100, or 300 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30 parts of R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof. 30 parts of I:C or its pharmaceutically acceptable salt;

[0090] (b) The tumor antigen is 300 or 400 samples;

[0091] (c) 50, 330, or 500 doses of anti-CTLA4 antibody; or 10, 50, or 100 doses of anti-CD40 antibody; or 100, 330, or 1000 doses of anti-PD-1 antibody, and 50, 330, or 500 doses of anti-CTLA4 antibody; or 10, 50, or 100 doses of anti-CD40 antibody, and 50, 330, or 500 doses of anti-CTLA4 antibody; or 100, 330, or 1000 doses of anti-PD-1 antibody. 000 doses, 50, 330 or 500 doses of anti-CTLA4 antibody, and 167 doses of anti-TIGIT antibody; or 100, 330 or 1000 doses of anti-PD1 antibody, 50, 330 or 500 doses of anti-CTLA4 antibody, and 10, 50 or 100 doses of anti-CD40 antibody; or 40 doses of anti-OX40 antibody, 50, 330 or 500 doses of anti-CTLA4 antibody, and 167 doses of anti-TIGIT antibody.

[0092] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 30 parts of R848 or a pharmaceutically acceptable salt thereof; or 30 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30 parts of R848 or a pharmaceutically acceptable salt thereof; or 30 parts of R848 or a pharmaceutically acceptable salt thereof, and 30 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30 parts of R848 or a pharmaceutically acceptable salt thereof, and 30 parts of Poly I:C or a pharmaceutically acceptable salt thereof;

[0093] (b) The tumor antigen is 300 or 400 samples;

[0094] (c) 330 anti-CTLA4 antibody samples; or 50 anti-CD40 antibody samples; or 330 anti-PD-1 antibody samples and 330 anti-CTLA4 antibody samples; or 50 anti-CD40 antibody samples and 330 anti-CTLA4 antibody samples; or 330 anti-PD-1 antibody samples, 330 anti-CTLA4 antibody samples and 167 anti-TIGIT antibody samples; or 330 anti-PD1 antibody samples, 330 anti-CTLA4 antibody samples and 50 anti-CD40 antibody samples; or 40 anti-OX40 antibody samples, 330 anti-CTLA4 antibody samples and 167 anti-TIGIT antibody samples.

[0095] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 10-100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 30-50 parts of R848 or a pharmaceutically acceptable salt thereof; or 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 10-100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of R848 or a pharmaceutically acceptable salt thereof; or 30-50 parts of R848 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 10-100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 10-100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of R848 or a pharmaceutically acceptable salt thereof, and Poly 30-50 parts of I:C or its pharmaceutically acceptable salt;

[0096] (b) The tumor antigen is 300-400 samples;

[0097] (c) 20-400 doses of anti-CTLA4 antibody; or 5-100 doses of anti-CD40 antibody; or 50-1000 doses of anti-PD-1 antibody and 20-400 doses of anti-CTLA4 antibody; or 5-100 doses of anti-CD40 antibody and 20-400 doses of anti-CTLA4 antibody; or 50-1000 doses of anti-PD-1 antibody, 20-400 doses of anti-CTLA4 antibody and 100-200 doses of anti-TIGIT antibody; or 50-1000 doses of anti-PD1 antibody, 20-400 doses of anti-CTLA4 antibody and 5-100 doses of anti-CD40 antibody; or 20-50 doses of anti-OX40 antibody, 20-400 doses of anti-CTLA4 antibody and 100-200 doses of anti-TIGIT antibody.

[0098] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 10, 40, or 100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 30 parts of R848 or a pharmaceutically acceptable salt thereof; or 50 parts of Poly I:C or a pharmaceutically acceptable salt thereof.

[0099] (b) The tumor antigen is 300 or 400 samples;

[0100] (c) 50, 100 or 1000 doses of anti-PD1 antibody, 20, 40 or 400 doses of anti-CTLA4 antibody, and 5, 30 or 100 doses of anti-CD40 antibody.

[0101] In some embodiments, the pharmaceutical composition or combination formulation comprises 100 parts of (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0102] (b) The tumor antigen is 300 or 400 samples;

[0103] (c) 330 samples of anti-CTLA4 antibody.

[0104] In some embodiments, the pharmaceutical composition or combination formulation comprises 100 parts of (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0105] (b) The tumor antigen is 300 or 400 samples;

[0106] (c) 330 samples of anti-PD-1 antibody and 330 samples of anti-CTLA4 antibody.

[0107] In some embodiments, the pharmaceutical composition or combination formulation comprises 100 parts of (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0108] (b) The tumor antigen is 300 or 400 samples;

[0109] (c) 330 samples of anti-PD-1 antibody, 330 samples of anti-CTLA4 antibody, and 167 samples of anti-TIGIT antibody.

[0110] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) 100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30 parts of R848 or a pharmaceutically acceptable salt thereof.

[0111] (b) The tumor antigen is 300 or 400 samples;

[0112] (c) 330 samples of anti-PD-1 antibody and 330 samples of anti-CTLA4 antibody.

[0113] In some embodiments, the pharmaceutical composition or combination formulation comprises 10 parts of (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0114] (b) The tumor antigen is 300 or 400 samples;

[0115] (c) 50 samples of anti-PD1 antibody, 20 samples of anti-CTLA4 antibody, and 50 samples of anti-CD40 antibody.

[0116] In some embodiments, the pharmaceutical composition or combination formulation comprises 40 parts of (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0117] (b) The tumor antigen is 300 or 400 samples;

[0118] (c) 100 samples of anti-PD1 antibody, 40 samples of anti-CTLA4 antibody, and 30 samples of anti-CD40 antibody.

[0119] In some embodiments, the pharmaceutical composition or combination formulation comprises 100 parts of (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0120] (b) The tumor antigen is 300 or 400 samples;

[0121] (c) 1000 anti-PD1 antibody samples, 400 anti-CTLA4 antibody samples, and 100 anti-CD40 antibody samples.

[0122] In some embodiments, the pharmaceutical composition or combination formulation comprises 30 parts of (a) R848 or a pharmaceutically acceptable salt thereof;

[0123] (b) The tumor antigen is 300 or 400 samples;

[0124] (c) 100 samples of anti-PD1 antibody, 40 samples of anti-CTLA4 antibody, and 30 samples of anti-CD40 antibody.

[0125] In some embodiments, the pharmaceutical composition or combination formulation comprises 50 parts of (a) Poly I:C or a pharmaceutically acceptable salt thereof;

[0126] (b) The tumor antigen is 300 or 400 samples;

[0127] (c) 100 samples of anti-PD1 antibody, 40 samples of anti-CTLA4 antibody, and 30 samples of anti-CD40 antibody.

[0128] In a third aspect, the pharmaceutical composition described in the first aspect and the combination formulation described in the second aspect of this application are used for the prevention and / or treatment of cancer.

[0129] In a fourth aspect, this application provides pharmaceutical use of the pharmaceutical composition described in the first aspect and the combination formulation described in the second aspect for the preparation of a medicament for the prevention and / or treatment of cancer.

[0130] In a fifth aspect, this application provides a method for preventing and / or treating cancer, comprising the steps of administering to a subject an effective amount of the pharmaceutical composition described in the first aspect or the combination preparation described in the second aspect.

[0131] In some embodiments, the cancer is a solid tumor, including but not limited to colorectal cancer, melanoma, breast cancer, lung cancer, stomach cancer, head and neck tumors, prostate cancer, liver cancer, nasopharyngeal carcinoma, esophageal cancer, and cervical cancer.

[0132] In some embodiments, the method for preventing and / or treating cancer includes the step of administering an effective amount of the pharmaceutical composition described in the first aspect to a subject.

[0133] In some embodiments, the pharmaceutical composition can be administered to the subject parenterally, including but not limited to intratumoral injection (e.g., multiple intratumoral injection), subcutaneous injection, intravenous injection, intramuscular injection, nasal inhalation, or administration via an interventional device. Preferably, the pharmaceutical composition is administered to the subject via intratumoral injection. Intratumoral administration significantly reduces the dosage of each active ingredient, thereby increasing the breadth of pharmacological action to enhance antitumor efficacy, reducing toxicity, and decreasing the likelihood of acquired immune resistance.

[0134] In some embodiments, the method for preventing and / or treating cancer includes the step of administering an effective amount of the combination preparation described in the first aspect to a subject.

[0135] In some embodiments, the combination formulation can be administered to the subject parenterally, preferably via intratumoral injection, subcutaneous injection, intravenous injection, intramuscular injection, nasal inhalation, or through an interventional device. The combination formulation can be administered to the subject simultaneously, sequentially, or at intervals.

[0136] In some embodiments, the routes of administration of (a) and (b) in the combined formulation are the same or different; preferably, (a) and (b) are administered via the same route, such as intratumoral injection or subcutaneous injection to the subject.

[0137] In some embodiments, the routes of administration for (a) and (c) in the combined formulation are the same or different. Preferably, (a) and (c) are administered via the same route, such as intratumoral injection or subcutaneous injection; preferably, (a) and (c) are administered via different routes, such as intratumoral injection or intravenous injection; preferably, a portion of (a) and (c) are administered via the same route, while the remaining portion of (c) is administered via a different route, such as a portion of (a) and (c) being administered to the subject via intratumoral injection, and the remaining portion of (c) being administered to the subject via intravenous injection.

[0138] In some implementations, (a) and (b) are administered to the subject via intratumoral injection; (c) is administered to the subject via intravenous injection; or (a), (b) and (c) are administered via the same route, via intratumoral injection or subcutaneous injection; or (a) and (b), and a portion of (c) are administered to the subject via intratumoral injection, and the remainder of (c) is administered to the subject via intravenous injection.

[0139] In some embodiments, the combined formulation can be administered to the subject simultaneously after mixing, such as by intratumoral injection or subcutaneous injection; the combined formulation can be administered to the subject simultaneously by different administration methods, such as by intratumoral injection, intravenous injection or subcutaneous injection.

[0140] In some embodiments, the combination formulation may be administered to the subject sequentially or at intervals via the same or different administration methods, preferably via different administration methods; more preferably via intratumoral injection, intravenous injection or subcutaneous injection.

[0141] In some embodiments, when (a) there are two or more TLR agonists in the combination formulation, the administration routes of the different TLR agonists are the same or different; when (c) there are two or more antibodies for treating cancer, the administration routes of the different antibodies are the same or different; preferably, the administration routes of the different TLR agonists in (a) are the same (more preferably, administered to the subject by intratumoral injection or subcutaneous injection); and the administration routes of the different antibodies in (c) are the same or different (more preferably, both are administered to the subject by intravenous injection or subcutaneous injection, or administered to the subject by intratumoral injection or intravenous injection respectively).

[0142] In some embodiments, when the combination formulation comprises two or more TLR agonists, the TLR agonists are mixed and administered to the subject simultaneously, or the TLR agonists are administered to the subject as a single drug composition, such as by intratumoral injection or subcutaneous injection.

[0143] In some embodiments, when (c) of the combination formulation consists of two or more antibodies for treating cancer, the antibodies are mixed and administered to the subject simultaneously, or the antibodies are administered to the subject as a single drug composition, preferably by intravenous injection, intratumoral injection, or subcutaneous injection; or a portion of the antibodies are administered to the subject by intratumoral injection, and the remaining portion of the antibodies are administered to the subject by intravenous injection.

[0144] In some embodiments, when (a) of the combination formulation consists of two or more TLR agonists, the TLR agonists may be administered to the subject sequentially and at intervals via the same or different administration methods, preferably via the same administration method; more preferably via intratumoral injection or subcutaneous injection.

[0145] In some embodiments, when (c) of the combination formulation consists of two or more antibodies for treating cancer, the antibodies may be administered to the subject sequentially and at intervals via the same or different administration methods; preferably, the antibodies are administered to the subject via the same administration method; more preferably, the antibodies are administered to the subject via intravenous injection, intratumoral injection, or subcutaneous injection; preferably, a portion of the antibodies are administered to the subject via intratumoral injection, and the remaining portion of the antibodies are administered to the subject via intravenous injection.

[0146] In some embodiments, (a) and (b) of the combination formulation are administered to the subject simultaneously after mixing, or (a) and (b) are administered to the subject as a single pharmaceutical composition, and (c) of the combination formulation is administered to the subject via a different administration method. Preferably, (a) and (b) are administered to the subject via intratumoral injection; and (c) is administered to the subject via intravenous injection.

[0147] In some embodiments, portions of (a) and (b) and (c) of the combined formulation are mixed and administered to the subject simultaneously, while the remaining portion of (c) is administered to the subject via a different administration method. Preferably, portions of (a) and (b) and (c) are mixed and administered to the subject via intratumoral injection, while the remaining portion of (c) is administered to the subject via intravenous injection.

[0148] In some embodiments, the combined formulation,

[0149] (a) CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; by intratumoral injection;

[0150] (b) is a tumor antigen; administered via intratumoral injection;

[0151] (c) The antibody is an anti-CTLA4 antibody administered intravenously; or an anti-CD40 antibody administered intratumorally; or an anti-PD-1 antibody and an anti-CTLA4 antibody administered intravenously; or an anti-CD40 antibody and an anti-CTLA4 antibody administered intratumorally, with the anti-CD40 antibody and the anti-CTLA4 antibody administered intravenously; or an anti-PD-1 antibody, an anti-CTLA4 antibody, and an anti-TIGIT antibody administered intravenously; or an anti-PD1 antibody, an anti-CTLA4 antibody, and an anti-CD40 antibody, wherein the anti-PD1 antibody and the anti-CTLA4 antibody are administered intravenously, and the anti-CD40 antibody is administered intratumorally; or an anti-OX40 antibody, an anti-CTLA4 antibody, and an anti-TIGIT antibody, wherein the anti-OX40 antibody is administered intratumorally, and the anti-CTLA4 antibody and the anti-TIGIT antibody are administered intravenously. The antibody is preferably a monoclonal antibody.

[0152] In some implementations, the administration or application can be performed by a single or multiple administrations; preferably, administration can be performed by 1 to 12 administrations.

[0153] In some embodiments, in the pharmaceutical composition or combination formulation, (a) has a single dose of 0.01-10 mpk, (b) has a single dose of 0.1-10 mpk, and (c) has a single dose of 0.01-60 mpk; preferably, (a) has a single dose of 0.02-5 mpk, (b) has a single dose of 1-5 mpk, and (c) has a single dose of 0.02-30 mpk. More preferably, (a) has a single dose of 0.2-5 mpk, (b) has a single dose of 2-5 mpk, and (c) has a single dose of 0.1-30 mpk. Most preferably, (a) has a single dose of 0.3-2 mpk, (b) has a single dose of 3-4 mpk, and (c) has a single dose of 0.2-15 mpk.

[0154] In some embodiments, the single-dose dose of the tumor antigen in the pharmaceutical composition or combination formulation is 0.1-10 mpk, such as 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10 mpk, preferably 1-5 mpk, more preferably 2-5 mpk, and most preferably 3-4 mpk; a TLR9 agonist (such as CpG) When ODN or its pharmaceutically acceptable salt is present, the single dose is 0.01-10 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.5, 3, 4, 5, 6, 7, 8, 9, 10 mpk, preferably 0.02-5 mpk, more preferably 0.2-5 mpk, most preferably 1-2 mpk; TLR7 When a dual agonist (such as R848 or a pharmaceutically acceptable salt thereof) is present, the single dose is 0.01-2 MPa, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55 MPa. 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 mpk, preferably 0.02-1 mpk, more preferably 0.2-1 mpk, most preferably 0.3-0.5 mpk; TLR3 agonists (such as Poly... When I:C or its pharmaceutically acceptable salt is present, the single dose is 0.01-2 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 mpk, preferably 0.02-1 mpk, more preferably 0.2-1 mpk, and most preferably 0.3-0.5 MPk; when anti-CTLA4 antibody is present, the single dose is 0.01-20 MPk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 mpk, preferably 0.02-10 mpk; more preferably 0.1-10 mpk, most preferably 3-5 mpk; when anti-CD40 antibody is present, its single dose is 0.01-5 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 2, 3, 4, 5 mpk, preferably 0.02-2 mpk; More preferably 0.1-2 MPk, most preferably 0.2-0.5 MPk; when anti-PD1 antibody is present, the single dose is 0.01-20 MPk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7 0.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 mpk, preferably 0.02-10 mpk, more preferably 0.1-10 mpk, and most preferably 3-5 mpk; when anti-OX40 antibody is present, its single-dose dose is 0.01-5 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.The dosage is 0, 2, 3, 4, or 5 mpk, preferably 0.02-2 mpk; more preferably 0.1-2 mpk, and most preferably 0.2-0.5 mpk; when anti-TIGIT antibody is present, the single dose is 0.01-10 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, or 0.2 mpk. 5, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1, 1.5, 1.67, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10 mpk, preferably 0.02-5 mpk, more preferably 0.1-5 mpk, and most preferably 1-2 mpk.

[0155] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk;

[0156] (b) Tumor antigen, with a single dose of 0.1-10 mpk;

[0157] (c) Anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-CD40 antibody, single dose 0.01-5 mpk; or anti-PD-1 antibody, single dose 0.01-20 mpk, and anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-CD40 antibody, single dose 0.01-5 mpk, and anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-PD-1 antibody, single dose 0.01-20 mpk, and anti-CTLA4 antibody, A single dose of 0.01-20 mpk and anti-TIGIT antibody, a single dose of 0.01-10 mpk; or anti-PD1 antibody, a single dose of 0.01-20 mpk, anti-CTLA4 antibody, a single dose of 0.01-20 mpk, and anti-CD40 antibody, a single dose of 0.01-5 mpk; or anti-OX40 antibody, a single dose of 0.01-5 mpk, anti-CTLA4 antibody, a single dose of 0.01-20 mpk, and anti-TIGIT antibody, a single dose of 0.01-10 mpk.

[0158] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-5 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-5 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk;

[0159] (b) Tumor antigen, with a single dose of 1-5 mpk;

[0160] (c) Anti-CTLA4 antibody, single dose 0.02-10 mpk; or anti-CD40 antibody, single dose 0.02-2 mpk; or anti-PD-1 antibody, single dose 0.02-10 mpk, and anti-CTLA4 antibody, single dose 0.02-10 mpk; or anti-CD40 antibody, single dose 0.02-2 mpk, and anti-CTLA4 antibody, single dose 0.02-10 mpk; or anti-PD-1 antibody, single dose 0.02-10 mpk, and anti-CTLA4 antibody A single dose of 0.02-10 mpk and anti-TIGIT antibody, with a single dose of 0.02-5 mpk; or anti-PD1 antibody, with a single dose of 0.02-10 mpk, anti-CTLA4 antibody, with a single dose of 0.02-10 mpk, and anti-CD40 antibody, with a single dose of 0.02-2 mpk; or anti-OX40 antibody, with a single dose of 0.02-2 mpk, anti-CTLA4 antibody, with a single dose of 0.02-10 mpk, and anti-TIGIT antibody, with a single dose of 0.02-5 mpk.

[0161] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-5 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2–1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2–5 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2–1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2–1 mpk;

[0162] (b) Tumor antigen, with a single dose of 2-5 mpk;

[0163] (c) Anti-CTLA4 antibody, single dose 0.1-10 MPk; or anti-CD40 antibody, single dose 0.1-2 MPk; or anti-PD-1 antibody, single dose 0.1-10 MPk, and anti-CTLA4 antibody, single dose 0.1-10 MPk; or anti-CD40 antibody, single dose 0.1-2 MPk, and anti-CTLA4 antibody, single dose 0.1-10 MPk; or anti-PD-1 antibody, single dose 0.1-10 MPk, and anti-CTLA4 antibody The dosage is 0.1-10 mpk for a single dose, and anti-TIGIT antibody, with a single dose of 0.1-5 mpk; or anti-PD1 antibody, with a single dose of 0.1-10 mpk, anti-CTLA4 antibody, with a single dose of 0.1-10 mpk, and anti-CD40 antibody, with a single dose of 0.1-2 mpk; or anti-OX40 antibody, with a single dose of 0.1-2 mpk, anti-CTLA4 antibody, with a single dose of 0.1-10 mpk, and anti-TIGIT antibody, with a single dose of 0.1-5 mpk.

[0164] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk, and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or its pharmaceutically acceptable salts, administered at a single dose of 1-2 mpk; and R848 or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk.

[0165] (b) Tumor antigen, with a single dose of 3-4 MPa;

[0166] (c) Anti-CTLA4 antibody, single dose 3-5 MPk; or anti-CD40 antibody, single dose 0.2-0.5 MPk; or anti-PD-1 antibody, single dose 3-5 MPk, and anti-CTLA4 antibody, single dose 3-5 MPk; or anti-CD40 antibody, single dose 0.2-0.5 MPk, and anti-CTLA4 antibody, single dose 3-5 MPk; or anti-PD-1 antibody, single dose 3-5 MPk, and anti-CTLA4 antibody For the body, a single dose of 3-5 mpk is given, along with anti-TIGIT antibody, a single dose of 1-2 mpk; or anti-PD1 antibody, a single dose of 3-5 mpk, anti-CTLA4 antibody, a single dose of 3-5 mpk, and anti-CD40 antibody, a single dose of 0.2-0.5 mpk; or anti-OX40 antibody, a single dose of 0.2-0.5 mpk, anti-CTLA4 antibody, a single dose of 3-5 mpk, and anti-TIGIT antibody, a single dose of 1-2 mpk.

[0167] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25, 1, or 3 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25, 1, or 3 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25, 1, or 3 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25, 1, or 3 mpk; and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk.

[0168] (b) Tumor antigen, administered at a single dose of 3 or 4 mpk;

[0169] (c) Anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 MPk; or anti-CD40 antibody, single dose of 0.1, 0.5, or 1 MPk; or anti-PD-1 antibody, single dose of 1, 3.3, or 10 MPk, and anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 MPk; or anti-CD40 antibody, single dose of 0.1, 0.5, or 1 MPk, and anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 MPk; or anti-PD-1 antibody, single dose of 1, 3.3, or 10 MPk, anti-CT... LA4 antibody, single dose of 0.5, 3.3, or 5 mpk, and anti-TIGIT antibody, single dose of 1.67 mpk; or anti-PD1 antibody, single dose of 1, 3.3, or 10 mpk, anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 mpk, and anti-CD40 antibody, single dose of 0.1, 0.5, or 1 mpk; or anti-OX40 antibody, single dose of 0.4 mpk, anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 mpk, and anti-TIGIT antibody, single dose of 1.67 mpk.

[0170] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk; R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk.

[0171] (b) Tumor antigen, administered at a single dose of 3 or 4 mpk;

[0172] (c) Anti-CTLA4 antibody, single dose 3.3 MPk; or anti-CD40 antibody, single dose 0.5 MPk; or anti-PD-1 antibody, single dose 3.3 MPk, and anti-CTLA4 antibody, single dose 3.3 MPk; or anti-CD40 antibody, single dose 0.5 MPk, and anti-CTLA4 antibody, single dose 3.3 MPk; or anti-PD-1 antibody, single dose 3.3 MPk, and anti-CTLA4 antibody. A single dose of 3.3 MPk and an anti-TIGIT antibody, administered at a single dose of 1.67 MPk; or an anti-PD1 antibody, administered at a single dose of 3.3 MPk, an anti-CTLA4 antibody, administered at a single dose of 3.3 MPk, and an anti-CD40 antibody, administered at a single dose of 0.5 MPk; or an anti-OX40 antibody, administered at a single dose of 0.4 MPk, an anti-CTLA4 antibody, administered at a single dose of 3.3 MPk, and an anti-TIGIT antibody, administered at a single dose of 1.67 MPk.

[0173] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.1-1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.1-1 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.1-1 mpk, and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or its pharmaceutically acceptable salts, administered at a single dose of 0.1-1 mpk; and R848 or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk.

[0174] (b) Tumor antigen, with a single dose of 3-4 MPa;

[0175] (c) Anti-CTLA4 antibody, single dose 0.2-4 MPk; or anti-CD40 antibody, single dose 0.05-1 MPk; or anti-PD-1 antibody, single dose 0.5-10 MPk, and anti-CTLA4 antibody, single dose 0.2-4 MPk; or anti-CD40 antibody, single dose 0.05-1 MPk, and anti-CTLA4 antibody, single dose 0.2-4 MPk; or anti-PD-1 antibody, single dose 0.5-10 MPk, and anti-CTLA4 Antibodies, single dose 0.2-4 MPk, and anti-TIGIT antibody, single dose 1-2 MPk; or anti-PD1 antibody, single dose 0.5-10 MPk, anti-CTLA4 antibody, single dose 0.2-4 MPk, and anti-CD40 antibody, single dose 0.05-1 MPk; or anti-OX40 antibody, single dose 0.2-0.5 MPk, anti-CTLA4 antibody, single dose 0.2-4 MPk, and anti-TIGIT antibody, single dose 1-2 MPk.

[0176] In some embodiments, the pharmaceutical composition or combination formulation comprises, (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.1, 0.4, or 1 mpk;

[0177] (b) Tumor antigen, administered at a single dose of 3 or 4 mpk;

[0178] (c) Anti-PD1 antibody, single dose of 0.5, 1, or 10 mpk; anti-CTLA4 antibody, single dose of 0.2, 0.4, or 4 mpk; and anti-CD40 antibody, single dose of 0.05, 0.3, or 1 mpk.

[0179] In some embodiments, the pharmaceutical composition or combination formulation comprises, (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.4 mpk;

[0180] (b) Tumor antigen, administered at a single dose of 3 or 4 mpk;

[0181] (c) Anti-PD1 antibody, single dose 1 mpk, anti-CTLA4 antibody, single dose 0.4 mpk, and anti-CD40 antibody, single dose 0.3 mpk.

[0182] In some embodiments, the pharmaceutical composition or combination formulation comprises, (a) R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk;

[0183] (b) Tumor antigen, administered at a single dose of 3 or 4 mpk;

[0184] (c) Anti-PD1 antibody, single dose 1 mpk, anti-CTLA4 antibody, single dose 0.4 mpk, and anti-CD40 antibody, single dose 0.3 mpk.

[0185] In some embodiments, the pharmaceutical composition or combination formulation comprises (a) Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.5 mpk;

[0186] (b) Tumor antigen, administered at a single dose of 3 or 4 mpk;

[0187] (c) Anti-PD1 antibody, single dose 1 mpk, anti-CTLA4 antibody, single dose 0.4 mpk, and anti-CD40 antibody, single dose 0.3 mpk.

[0188] In some embodiments, the pharmaceutical composition or combination formulation comprises, (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.1 mpk;

[0189] (b) Tumor antigen, administered at a single dose of 3 or 4 mpk;

[0190] (c) Anti-PD1 antibody, single dose 0.5 mpk, anti-CTLA4 antibody, single dose 0.2 mpk, and anti-CD40 antibody, single dose 0.05 mpk.

[0191] In some embodiments, the pharmaceutical composition or combination formulation comprises, (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk.

[0192] (b) Tumor antigen, administered at a single dose of 3 or 4 mpk;

[0193] (c) Anti-PD1 antibody, single dose 10 mpk, anti-CTLA4 antibody, single dose 4 mpk, and anti-CD40 antibody, single dose 1 mpk.

[0194] In some implementations, the subject is a mammal.

[0195] In some implementations, the subjects are humans, dogs, or cats.

[0196] In some embodiments, the methods for preventing and / or treating cancer may be used in combination with surgery, radiotherapy, chemotherapy, and / or electromagnetic therapy.

[0197] In a sixth aspect, this application provides a method for preventing and / or treating cancer, comprising the steps of administering a therapy to a subject to release a tumor antigen; and administering to the subject an effective amount of (a) one, two, or more TLR agonists, and (c) one, two, or more antibodies for treating cancer.

[0198] In some implementations, therapies that release tumor antigens include:

[0199] (1) Use physical means, such as cryolysis, microwave, thermal ablation, irreversible electroporation, or any other device that mediates in situ physical destruction of the patient’s tumor cells;

[0200] (2) Using chemical or biochemical methods, such as intratumoral injection of exfoliating agents, denaturing agents, toxins, lysing peptides, detergents, etc.; or,

[0201] (3) Use biological agents, such as intratumoral injection of oncolytic viruses or intratumoral injection of genetically engineered cytotoxic cells (CAR-T, CAR-M, NK, etc.);

[0202] (4) Using radiotherapy, administering radiotherapy to the subject to release tumor antigens, preferably, administering radiotherapy to the subject to release tumor antigens.

[0203] In some embodiments, the cancer in the method of preventing and / or treating cancer is a solid tumor, preferably selected from colorectal cancer, melanoma, breast cancer, lung cancer, gastric cancer, head and neck tumors, prostate cancer, liver cancer, nasopharyngeal carcinoma, esophageal cancer, and cervical cancer.

[0204] In some embodiments, the TLR agonist is selected from TLR1 agonists, TLR2 agonists, TLR3 agonists, TLR4 agonists, TLR5 agonists, TLR6 agonists, TLR7 agonists, TLR8 agonists, TLR7 / 8 dual agonists, TLR9 agonists, and TLR10 agonists. Preferably, the TLR agonist is selected from TLR3 agonists, TLR7 agonists, TLR8 agonists, TLR7 / 8 dual agonists, and TLR9 agonists.

[0205] In some embodiments, the TLR agonist is selected from CpG ODN, R848, Poly I:C, BDB001, BDB018, NKTR-262, VTX-2337, imiquimod, RO7119929, SHR2150, TQ-A3334, LHC165, CBLB502, GLA-SE, GSK1795091, Rintatolimod, poly-ICLC, and pharmaceutically acceptable salts thereof. The CpG ODN includes types A, B, and C.

[0206] In some embodiments, the TLR9 agonist is CpG ODN or a pharmaceutically acceptable salt thereof, including type A, type B, and type C. Preferably, the TLR9 agonist is type C CpG ODN or a pharmaceutically acceptable salt thereof.

[0207] In some embodiments, the CpG ODN includes chemical modifications. In some embodiments, the CpG ODN includes modification with one or more phosphate ester groups. In some embodiments, the modification with one or more phosphate ester groups is a thiophosphate bond. In some embodiments, the phosphate backbone of the CpG ODN is fully modified with thiophosphate.

[0208] In some embodiments, the CpG ODN is CpG HP007, CpG HP021, CpG HP026, CpG 684, SD-101, G10 CpG ODN, CpG 1018, CpG 7909, IMO-2125, and pharmaceutically acceptable salts thereof.

[0209] In some embodiments, the TLR9 agonist is a pharmaceutically acceptable salt of CpG HP007. In some embodiments, the TLR7 / 8 dual agonist is R848 or a pharmaceutically acceptable salt thereof. The TLR3 agonist is Poly I:C or a pharmaceutically acceptable salt thereof.

[0210] In some embodiments, (a) is a TLR9 agonist; or a TLR7 / 8 agonist; or a TLR3 agonist; or a combination of a dual TLR7 / 8 agonist and a TLR9 agonist; or a combination of a TLR7 / 8 agonist and a TLR3 agonist; or a combination of a TLR3 agonist and a TLR9 agonist; or a combination of a TLR7 / 8 agonist, a TLR3 agonist, and a TLR9 agonist; preferably, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof.

[0211] In some embodiments, the cancer-treating antibody targets PD-1, CTLA-4, TIGIT, PD-L1, PD-L2, OX40, 4-1BB, ICOS, LAG3, TIM3, IDO, HER2, CD20, CD22, CD27, CD40, CD40L, GITR, EGFR, CD2, CD19, CD33, CD37, CD38, CD44, CD45, CD47, CD52, CD56, CD70, CD79, 5T4, AGS-5, AGS-16, angiopoietin 2, B7.1, and B7. 2. B7DC, B7H1, B7H2, B7H3, BT-062, BTLA, CAIX, carcinoembryonic antigen, Cripto, ED-B, ErbB1, ErbB2, ErbB3, ErbB4, EGFL7, EpCAM, EphA2, EphA3, EphB2, FAP, fibronectin, folate receptor, ganglioside GM3, GD2, glucocorticoid-induced tumor necrosis factor receptor GITR, gp100, gpA33, GPNMB, ICOS, IGF1R, integrin αν, integrin ανβ, KIR, Lewis Y, mesothelin, c-MET, MN carbonic anhydrase IX, MUC1, MUC16, cohesin-4, NKGD2, NOTCH, OX40L, PSCA, PSMA, RANKL, ROR1, ROR2, SLC44A4, STING, multiligand proteoglycan-1, TACI, TAG-72, tendinin, TIM3, TRAILR1, TRAILR2, VEGFR-1, VEGFR-2, VEGFR-3, SSTR, SSTR2, CXCR4, NTR, GRPR. Preferably, the antibody for treating cancer targets one, two, or more (e.g., three) of PD-1, CTLA-4, TIGIT, OX40, and CD40. More preferably, the antibody for treating cancer or a combination thereof targets one, two, or more (e.g., three) of PD-1, CTLA-4, TIGIT, and CD40.

[0212] In some embodiments, the antibody used to treat cancer is a monoclonal antibody, a bispecific antibody, a trispecific antibody, an antibody-drug conjugate (ADC), or a radionuclide conjugate (RDC).

[0213] In some embodiments, (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody.

[0214] In some embodiments, the anti-PD-1 antibody is a monoclonal antibody selected from Keytruda, Opdivo, cimiprimab, toripalimab, sintilimab, camrelizumab, patelilimab, sugelilimab, penapralimab, cepalimumab, and slulilimab; the anti-CTLA4 antibody is a monoclonal antibody selected from ipilimumab and tremelimumab; and the anti-TIGIT antibody is a monoclonal antibody selected from domvanalimab, belrestotug, and tiragol. The following are listed: umab, ospermumab, JS-006, vibostolimab, Lancastotug, Renvistobart, Etigilimab, HB-0030, PM-1021, AB-308, COM-902, M6223, ASP-8374, BAT-6005, Tamgiliblimab; the anti-CD40 antibody is a monoclonal antibody selected from SEA-CD40, Sotigalimab, BSI-038, and Anti-CD40. ChiLob7 / 4 (BioNTech SE), 2141-V11, CDX-1140, YH-003, Selicrelumab, Mitazalimab, ILB-2101, ABBV-927, LVGN-7409; the anti-OX40 antibody is a monoclonal antibody selected from BGB-A445, INBRX-106, YH-002, INCAGN-1949, MEDI-6469, GEN-1055, BAT-6026, IBI-101, ABBV-368, MEDI-0562, HFB-3010.

[0215] In some embodiments, the bispecific antibody is selected from: epalolimbab / tovoretalimab (CTLA4 inhibitor / PD1 inhibitor), candunimbab / tovoretalimab (CTLA4 inhibitor / PD1 inhibitor), Volrustomig (CTLA4 inhibitor / PD1 inhibitor), Danvilostomig (CTLA4 inhibitor / PD1 inhibitor), Lorigerlimab (CTLA4 inhibitor / PD1 inhibitor), Vudalimab (CTLA4 inhibitor / PD1 inhibitor), SHS-010 (CTLA4 inhibitor / PD1 inhibitor), REMD-511 (CTLA4 inhibitor / PD1 inhibitor), YH-008 (CD40 inhibitor / PD1 inhibitor), FS-120 (4-1BB agonist / OX40 agonist), YH-006 (CTLA4 inhibitor / OX40 agonist), ATOR-1015 (CTLA4 inhibitor / OX40 agonist), APVO-603 (4-1BB agonist / OX40 agonist). The following are selected as inhibitors: 0 agonist), GEN-1042 (4-1BB agonist / CD40 agonist), IMB-071703 (4-1BB agonist / CD40 agonist), SM2256 (4-1BB agonist / CD40 agonist), Rilvegostomig (PD1 inhibitor / TIGIT inhibitor), ZG-005 (PD1 inhibitor / TIGIT inhibitor), REMD-532 (PD1 inhibitor / TIGIT inhibitor), IBI-321 (PD1 inhibitor / TIGIT inhibitor), BC008-1A (PD1 inhibitor / TIGIT inhibitor), ZGGS15 (LAG3 inhibitor / TIGIT inhibitor); the trispecific antibody is selected from GBD209 (CTLA4 inhibitor / PD1 inhibitor / TIGIT inhibitor), FG-T903 (CD112R antagonist / PD1 inhibitor / TIGIT inhibitor), and HC010 (hcbiopharma) (CTLA4 inhibitor / PD1 inhibitor / VEGF inhibitor).

[0216] In some embodiments, the ADC drug is selected from: Mylotarg, Besponsa, Kadcyla, Polivy, Lumoxiti, Enhertu, Adcetris, Padcev, Tivdak, Blenrep, Trodelvy, Akalux, Zynlonta, Adrenaline, Luconsartuzumab, and Elahere.

[0217] In some embodiments, the radionuclide conjugate drug RDC is selected from: Pluvicto and Lutathera.

[0218] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof;

[0219] c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody. The antibody is preferably a monoclonal antibody.

[0220] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof;

[0221] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; wherein the antibody is preferably a monoclonal antibody.

[0222] In some embodiments, the pharmaceutical composition or combination formulation comprises:

[0223] (a) CpG HP007 or a pharmaceutically acceptable salt thereof;

[0224] (c) Anti-CD40 antibody. The antibody is preferably a monoclonal antibody.

[0225] In some embodiments, (a) is R848 or a pharmaceutically acceptable salt thereof;

[0226] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody. The antibody is preferably a monoclonal antibody.

[0227] In some embodiments, (a) is Poly I:C or a pharmaceutically acceptable salt thereof;

[0228] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody. The antibody is preferably a monoclonal antibody.

[0229] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, and R848 or a pharmaceutically acceptable salt thereof;

[0230] (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; wherein the antibody is preferably a monoclonal antibody.

[0231] In some embodiments, the subject is given radiotherapy to release tumor antigens. Preferably, the subject is given radiotherapy using rays, preferably selected from alpha, beta, gamma rays, and X-rays, electron beams, proton beams and other particle beams generated by various X-ray therapy machines or accelerators.

[0232] In some implementations, radiotherapy is administered to the subject using radiopharmaceuticals.

[0233] In some implementations, stereotactic radiotherapy (SRT) is used to administer radiotherapy to the subject, preferably three-dimensional conformal radiotherapy (3DCRT), intensity-modulated radiotherapy (IMRT), or image-guided radiotherapy (IGRT).

[0234] In some implementations, stereotactic radiosurgery (SRS) is used to administer radiation therapy to the subject, preferably using X-knife, gamma knife, or CyberKnife.

[0235] In some embodiments, the radiation dose of the radiotherapy is 0.1-4gy, preferably 1gy.

[0236] In some embodiments, (a) and (c) are administered to the subject parenterally, preferably via intratumoral injection (e.g., multiple intratumoral injection), subcutaneous injection, intravenous injection, intramuscular injection, nasal inhalation, or via an interventional device. (a) and (c) may be administered to the subject simultaneously, sequentially, or at intervals.

[0237] In some embodiments, the administration routes of (a) and (c) are the same or different; preferably, (a) and (c) are administered by the same route, such as by intratumoral injection or subcutaneous injection to the subject; (a) and (c) are administered by different routes, such as by intratumoral injection or intravenous injection to the subject, respectively; (a) and (c) are administered by the same route for a portion of their components, and (c) is administered by different routes for the remaining portion, such as by intratumoral injection of a portion of (a) and (c) to the subject, and by intravenous injection of the remaining portion of (c) to the subject.

[0238] In some embodiments, (a) and (c) are administered to the subject as a single pharmaceutical composition, or (a) and (c) are mixed and administered to the subject simultaneously, such as by intratumoral injection or subcutaneous injection; or (a) and (c) are administered to the subject simultaneously by different administration methods, such as by intratumoral injection or intravenous injection respectively; or a portion of (a) and (c) are mixed and administered to the subject by intratumoral injection, and the remaining portion of (c) is administered to the subject by intravenous injection.

[0239] In some embodiments, (a) and (c) may be administered to the subject sequentially or at intervals via the same or different administration methods; preferably, the administration methods are the same, administered to the subject via intratumoral injection, or administered to the subject via intratumoral injection and intravenous injection, respectively.

[0240] In some embodiments, when (a) consists of two or more TLR agonists, the administration routes of the different TLR agonists are the same or different; when (c) consists of two or more antibodies for treating cancer, the administration routes of the different antibodies are the same or different. Preferably, the administration routes of the different TLR agonists in (a) are the same (more preferably, administered to the subject via intratumoral injection or subcutaneous injection); the administration routes of the different antibodies in (c) are the same or different (more preferably, both are administered to the subject via intravenous injection, intratumoral injection or subcutaneous injection, or administered to the subject via intratumoral injection or intravenous injection, respectively).

[0241] In some embodiments, when (a) comprises two or more TLR agonists, the TLR agonists are mixed and administered to the subject simultaneously, or the TLR agonists are administered to the subject as a single pharmaceutical composition, such as by intratumoral injection or subcutaneous injection.

[0242] In some embodiments, when (c) comprises two or more antibodies for treating cancer, the antibodies are mixed and administered to the subject simultaneously, or the antibodies are administered to the subject as a single pharmaceutical composition; preferably, the antibodies are administered to the subject via intravenous injection, intratumoral injection, or subcutaneous injection; or a portion of the antibodies are administered to the subject via intratumoral injection, and the remaining portion is administered to the subject via intravenous injection.

[0243] In some embodiments, when (a) comprises two or more TLR agonists, the TLR agonists may be administered to the subject sequentially or at intervals via the same or different administration methods. Preferably, the subject is administered via the same administration method, either via intratumoral injection or subcutaneous injection.

[0244] In some embodiments, when (c) comprises two or more antibodies for treating cancer, the antibodies may be administered to the subject sequentially or at intervals via the same or different administration methods; preferably, the subject may be administered via the same administration method, such as intravenous injection, intratumoral injection, or subcutaneous injection; or a portion of the antibodies may be administered to the subject via intratumoral injection, and the remaining portion of the antibodies may be administered to the subject via intravenous injection.

[0245] In some embodiments, a portion of (a) and (c) is mixed and administered to the subject simultaneously, while the remaining portion of (c) is administered to the subject via a different method. Preferably, a portion of (a) and (c) is mixed and administered to the subject via intratumoral injection, while the remaining portion of (c) is administered to the subject via intravenous injection.

[0246] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; by intratumoral injection;

[0247] (c) The antibody is an anti-CTLA4 antibody administered intravenously; or an anti-CD40 antibody administered intratumorally; or an anti-PD-1 antibody and an anti-CTLA4 antibody administered intravenously; or an anti-CD40 antibody and an anti-CTLA4 antibody administered intratumorally, with the anti-CD40 antibody and the anti-CTLA4 antibody administered intravenously; or an anti-PD-1 antibody, an anti-CTLA4 antibody, and an anti-TIGIT antibody administered intravenously; or an anti-PD1 antibody, an anti-CTLA4 antibody, and an anti-CD40 antibody, wherein the anti-PD1 antibody and the anti-CTLA4 antibody are administered intravenously, and the anti-CD40 antibody is administered intratumorally; or an anti-OX40 antibody, an anti-CTLA4 antibody, and an anti-TIGIT antibody, wherein the anti-OX40 antibody is administered intratumorally, and the anti-CTLA4 antibody and the anti-TIGIT antibody are administered intravenously. The antibody is preferably a monoclonal antibody.

[0248] In some implementations, after the subject receives continuous radiotherapy for 1-5 days (e.g., 1, 2, 3, 4, 5 days), an effective dose of (a) and (c) is given, followed by a 1-2 day interval before another round of radiotherapy. One round of radiotherapy plus administration is repeated more than once (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 times). Preferably, after the subject receives continuous radiotherapy for 5 days, an effective dose of (a) and (c) is given again, followed by a 2-day interval before another round of radiotherapy. More preferably, one treatment cycle is 1-12 weeks (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks), and there can be multiple cycles (e.g., 2-20 cycles, such as 2, 3, 4, 5, 6, 7, 8, 9, 10 cycles).

[0249] In some implementations, the subject is first given radiotherapy, followed by effective doses of (a) and (c) every 0–96 hours (e.g., 0, 1, 2, 3, 4, 5, 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72, 78, 84, 90, 96 hours). The above treatment is repeated once at 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, and 28 days; preferably, an effective dose of (a) and (c) is given again; more preferably, a treatment cycle is 1-12 weeks (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 weeks), and can be multiple cycles (e.g., 2-20 cycles, such as 2, 3, 4, 5, 6, 7, 8, 9, and 10 cycles).

[0250] In some embodiments, effective doses of (a) and (c) are administered at intervals while the subject is continuously treated with radiotherapy. Preferably, effective doses of (a) and (c) are administered every 1-28 days (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 days). More preferably, a treatment cycle is 1-12 weeks (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks), and there may be multiple cycles (e.g., 2-20 cycles, such as 2, 3, 4, 5, 6, 7, 8, 9, 10 cycles).

[0251] In some implementations, the prevention or treatment may include one or more treatment cycles.

[0252] In some embodiments, the single dose of (a) is 0.01-10 mpk, and the single dose of (c) is 0.01-60 mpk; preferably, the single dose of (a) is 0.02-5 mpk, and the single dose of (c) is 0.02-30 mpk. More preferably, the single dose of (a) is 0.2-5 mpk, and the single dose of (c) is 0.1-30 mpk. Most preferably, the single dose of (a) is 0.3-2 mpk, and the single dose of (c) is 0.2-15 mpk.

[0253] In some embodiments, when a TLR9 agonist (such as CpG ODN or a pharmaceutically acceptable salt thereof) is present, the single-dose administration is 0.01-10 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.5, 3, 4, 5, 6, 7, 8, 9, 10 mpk, preferably 0.02-5 mpk, more preferably 0.2-5 mpk, most preferably 1-2 mpk; TLR7 When a dual agonist (such as R848 or a pharmaceutically acceptable salt thereof) is present, the single dose is 0.01-2 MPa, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55 MPa. 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 mpk, preferably 0.02-1 mpk, more preferably 0.2-1 mpk, most preferably 0.3-0.5 mpk; TLR3 agonists (such as Poly... When I:C or its pharmaceutically acceptable salts are present, the single dose is 0.01-2 MPa, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1. 9. 2.0 mpk, preferably 0.02-1 mpk, more preferably 0.2-1 mpk, and most preferably 0.3-0.5 mpk; when anti-CTLA4 antibody is present, the single dose is 0.01-20 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.The dosages are 90, 0.95, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 mpk, preferably 0.02-10 mpk; more preferably 0.1-10 mpk, and most preferably 3-5 mpk; when anti-CD40 antibody is present, the single dose is 0.01-5 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45. The dosage ranges are 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 2, 3, 4, 5 mpk, preferably 0.02-2 mpk; more preferably 0.1-2 mpk, and most preferably 0.2-0.5 mpk; when anti-PD1 antibody is present, the single dose is 0.01-20 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 mpk, preferably 0.02-10 mpk, more preferably 0.1-10 mpk, and most preferably 3-5 mpk; when anti-OX40 antibody is present, its single-dose dose is 0.01-5 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.15, 0.20, 0. 25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 2, 3, 4, 5 mpk, preferably 0.02-2 mpk; more preferably 0.1-2 mpk, most preferably 0.2-0.5 mpk; when anti-TIGIT antibody is present, its single dose is 0.01-10 mpk, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.The preferred values ​​are 0.02-5 mpk, more preferably 0.1-5 mpk, and most preferably 1-2 mpk. (The values ​​are listed as 50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1, 1.5, 1.67, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10 mpk, 0.02-5 mpk, 0.1-5 mpk, and 1-2 mpk.)

[0254] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk;

[0255] (c) Anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-CD40 antibody, single dose 0.01-5 mpk; or anti-PD-1 antibody, single dose 0.01-20 mpk, and anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-CD40 antibody, single dose 0.01-5 mpk, and anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-PD-1 antibody, single dose 0.01-20 mpk, and anti-CTLA4 antibody, A single dose of 0.01-20 mpk and anti-TIGIT antibody, a single dose of 0.01-10 mpk; or anti-PD1 antibody, a single dose of 0.01-20 mpk, anti-CTLA4 antibody, a single dose of 0.01-20 mpk, and anti-CD40 antibody, a single dose of 0.01-5 mpk; or anti-OX40 antibody, a single dose of 0.01-5 mpk, anti-CTLA4 antibody, a single dose of 0.01-20 mpk, and anti-TIGIT antibody, a single dose of 0.01-10 mpk.

[0256] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-5 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-5 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.02-1 mpk;

[0257] (c) Anti-CTLA4 antibody, single dose 0.02-10 mpk; or anti-CD40 antibody, single dose 0.02-2 mpk; or anti-PD-1 antibody, single dose 0.02-10 mpk, and anti-CTLA4 antibody, single dose 0.02-10 mpk; or anti-CD40 antibody, single dose 0.02-2 mpk, and anti-CTLA4 antibody, single dose 0.02-10 mpk; or anti-PD-1 antibody, single dose 0.02-10 mpk, and anti-CTLA4 antibody The dosage is 0.02-10 mpk for a single dose, and anti-TIGIT antibody, with a single dose of 0.1-5 mpk; or anti-PD1 antibody, with a single dose of 0.02-10 mpk, anti-CTLA4 antibody, with a single dose of 0.02-10 mpk, and anti-CD40 antibody, with a single dose of 0.02-2 mpk; or anti-OX40 antibody, with a single dose of 0.02-2 mpk, anti-CTLA4 antibody, with a single dose of 0.02-10 mpk, and anti-TIGIT antibody, with a single dose of 0.02-5 mpk.

[0258] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-5 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2-5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2–1 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2–5 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2–1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.2–1 mpk;

[0259] (c) Anti-CTLA4 antibody, single dose 0.1-10 MPk; or anti-CD40 antibody, single dose 0.1-2 MPk; or anti-PD-1 antibody, single dose 0.1-10 MPk, and anti-CTLA4 antibody, single dose 0.1-10 MPk; or anti-CD40 antibody, single dose 0.1-2 MPk, and anti-CTLA4 antibody, single dose 0.1-10 MPk; or anti-PD-1 antibody, single dose 0.1-10 MPk, and anti-CTLA4 antibody The dosage is 0.1-10 mpk for a single dose, and anti-TIGIT antibody, with a single dose of 0.1-5 mpk; or anti-PD1 antibody, with a single dose of 0.1-10 mpk, anti-CTLA4 antibody, with a single dose of 0.1-10 mpk, and anti-CD40 antibody, with a single dose of 0.1-2 mpk; or anti-OX40 antibody, with a single dose of 0.1-2 mpk, anti-CTLA4 antibody, with a single dose of 0.1-10 mpk, and anti-TIGIT antibody, with a single dose of 0.1-5 mpk.

[0260] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk, and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or its pharmaceutically acceptable salts, administered at a single dose of 1-2 mpk; and R848 or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk.

[0261] (c) Anti-CTLA4 antibody, single dose 3-5 MPk; or anti-CD40 antibody, single dose 0.2-0.5 MPk; or anti-PD-1 antibody, single dose 3-5 MPk, and anti-CTLA4 antibody, single dose 3-5 MPk; or anti-CD40 antibody, single dose 0.2-0.5 MPk, and anti-CTLA4 antibody, single dose 3-5 MPk; or anti-PD-1 antibody, single dose 3-5 MPk, and anti-CTLA4 antibody For the body, a single dose of 3-5 mpk is given, along with anti-TIGIT antibody, a single dose of 1-2 mpk; or anti-PD1 antibody, a single dose of 3-5 mpk, anti-CTLA4 antibody, a single dose of 3-5 mpk, and anti-CD40 antibody, a single dose of 0.2-0.5 mpk; or anti-OX40 antibody, a single dose of 0.2-0.5 mpk, anti-CTLA4 antibody, a single dose of 3-5 mpk, and anti-TIGIT antibody, a single dose of 1-2 mpk.

[0262] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25, 1, or 3 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25, 1, or 3 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25, 1, or 3 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25, 1, or 3 mpk; R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk.

[0263] (c) Anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 MPk; or anti-CD40 antibody, single dose of 0.1, 0.5, or 1 MPk; or anti-PD-1 antibody, single dose of 1, 3.3, or 10 MPk, and anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 MPk; or anti-CD40 antibody, single dose of 0.1, 0.5, or 1 MPk, and anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 MPk; or anti-PD-1 antibody, single dose of 1, 3.3, or 10 MPk, anti-CT... LA4 antibody, single dose of 0.5, 3.3, or 5 mpk, and anti-TIGIT antibody, single dose of 1.67 mpk; or anti-PD1 antibody, single dose of 1, 3.3, or 10 mpk, anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 mpk, and anti-CD40 antibody, single dose of 0.1, 0.5, or 1 mpk; or anti-OX40 antibody, single dose of 0.4 mpk, anti-CTLA4 antibody, single dose of 0.5, 3.3, or 5 mpk, and anti-TIGIT antibody, single dose of 1.67 mpk.

[0264] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk; R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk.

[0265] (c) Anti-CTLA4 antibody, single dose 3.3 MPk; or anti-CD40 antibody, single dose 0.5 MPk; or anti-PD-1 antibody, single dose 3.3 MPk, and anti-CTLA4 antibody, single dose 3.3 MPk; or anti-CD40 antibody, single dose 0.5 MPk, and anti-CTLA4 antibody, single dose 3.3 MPk; or anti-PD-1 antibody, single dose 3.3 MPk, and anti-CTLA4 antibody. A single dose of 3.3 MPk and an anti-TIGIT antibody, administered at a single dose of 1.67 MPk; or an anti-PD1 antibody, administered at a single dose of 3.3 MPk, an anti-CTLA4 antibody, administered at a single dose of 3.3 MPk, and an anti-CD40 antibody, administered at a single dose of 0.5 MPk; or an anti-OX40 antibody, administered at a single dose of 0.4 MPk, an anti-CTLA4 antibody, administered at a single dose of 3.3 MPk, and an anti-TIGIT antibody, administered at a single dose of 1.67 MPk.

[0266] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk.

[0267] (c) Anti-CD40 antibody, single dose of 0.5 mpk.

[0268] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk;

[0269] (c) Anti-PD-1 antibody, single dose 3.3mpk; anti-CTLA4 antibody, single dose 3.3mpk.

[0270] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3 mpk.

[0271] (c) Anti-PD1 antibody, single dose 3.3 mpk, anti-CTLA4 antibody, single dose 3.3 mpk, and anti-CD40 antibody, single dose 0.5 mpk.

[0272] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.25 mpk;

[0273] (c) Anti-PD1 antibody, single dose 1 mpk, anti-CTLA4 antibody, single dose 0.5 mpk, and anti-CD40 antibody, single dose 0.1 mpk.

[0274] In some embodiments, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 3 mpk;

[0275] (c) Anti-PD1 antibody, single dose 10 mpk, anti-CTLA4 antibody, single dose 5 mpk, and anti-CD40 antibody, single dose 1.5 mpk.

[0276] In some embodiments, the subject is a mammal; preferably, the subject is a human, dog, or cat.

[0277] Compared with the prior art, this application has the following beneficial effects:

[0278] This application uses different active pharmaceutical ingredients in combination, including TLR agonists, tumor antigens, and antibodies for treating cancer. Each active pharmaceutical ingredient has different anti-tumor immune activation effects and has a synergistic effect.

[0279] This application also provides a combination therapy of releasing tumor antigens and using TLR agonists and antibodies to treat cancer. The combination of radiotherapy to release tumor antigens and the anti-tumor immune activation effects of TLR agonists and antibodies has a synergistic effect.

[0280] This application, by combining different active pharmaceutical ingredients or combination therapies, can improve efficacy compared to therapies that rely on a single drug class, and can reduce dosage, thereby reducing the likelihood of toxicity and adverse events, and reducing the likelihood of acquired drug resistance.

[0281] Specifically, TLR agonists have immunostimulatory activity, and intratumoral administration can significantly reduce their dosage, which allows them to be combined with other therapeutic agents to increase the breadth of their pharmacological effects and enhance their antitumor efficacy.

[0282] It is known that a major reason why tumor cells cannot be cleared by an individual's immune system is that tumor cells originate from the individual's own cells and therefore have weak immunogenicity. This application mixes a TLR receptor agonist with a tumor antigen, such as a tumor cell lysate, which can enhance the immunogenicity of tumor cells; or releases the tumor antigen through a therapy such as radiotherapy and then administers the TLR receptor agonist intratumorally; this can stimulate an effective anti-tumor immune response, thereby inhibiting or clearing the corresponding tumor cells in the individual.

[0283] TLR-like receptor agonists have synergistic and toxicity-reducing effects and can be used in conjunction with immunotherapy. When combined with multiple antibodies, they not only enhance anti-tumor effects but also reduce the systemic dosage of antibodies, thereby reducing the likelihood of toxicity and adverse events and improving tolerability.

[0284] This application improves therapeutic efficacy by using lower doses of the active pharmaceutical ingredient and increasing the number of therapeutic drugs with synergistic mechanisms; or by combining a therapy that releases tumor antigens with a drug therapy, while reducing the frequency and severity of adverse events and the likelihood of acquired immune resistance. Attached Figure Description

[0285] To more clearly illustrate the technical solutions of the specific embodiments of this application, the drawings used in the specific embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0286] Figure 1 shows the tumor volume in mice after administration of drugs to groups G1, G3, G4, G5, G6, G7, G8, G9, and G10 in Table 1 of Example 2. It illustrates the tumor-suppressing effect of CpG C / lysate in combination with anti-mCTLA-4 iv, or in combination with anti-mCTLA-4 / anti-mPD-1 iv, or in combination with anti-mCTLA-4 / anti-mPD-1 / anti-TIGIT iv.

[0287] Figure 2 shows the tumor volume in mice after administration to groups G1, G4, G6, G12, G15, and G16 in Table 1 of Example 2, demonstrating the tumor-suppressing effect of CpG C / R848 / lysate in combination with anti-mCTLA-4 / anti-mPD-1 IV.

[0288] Figure 3 shows the tumor volume in mice after administration of drugs in groups G1, G2, G5, G6, G11, G12, G14, and G15 in Table 1 of Example 2, demonstrating the tumor-suppressing effect of CpG C / R848it combined with anti-mCTLA-4 iv, or combined with anti-mCTLA-4 / anti-mPD-1 iv.

[0289] Figure 4 shows the tumor volume in mice after administration in groups G1, G4, G6, G10, and G17 in Table 3 of Example 3, demonstrating the tumor-suppressing effect of combined administration of CpG C+lysate+anti-mCD40+anti-mPD-1+anti-mCTLA-4 and it.

[0290] Figure 5 shows the tumor volume in mice after administration in groups G2, G5, G7, G11, and G16 of Example 3, illustrating the tumor-suppressing effect of CpG C+lysate+anti-mCD40+anti-mPD-1+anti-mCTLA-4 combined with subcutaneous administration.

[0291] Figure 6 shows the tumor volume in mice after administration to groups G3, G4, G6, G12, G13, G14, and G15 in Table 3 of Example 3, demonstrating the tumor-suppressing effect of CpG C / lysate / anti-mCD40it combined with anti-mPD-1 / anti-mCTLA-4iv at different doses.

[0292] Figure 7 shows the tumor volume in mice after administration to groups G1, G4, G8, G10, G18, and G19 in Table 3 of Example 3, demonstrating the tumor-suppressing effect of combined iterative administration (it) or subcutaneous administration (sc) of R848+lysate+anti-mCD40+anti-mPD-1+anti-mCTLA-4.

[0293] Figure 8 shows the tumor volume in mice after administration to groups G1, G4, G9, G10, G20, and G21 in Table 3 of Example 3, demonstrating the tumor-suppressing effect of Poly I:C+lysate+anti-mCD40+anti-mPD-1+anti-mCTLA-4 combined with itinerant (it) or subcutaneous (sc) administration.

[0294] Figure 9 shows the tumor volume in mice after administration of drugs in groups G1, G2, G3, G6, G7, G8, G9, G11, G13, G15, and G22 in Table 5 of Example 4. It shows the tumor-suppressing effect of radiotherapy + CpG C it combined with anti-mCD40 it, or combined with anti-mCTLA-4 / anti-mPD-1 iv, or combined with anti-mCD40 it + anti-mCTLA-4 / anti-mPD-1 i.v.

[0295] Figure 10 shows the tumor volume in mice after administration in groups G1, G2, G3, G9, G11, G17, G18, and G19 in Table 5 of Example 4, demonstrating the tumor-suppressing effect of radiotherapy + CpG C + anti-mCD40 + anti-mCTLA-4 + anti-mPD-1 combined with it administration at different doses.

[0296] Figure 11 shows the tumor volume in mice after administration in groups G1, G2, G8, G9, G11, G18, and G22 in Table 5 of Example 4, demonstrating the tumor-suppressing effect of radiotherapy + CpG C it combined with anti-mCD40 it + anti-mCTLA-4 / anti-mPD-1 i.v., or combined with anti-mCD40 / anti-mCTLA-4 / anti-mPD-1 it.

[0297] Figure 12 shows the tumor volume in mice after administration of drugs in groups G1, G2, G5, G7, G9, G12, G16, and G21 in Table 5 of Example 4, demonstrating the tumor-suppressing effect of radiotherapy + Poly I:C it combined with anti-mPD-1 / anti-mCTLA-4 iv, or combined with anti-mCD40 / anti-mPD-1 / anti-mCTLA-4 it.

[0298] Figure 13 shows the tumor volume in mice after administration of drugs in groups G1, G2, G4, G6, G9, G10, G14, and G20 in Table 5 of Example 4, demonstrating the tumor-suppressing effect of radiotherapy + R848 it combined with anti-mCD40 it, or combined with anti-mCD40 / anti-mPD-1 / anti-mCTLA-4 it. Detailed Implementation

[0299] Reference will now be made to detailed embodiments of this application, one or more of which are described below. Each example is provided for explanation and not for limitation of the invention. In fact, it will be apparent to those skilled in the art that various modifications and variations can be made to the invention without departing from its scope or spirit. For example, features described or illustrated as part of some embodiments may be used in other embodiments to produce further embodiments.

[0300] Unless otherwise stated, all terms used to disclose this invention (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Further guidance is provided below for a better understanding of the teachings of this invention. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0301] the term

[0302] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND," and also undoubtedly includes technical solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0303] The terms “containing,” “comprising,” and “including” as used in this invention are synonyms and are inclusive or open-ended, not excluding additional, uncited members, elements, or method steps.

[0304] The term "two or more" as used in this invention refers to more than two types, such as three or four types.

[0305] In this invention, the numerical range represented by endpoints includes all numerical values ​​and fractions contained within that range, as well as the endpoints mentioned.

[0306] This invention relates to concentration values, which include fluctuations within a certain range. For example, fluctuations are allowed within a corresponding precision range. For instance, 2% may fluctuate within ±0.1%. For larger values ​​or values ​​that do not require overly precise control, even greater fluctuations are permitted. For example, 100mM may fluctuate within ranges of ±1%, ±2%, ±5%, etc. Regarding molecular weight, fluctuations of ±10% are allowed.

[0307] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0308] TLR agonists:

[0309] Toll-like receptors (TLRs) are a family of receptors that recognize microbial-derived molecular structures (pathogen-associated molecular models). Immune cells expressing TLRs are activated by binding to these pathogen-associated molecular models. TLRs are a class of naturally occurring pattern recognition receptors, highly expressed in both humans and mice, and their family includes a series of receptors such as TLR1-TLR10. TLRs can recognize many substances derived from pathogenic microorganisms, which are all TLR agonists. Activation of TLR ligands can activate downstream signaling pathways, triggering the release of various bioactive factors. TLR ligands can be used as adjuvants or immune activators in cancer therapy.

[0310] TLR3 agonists:

[0311] TLR3 is an intracellular recognition system that responds to viral nucleic acids (dsRNA, ssRNA, and ssDNA) as well as endogenous double-stranded RNA. Many TLR3 agonists have been developed, such as RGC100 and ARNAX, a synthetic DNA-RNA hybrid compound. However, in the past two years, researchers have returned to using traditional dsRNA mimicry ligands, such as poly I:C, focusing on improving their delivery modalities and novel disease applications.

[0312] TLR4 agonists:

[0313] TLR4 is the most studied member of the TLR family, recognizing lipopolysaccharide (LPS). Located on the plasma membrane, TLR4 is primarily expressed on myeloid cells, but not on pDCs and naive B cells. TLR4 recognizes LPS through its co-receptors myeloid differentiation factor-2 (MD-2) and CD14. Recent work on TLR4 agonists has focused on the development and evaluation of modified products, such as lipid monophosphate A (MPLA) and glucopyranolipide A (GLA), which are structurally related to LPS but lack high pyrogenicity while retaining strong immunomodulatory properties, thus increasing their feasibility for clinical application.

[0314] TLR5 agonists:

[0315] TLR5 recognizes flagellin and is expressed on epithelial cells and immune cells (such as macrophages and immature dendritic cells), generating an immune response via the MyD88-dependent signaling pathway. Recent work on TLR5 agonists has primarily focused on improving tolerability. Unfortunately, flagellin can induce unwanted hyperreactivity; as a protein, it can induce self-antibodies, interfering with its function as an adjuvant. Studies have addressed this issue by removing the B-cell epitope region from flagellin; deimmunized flagellin retains its TLR5 adjuvant activity.

[0316] TLR7 agonists:

[0317] TLR7 molecules are expressed on the endosomal membrane and form heterodimers with TLR8 to recognize viral-derived single-stranded RNA and synthetically produced ligands such as CpG-A and Poly G3, promoting type I interferon expression. Exemplary TLR7 agonists include, but are not limited to, imiquimod and loxoribin.

[0318] TLR8 agonists:

[0319] TLR8 is expressed on the endosome membrane, and its recognized ligands and downstream signals are consistent with those of TLR7. Only human TLR8 recognizes single-stranded RNA; mouse TLR8 cannot. Exemplary TLR8 agonists include, but are not limited to, VTX-2337.

[0320] TLR9 agonists:

[0321] TLR9 is an intracellular pattern recognition receptor found in the endosomal compartments of mouse plasmacytoid dendritic cells, macrophages, natural killer cells, and other APCs. In humans, TLR9 is primarily expressed in B cells and pDCs. The physiological ligand of TLR9 is a bacterial dsDNA fragment with an unmethylated CpG dinucleotide. TLR9 agonists are artificial oligonucleotides with an unmethylated CpG motif. They can induce signaling cascades, leading to transcriptional programs that result in inflammatory processes, enhanced killing of cancer cells, and the generation of adaptive immune responses.

[0322] CpG ODN:

[0323] CpG ODN refers to CpG oligodeoxynucleotides, which are at least about ten nucleotides in length and include one unmethylated CpG. CpG ODN is single-stranded. The entire CpG ODN can be unmethylated or partially unmethylated. CpG ODN includes D-type (also known as A-type), K-type (also known as B-type), C-type, and P-type ODN.

[0324] Commonly used CpG ODNs include CpG HP007 (SEQ ID NO.1: 5'-tcgcg aacgt tcgcc gcgta cgtacgcgg-3'), CpG HP021 (SEQ ID NO.2: 5'-tcgca acgtt gcctt cgaag g-3'), CpG HP026 (SEQ ID NO.3: 5'-tcgcg acgtt cgccg acgtt cgta-3'), CpG 684 (SEQ ID NO.4: 5'-tcgac gttcg tcgtt cgtcg ttc-3'), SD-101 (SEQ ID NO.5: 5'-tcgaa cgttc gaacg ttcga acgtt cgaat-3'), G10 CpG ODN (SEQ ID NO.6: 5'-ggggg ggggg gacga tcgtc ggggg ggggg-3'), CpG 1018 (SEQ ID NO.7: 5'-tgact gtgaa cgttc gagatga-3'), CpG 7909 (SEQ ID NO.8: 5'-tcgtc gtttt gtcgt tttgt cgtt-3'), IMO-2125 (SEQ ID NO.8: 5'-tcg*aacg*tcg*-Xg*cttg*caag*ct-5', where g* represents 2'-deoxy-7-denitro-guanosine, and X is the glycerol linker).

[0325] R848:

[0326] R848, or rectimote, is an analogue of imiquimod and exhibits immunomodulatory effects both in vitro and in vivo. As a dual agonist of TLR7 and TLR8, R848 stimulates immune cells to produce large amounts of inflammatory cytokines, promoting the Th1 response while inhibiting the Th2 response, thus exerting antiviral, antitumor, and anti-allergic effects.

[0327] Poly I:C:

[0328] Poly I:C is a synthetic double-stranded ribonucleic acid (BSA) and an interferon inducer with antiviral and immunomodulatory functions.

[0329] Combination TLR agonist adjuvant:

[0330] Combining agonists of different TLRs in a single vaccine can produce synergistic effects, driving robust vaccine-mediated immune responses. For example, a liposomal adjuvant containing 1V270 (a TLR7 agonist) and 2B182C (a TLR4 agonist) induced balanced anti-HA and anti-NA IgG1 and IgG2a responses against influenza without the overreactivity common in Th1 pro-inflammatory responses. Similarly, one study evaluated the co-encapsulation of ovalbumin (OVA) with ten unique combinations of two to three TLR ligands, including Pam3CSK4 (a TLR2 agonist), MPLA (a TLR4 agonist), imiquimod (a TLR7 / 8 agonist), and CpG (a TLR9 agonist), with triple combinations promoting antigen-specific antibody titers with an overall balanced Th1 / Th2 response. Therefore, combinations of TLR agonist adjuvants can provide a broad range of customized immune responses.

[0331] Cancer and tumor:

[0332] Cancer and tumor are interchangeable terms that express the same meaning. Cancer refers to a pathological condition in the body characterized by uncontrolled cell proliferation. Cancer cells grow uncontrollably, can invade surrounding tissues and metastasize, endangering an individual's life. Examples of cancer include, but are not limited to: malignant epithelial tumors, lymphomas, blastomas, and leukemia. More specific examples of cancer include solid tumors and hematologic malignancies, including but not limited to: lung (small cell and non-small cell) cancer, breast cancer, prostate cancer, carcinoid tumors, bladder cancer, stomach cancer, pancreatic cancer, liver cancer (hepatocellular carcinoma), hepatoblastoma, colorectal cancer, head and neck cancer, nasopharyngeal carcinoma, squamous cell carcinoma, esophageal cancer, ovarian cancer, uterine cancer, endometrial cancer, cervical cancer, mesothelioma, melanoma, sarcoma, osteosarcoma, liposarcoma, thyroid cancer, desmoidoma, acute myeloid leukemia (AML), and chronic myeloid leukemia (CML).

[0333] Tumor antigen:

[0334] There are three types of tumor antigens: tumor-associated antigens (TAAs), oncogenic virus-derived antigens, and tumor-specific antigens (TSAs, neoantigens). Tumor antigens are antigenic substances produced by tumor cells, that is, they trigger an immune response in the host. Tumor antigens can stimulate an anti-tumor immune response, and an immune response against a tumor antigen can inhibit the growth of tumor cells expressing that tumor antigen or kill them.

[0335] Whole tumor lysate:

[0336] Whole tumor cell lysates, also known as tumor lysates, refer to lysates prepared by disrupting tumor cells. Methods for disrupting tumor cells include, but are not limited to, repeated freeze-thaw cycles, ultrasonic treatment, and mechanical disruption.

[0337] Tumor cell lines can be obtained by culturing primary tumor cells and secondary or metastatic tumor cells in vitro. These tumor cell lines are then used to prepare tumor cell lysates. The cells used to prepare tumor cell lysates can be autologous, allogeneic, or xenogeneic tumor cells. These tumor cell lysates contain a wide variety of tumor antigens.

[0338] Tumor neoantigens:

[0339] Tumor neoantigens are abnormal proteins encoded by mutated genes in tumor cells that can be recognized by immune cells and activate the body's immune response. These antigenic mutant proteins are called tumor neoantigens. The mechanism by which T cells specifically recognize tumor cells lies in the presence of gene mutations in tumor cells that distinguish them from normal cells. These mutations, when translated into proteins, differ from normal proteins. Some of these mutant proteins, after ubiquitination and degradation, form polypeptides that can be recognized by the immune system—this is called polypeptide antigenicity. These antigenic polypeptides are presented to the cell surface by the tumor cell's major histocompatibility complex, and are subsequently recognized as foreign substances by the immune system, leading to the attack and killing of tumor cells by immune cells. Tumor neoantigens are tumor-specific antigens, unique to tumor cells, absent in normal tissues, and do not easily induce autoimmunity. They possess strong tumor specificity and immunogenicity and are unaffected by central and peripheral immune tolerance.

[0340] To proactively identify tumor neoantigens, two conditions must be met: first, high-throughput DNA sequencing of each patient's individual tumor genome is required to identify all tumor-specific mutations; second, antigenicity prediction of proteins and peptides expressed by these mutations is necessary to synthesize potential neoantigen peptides in vitro.

[0341] Antibody:

[0342] Antibodies include polyclonal antibodies, monoclonal antibodies, and antibody fragments. "Antibody fragments" include the antigen-binding fragments of these antibodies, including Fab, F(ab')2, Fd, Fv, Fab'-SH, scFv, bispecific antibodies, and the smallest antibody recognition unit, as well as single-chain derivatives of these antibodies and fragments, such as scFv-Fc. Antibody types can be selected from IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, and IgD. Furthermore, antibodies include naturally occurring antibodies and non-naturally occurring antibodies, including, for example, chimeric, bifunctional, humanized antibodies and human antibodies, as well as related synthetic isoforms. The term "antibody" is used interchangeably with "immunoglobulin."

[0343] PD-1:

[0344] PD-1 is expressed on T cells activated after TCR-mediated antigen stimulation, as well as on B cells, NK cells, and monocytes. The function of PD-1 is to control T cell activation in peripheral tissues to prevent tissue damage caused by persistent inflammation. Prolonged or high-level antigen exposure (e.g., in advanced cancer) leads to PD-1-mediated T cell exhaustion. During T cell sensitization, IFNγ induces PD-L1 in APCs as a counterregulatory mechanism after T cell activation. Under TCR activation, PD-1 ligation with PD-L1 leads to phosphorylation of PD1 by Lek tyrosine kinase, which subsequently recruits SHP2 phosphatase to the intracellular domain of PD-1, thereby dephosphorylating and inactivating TCR and CD28, thus blocking downstream activation signals from stimulation and co-stimulation during T cell sensitization. Similarly, in the effector phase of tumors, PD-L1 is induced by IFNγ and expressed by tumor cells. PD-L1 attaches to PD1+ Teff cells (PD1+ Teff cells recognize MHC-I-presented tumor antigens via TCR), recruits SHP2, thereby inactivating TCR signaling, reducing cytokine production, and inducing T cell apoptosis or PD1+TIL depletion and dysfunction, thus preventing Teff cell infiltration into tumors. Blocking PD1 with antagonistic mAbs works by restoring the immune function of depleted T cells in the periphery and can enhance tumor killing through various immune mechanisms, including directly inhibiting T cell apoptosis, leading to the restoration of T cell effector function, enhancing APC-mediated T cell activation, downregulating Treg-mediated suppression, downregulating immunosuppressive cytokines IL-10 and TGFβ, and upregulating IL-2 and IFN-γ, while simultaneously resisting PD-L1-induced expression in tumors. Clinical evidence supports a model that blocking the PD1 signaling axis is most effective for tumors that have already elicited an endogenous T cell response but are suppressed by the binding of PD-1 to ligands PD-L1 and PD-L2. However, the response to αPD1 mAb in PD-L1(-) tumors suggests that a pre-existing immune response is not always an absolute necessity for tumor rejection after αPD1 treatment.

[0345] CTLA-4:

[0346] As T cell activation progresses, counter-regulatory mechanisms are induced, preventing the activation and survival of T cells with excessively weak or strong TCR / MHC / Ag interactions. One of the inducing signals expressed on Teff cells is CTLA-4, which is induced when TCR / MHC / Ag interactions are too strong or prolonged. CTLA-4 has functions beyond CD28, thereby blocking the co-stimulatory signaling provided by CD80 / 86. This limits the polyclonal nature of TCR variants and the selection, proliferation, and survival of T cell clones. Furthermore, CTLA-4 is constitutively highly expressed by Tregs, which can also inhibit CD80 / 86 co-stimulatory signaling by directly blocking interactions with CD28 and by trans-endocytosis through membrane flaps of CD80 / 86-containing DCs. Increased T cell stimulation leads to increased secretion of Th1 cytokines such as TNFα, IL2, and IFNylate, as well as increased T cell proliferation (↑CD3 / Ki67). Therefore, blocking CTLA4 with αCTLAmAb will increase the co-stimulatory signal provided by CD28 to continue, ultimately enabling T cell clones with strong TCR / MHC / Ag affinity to survive and proliferate, and increasing TCR variant diversity.

[0347] TIGIT:

[0348] TIGIT, a relatively new molecule discovered recently, belongs to the class of cell surface proteins with immunosuppressive functions. Its full name is T cell immune receptor with Ig and ITIM domains, also known as WUCAM, Vstm3, or VSIG9. It is an inhibitory receptor belonging to the Ig superfamily and possesses many characteristic structures of immunoregulatory proteins, such as extracellular immunoglobulin domains, type I transmembrane structures, and intracellular immune receptor tyrosine inhibitory / activating motifs (ITIM / ITAM).

[0349] CD40:

[0350] CD40 (Bp50) is a surface antigen associated with T cell and B cell function. Its precursor contains 297 amino acids and is a type I transmembrane glycoprotein composed of an N-terminal signal peptide (20 amino acids), an extracellular domain (193 amino acids), a transmembrane domain (22 amino acids), and a cytoplasmic domain (62 amino acids). The extracellular domain contains four CDRs with a total of 22 cysteine ​​residues. Within the 71 amino acids of the hinge domain, there are 9 serine residues and 7 threonine residues, which are potential O-glycosylation sites, as well as 2 potential N-glycosylation sites. Therefore, CD40 has a high degree of glycosylation. The molecular weight estimated from the amino acid sequence is 28 kDa, but the molecular weight after glycosylation is 40-50 kDa. CD40 is expressed on B cells, thymic epithelial cells, activated monocytes / macrophages, dendritic cells, hematopoietic progenitor cells, epithelial cells, endothelial cells, and some tumor cell lines, such as the HepG2 liver cancer cell line and the HS294T melanoma cell line. PMA, anti-IgM antibodies, anti-CD20 antibodies, and IFN-γ can all stimulate CD40 expression on B cells, and IFN-γ can also stimulate CD40 expression on tumor cells. CD40 signal transduction mainly regulates the activity of non-receptor tyrosine protein kinases, such as Lyn, Fyn, and Syk, and can also activate PI3K, phospholipase Cg2, Rel / NF-κB transcription factors, and induce Bcl-xL, Cdk4, and Cdk6 proteins.

[0351] OX40:

[0352] OX40 (also known as CD134) is an important T cell costimulatory molecule and a member of the tumor necrosis factor receptor (TNF) superfamily, primarily expressed on activated T cells. The ligand OX40L (also known as CD252) binds to three molecules of OX40 protein in a trimeric form, forming a hexamer complex. This complex activates downstream signaling pathways such as NF-κB, PI3K, and AKT, thereby regulating genes involved in T cell division and survival, promoting the transcription of cytokine genes, and enhancing the expression of cytokine receptors—all crucial for cell survival.

[0353] Effective dosage:

[0354] An effective dose refers to the amount of medication that is effective in treating or preventing the recurrence of a condition or its signs or symptoms. A single dose may not produce the full therapeutic effect; a series of doses may be required for the full therapeutic effect. Therefore, an effective therapeutic dose can be administered in single or multiple applications of a single unit. The dosage depends on the individual being treated, the individual's immune system's ability to synthesize antibodies, and the expected level of protection. The exact amount of active ingredient required depends on the physician's judgment and varies from person to person.

[0355] Anti-tumor effects:

[0356] In this invention, anti-tumor effects refer to the inhibition of tumor cell growth, reduction of tumor cell number, tumor shrinkage, tumor disappearance, improvement of individual clinical symptoms, slowing of tumor growth, extension of individual life, improvement of individual quality of life, and suppression of tumor metastasis and recurrence. Anti-tumor effects include both the treatment of tumors and the prevention of tumor occurrence and recurrence.

[0357] Pharmaceutically acceptable salts:

[0358] Pharmaceutically acceptable salts are those that, within the bounds of reasonable medical judgment, are suitable for contact with the tissues of mammals, particularly humans, without excessive toxicity, irritation, allergic reactions, etc., and whose benefits / risks are proportionate to a reasonable ratio. If the compound is basic, pharmaceutically acceptable salts include salts prepared from inorganic acids as well as salts prepared from organic acids. If the compound is acidic, pharmaceutically acceptable salts include salts prepared from inorganic bases and / or organic bases.

[0359] Pharmaceutically acceptable salts of exemplary CpG oligonucleotides include: ammonium salts, alkali metal salts such as sodium, lithium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, zinc salts, salts formed with organic bases (e.g., organic amines), such as N-Me-D-reduced glucosamine, N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride, choline, tromethamine, dicyclohexylamine, tert-butylamine, and salts formed with amino acids such as arginine, lysine, etc.

[0360] Pharmaceutically acceptable salts of R848 include salts that form with a base or an acid, with exemplary salts including hydrochloride, toluenesulfonate, or citrate of R848.

[0361] Pharmaceutically acceptable carriers or excipients:

[0362] Pharmaceutically acceptable means any substance permitted by the relevant government regulatory authority as acceptable for human or livestock use. A carrier refers to one or more solid or liquid fillers, diluents, or encapsulating substances, which may be organic, inorganic, natural, or synthetic. This includes various solutions, diluents, solvents, dispersants, liposomes, emulsifiers, antibacterial agents, antifungal agents, isotonic and de-absorption agents, and other carriers suitable for use in the pharmaceutical combinations of this invention. Excipients include, for example, solvents, fillers, buffers, tension modifiers, and preservatives. In some embodiments, the pharmaceutical composition may contain one or more excipients acting as solvents, fillers, buffers, and tension modifiers. The choice of pharmaceutically acceptable carriers and excipients depends on the manner of application of the pharmaceutical combinations provided by this invention.

[0363] Subjects:

[0364] In this invention, "subject" and "patient" refer to mammals. Exemplary subjects include humans, dogs, and cats.

[0365] Radiation therapy:

[0366] Radiation therapy for tumors, or simply radiotherapy, is the treatment of cancer using radiation. It is a localized treatment method that utilizes radiation, such as alpha, beta, and gamma rays produced by radioactive isotopes, and X-rays, electron beams, proton beams, and other particle beams produced by various X-ray therapy machines or accelerators, to treat malignant tumors.

[0367] Radiation is a beam of particles or energy-carrying waves. It can damage genes (DNA) and some molecules within cells. Genes control cell growth and differentiation. Radiation damages the genes of cancer cells, preventing them from regrowing and differentiating. In other words, radiation can be used to kill cancer cells and shrink tumor tissue.

[0368] Radiotherapy plays an increasingly prominent role in cancer treatment and has become one of the main methods for treating malignant tumors. Radiotherapy can be used alone or in combination with other therapies (surgery, chemotherapy, etc.) to shrink tumors before surgery or chemotherapy, or to kill cancer cells that may remain after other therapies.

[0369] While radiation therapy kills cancer cells, it inevitably causes some damage to surrounding normal tissues. The goal of radiation therapy is to kill as many cancer cells as possible while protecting surrounding normal tissues and organs from radiation. Therefore, radiation therapy requires careful planning to ensure that the radiation accurately targets the tumor while minimizing the impact on surrounding tissues.

[0370] The choice of radiation therapy depends on a variety of factors: tumor type, tumor size, age, previous cancer history, physical condition, and other factors. Radiation therapy includes conventional radiation therapy, stereotactic body radiotherapy (SBRT), three-dimensional conformal radiotherapy (3DCRT), intensity-modulated radiation therapy (IMRT), image-guided radiation therapy (IGRT), volumetric modulated arc radiation therapy (VMAT / RapidArc), proton therapy, and other treatment methods.

[0371] The embodiments of the present invention will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this invention, or follow experimental manuals or conventional conditions in the art, or other experimental methods known in the art, or follow the conditions recommended by the manufacturer.

[0372] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0373] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0374] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0375] In the examples below, PBS was purchased from Solarbio, catalog number P1010. Unless otherwise specified, the concentration of PBS solution was 0.01M and the pH value was 7.2-7.4.

[0376] Example 1: Tumor Suppression Experiment in MC38 Colon Cancer Model Mice

[0377] 1. Experimental Materials

[0378] TLR receptor agonists:

[0379] TLR9 agonist CpG C1: 5'-tcgcgaacgttcgccgcgtacgtacgcgg-3', SEQ ID NO.: 1, (CpG HP007), was provided by Huapu Biotechnology (Hebei) Co., Ltd.

[0380] TLR7 / 8 agonist R848 (Resiquimod), RCE, catalog number: 113202.

[0381] TLR3 agonist Poly I:C, Sigma, catalog number: P1350-25MG.

[0382] Antibody:

[0383] Anti-mPD-1 (clone number: RMP1-14), BioXcell, catalog number: 861023J2.

[0384] Anti-mCTLA4 (clone number: 9D9), BioXcell, catalog number: 809522M2.

[0385] Anti-mTIGIT (clone number: 1B4), Absolute, catalog number: Ab01258-1.1 / T2434B03.

[0386] Anti-mCD40 (clone number: FGK4.5 / FGK45), BioXcell, catalog number: BE0016-2.

[0387] Anti-mOX40 (clone number: OX-36), BioXcell, catalog number: BE0031.

[0388] Tumor antigen:

[0389] Tumor lysate 1 (Lysate-1): MC38 colon cancer tumor lysate, provided by Crown Bioscience (Beijing) Co., Ltd.

[0390] Mice:

[0391] Female C57BL / 6N mice aged 6-7 weeks, Beijing Vital River Biotechnology Co., Ltd.

[0392] Cell lines:

[0393] MC38 tumor cells. MC38 tumor cells were cultured in DMEM medium containing 10% fetal bovine serum. MC38 cells in the exponential growth phase were collected and resuspended in PBS to a suitable concentration for subcutaneous inoculation in C57BL / 6 mice.

[0394] 2. Experimental Methods

[0395] The experimental mice were subcutaneously inoculated with 1×10⁻⁶ spores on their right back 6 MC38 cells were resuspended in PBS (0.1 mL / cell), and tumor growth was observed regularly until the tumor reached an average volume of 150 mm². 3 Mice were randomly assigned to groups based on tumor size and body weight for drug administration. Treatment was administered every 3 days for 5 consecutive days. Before administration, all animals were weighed, and tumor size was measured using calipers. Tumor volume was estimated using the following formula: Volume = Length × Width 2 ×0.5. All data are expressed as averages in mm. 3 +SEM. The day of grouping is designated as Day 0 (D0), and the administration begins on Day 1 (D1) according to the experimental design.

[0396] Calculate the relative tumor volume increase ratio T / C (%), T / C (%) = (mean of relative tumor volume in the treatment group / mean of relative tumor volume in the control group) × 100, and then calculate the tumor growth inhibition rate (TGI, %) = (1-T / C) × 100.

[0397] During the experiment, mice whose body weight decreased by more than 20% compared to the first day of administration, or mice deemed unsuitable by a veterinarian, were euthanized. Total tumor volume exceeding 2000 mm² was also considered. 3 The mice will be euthanized.

[0398] Six mice were in each experimental group. They were administered either systemically (iv) at a volume of 10 μL / g or intratumorally (it) at a volume of 50 μL / mouse. The administration cycle was Q3D for a total of 5 administrations.

[0399] Example 2. Efficacy of TLR agonists combined with anti-PD1, anti-CTLA4, and anti-TIGIT in a murine MC38 colon cancer model.

[0400] The animals are grouped as shown in Table 1:

[0401] Table 1 Note: 1) Antibody drugs are administered intravenously, with all antibodies dissolved together in the same system. CpGC, R848, and Lysate1 are administered intratumorally, with a maximum volume of 50 μL per mouse. 2) When multiple drugs are prepared in the same system, they should be thoroughly mixed before administration. 3) For intratumoral administration, the total volume is 50 μL per mouse. CpGC and R848 can be mixed with tumor lysate in the same system. The concentrations in the table are calculated based on a mouse weight of 20 g. 4) For the route-specific administration groups, intratumoral administration is performed first, followed by intravenous administration.

[0402] The experimental results are shown in Table 2 and Figures 1 to 3.

[0403] Table 2 shows the antitumor effects of different treatment groups for D13.

[0404] Table 2: Antitumor effects of different treatment groups for D13

[0405] Figures 1, 2, and 3 show the tumor volumes in mice after administration of the drugs in groups G1, G3, G4, G5, G6, G7, G8, G9, G10; groups G1, G4, G6, G12, G15, G16; and groups G1, G2, G5, G6, G11, G12, G14, G15, as shown in Table 1. Figure 1 shows the tumor-suppressing effect of CpG C / lysate it combined with anti-mCTLA-4 iv, or combined with anti-mCTLA-4 / anti-mPD-1 iv, or combined with anti-mCTLA-4 / anti-mPD-1 / anti-TIGIT iv. Figure 2 shows the tumor-suppressing effect of CpG C / R848 / lysate it combined with anti-mCTLA-4 / anti-mPD-1 iv. Figure 3 shows the antitumor effect of CpG C / R848 in combination with anti-mCTLA-4 IV, or in combination with anti-mCTLA-4 / anti-mPD-1 IV.

[0406] As shown in Figure 1, intratumoral administration of tumor lysate 1 alone has no antitumor effect; intratumoral administration of both CpG C and tumor lysate 1 enhances the tumor-suppressive effect.

[0407] Intratumoral administration of CpG C and tumor lysate 1 and intravenous administration of anti-mCTLA4 alone had comparable antitumor effects. Intratumoral administration of CpG C and tumor lysate 1 combined with intravenous administration of anti-mCTLA4 alone further enhanced the antitumor effect, which was stronger than intravenous administration of anti-mCTLA4+anti-mPD-1 or anti-mCTLA4+anti-mPD-1+anti-mTIGIT. Intratumoral administration of CpG C and tumor lysate 1 combined with intravenous administration of anti-mCTLA4+anti-mPD-1 or anti-mCTLA4+anti-mPD-1+anti-mTIGIT showed a strong antitumor effect, significantly reducing the tumor volume in mice. Moreover, the antitumor effect was even stronger when all three monoclonal antibodies were administered intravenously at the same time.

[0408] As shown in Figure 2, intratumoral administration of CpG C and R848, combined with intravenous injection of anti mCTLA4+anti mPD-1, and simultaneous intratumoral injection of the same tumor lysate can further enhance the antitumor effect.

[0409] As shown in Figure 3, both intratumoral administration of CpG C and intratumoral administration of R848 have certain antitumor effects, and their combined use further enhances the antitumor effect. Intratumoral administration of CpG C and R848, along with intravenous injection of anti-mCTLA4 + anti-mPD-1, has comparable antitumor effects. Intratumoral administration of CpG C and R848, combined with intravenous injection of anti-mCTLA4 or intravenous injection of anti-mCTLA4 + anti-mPD-1, exhibits strong antitumor effects, and the simultaneous intravenous administration of both monoclonal antibodies further enhances the antitumor effect.

[0410] It is known that intratumoral injection of TLR receptor agonists and tumor lysates, combined with intravenous injection of monoclonal antibodies, has a strong tumor-suppressive effect, while the simultaneous administration of TLR9 receptor agonist CpG C and TLR7 / 8 receptor agonist R848 has the strongest antitumor activity.

[0411] In summary, TLR receptor agonists, tumor antigens, and low-dose antibodies, especially when two or three monoclonal antibodies are administered in combination, exhibit excellent antitumor effects. Furthermore, the combined administration of TLR9 and TLR7 / 8 receptor agonists enhances the antitumor effect. In particular, TLR receptor agonists and tumor antigens demonstrate strong antitumor activity even with significantly reduced monoclonal antibody dosages.

[0412] Example 3: The therapeutic effects of different TLR agonists combined with different antibodies in a murine MC38 colon cancer model

[0413] The experimental materials and methods were the same as above. The tumors were allowed to grow to an average volume of approximately 100 mm². 3 The mice were randomly grouped and given medication based on tumor size and body weight.

[0414] The animals are grouped as shown in Table 3:

[0415] Table 3

[0416] The dosage concentrations in the table are calculated based on a mouse weight of 20g.

[0417] For the route-specific administration group, intratumoral administration was performed first, followed by intravenous administration.

[0418] The experimental results are shown in Table 4 and Figures 4 to 8.

[0419] Table 4: Antitumor effects of different treatment groups in Example 3 a VS G1; b VS G2; c VS G3.

[0420] Figures 4 to 8 show the tumor volume in mice after drug administration for the following groups (G1, G4, G6, G10, G17; G2, G5, G7, G11, G16; G3, G4, G6, G12, G13, G14, G15; G1, G4, G8, G10, G18, G19; G1, G4, G9, G10, G20, G21) listed in Table 3. Figure 4 shows the tumor-suppressing effect of combined itching (it) administration of CpG C+lysate+anti-mCD40+anti-mPD-1+anti-mCTLA-4. Figure 5 shows the tumor-suppressing effect of combined subcutaneous (sc) administration of CpG C+lysate+anti-mCD40+anti-mPD-1+anti-mCTLA-4. Figure 6 shows the antitumor effect of CpG C / lysate / anti-mCD40 it combined with anti-mPD-1 / anti-mCTLA-4 iv administration at different doses. Figure 7 shows the antitumor effect of R848+lysate+anti-mCD40+anti-mPD-1+anti-mCTLA-4 combined it or subcutaneous administration. Figure 8 shows the antitumor effect of Poly I:C + lysate + anti-mCD40 + anti-mPD-1 + anti-mCTLA-4 combined itinerant (IT) or subcutaneous (SC) administration (Statistical methods for Figures 4 and 5: First, Bartlett's test was performed to check the consistency of variance among groups. P > 0.05 indicates that the data conforms to a normal distribution. Therefore, ANOVA was used to test whether the means of all groups were equal, and Tukey's HSD test was run for pairwise comparisons among all groups. Statistical methods for Figures 6, 7, and 8: First, Bartlett's test was performed to check the consistency of variance among groups. P < 0.05 indicates that the data does not conform to a normal distribution. Therefore, the Kruskal-Wallis test was used to check whether there were groups with different medians, and Conover's non-parametric all-pairs comparison test was run for pairwise comparisons).

[0421] As shown in Figure 4 and Table 4, compared with the PBS group (G1), intratumoral administration of tumor lysate had almost no antitumor effect (G4, TGI = 10.07%). Intratumoral administration of tumor lysate and low-dose CpGC showed significant antitumor effects (G6, TGI = 42.33%). Intratumoral administration of low-dose anti-mCD40 + anti-mCTLA4 + anti-mPD1 showed even better antitumor effects (G10, TGI = 65.67%). Furthermore, intratumoral administration of tumor lysate + low-dose CpGC + low-dose anti-mCD40 + anti-mCTLA4 + anti-mPD1 further enhanced the antitumor effect (G17, TGI = 84.69%). The difference between group G17 and group G6 was statistically significant (P < 0.05).

[0422] As shown in Figure 5 and Table 4, compared with the PBS group (G2), subcutaneous administration of tumor lysate showed a weak antitumor effect (G5, TGI = 22.31%). When subcutaneous administration of tumor lysate and low-dose CpGC was performed, the antitumor effect was not further enhanced (G7, TGI = 17.36%). When subcutaneous administration of low-dose anti-mCD40 + anti-mCTLA4 + anti-mPD1 was performed, a significant antitumor effect was shown (G11, TGI = 60.84%). However, when subcutaneous administration of these three antibodies was combined with subcutaneous administration of tumor lysate + low-dose CpGC, the antitumor effect was not further enhanced (G16, TGI = 52.61%).

[0423] As shown in Figure 6 and Table 4, compared with the PBS group (G3), intratumoral administration of tumor lysate showed a weak antitumor effect (G4, TGI = 22.89%). When intratumoral administration of tumor lysate and low-dose CpGC was performed, a significant antitumor effect was shown (G6, TGI = 50.55%). When low-dose anti-mCD40 was administered intratumorally and low-dose anti-mCTLA4+anti-mPD1 was administered intravenously, a strong antitumor effect was shown (G12, TGI = 76.05%). However, when this group (anti-mCD40 it + anti-mCTLA-4 / anti-mPD-1 iv combined administration group) was further combined with intratumoral administration of tumor lysate and low-dose CpGC, the antitumor effect was not further enhanced (G14, TGI = 75.26%).

[0424] As shown in Figure 6, the antitumor effect was dose-dependent when tumor lysate, CpGC, and anti-mCD40 were administered intratumorally, followed by intravenous administration of anti-mCTLA4 and anti-mPD1. The antitumor effect was further enhanced with increasing doses of CpGC, anti-mCD40, anti-mCTLA4, and anti-mPD1 (TGIs of groups G13, G14, and G15 were 26.63%, 75.26%, and 91.33%, respectively). Groups G14 and G15 showed statistically significant differences compared to group G13 (P < 0.001).

[0425] As shown in Figure 7 and Table 4, compared with the PBS group (G1), intratumoral administration of tumor lysate had almost no antitumor effect (G4, TGI = 10.07%). Intratumoral administration of tumor lysate and low-dose R848 showed some antitumor effect (G8, TGI = 39.67%). Intratumoral administration of low-dose anti-mCD40 + anti-mCTLA4 + anti-mPD1 showed significant antitumor effect (G10, TGI = 65.67%). The antitumor effect was further enhanced when intratumoral administration of tumor lysate + low-dose R848 + low-dose anti-mCD40 + anti-mCTLA4 + anti-mPD1 (G18, TGI = 75.88%). The difference between the G18 group and the G8 group was statistically significant (P < 0.05).

[0426] As shown in Figure 8 and Table 4, compared with the PBS group (G1), intratumoral administration of tumor lysate had almost no antitumor effect (G4, TGI = 10.07%). Intratumoral administration of tumor lysate and low-dose Poly I:C did not significantly enhance the antitumor effect (G9, TGI = 16.00%). Intratumoral administration of low-dose anti-mCD40 + anti-mCTLA4 + anti-mPD1 showed a significant antitumor effect (G10, TGI = 65.67%). Furthermore, intratumoral administration of tumor lysate + low-dose Poly I:C + low-dose anti-mCD40 + anti-mCTLA4 + anti-mPD1 further enhanced the antitumor effect (G20, TGI = 90.13%). The G20 group showed statistically significant differences compared to both the G9 and G10 groups (P < 0.05).

[0427] Furthermore, as shown in Figures 7 and 8 and Table 4, for the same TLRA, the intratumoral administration route has a superior antitumor effect compared to the subcutaneous administration route. This is evident in the fact that intratumoral administration of R848 + tumor lysate + anti-mCD40 + anti-mCTLA4 + anti-mPD1 (G18, TGI = 75.88%) is superior to subcutaneous administration of R848 + tumor lysate + anti-mCD40 + anti-mCTLA4 + anti-mPD1 (G19, TGI = 65.11%). Similarly, intratumoral administration of Poly I:C + tumor lysate + anti-mCD40 + anti-mCTLA4 + anti-mPD1 (G20, TGI = 90.13%) is superior to subcutaneous administration of Poly I:C + tumor lysate + anti-mCD40 + anti-mCTLA4 + anti-mPD1 (G21, TGI = 74.79%).

[0428] In summary, different TLR agonists and tumor lysates, when combined with low doses of the three antibodies administered intratumorally, all exhibited excellent antitumor effects. Furthermore, altering the route of administration of some or all drug components affected the antitumor efficacy, with intratumoral administration showing the best tumor-suppressive effect. Moreover, the tumor-suppressive effect showed a dose-dependent relationship with the dosage of both the TLR receptor agonist and the antibody; within the dose range of this experiment, higher doses demonstrated the most superior antitumor effect.

[0429] Example 4: Efficacy of radiotherapy combined with different TLR agonists and antibodies in a murine MC38 colon cancer model

[0430] MC38 tumor cells resuspended in serum-free DMEM medium were inoculated at 1×10⁻⁶. 6 0.1 mL (4:1 Matrigel) was subcutaneously injected into the right side of 6-8 week old female C57BL / 6J mice (Beijing Vital River Biotechnology Co., Ltd.), until the tumor grew to 91 mm. 3 Treatment was divided into left and right groups.

[0431] On the day treatment began, the tumor sites of mice were irradiated with X-rays using a precision X-ray machine (model X-RAD 225, cabinet voltage 225kV), and the drugs were administered 24 hours after irradiation.

[0432] The remaining experimental materials and methods are the same as above.

[0433] The animals are grouped as shown in Table 5:

[0434] Table 5 Note: In the radiotherapy combined with drug treatment group, X-ray irradiation was performed first, followed by drug administration 24 hours later. The drug concentrations in the table are calculated based on a mouse weight of 20g. For the multi-route administration group, intratumoral administration was performed first, followed by intravenous administration.

[0435] The experimental results are shown in Figures 9 to 13.

[0436] Figures 9, 10, 11, 12, and 13 show the tumor volume in mice after drug administration in the following groups (G1, G2, G3, G6, G7, G8, G9, G11, G13, G15, G22; G1, G2, G3, G9, G11, G17, G18, G19; G1, G2, G8, G9, G11, G18, G22; G1, G2, G5, G7, G9, G12, G16, G21; G1, G2, G4, G6, G9, G10, G14, G20) as described in Table 5 of Example 4. Figure 9 shows the antitumor effects of radiotherapy + CpG C it combined with anti-mCD40 it, or combined with anti-mCTLA-4 / anti-mPD-1 iv, or combined with anti-mCD40 i.t. + anti-mCTLA-4 / anti-mPD-1 iv. Figure 10 shows the antitumor effects of radiotherapy + CpG C + anti-mCD40 + anti-mCTLA-4 + anti-mPD-1 combined it at different doses. Figure 11 shows the antitumor effects of radiotherapy + CpG C it combined with anti-mCD40 it + anti-mCTLA-4 / anti-mPD-1 iv, or combined with anti-mCD40 / anti-mCTLA-4 / anti-mPD-1 it. Figure 12 shows the antitumor effect of radiotherapy + Poly I:C it combined with anti-mPD-1 / anti-mCTLA-4 iv, or combined with anti-mCD40 / anti-mPD-1 / anti-mCTLA-4 i.t. Figure 13 shows the antitumor effect of radiotherapy + R848 it combined with anti-mCD40 it, or combined with anti-mCD40 / anti-mPD-1 / anti-mCTLA-4 it.

[0437] Figure 9 shows the tumor growth curves in animals treated with radiotherapy + CpG + different antibody combinations. Compared with the PBS group (G2), the radiotherapy + CpG Cit group (G11) showed stronger tumor inhibition (TGI = 74.2%); the anti-mCD40 it group (G6) had a TGI of 61.1%; the anti-mCTLA-4 / anti-mPD-1 IV combination group (G7) had a TGI of 13.1%; and the anti-mCD40 it + anti-mCTLA-4 / anti-mPD-1 IV combination group (G8) had a TGI of 71.3%.

[0438] When radiotherapy combined with CpG C and antibody administration, the tumor growth inhibition effect was further enhanced in the radiotherapy + CpG C + anti-mCD40 combined itinerary administration group (G13, TGI = 86.6%), the radiotherapy + CpG C itinerary + anti-mCTLA-4 / anti-mPD-1 IV combined itinerary administration group (G15, TGI = 93.1%), and the radiotherapy + CpG C / anti-mCD40 itinerary + anti-mCTLA-4 / anti-mPD-1 IV combined itinerary administration group (G22, TGI = 96.6%).

[0439] The above data indicate that, compared with the combined use of different antibody combinations, the addition of radiotherapy + CpG C can further enhance the anti-tumor effect of different antibody combinations.

[0440] As shown in Figure 10, radiotherapy combined with different doses of CpG C and three antibodies exhibited strong antitumor effects. Compared with the PBS group (G2), the TGI of groups G17, G18, and G19 were 87.4%, 94.3%, and 90.7%, respectively. In particular, within the dose range of this experiment, radiotherapy combined with a medium dose of a TLR receptor agonist and a medium dose of an antibody (G18: X-ray combined with intratumoral administration of 1 mg / kg CpG C + 0.5 mg / kg anti-mCD40 + 3.3 mg / kg anti-mCTLA4 + 3.3 mg / kg anti-mPD-1) showed the best antitumor effect.

[0441] Figure 11 shows the tumor growth curves in animals treated with radiotherapy, CpG C, and three antibodies via different routes. Compared with the PBS group (G2), the radiotherapy + CpG C / anti-mCD40 / anti-mCTLA-4 / anti-mPD-1 combined ital administration group (G18, TGI = 94.3%) and the radiotherapy + CpG C / anti-mCD40 it + anti-mCTLA-4 / anti-mPD-1 iv combined administration group (G22, TGI = 96.6%) showed similar tumor-suppressing effects. This indicates that within the dose range of this experiment, changing the administration route of some active ingredients can still maintain excellent anti-tumor effects.

[0442] Figure 12 shows the tumor growth curves in animals treated with radiotherapy + PolyIC + different antibody combinations. Compared with the PBS group (G2), the TGI of the radiotherapy + PolyIC group (G12) was 62.9%, while the TGI of the radiotherapy + PolyIC + anti-mCTLA-4 / anti-mPD-1 IV combination group (G16) was 88.3%; and the TGI of the radiotherapy + PolyIC / anti-CD40 / anti-mCTLA-4 / anti-mPD-1 IV combination group (G21) was 94.6%.

[0443] The above data indicate that radiotherapy combined with Poly IC and different antibody combinations can further enhance the anti-tumor effect.

[0444] Figure 13 shows the tumor growth curves in animals treated with radiotherapy + R848 + different antibody combinations. Compared with the PBS group (G2), the TGI of the radiotherapy + R848 it group (G10) was 73.7%, the TGI of the radiotherapy + R848 / anti-mCD40 it combination group (G14) was 97.0%, and the TGI of the radiotherapy + R848 / anti-CD40 / anti-mCTLA-4 / anti-mPD-1 it combination group (G20) was 87.1%.

[0445] The above data indicate that radiotherapy combined with R848 and different antibody combinations can further enhance the anti-tumor effect.

[0446] In summary, radiotherapy combined with TLR receptor agonists and low-dose antibodies (especially one to three monoclonal antibodies) exhibits excellent antitumor activity. Furthermore, within the dose range tested, altering the route of antibody administration maintained excellent antitumor activity. Moreover, the tumor-suppressive effect showed a dose-dependent relationship with the dosage of both the TLR receptor agonist and the antibody; within the dose range tested, the medium dose demonstrated the most superior antitumor effect.

[0447] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. A pharmaceutical composition, characterized in that, Include: (a) One, two, or more TLR agonists; (b) Tumor antigens; (c) One, two, or more antibodies for treating cancer; (d) Pharmaceutically acceptable carriers or excipients.

2. The pharmaceutical composition according to claim 1, characterized in that, The pharmaceutical composition does not contain any other active ingredients.

3. A combination formulation, characterized in that, It includes: (a) One, two, or more TLR agonists; (b) Tumor antigens; (c) One, two, or more antibodies for treating cancer.

4. The combination formulation according to claim 3, characterized in that, The combined formulation does not contain any other active ingredients.

5. The pharmaceutical composition according to claim 1 or 2, or the combination formulation according to any one of claims 3-4, characterized in that, The TLR agonist is selected from TLR3 agonists, TLR4 agonists, TLR5 agonists, TLR7 agonists, TLR8 agonists, TLR7 / 8 dual agonists, and TLR9 agonists.

6. The pharmaceutical composition or combination formulation according to claim 5, characterized in that, The TLR9 agonist is selected from CpG ODN and pharmaceutically acceptable salts thereof, preferably from CpG HP007, CpG HP021, CpG HP026, CpG 684, SD-101, G10 CpG ODN, CpG 1018, CpG 7909, IMO-2125 and pharmaceutically acceptable salts thereof; The TLR7 / 8 dual agonist is R848 or a pharmaceutically acceptable salt thereof; The TLR3 agonist is Poly I:C or a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition or combination formulation according to claim 5 or 6, characterized in that, (a) is a TLR9 agonist; or a TLR7 / 8 agonist; or a TLR3 agonist; or a combination of a dual TLR7 / 8 agonist and a TLR9 agonist; or a combination of a TLR7 / 8 agonist and a TLR3 agonist; or a combination of a TLR3 agonist and a TLR9 agonist; or a combination of a TLR7 / 8 agonist, a TLR3 agonist, and a TLR9 agonist; Preferably, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof.

8. The pharmaceutical composition or combination formulation according to any one of claims 1-7, characterized in that, The tumor antigen is a whole tumor lysate, a neotumor antigen, a genetically engineered protein, polypeptide, nucleic acid, or DNA; preferably a whole tumor lysate.

9. The pharmaceutical composition or combination formulation according to any one of claims 1-8, characterized in that, The antibody used to treat cancer targets one, two, or more of PD-1, CTLA-4, TIGIT, OX40, and CD40.

10. The pharmaceutical composition or combination formulation according to any one of claims 1-9, characterized in that, The antibodies used to treat cancer are monoclonal antibodies, bispecific antibodies, trispecific antibodies, antibody-drug conjugates (ADCs), or radionuclide conjugates (RDCs).

11. The pharmaceutical composition or combination formulation according to claim 9 or 10, characterized in that, (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody.

12. The pharmaceutical composition or combination formulation according to claim 11, characterized in that, Include: (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; (b) is a tumor antigen; (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody.

13. The pharmaceutical composition or combination formulation according to any one of claims 1-12, characterized in that, In the pharmaceutical composition or combination preparation, the mass ratio of (a):(b):(c) is (1-1000):(10-1000):(1-6000), preferably (2-500):(100-500):(2-3000), more preferably (20-500):(200-500):(10-3000), and most preferably (30-200):(300-400):(20-1500).

14. The pharmaceutical composition or combination formulation according to claim 13, characterized in that, Contains (a) 1-1000 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 1-200 parts of R848 or a pharmaceutically acceptable salt thereof; or 1-200 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 1-1000 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 1-200 parts of R848 or a pharmaceutically acceptable salt thereof; or 1-1000 parts of R848 or a pharmaceutically acceptable salt thereof, and 1-200 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 1-1000 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 1-200 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 1-1000 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 1-200 parts of R848 or a pharmaceutically acceptable salt thereof, and 1-200 parts of Poly I:C or a pharmaceutically acceptable salt thereof; (b) Tumor antigens are 10-1000 samples; (c) 1-2000 doses of anti-CTLA4 antibody; or 1-500 doses of anti-CD40 antibody; or 1-2000 doses of anti-PD-1 antibody and 1-2000 doses of anti-CTLA4 antibody; or 1-500 doses of anti-CD40 antibody and 1-2000 doses of anti-CTLA4 antibody; or 1-2000 doses of anti-PD-1 antibody, 1-2000 doses of anti-CTLA4 antibody and 1-1000 doses of anti-TIGIT antibody; or 1-2000 doses of anti-PD1 antibody, 1-2000 doses of anti-CTLA4 antibody and 1-500 doses of anti-CD40 antibody; or 1-500 doses of anti-OX40 antibody, 1-2000 doses of anti-CTLA4 antibody and 1-1000 doses of anti-TIGIT antibody; Preferably, it comprises (a) 100-200 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 30-50 parts of R848 or a pharmaceutically acceptable salt thereof; or 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100-200 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of R848 or a pharmaceutically acceptable salt thereof; or 30-50 parts of R848 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100-200 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 100-200 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of R848 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof. (b) The tumor antigen is 300-400 samples; (c) 300-500 doses of anti-CTLA4 antibody; or 20-50 doses of anti-CD40 antibody; or 300-500 doses of anti-PD-1 antibody and 300-500 doses of anti-CTLA4 antibody; or 20-50 doses of anti-CD40 antibody and 300-500 doses of anti-CTLA4 antibody; or 300-500 doses of anti-PD-1 antibody, 300-500 doses of anti-CTLA4 antibody and 100-200 doses of anti-TIGIT antibody; or 300-500 doses of anti-PD1 antibody, 300-500 doses of anti-CTLA4 antibody and 20-50 doses of anti-CD40 antibody; or 20-50 doses of anti-OX40 antibody, 300-500 doses of anti-CTLA4 antibody and 100-200 doses of anti-TIGIT antibody; Alternatively, preferably, it comprises (a) 10-100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof; or 30-50 parts of R848 or a pharmaceutically acceptable salt thereof; or 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 10-100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of R848 or a pharmaceutically acceptable salt thereof; or 30-50 parts of R848 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 10-100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof; or 10-100 parts of CpG HP007 or a pharmaceutically acceptable salt thereof, and 30-50 parts of R848 or a pharmaceutically acceptable salt thereof, and 30-50 parts of Poly I:C or a pharmaceutically acceptable salt thereof. (b) The tumor antigen is 300-400 samples; (c) 20-400 doses of anti-CTLA4 antibody; or 5-100 doses of anti-CD40 antibody; or 50-1000 doses of anti-PD-1 antibody and 20-400 doses of anti-CTLA4 antibody; or 5-100 doses of anti-CD40 antibody and 20-400 doses of anti-CTLA4 antibody; or 50-1000 doses of anti-PD-1 antibody, 20-400 doses of anti-CTLA4 antibody and 100-200 doses of anti-TIGIT antibody; or 50-1000 doses of anti-PD1 antibody, 20-400 doses of anti-CTLA4 antibody and 5-100 doses of anti-CD40 antibody; or 20-50 doses of anti-OX40 antibody, 20-400 doses of anti-CTLA4 antibody and 100-200 doses of anti-TIGIT antibody.

15. The pharmaceutical composition or combination preparation according to any one of claims 1-14, for the prevention and / or treatment of cancer; preferably, the cancer is a solid tumor; more preferably, the cancer is selected from colorectal cancer, melanoma, breast cancer, lung cancer, gastric cancer, head and neck tumors, prostate cancer, liver cancer, nasopharyngeal carcinoma, esophageal cancer, and cervical cancer.

16. The pharmaceutical use of the pharmaceutical composition or combination preparation according to any one of claims 1-14, for the preparation of a medicament for the prevention and / or treatment of cancer; preferably, the cancer is a solid tumor; more preferably, the cancer is selected from colorectal cancer, melanoma, breast cancer, lung cancer, gastric cancer, head and neck tumors, prostate cancer, liver cancer, nasopharyngeal carcinoma, esophageal cancer, and cervical cancer.

17. A method for preventing and / or treating cancer, characterized in that, The method includes the following steps: administering to a subject an effective amount of the pharmaceutical composition or combination formulation according to any one of claims 1-14; preferably, the cancer is a solid tumor; more preferably, the cancer is selected from colorectal cancer, melanoma, breast cancer, lung cancer, gastric cancer, head and neck tumors, prostate cancer, liver cancer, nasopharyngeal carcinoma, esophageal cancer, and cervical cancer.

18. The method according to claim 17, characterized in that, The pharmaceutical composition or combination formulation is administered to the subject via parenteral administration, preferably via intratumoral injection, subcutaneous injection, intravenous injection, intramuscular injection, nasal inhalation, or via an interventional device.

19. The method according to claim 18, characterized in that, The combined formulation can be administered to the subject simultaneously, sequentially, or at intervals.

20. The method according to claim 18 or 19, characterized in that, In the combined formulation, (a) and (b) are administered via the same route; (a) and (c) are administered via the same or different routes. Preferably, (a) and (b) are administered to the subject via intratumoral injection, and (c) are administered to the subject via intravenous injection; Alternatively, (a), (b), and (c) may be administered via the same route, either by intratumoral injection or subcutaneous injection; or (a) and (b), and a portion of (c) may be administered to the subject by intratumoral injection, and the remainder of (c) may be administered to the subject by intravenous injection.

21. The method according to claim 19 or 20, characterized in that, The combined formulation is administered to the subject simultaneously after mixing; or the combined formulation is administered to the subject simultaneously via different administration methods; preferably, the combined formulation is administered to the subject simultaneously via intratumoral injection or subcutaneous injection, or the combined formulation is administered to the subject simultaneously via different methods selected from intratumoral injection and intravenous injection.

22. The method according to claim 19 or 20, characterized in that, The combined formulation can be administered to the subject sequentially or at intervals via the same or different administration methods, preferably via intratumoral injection or intravenous injection.

23. The method according to claim 18 or 19, characterized in that, In the combined formulation, (a) when there are two or more TLR agonists, the administration routes of the different TLR agonists are the same or different; and (c) when there are two or more antibodies for treating cancer, the administration routes of the different antibodies are the same or different. Preferably, the administration routes of the different TLR agonists in (a) are the same.

24. The method according to claim 23, characterized in that, In the combination formulation, (a) when there are two or more TLR agonists, the TLR agonists are mixed and administered to the subject simultaneously, or the TLR agonists are administered to the subject as a single drug composition; preferably, the subject is administered via intratumoral injection or subcutaneous injection. (c) When there are two or more antibodies for treating cancer, the antibodies are mixed and administered to the subject simultaneously, or the antibodies are administered to the subject as a single drug composition, preferably by intravenous injection, intratumoral injection or subcutaneous injection; or a portion of the antibodies are administered to the subject by intratumoral injection and the remaining portion of the antibodies are administered to the subject by intravenous injection.

25. The method according to claim 23, characterized in that, In the combination formulation, (a) when there are two or more TLR agonists, the TLR agonists can be administered to the subject sequentially and at intervals by the same or different administration methods. Preferably, the TLR agonists are administered in the same manner, by intratumoral injection or subcutaneous injection. (c) When there are two or more antibodies for treating cancer, the antibodies may be administered to the subject sequentially or at intervals by the same or different administration methods; preferably, the antibodies are administered in the same way, by intratumoral injection, intravenous injection or subcutaneous injection; or a portion of the antibodies are administered to the subject by intratumoral injection and the remaining portion of the antibodies are administered to the subject by intravenous injection.

26. The method according to any one of claims 21-25, characterized in that, The combination formulation (a) and (b) are administered to the subject simultaneously or (a) and (b) are administered to the subject as a single drug composition, and the combination formulation (c) is administered to the subject in different ways; Alternatively, (a) and (b), and a portion of (c) may be mixed and given to the subject simultaneously, while the remainder of (c) may be given to the subject in a different manner.

27. The method according to claim 26, characterized in that, In the aforementioned combination formulation (a) CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; by intratumoral injection; (b) is a tumor antigen; administered via intratumoral injection; (c) Anti-CTLA4 antibody, administered intravenously; or anti-CD40 antibody, administered intratumorally; or anti-PD-1 antibody and anti-CTLA4 antibody, administered intravenously; or anti-CD40 antibody and anti-CTLA4 antibody, administered intratumorally with anti-CD40 antibody and intravenously with anti-CTLA4 antibody; or anti-PD-1 antibody, anti-CTLA4 antibody and anti-TIGIT antibody, administered intravenously; or anti-PD1 antibody, anti-CTLA4 antibody and anti-CD40 antibody, wherein anti-PD1 antibody and anti-CTLA4 antibody are administered intravenously and anti-CD40 antibody is administered intratumorally; or anti-OX40 antibody, anti-CTLA4 antibody and anti-TIGIT antibody, wherein anti-OX40 antibody is administered intratumorally and anti-CTLA4 antibody and anti-TIGIT antibody are administered intravenously.

28. The method according to any one of claims 17-27, characterized in that, Giving can be done in one instance or multiple instances.

29. The method according to any one of claims 17-28, characterized in that, In the pharmaceutical composition or combination formulation, (a) has a single dose of 0.01-10 mpk, (b) has a single dose of 0.1-10 mpk, and (c) has a single dose of 0.01-60 mpk; preferably, (a) has a single dose of 0.02-5 mpk, (b) has a single dose of 1-5 mpk, and (c) has a single dose of 0.02-30 mpk; more preferably, (a) has a single dose of 0.2-5 mpk, (b) has a single dose of 2-5 mpk, and (c) has a single dose of 0.1-30 mpk; most preferably, (a) has a single dose of 0.3-2 mpk, (b) has a single dose of 3-4 mpk, and (c) has a single dose of 0.2-15 mpk.

30. The method according to claim 29, characterized in that, The drug or combination formulation comprises, (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; (b) Tumor antigen, with a single dose of 0.1-10 mpk; (c) Anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-CD40 antibody, single dose 0.01-5 mpk; or anti-PD-1 antibody, single dose 0.01-20 mpk, and anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-CD40 antibody, single dose 0.01-5 mpk, and anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-PD-1 antibody, single dose 0.01-20 mpk, and anti-CTLA4 antibody, A single dose of 0.01-20 mpk and anti-TIGIT antibody, a single dose of 0.01-10 mpk; or anti-PD1 antibody, a single dose of 0.01-20 mpk, anti-CTLA4 antibody, a single dose of 0.01-20 mpk, and anti-CD40 antibody, a single dose of 0.01-5 mpk; or anti-OX40 antibody, a single dose of 0.01-5 mpk, anti-CTLA4 antibody, a single dose of 0.01-20 mpk, and anti-TIGIT antibody, a single dose of 0.01-10 mpk. Preferably, the drug or combination formulation comprises (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk, and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or its pharmaceutically acceptable salts, administered at a single dose of 1-2 mpk; and R848 or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk. (b) Tumor antigen, with a single dose of 3-4 MPa; (c) Anti-CTLA4 antibody, single dose 3-5 MPk; or anti-CD40 antibody, single dose 0.2-0.5 MPk; or anti-PD-1 antibody, single dose 3-5 MPk, and anti-CTLA4 antibody, single dose 3-5 MPk; or anti-CD40 antibody, single dose 0.2-0.5 MPk, and anti-CTLA4 antibody, single dose 3-5 MPk; or anti-PD-1 antibody, single dose 3-5 MPk, and anti-CTLA4 antibody For the body, a single dose of 3-5 MPk is given, along with anti-TIGIT antibody, a single dose of 1-2 MPk; or anti-PD1 antibody, a single dose of 3-5 MPk, anti-CTLA4 antibody, a single dose of 3-5 MPk, and anti-CD40 antibody, a single dose of 0.2-0.5 MPk; or anti-OX40 antibody, a single dose of 0.2-0.5 MPk, anti-CTLA4 antibody, a single dose of 3-5 MPk, and anti-TIGIT antibody, a single dose of 1-2 MPk. Alternatively, preferably, the drug or combination formulation comprises, (a) CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.1-1 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.1-1 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.1-1 mpk, and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or its pharmaceutically acceptable salts, administered at a single dose of 0.1-1 mpk; and R848 or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk. (b) Tumor antigen, with a single dose of 3-4 MPa; (c) Anti-CTLA4 antibody, single dose 0.2-4 MPk; or anti-CD40 antibody, single dose 0.05-1 MPk; or anti-PD-1 antibody, single dose 0.5-10 MPk, and anti-CTLA4 antibody, single dose 0.2-4 MPk; or anti-CD40 antibody, single dose 0.05-1 MPk, and anti-CTLA4 antibody, single dose 0.2-4 MPk; or anti-PD-1 antibody, single dose 0.5-10 MPk, and anti-CTLA4 Antibodies, single dose 0.2-4 MPk, and anti-TIGIT antibody, single dose 1-2 MPk; or anti-PD1 antibody, single dose 0.5-10 MPk, anti-CTLA4 antibody, single dose 0.2-4 MPk, and anti-CD40 antibody, single dose 0.05-1 MPk; or anti-OX40 antibody, single dose 0.2-0.5 MPk, anti-CTLA4 antibody, single dose 0.2-4 MPk, and anti-TIGIT antibody, single dose 1-2 MPk.

31. The method according to claims 17-30, characterized in that, The subjects are mammals; preferably, the subjects are humans, dogs, or cats.

32. A method for preventing and / or treating cancer, characterized in that, The procedure includes the following steps: administering a therapy to a subject to release tumor antigens; and administering to the subject an effective amount of (a) one, two, or more TLR agonists, and (c) one, two, or more antibodies for treating cancer.

33. The method according to claim 32, characterized in that, The cancer is a solid tumor, preferably selected from colorectal cancer, melanoma, breast cancer, lung cancer, stomach cancer, head and neck tumors, prostate cancer, liver cancer, nasopharyngeal carcinoma, esophageal cancer, and cervical cancer.

34. The method according to claim 32 or 33, characterized in that, The TLR agonist is selected from TLR3 agonists, TLR4 agonists, TLR5 agonists, TLR7 agonists, TLR8 agonists, TLR7 / 8 dual agonists, and TLR9 agonists.

35. The method according to claim 34, characterized in that, The TLR9 agonist is selected from CpG ODN and its pharmaceutically acceptable salts, preferably from CpG HP007, CpG HP021, CpG HP026, CpG 684, SD-101, G10 CpG ODN, CpG 1018, CpG 7909, IMO-2125, and their pharmaceutically acceptable salts; the TLR7 / 8 dual agonist is R848 or its pharmaceutically acceptable salt; the TLR3 agonist is Poly I:C or its pharmaceutically acceptable salt.

36. The method according to claim 34 or 35, characterized in that, (a) is a TLR9 agonist; or a TLR7 / 8 agonist; or a TLR3 agonist; or a combination of a dual TLR7 / 8 agonist and a TLR9 agonist; or a combination of a TLR7 / 8 agonist and a TLR3 agonist; or a combination of a TLR3 agonist and a TLR9 agonist; or a combination of a TLR7 / 8 agonist, a TLR3 agonist, and a TLR9 agonist; preferably, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof, R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof.

37. The method according to any one of claims 32-36, characterized in that, The antibody used to treat cancer targets one, two, or more of PD-1, CTLA-4, TIGIT, OX40, and CD40.

38. The method according to any one of claims 32-37, characterized in that, The antibodies used to treat cancer are monoclonal antibodies, bispecific antibodies, trispecific antibodies, antibody-drug conjugates (ADCs), or radionuclide conjugates (RDCs).

39. The method according to claim 37 or 38, characterized in that, (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody.

40. The method according to claim 39, characterized in that, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof, and Poly I:C or a pharmaceutically acceptable salt thereof; (c) is an anti-CTLA4 antibody; or an anti-CD40 antibody; or an anti-PD-1 antibody and an anti-CTLA4 antibody; or an anti-CD40 antibody and an anti-CTLA4 antibody; or an anti-PD-1 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody; or an anti-PD1 antibody, an anti-CTLA4 antibody and an anti-CD40 antibody; or an anti-OX40 antibody, an anti-CTLA4 antibody and an anti-TIGIT antibody.

41. The method according to any one of claims 32-40, characterized in that, The subject is given radiotherapy to release tumor antigens. Preferably, the subject is given radiotherapy using radiation or radiopharmaceuticals. More preferably, the radiation is selected from alpha, beta, gamma rays, and X-rays, electron beams, proton beams and other particle beams generated by various X-ray therapy machines or accelerators.

42. The method according to any one of claims 32-41, characterized in that, The subject is given radiotherapy using stereotactic radiotherapy (SRT), preferably using three-dimensional conformal radiotherapy (3DCRT), intensity-modulated radiotherapy (IMRT), or image-guided radiotherapy (IGRT).

43. The method according to any one of claims 32-41, characterized in that, The subject is given radiation therapy using stereotactic radiosurgery (SRS), preferably using X-knife, gamma knife, or CyberKnife.

44. The method according to any one of claims 32-43, characterized in that, The radiation dose for radiotherapy is 0.1-4 Gy.

45. The method according to any one of claims 32-44, characterized in that, The (a) and (c) are administered to the subject via parenteral administration, preferably via intratumoral injection, subcutaneous injection, intravenous injection, intramuscular injection, nasal inhalation, or via an interventional device.

46. ​​The method according to any one of claims 32-45, characterized in that, (a) and (c) can be administered to the subject simultaneously, sequentially, or at intervals.

47. The method according to claim 45 or 46, characterized in that, The routes of application for (a) and (c) are the same or different; Preferably, (a) and (c) are administered via the same route, by intratumoral injection or subcutaneous injection to the subject; or (a) and (c) are administered via different routes, by intratumoral injection and intravenous injection to the subject, respectively; or (a) and (c) are administered via a portion of the same route, while the remaining portion of (c) is administered via a different route. More preferably, a portion of (a) and (c) is administered to the subject via intratumoral injection, and the remainder of (c) is administered to the subject via intravenous injection.

48. The method according to claim 47, characterized in that, The drug (a) and (c) are administered to the subject as a single drug composition, or (a) and (c) are mixed and administered to the subject simultaneously, preferably by intratumoral injection or subcutaneous injection. Alternatively, (a) and (c) may be administered to the subject simultaneously via different administration methods; preferably, the subject may be administered via intratumoral injection or intravenous injection, respectively. Alternatively, a portion of (a) and (c) may be mixed and administered to the subject via intratumoral injection, while the remainder of (c) may be administered to the subject via intravenous injection.

49. The method according to claim 47, characterized in that, (a) and (c) can be administered to the subject sequentially or at intervals using the same or different administration methods; preferably, the administration methods are the same, administered to the subject via intratumoral injection, or administered to the subject via intratumoral injection and intravenous injection respectively.

50. The method according to claim 45 or 46, characterized in that, When (a) consists of two or more TLR agonists, the administration routes of the different TLR agonists are the same or different; when (c) consists of two or more antibodies for treating cancer, the administration routes of the different antibodies are the same or different; preferably, the administration routes of the different TLR agonists in (a) are the same.

51. The method according to claim 50, characterized in that, When (a) comprises two or more TLR agonists, the TLR agonists are mixed and administered to the subject simultaneously, or the TLR agonists are administered to the subject as a single drug composition; preferably, (a) is administered to the subject by intratumoral injection or subcutaneous injection. (c) When there are two or more antibodies for treating cancer, the antibodies are mixed and administered to the subject simultaneously, or the antibodies are administered to the subject as a single drug composition, preferably by intravenous injection, intratumoral injection or subcutaneous injection; or a portion of the antibodies are administered to the subject by intratumoral injection and the remaining portion of the antibodies are administered to the subject by intravenous injection.

52. The method according to claim 50, characterized in that, When (a) consists of two or more TLR agonists, the TLR agonists may be administered to the subject sequentially or at intervals via the same or different administration methods. Preferably, the subject may be administered via the same administration method, either via intratumoral injection or subcutaneous injection. (c) When there are two or more antibodies for treating cancer, the antibodies may be administered to the subject sequentially or at intervals by the same or different administration methods; preferably, the antibodies may be administered to the subject by the same administration method, such as intravenous injection, intratumoral injection or subcutaneous injection; or a portion of the antibodies may be administered to the subject by intratumoral injection and the remaining portion of the antibodies may be administered to the subject by intravenous injection.

53. The method according to claim 48, characterized in that, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof; or Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof; or R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; or CpG HP007 or a pharmaceutically acceptable salt thereof and R848 or a pharmaceutically acceptable salt thereof and Poly I:C or a pharmaceutically acceptable salt thereof; by intratumoral injection; (c) Anti-CTLA4 antibody, administered intravenously; or anti-CD40 antibody, administered intratumorally; or anti-PD-1 antibody and anti-CTLA4 antibody, administered intravenously; or anti-CD40 antibody and anti-CTLA4 antibody, administered intratumorally with anti-CD40 antibody and intravenously with anti-CTLA4 antibody; or anti-PD-1 antibody, anti-CTLA4 antibody and anti-TIGIT antibody, administered intravenously; or anti-PD1 antibody, anti-CTLA4 antibody and anti-CD40 antibody, wherein anti-PD1 antibody and anti-CTLA4 antibody are administered intravenously and anti-CD40 antibody is administered intratumorally; or anti-OX40 antibody, anti-CTLA4 antibody and anti-TIGIT antibody, wherein anti-OX40 antibody is administered intratumorally and anti-CTLA4 antibody and anti-TIGIT antibody are administered intravenously.

54. The method according to any one of claims 32-53, characterized in that, After administering radiotherapy to the subject for 1-5 consecutive days, an effective dose of (a) and (c) is given. Radiotherapy is then administered again after an interval of 1-2 days. One round of radiotherapy plus administration is repeated more than once. Preferably, after administering radiotherapy to the subject for 5 consecutive days, an effective dose of (a) and (c) is given again. Radiotherapy is then administered again after an interval of 2 days. More preferably, one treatment cycle is 1-12 weeks, and there may be multiple cycles. Alternatively, the subject may be given radiotherapy first, followed by an effective dose of (a) and (c) after 0-96 hours, and the above treatment may be repeated every 1-28 days; preferably, an effective dose of (a) and (c) may be given again; more preferably, one treatment cycle is 1-12 weeks, and there may be multiple cycles; Alternatively, while continuously administering radiotherapy to the subject, effective doses of (a) and (c) may be given at intervals, preferably every 1-28 days; more preferably, a treatment cycle is 1-12 weeks, and there may be multiple cycles.

55. The method according to any one of claims 32-54, characterized in that, The single dose of (a) is 0.01-10 mpk, and the single dose of (c) is 0.01-60 mpk; preferably, the single dose of (a) is 0.02-5 mpk, and the single dose of (c) is 0.02-30 mpk; more preferably, the single dose of (a) is 0.2-5 mpk, and the single dose of (c) is 0.1-30 mpk. Most preferably, the single dose of (a) is 0.3-2 mpk, and the single dose of (c) is 0.2-15 mpk.

56. The method according to any one of claims 55, characterized in that, (a) refers to CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-10 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.01-2 mpk; (c) Anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-CD40 antibody, single dose 0.01-5 mpk; or anti-PD-1 antibody, single dose 0.01-20 mpk, and anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-CD40 antibody, single dose 0.01-5 mpk, and anti-CTLA4 antibody, single dose 0.01-20 mpk; or anti-PD-1 antibody, single dose 0.01-20 mpk, and anti-CTLA4 antibody, A single dose of 0.01-20 mpk and anti-TIGIT antibody, a single dose of 0.01-10 mpk; or anti-PD1 antibody, a single dose of 0.01-20 mpk, anti-CTLA4 antibody, a single dose of 0.01-20 mpk, and anti-CD40 antibody, a single dose of 0.01-5 mpk; or anti-OX40 antibody, a single dose of 0.01-5 mpk, anti-CTLA4 antibody, a single dose of 0.01-20 mpk, and anti-TIGIT antibody, a single dose of 0.01-10 mpk. Preferably, (a) is CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk, and R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or R848 or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk, and Poly I:C or a pharmaceutically acceptable salt thereof, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or a pharmaceutically acceptable salt thereof, administered at a single dose of 1-2 mpk, and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; or CpG HP007 or its pharmaceutically acceptable salts, administered at a single dose of 1-2 mpk; and R848 or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk; and Poly I:C or its pharmaceutically acceptable salts, administered at a single dose of 0.3-0.5 mpk. (c) Anti-CTLA4 antibody, single dose 3-5 MPk; or anti-CD40 antibody, single dose 0.2-0.5 MPk; or anti-PD-1 antibody, single dose 3-5 MPk, and anti-CTLA4 antibody, single dose 3-5 MPk; or anti-CD40 antibody, single dose 0.2-0.5 MPk, and anti-CTLA4 antibody, single dose 3-5 MPk; or anti-PD-1 antibody, single dose 3-5 MPk, and anti-CTLA4 antibody For the body, a single dose of 3-5 mpk is given, along with anti-TIGIT antibody, a single dose of 1-2 mpk; or anti-PD1 antibody, a single dose of 3-5 mpk, anti-CTLA4 antibody, a single dose of 3-5 mpk, and anti-CD40 antibody, a single dose of 0.2-0.5 mpk; or anti-OX40 antibody, a single dose of 0.2-0.5 mpk, anti-CTLA4 antibody, a single dose of 3-5 mpk, and anti-TIGIT antibody, a single dose of 1-2 mpk.

57. The method according to claims 32-56, characterized in that, The subjects are mammals; preferably, the subjects are humans, dogs, or cats.

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